Aspects of the molecular and functional genetics in T1DM

This review has been accepted as a thesis together with six previously published papers, by the University of Copenhagen, January 24, 2006 and defended on March 24, 2006.

Hagedorn Research Institute, Steno Diabetes Center, Gentofte.

Correspondence: Ørholmvej 10, 2800 Lyngby, Denmark.

E-mail: johannesen@dadlnet.dk

Official opponents: Stellan Sandler, Sweden, Klaus Badenhoop, Germany, and Jens Højriis Nielsen.

2. Putative predisposing genes to T1DM

This chapter briefly reviews some general aspects regarding genetic studies in T1DM, various ways and approaches to identify susceptibility genes as well as genetic areas of interest within the genome (e.g. genome scans). Different genetic tests of such genes/genetic markers are briefly touch upon and the main findings from these studies are presented in tabulated form.

2.1. General aspects

Over the years, many genes have been investigated as predisposing genes to T1DM. Today it is generally considered that the HLA region is the only major genetic contributor along with minor contributions by other genes. However, no gene is neither sufficient nor necessary for T1DM development (Pociot et al., 2002).

In general, at least three aspects need to be considered when conducting genetic studies: (i) identification, characterisation and collection of the population to be studied, (ii) identification of genes or genetic regions to be investigated and, (iii) methods to analyse data.

Ad (i). The study population investigated in the papers included in this thesis is derived partly from a national survey obtained in 1990-1991 describing epidemiological parameters of T1DM individuals ageing less than 18 years - a study performed in collaboration with The Danish Society of Diabetes in Childhood and Adolescent (DSBD) (Pociot et al., 1993) - and partly from The Danish Insulin-Dependent Diabetes Mellitus Epidemiology and Genetics Group (DIEGG) in 1994-1999 (Lorenzen et al., 1998) identifying all T1DM probands below the age of 30 years. Population based sampling of probands (the T1DM individual through which the family was identified) and their families in racial/ethnically uniform populations in large sample sizes are important to identify genes with minor contributions (Risch et al., 1998; Altmuller et al., 2001; Cox et al., 2001; Risch et al., 2002). Sample size is particular important when the original data set is stratified for various parameters in order to test for association or linkage in relevant sub-fractions. When the candidate gene approach is undertaken - using either a case-control design or the design using Transmission Disequelibrium Testing of family based data - calculations regarding the power and size of the study population can be performed. The power of the study is determined by e.g.:

the different allele frequencies of the tested gene(s)
penetrance of the disease
the relative disease risk of a given polymorphism

- parameters often unknown beforehand, when testing new polymorphisms. However, papers have been published comparing the power using different analytical approaches e.g. using different sub-tests of TDT (Deng et al., 2001), and the number needed in TDT testing under various permissions (McGinnis, 2000).

Hence, the power calculation in our negative findings has been performed as follows: Given OR = 1.25 leads to P 1 = 0.5 and P 2 = 0.625 and hence, p(average): 0.5625.

Standard difference can then be calculated to 0.252.

N = 500/power 80 at 5% level and, N = 350/power 70 at 5% level (Altmann, 1993).

The collected multiplex families are characterised as being either affected sibs, including parents (n = 154) or parent/offspring families (trios) (n = 103) - in total 1143 family members.

Phenotypic characterisation is important to reduce genetic heterogeneity in the population studied. Hence, subsequent stratification of the patient material i.e. by onset of age or HLA-status may furthermore result in more homogeneous classes studied. In T1DM, variation in phenotype may not be a major problem as the clinical presentation of the disease is quite unique. However, the clinical presentation in very young childhood may clinically be slightly different, as the length of the remission period may be shorter or even absent (Bonfanti et al., 1998; Muhammad et al., 1999). This difference could hold a genetic component (Veijola et al., 1995). Age of onset has also been suggested to possess a genetic component (Fava et al., 1998). Another recent study found a lower MZ concordance rate when the index case was diagnosed at 25 years of age or older, suggesting a role for age-related non-genetic dependent factors (Redondo et al., 2001). The probands in the present material are identified in accordance to WHO criteria for T1DM (WHO, 1999). Data have subsequently been stratified according to e.g. HLA status or age of onset.

Ad (ii). Two different forms of genetic variation have been used in most studies of genetics in T1DM: (i) single nucleotide polymorphisms (SNP) being one nucleotide substituted by another at the same genomic position, when located in the coding regions may alter the "triplet" and give rise to an amino acid shift. (ii) variable numbers of tandem repeats (VNTR) also called "microsatelites" or "minisatelites" being two (or more) nucleotides repeated for a variable number of times. Microsatelites are typically located in genomic regions between genes and are widely distributed throughout the entire genome. The SNP provides two genetic variants whereas the VNTR may lead to typically 10-15 alleles, thereby being more informative than the SNPs in genetic testing. Today, the localisation and nature of many microsatelites are public available in various databases and have been generated from the world-wide efforts in sequencing the entire human genome. SNPs are also public available e.g NCBI dbSNP database (Sherry et al., 2000) but much of this information is based on comparisons of various submitted base-pair sequences, and many SNPs have not been confirmed (Taillon-Miller et al., 1998; Marth et al., 2001). In the studies included in this thesis we have screened the coding regions for polymorphisms. When doing so, two issues need to be considered:

Number of chromosomes tested: we have tested in the range of 34 and 40 persons equalling a frequency of minimum 1.25% for the most rare allele if only one copy was identified. Allele frequencies less than 5% were not studied further due to the low chance of detecting such a gene to influence T1DM susceptibility.
The method used for identification of the polymorphism: as direct sequencing is automated for most procedures today, this would be the method of choice. Previously, we did not have the capacity needed for such an approach, hence we used the technique of Single Stranded Conformational Polymorphism (SSCP), which in our hands had a sensitivity and specificity of 91% and 92%, respectively (Johannesen et al., 2001a).

Finally, when a candidate gene has been screened for polymorphisms the identified SNPs should be prioritised according to their putative functional impact of the protein before selecting of which SNP should be tested for association and/or linkage to disease (Tabor et al., 2002). However, even silent mutations may confer considerable impact on protein function due to e.g. involvement in mRNA splicing (Cartegni et al., 2002).

Ad (iii). Two analytical approaches have been undertaken in the analysis of T1DM genetics: association of a polymorphism to T1DM tested in case-control designs and association/linkage analysis applied to data generated from T1DM family collections. Linkage means that a marker allele co-segregate with the disease within each family - different families can have different marker alleles segregating with disease - in contrast to association where different families have the same specific marker co-occurring with disease (Field, 2002).

The association test in case-control designs is typically a chi-square test simply testing whether there is a difference in allele or genotype frequencies between cases and controls. The case-control design is straightforward in the sense that only genetic testing of the proband is required and the statistics are simple. When positive association is identified it is considered to be due to linkage disequilibrium between the disease and marker loci. However, especially the control population should be carefully selected in order to mirror the general population best possible and obviously the case population should be phenotypically well characterised and randomly included to avoid selection bias. The case-control design is typically used in the candidate gene approach.

The genetic analyses used in T1DM family collections have either been linkage or association based tests. Linkage analysis is a method to determine whether there is evidence for co-segregation - due to physical linkage on the chromosome - of alleles at a hypothetical disease-susceptibility locus and alleles at a marker locus in families with multiple affected members. Classical linkage requires the collection of families comprising affected and unaffected members in consecutive generations and a defined hypothesis for heredity to be tested. As T1DM is considered a genetic multiplex disease without a known mode of heredity, model-free methods testing linkage has been used in T1DM. The most common model-independent method is the affected sib-pair (ASP) linkage analysis - used in genome scans (see Chapter 2.3). The average proportion of alleles shared in affected sibs is tested against the 50% sharing expected by chance. A higher sharing is indicative of the marker locus also contains a disease locus - hence being linked. The relatively low frequency of affected sib-pair families lead to the development of the Linkage Disequelibrium Test (TDT), as this test uses the information obtained from simplex families ("Trios"). The TDT compares the number of transmitted alleles to non-transmitted alleles from heterozygous parents to affected offspring and is an association test (Spielman et al., 1993). This method was extended to handle multiallelic marker systems (ETDT) (Sham et al., 1995). TDT statistics have become the golden standard for testing candidate genes in family collections for linkage disequilibrium (Spielman et al., 1996), and have been used for testing the candidate genes in the papers included in this thesis.

As transmission distortion in general seems to be evident in humans, all polymorphisms tested by TDT have been performed for affected as well as non-affected individuals, to ensure random transmission to non-affected individuals (Zollner, 2004).

Hence, initially the classical candidate gene approach testing for association in a case-control design was taken - later, ASP linkage analysis of genome scan data and TDT analysis were applied to T1DM family collections.

2.2. The candidate gene approach

The candidate gene approach is a classical strategy. Based upon a pathophysiologically relevant indication allelic variants of such selected genes are tested for either association or linkage to T1DM.

The strength of the candidate gene approach depends upon the model in which it is a candidate. In favour of the candidate gene approach is the testing of the gene encoding the relevant protein in contrast to genomic markers of chromosomal loci as in genome scans. Using the candidate gene approach in a classical association study design, the identification and collection of a large T1DM population are more easily achieved than for a large T1DM family material, whereas the draw back is the risk of selection bias and confounding. Thus, by using the family based design testing candidate genes this potential bias is eliminated. In order to exclude a candidate gene as a susceptibility or protective gene, the search for polymorphisms to be tested can be quite extensive. In addition to the coding region, the functional regulation of the gene can be found 5' in the proximal promoter region and 3' UTR's distant regulatory regions as well as within introns (intron/exon splicing sites) (Cartegni et al., 2002). In a recent review more than 600 positive association studies were reviewed of which only 6 were considered consistently replicated. It was concluded that in order to substantiate association, case-control studies should contain large number of cases and controls tested of uniform ethnical origin and that replication studies seem mandatory (Hirschhorn et al., 2002). However, a rejection of genetic association of a protein does not exclude a pathogenetical relevance of the protein.

This thesis will review the current status of candidate genes tested in T1DM at two levels:

Candidate genes subdivided into categories based upon "The Copenhagen Model":
- T-cell regulation and inflammation
- cytokine genes
- genes relating to deleterious and protective mechanisms in the beta-cell and, finally
- other tested classical candidate genes in T1DM. The HLA region is described separately, the remaining tabulated and categorised as above described, see Table 2 .
Encoding genes for proteins identified upon the "combined approach to select functionally focused candidate genes" - as previously defined and reviewed in Chapter 4.

2.2.1. HLA genes

The HLA region has been proposed to account for 40-50% of the genetic susceptibility to T1DM (Risch, 1987; Noble et al., 1996). The HLA class II mediated susceptibility/protection seems to be mediated through class II antigen presentation in the islets as well as through the development of central and peripheral tolerance (Lee et al., 2001; Todd et al., 2001).

The human leukocyte antigen (HLA) region is located at the short arm of chromosome 6, 6p21. Its organisation is shown in Figure 2 (HLA overview incl. genes).

The current understanding of HLA-DQ association shows the strongest association for individuals being heterozygous carrying the genotype DQA1*0501-DQB1*0201/DQA1*0301-DQB1*0302 (encoding the DQ2 and DQ8 molecules, respectively) conferring a relative risk of ≥ 10. Likewise, protection from developing T1DM is conferred by the haplotype DRB1*1501-DQA1*0102-DQB1*0602 (DQ6 molecule), which may provide dominant protection over the susceptibility conferred by other HLA genes. Finally, the risk conferred by DQ2 and DQ8 molecules is modified by DR (Thorsby et al., 1993; Boitard et al., 1997; Undlien et al., 1999) which points to a role of the DR locus in susceptibility to T1DM.

The observation that the highest susceptibility are seen for DR3/4 heterozygous, has lead to the hypothesis of transcomplementation allowing for the construction of DQA1*0501-DQB1*0302/DQA1* 0301-DQB1*0201 molecules. Linkage studies have also shown the existence of susceptibility genes in the HLA region: of 538 diabetic sib-pairs 54% shared two HLA haplotypes and only 7.3% shared no haplotypes, both frequencies being significantly different from the 25% expected (Payami et al., 1985; Robinson et al., 1993). Recently, the genome scans within T1DM have all demonstrated highly significant LOD-scores for the HLA region, demonstrating linkage to T1DM of the HLA region (see Chapter 2.3 for references). The above listed associations are primarily found in Caucasians, see review by (She, 1996; Zamani et al., 1998) for further explorations into inter-racial differences.

Recently, changes in the frequencies of HLA genotypes over time in Finnish T1DM patients have been reported: the frequency of high risk HLA genotype has decreased from 25.3% to 18.2% while the protective HLA genotypes have doubled comparing data from patients diagnosed before 1965 and after 1990, despite an increase in incidence of 2.5 times during the period from 1966 to 2000 in Finland (Hermann et al., 2003). It is concluded that the environmental pressure has increased resulting in higher penetrance of disease, especially in individuals with protective HLA genotypes.

The functional basis of the HLA class II molecule in T1DM has been related to peptide/antigen binding of the molecule, for review please see (Nepom et al., 1998).

2.2.2. HLA non-DQ/DR genes

Within the HLA region, other genes - apart from the HLA-DQ and DR - have been tested for genetic susceptibility to T1DM.

Based upon a review of the literature, genes tested for genetic susceptibility are listed in Table 1 ( Tabel 1A , Tabel 1 B , Tabel 1C ) and Table 2 ( Tabel 2A , Tabel 2B , Tabel 2C ) . An evaluation of the genes being demonstrated or rejected as risk modifying genes is based upon the the following criteria:

The study of a candidate gene must have been consistently replicated at least once, in order to minimize the risk of false positive reports (Lohmueller, 2003).
A single case/control study should comprise approximately 200 or more cases and a matching number of controls. This number is required to have a power of 80, at the significance level of 0.05, identifying a relative risk of 1.5-2.0, given the frequency of the associated allele in the control group is 0.15-0.60 (Breslow et al., 1987). However, the finding of several minor case/control studies (n: 4-5) uniformly indicating the same result has also been taken into account within the overall evaluation of a gene influencing the risk of T1DM.
All family based studies are included, as the number of qualified transmissions within the family collection depends on the allelic frequencies of the tested polymorphism and the analysis used.

These simple criteria represent one way to select the more robust candidate genes in T1DM, as a huge number of genes have been tested as candidate genes in T1DM.

However, in some cases even replicated results from independent studies of tested candidate genes are contradictory. Hence, in such cases it can be difficult to determine whether the candidate gene is truly associated to T1DM or not - the genetic risk modification of the candidate gene being inconclusive. Within the column "Confirmed replication" (Conf. Rep) these genes are marked "Yes for both outcomes". An explanation of these apparent contradictory results could be due to genetic heterogeneity of disease susceptibility between and within populations e.g. (Metcalfe et al., 1996). The risk modifying effect is considered minimal for these genes. Furthermore, in some cases different genetic variants have been tested within the gene, hence no meta-analysis has been performed.

Table 1 lists the HLA non-DQ/DR candidate genes. As strong linkage disequilibrium (LD) exists within the HLA region - strong LD exists between studied non-DR/DQ genes in the HLA region and the high risk HLA DR/DQ genes - different strategies have been used to evaluate the independent effect of the studied non-DR/DQ genes. Within the case/control design the use of HLA haplo-identical control subjects and diabetic patients have been used (Deng et al., 1995). Furthermore in the case/control study by Gambelunghe testing the MIC-A gene polymorphism (Gambelunghe et al., 2000), a test for the strongest HLA association was performed as described by (Svejgaard et al., 1994).

Within family studies subset TDT analyses have been performed e.g. (i) comparing the risk conferred by HLA-DQ8 and HLA-DQ2 in the presence/absence of the tested genetic variation as illustrated for HERV-K(C4) (Pani et al., 2002) or (ii) by testing the transmission of parents being homozygous for the high risk DR/DQ and heterozygous for the variant in question to affected offspring as illustrated for LMP2 and LMP7 (Undlien et al., 1997).

As previously described, strong linkage disequilibrium exists within the HLA region, making identification of DR/DQ independent contributions of other genes within the HLA region difficult. A pathogenically interesting observation is the association of the diabetogenic TNF haplotype, TNFa2/TNFB*2/HLA-B15 to high TNF α production from macrophages (Pociot et al., 1993). This TNF microsatelite has been shown to be associated to age of onset of T1DM (Obayashi et al., 1999). Furthermore, a retroviral long terminal repeat adjacent to the HLA-DQB1 gene (DQ-LTR13) has been shown to modify T1DM susceptibility on high risk DQ haplotypes (Bieda et al., 2002). Recently, the random marker approach has been applied to the HLA region, identifying susceptibility regions outside HLA class II (Lie et al., 1999; Undlien et al., 2001), and Noble has shown an importance of class I antigens in modulating susceptibility to T1DM (Noble et al., 2002). Support for additional susceptibility genes in the HLA class III region, close to the TNF genes, has been provided by an analysis of the Belgian diabetes registry (Moghaddam et al., 1998).

Apart from determining T1DM risk, the HLA genes have been associated to modulation of clinical features of the disease, e.g. age of onset or outcome of active cellular autoimmunity, see (Bach et al., 2001).

2.2.3. Candidate genes outside the HLA region
catagorised according to "The Copenhagen Model"

The major genetic contribution of the HLA region in T1DM has been assessed to approximately 40-50% (Risch, 1987; Noble et al., 1996). Hence, the remaining genetic susceptibility comes from several other minor contributions outside the HLA region. Many different genes have been tested for association and linkage to T1DM. In Table 2 are listed putative candidate genes tabulated according to - but not identified by - "The Copenhagen Model" of pathogenesis to T1DM, as other strategies naturally have been advocated to qualify candidate genes in T1DM than based upon "The Copenhagen Model".

The idea has not been to provide the reader with a complete list of published papers in the field, as a meaningful review of a specific gene in T1DM would require a separate up to date search of the literature, but to illustrate the huge effort world wide that has been put into this field - and the relatively sparse outcome.

The criteria for selection of genes in Table 2 are identical to those listed for the HLA non-DQ/DR genes in Table 1.

In conclusion: The success of the candidate gene approach in identifying the HLA region is evident, since the major genetic predisposition to T1DM resides in the HLA region. However, the identification of specific genes inside the HLA region associated and/or linked to T1DM is complicated by strong linkage disequilibrium within this region. Genes outside the HLA region each contributing to a minor degree of the overall genetic predisposition to T1DM have also been identified by means of the candidate gene approach - however, the number of genes and their significance as well as interactions need further exploration. The functional implications of the genetic contributors to T1DM identified so far (HLA, CTLA4 and INS) do not reject "The Copenhagen Model" as a pathogenetic model of T1DM as the immune system as well as the beta-cell are considered to be important in this model. Neither has the identification of genes not influencing the genetic risk of T1DM lead to the rejection of "The Copenhagen Model". The encoding gene to a pathological important protein does not need to be genetically associated to the disease.

In the search for identifying the genetic predisposition of T1DM supplementary model-independent approaches has been initiated, e.g. genome scans.

2.3. Genome scans

As a consequence of T1DM being a polygenetic disorder and the failure of the candidate gene approach to identify all the genetic components conferring increased or decreased risk of T1DM development, new approaches to solve the T1DM genetic puzzle were sought. In the early 1990's, an alternative to the classical candidate gene approach emerged: complete and partial genome scans using polymorphic microsatelite markers spread over the entire genome or specific parts of the genome, in order to identify chromosomal regions linked to the disease.

The obvious strength of using polymorphic markers widely spread over the entire genome is that no a priori considerations regarding interesting regions are required; hence, the opportunity of identifying unknown regions of putative importance exists. Furthermore, as for the case/control design testing for association the linkage analysis (examining identity by descent) in affected sibs pairs overcomes the lack of knowledge regarding the mode of inherence of T1DM. Drawbacks, however, are that after identifying a region of interest, a major effort has to be put into identifying the pathogenically relevant gene(s) (fine mapping) and subsequent cloning and functional characterisation (positional cloning) as the chosen polymorphic markers used in genome scans typically are located in genetic areas between the coding genes, see Table 3 . However, new strategies for positional cloning are continuously emerging, e.g. hierarchical genotyping design using successive rounds of genotyping and analysis by the haplotype pattern mining algorithm (Laitinen, 2004).

Experience from the first complete genome scans in T1DM has generated quite different results with only few consistently identified chromosomal regions contributing to the risk of T1DM. The HLA region (IDDM1) has been identified within all complete human genome scans within T1DM (Davies et al., 1994) (Hashimoto et al., 1994; Concannon et al., 1998; Mein et al., 1998; Nerup et al., 2001). The VNTR at the 5' end of the insulin gene (IDDM2) has also demonstrated to confer risk of T1DM in two genome scans and several association studies. As these two regions together only can account for a λ s of 4-5 (Todd et al., 1997) of the total λ s of 15, the remaining genetic susceptibility is located elsewhere (Lernmark et al., 1998).

A major problem in the genome scans using linkage analysis is the limited power to map genes with a weak genetic component, e.g. testing relatively small sample sizes only detect disease genes with a genotypic risk ratio of more than 4 (the increased chance that an individual with a particular genotype has the disease); - hence, increasing the chance of identifying minor genetic components would require very large family materials (1000s of ASPs) (Risch et al., 1996). This can partly explain the deviating results obtained in the different genome scans indicating the importance of sample size. Other possible explanations of the variation observed in the results between the different genome scans are (i) genetic heterogeneity (an apparently uniform phenotype being caused by two or more different genotypes), (ii) differences in disease phenotype: age of onset, presence/absence of IDDM-associated auto antibodies at onset, other autoimmune diseases, gender-specific effect (iii) ethnic origin, (iv) gene to gene and gene to environmental interactions being different in various populations and (v) variation due to random chance (She, 1998; Altmuller et al., 2001; Cox et al., 2001). In an attempt to overcome these drawbacks of complete genome scans, linkage disequelibrium analysis has been taken into use, as a tool to confirm and fine map susceptibility intervals. This approach has successfully been used for e.g. IDDM12/CTLA4 (Nistico et al., 1996; Marron et al., 1997; Kristiansen et al., 2000; Ueda et al., 2003).

On the other hand, besides suggesting chromosomal regions of importance in modifying genetic risk, the genome-scan data potentially exclude chromosomal regions as disease modifying.

In the future, there is an urgent need for collaboration world wide within this area in order to increase the number of tested families. This can be done by two separate approaches (i) pooling of existing data set and (ii) identification and sampling of new families. Finally, stratification of genome-scan data has been proposed to identify various interactions between different loci, as initially proposed by Cox (Cox et al., 2001; Nerup et al., 2001). This approach has lead to the identification of an increased LOD score on chromosome 6q27 from 0.94 to 3.69 when conditioned for age at onset less that 11 years in the combined UK and US family material (Cox et al., 2001), and in the Scandinavian genome scan evidence of heterogeneity was demonstrated with markers at 16p and in the HLA DR3/4 group (Nerup et al., 2001).

A spin-off from the genome scans has been the opportunity to compare genome scan data obtained from different autoimmune mediated diseases in order to identify shared loci within e.g. lupus erythematosus, multiple sclerosis and Crohn's Disease (Becker et al., 1998; Becker, 1999). Overlapping regions at chromosome 2q (CTLA4), 6p (HLA), 11p, and Xp have been reported and may lead to identification of common pathogenically pathways encoded by genes within these regions.

In conclusion: The initial high expectations of whole genome scans to enlighten the genetic puzzle of T1DM have not been fully met as only few genetic regions have been consistently identified. The limitations of genome scans possibly responsible for these findings e.g. population and ethnic differences and imperfect statistical and analytical methods have led to initiatives of large scale sampling of affected sib pair families and pooling of existing data. However, pooling of data worldwide does not exclude population based differences. Secondly, it may turn out, that the major benefit from T1DM genome scans is to exclude certain genomic areas as potential candidate gene containing areas. However, genome scans might exclude a chromosomal region being important in gene-to-gene interactions, hence, the analytical methods need to include such interaction analyses. New analytical methods should be introduced e.g. haplotype interactions (Zhang, 2003) and non-model based analytical methods e.g. data mining (Pociot et al., 2004) in which non-genetic factors may also be included in the analyses. Finally, as the genome scans generate data from affected individuals and do not include data from non-affected - no protective chromosomal regions are identified.

2.4. Other approaches

In order to limit the genetic variation in the study population, diabetes related genes have been searched for within: (i) populations with few founders and no mixing to other populations (e.g. Arab families, IDDM17), and (ii) T1DM encountered in rare genetic syndromes (e.g. mitochondrial disorders, Downs Syndrome, Friedreisch's ataxia, Wolcott Rallison Syndrome and Wolfram Syndrome (Watkins et al., 1998)) in order to examine common diabetes associated genes. Finally, animal models spontaneously developing disease as homologous genes/chromosomal regions may be of interest regarding human diabetes.

In order to study the effect of limited population mixing in a population with a common ancestor, a genome scan of an Bedouin Arab family with a high prevalence of T1DM has been performed (Verge et al., 1998), identifying a locus mapping to the long arm of chromosome 10 (10q25) (IDDM17) being in linkage to T1DM. At this locus, increased LOD scores were observed near the reported location of this putative IDDM17 locus when conditioning the analysis for DR3 positive individuals in the combined UK/US data set (Cox et al., 2001). So far no candidate gene has been identified within this region.

Wolfram's syndrome is an autosomal recessive disorder defined by the occurrence of young-onset diabetes mellitus and progressive bilateral optic atrophy; neurological symptoms and predisposition to psychiatric disease may also associate to the diagnosis (Swift et al., 1998). Linkage of the wolfram syndrome to the short arm of chromosome 4 (D4S431) was established in 1994 (Polymeropoulos et al., 1994). Within the Scandinavian genome scan of T1DM, evidence of linkage to chromosome 4p16.1 was found, particular in the subset of Danish families (Nerup et al., 2001). In a Danish study, additional markers to those used in the Scandinavian genome scan further confirmed linkage to this region, however the 15 new polymorphisms identified did not show linkage to T1DM in the Danish population (Larsen et al., 2004). These results are indicative of a role of yet unidentified polymorphisms of the WFS1 gene in the development of common T1DM.

Regarding the main candidate gene loci in the NOD mouse, please see the following reviews: (Wicker et al., 1995; Todd et al., 2001; Serreze et al., 2001). Special focus has been set upon loci of disease protection (Todd et al., 2001; Adorini et al., 2002). In the paper of Kloting et al., the disease associated chromosomal regions within the BB rat have been reviewed ( Iddm1, Iddm2 and Iddm3 ) as well as alleles within diabetes-resistant BB rats contributing to insulin-dependent type 1 diabetes mellitus (Kloting et al., 2003) are described. These studies have mainly confirmed association to the MHC complex.

In conclusion: The use of clinical syndromes comprising immune-mediated diabetes mellitus, the study of isolated populations and animal models of diabetes, have been used as a supplement to the candidate gene approach and genome scans within the general population in order to identify common genetic disposition to T1DM.

Conclusion from Chapter 2

The genetic predisposition to T1DM is complex and despite major efforts to identify the genetic disposition to T1DM many questions still remain. Both the candidate gene approach and whole genome scans have been applicated in the search for T1DM genetic predisposition, however the results so far have been incomplete. Inconsistency between the results obtained from the different genome scans and the partial overlap of the genome scan findings to the results generated by the candidate gene approach are future challenges. Putative explanations could be different markers used in the genome scans as well as the markers used in the genome scans being too far apart - hence, the small chromosomal regions harbouring the candidate genes are missed. In the future, there is a need for sampling large ethnically homogeneous population based T1DM family collections to expand the genome scans by using SNP's or haplotype Tag SNP's and to refine the statistical methods for evaluation of the candidate genes, e.g. to include interaction with other genes or environmental factors.

Finally, new approaches for candidate gene identification may supplement the search for T1DM modifying genes. In vitro data derived from a functional testing of the target organ based upon `The Copenhagen Model' will be proposed for selection of new candidate genes. In contrast to e.g. the genome scans, this approach allows the identification of protective candidate genes, as the functional testing will illuminate a putative race between deleterious and protective mechanisms in the target organ.

3. "The Copenhagen Model"
- a way to select candidate genes

3.1. A combined approach to select
candidate genes

As previous strategies to identify susceptibility genes in T1DM have not succeeded in clarifying the genetic predisposition to T1DM, new strategies may provide additional information. Due to the possibility of gaining more detailed information regarding intracellular processes by the protein and mRNA expressing profiling technologies, a broader understanding of the cytokine mediated beta-cell destruction has become possible.

Hence, a combination of various strategies, all pin-pointing towards the same candidate gene, increases the a priori chances of identifying genes affecting T1DM susceptibility. The different strategies used in this combined approach to select candidate genes are based upon:

Theoretical pathogenetical considerations derived from "The Copenhagen Model",
An in-vitro , testable model hereof - focusing at the beta-cell - using expressional profiling: As cytokine induced beta-cell destruction may play a role in the pathogenesis of T1DM (Bergholdt et al., 2003) IL-1 β induced altered protein expression in beta-cells reflects putative pathogenetic mechanisms involved in cytokine induced beta-cell destruction. It has been speculated, that in T1DM the beta-cell destruction is not only dependent upon an auto-aggressive immune response - the beta-cells themselves may also influence the outcome (Andersen, 1999). Hence, islet proteins identified as having a changed expression level due to cytokine exposure qualify as putative candidate genes: Firstly, in contrast to the classic candidate gene approach, where subsequent functional evaluation of novel genetic variations is standard, candidate genes identified by an altered expression profile after cytokine exposure have been selected upon a functional basis. However, to what extent such altered expression can influence the outcome of the cytokine exposed beta-cell needs to be evaluated in subsequent functional analyses, e.g. in over-expression studies. Secondly, such genes are focused, as only target organ proteins are considered.
Linkage analyses data derived from T1DM genome scans.

This approach has been advocated as a general way to identify susceptibility genes in genetically complex diseases (Hirschhorn et al., 2002) and specifically for T1DM (Pociot et al., 2002) (see Figure 3 ).

As this approach is based upon "The Copenhagen Model" - cytokine induced beta-cell destruction - and a functional evaluation hereof by use of expressing profiling, these two topics are summarised below. The data from T1DM genome scans are reviewed in the previous chapter. In the end of this chapter, the selection of three candidate genes based upon the combined approach and the strategy for their evaluation are outlined.

3.1.1. Expression profiling

The development of two different technologies provides the possibility to gain insight into the expression profiling of cellular systems at different levels: (i) proteome analysis, e.g 2D-protein gel analysis combined with mass spectrometry as protein identification and (ii) transcriptome analysis e.g. microarray or genechip array technology, for review plase see (Jungblut et al., 1999; Celis et al., 2000; Lockhart et al., 2000; Karlsen et al., 2001). In short, these two complementary technologies aim at identifying and quantifying gene transcripts at the mRNA expression (transcriptome analysis) or the protein level including posttranslational protein modifications (proteome analysis) in order to obtain further insight into pathological and pathogenetic mechanisms of different diseases and/or altered physiological conditions (e.g. toxicology). Examples of application areas within human diseases have been leukaemia, breast-, colorectal- and bladder cancers, and heart diseases, e.g. dilated cardiomyopathy and atherosclerosis. Within these diseases various prognostic markers and different transcription factors of putative pathogenetic relevance have been identified.

The technologies comprise obvious advantages as they mirror the intracellular changes in expression within the target organ or cellular system in much more detail than other methods are capable of. The microarray or gene chip arrays can display several thousands of Expressed Sequence Tags (EST) or known mRNA's at the same time making comparisons to different conditions possible by analysing the change in the expression level. A draw back of microarray compaired to 2-dimentional protein gel analysis is that not all mRNAs present in a cell are translated into protein (Gygi et al., 1999) and mRNAs encode for unmodified pre-forms of proteins. On the other hand, 2-dimentional protein gel analysis is able to detect the proteins as well as identifying post-transcriptional modified proteins which is very important, as (i) it is the proteins that initiate and run the cellular processes, not the mRNA - and (ii) posttranscriptional changes e.g. phosphorylation often activate inactivated cytosolic proteins. However, it is only a part of the total number of proteins present in a cell preparation that is actually displayed at a protein gel e.g. proteins with high and low molecular weight as well as membrane bound proteins are missed. General drawbacks of both methods are (i) they represent snap shots of processes that are dynamic in nature as they only reflect the cellular status at a defined time point or period, (ii) they do not allow for discrimination between primary and secondary events or elucidation of putative interactions.

Results from expressing profiling in insulin producing cells: So far, 7 papers have been published applying the proteome analysis at cytokine or NO-donor treated insulin producing cell lines or islets of Langerhans (Andersen et al., 1995; Andersen et al., 1997; Christensen et al., 2000; John et al., 2000; Mose-Larsen et al., 2001; Sparre et al., 2002; Nielsen, 2004). Based upon "The Copenhagen Model", it has been attempted to categorise the identifications from these studies into the following main areas: (i) cytokine-signalling, (ii) energy generation, (iii) NO-production, (iv) insulin production/beta-cell function, (v) apoptosis and, (vi) defence/repair. Transcriptome data have been obtained using either RINm5F cells, primary rat beta-cells, INS-1 cells or NHI-glu/NHI-ins cell lines exposed to various combinations of cytokines (Rieneck et al., 2000; Cardozo et al., 2001a; Cardozo et al., 2001b; Kutlu et al., 2003; Nielsen et al., 2004). Comparing the data generated using these two methods has revealed only partial overlap. Possible explanations for the different findings can be different cellular sources, variation in cell phenotype and experimental settings, and the biphasic effect of IL-1 - some cells may be stimulated other suppressed. Finally, not all mRNA changes lead to altered protein expressions. The results will be discussed in more detail in relevant chapters.

In conclusion: These novel and powerful technologies are promising and may add new valuable information to cytokine mediated beta-cell destruction and increase our understanding of biology in general. Naturally, there are obstacles: the generation of huge amounts of data requires development of new software, further insight in bioinformatics, standardisation of normal expression levels in various tissues e.g. target cells, and finally the limitation of the techniques of showing a static picture of a dynamic process. However, the approach of combining transcriptome analysis with serial experiments and cluster analysis has been attempted in order to include a dynamic dimension to this technology (Kutlu et al., 2003). Finally, it seems more relevant to select putative candidate genes based on the data generated from the changed protein expression in the target organ than using altered mRNA in the target as the altered protein expression pattern reflects the functional significant processes best.

3.2. Aspects of cytokine mediated
beta-cell destruction

This chapter reviews some aspects of cytokine mediated beta-cell destruction in order to give background (i) for the functional selection of parameters and expressed mRNA and/or protein transcripts when comparing cytokine exposed islets from two genetically different rat strains (a signalling factor, deleterious as well as protective molecules), and (ii) for the selection of the tested candidate genes. The apoptotic and necrotic process of beta-cells initiated by cytokines is depicted within the iNOS chapter.

3.2.1. Cytokines in beta-cell destruction

The observations by Gepts in 1965 (Gepts, 1965) of lymphocytic infiltration within the islets of Langerhans (insulitis) seen in newly diagnosed T1DM patients were demonstrated in vivo and in vitro to correlate to immune mediated beta-cell destruction (Nerup et al., 1971). In 1974 association of the HLA system to type 1 diabetes became evident (Nerup et al., 1974), and in 1985 it was suggested that soluble mediators of the immune system liberated during the inflammatory process were beta-cell cytotoxic (Mandrup-Poulsen et al., 1985). IL-1 β was identified as being the single cytokine which alone could impair beta-cell function (Bendtzen et al., 1986; Mandrup-Poulsen et al., 1986a; Mandrup-Poulsen et al., 1986b). IL-1 was shown to be selectively beta-cell cytotoxic (Mandrup-Poulsen et al., 1987a; Sandler et al., 1989; Helqvist et al., 1991b), an effect intensified by INF γ and TNF α (Mandrup-Poulsen et al., 1987b; Eizirik, 1988).

In animal models spontaneous developing diabetes, cytokines were identified in the insulitis lesion in the NOD mouse and BB rat (IL-1, TNF α , IFN α and IFN γ , IL-6 and IL-12). Transgenic NOD mice or BB rats expressing IL-4 under the rat insulin promoter (RIP) were protected against diabetes development, and expression of IL-4 and IL-10 was observed in NOD mice protected from diabetes development by various treatments such as oral administration of insulin, injection of CFA or intraperitoneal injections of long-lasting IL-10 preparation. Further, blocking cytokines by either anti-cytokine antibodies (against IFN γ , IL-6, TNF α ) or blocking cytokine receptors (by either soluble IL-1 receptor or IL-1Ra) or disrupting cytokine genes (IL-12 and INF γ ) have been reported to delay and/or decrease diabetes incidence in NOD mice. Recently, it has been shown that NOD mice being deficient of the IL-1R demonstrated slowed progression to diabetes (Thomas et al., 2004). Finally, over-expression of various cytokines in beta-cells (IFN γ , IFN α ) under the RIP in non-diabetic-prone mice resulted in severe lymphocytic islet infiltration and diabetes; whereas beta-cell expression of IL-2, IL-6, IL-10 and TNF α induced insulitis without causing overt diabetes. For more details, please see review (Rabinovitch et al., 1998b).

Histopathology of the islet has identified antigen presenting macrophages (MHC class II positive) and CD4 + T helper cells (Th) as the first cells to infiltrate the islet in the BB rat, NOD mouse and the low-dose streptozotocin animal models of diabetes (Kolb-Bachofen et al., 1988; Lee et al., 1988; Hanenberg et al., 1989). The end-stage infiltrate comprises large number of macrophages, CD8 + cytotoxic T-cells (Tc) and CD4+ T-helper cells (Th), as well as B-lymphocytes (Kay et al., 1991; O'Reilly et al., 1991; Thivolet et al., 1991; Bach, 1994). This lead to the hypothesis of an initiating phase, characterised by antigen presentation and recognition, followed by a perpetuation and amplification phase in which the infiltrate builds up during the insulitis process (Nerup et al., 1994). The role of the CD8 + T-cell in the initiating phase is controversial, as the CD8 + T-cell has been suggested to be necessary but not sufficiently early in the initiating phase (Serreze et al., 1997; DiLorenzo et al., 1998) whereas NOD mice lacking beta-cell class I expression show both initiation and progression of the benign insulitis process (Hamilton-Williams et al., 2003). However, recently a paper demonstrating over-expression of SOCS-1 leading to protection of CD8 + T-cell mediated beta-cell destruction indicates a role of cytokines in CD8 + function in beta-cells destruction (Chong et al., 2004).

Transplantation experiments using mixed syngenic and xenograft islets in C57BL/6 mice have been used for evaluating the effect of locally sustained exposure of islets to cytokines in vivo. The xenograft response elected a cellular infiltrate dominated by the presence of macrophages, CD4 + T-lymphocytes and eosinophils with only a small number of CD8 + cells. Within the mixed xenogenic/syngenic islet graft, irreversible impairment of first and second phase insulin response was seen, contrasting the observations of no structural or functional impairment in allogenic/syngenic islet grafts (Korsgren et al., 1994). The latter could be due to low intra-islet IL-1 production, as within the allograft rejection only very few macrophages are seen (Simeonovic et al., 1990).

At the time of the first reports of the proposed role of cytokines in beta-cell destruction, Mosmann et al suggested a dividing of the CD4 + T helper cells into two populations with contrasting and cross-regulating cytokine profiles Th1: secreting IL-2, TNF β and INF γ leading to a cell mediated (type IV) delayed hypersensitivity reaction, and Th2: secreting IL-4, IL-5, IL-6, and IL-10 mainly initiating antibody formation and inhibition of cell mediated immunity (Mosmann et al., 1986; Liblau et al., 1995). On the basis of the cytokine profiles identified above being able to either promote or inhibit diabetes development, these cytokine profiles have subsequently been characterised as being either (i) Th1 associated with a destructive insulitis process or (ii) Th2 associated with a benign insulitis process (Rabinovitch, 1994c; Charlton et al., 1995; Liblau et al., 1995; Kolb, 1997). Today, it is generally accepted that cytokine mediated beta-cell destruction is related to a Th1 associated cytokine profile. However, in MLD-STZ induced diabetes reduction and upregulation of Th2-type cytokines were more strongly associated to susceptibility and resistance, respectively, than upregulation of Th1-type cytokine levels (Müller et al., 2002).

3.2.2. Functional changes induced by cytokine exposure
in beta-cells

Rat islets exposed to cytokines were used in the initial in vitro studies. Exposing rat islets to IL-1 β as a single agent was initially shown to:

inhibit glucose stimulated insulin release (Mandrup-Poulsen et al., 1986a), (pro)insulin as well as total protein biosynthesis (Spinas et al., 1987)
decrease oxidative metabolism (Sandler et al., 1987) and glucose oxidation at the mitochondrial level and consequently decrease ATP production and Ca ++ uptake (Sandler et al., 1991)
increase DNA damage and reduce DNA content (Sandler et al., 1987; Johannesen et al., 1990; Delaney et al., 1993)

leading to the destruction and death of the beta-cells. The IL-1 mediated beta-cell destruction has been proposed to be due to production of toxic substances in the beta-cell (Thomas et al., 2002) and released within islets from bystander cells (activated MØ and endothelial cells) (Kroncke et al., 1991; Steiner et al., 1997). These effects of IL-1 were intensified by IFN γ and TNF α (Mandrup-Poulsen et al., 1987b; Eizirik, 1988). However, in purified single rat beta-cells: IL-1 failed to destruct the beta-cells (Ling et al., 1993; Hoorens et al., 2001) whereas a mixed cytokine exposure lead to destruction (Hoorens et al., 1999; Pavlovic et al., 1999b; Liu et al., 2000; Hoorens et al., 2001; Liu et al., 2001).

Initially, most studies using human islets describe neither any cytotoxic effect (Kawahara et al., 1991; Eizirik et al., 1993c; Rabinovitch et al., 1994a) nor decreased accumulated or glucose stimulated insulin release after exposure of IL-1 (Mandrup-Poulsen et al., 1987a; Vara et al., 1994). However, beta-cell destruction due to direct IL-1 exposure alone has been indicated (Giannoukakis et al., 2000) and glucose induced IL-1 β production in human islets reduced stimulated insulin secretion and increased apoptosis in beta-cells (Maedler et al., 2002). Finally, Zumsteg and co-workers have shown IL-1 mediated inhibition of glucose stimulated insulin release from human islets (Zumsteg et al., 1993), whereas other studies have shown the need of cytokine mixture to induce beta-cell destruction (Rabinovitch et al., 1994a; Delaney et al., 1997; Hoorens et al., 2001). Using monolayer human beta-cells/single cell preparations enriched in beta-cells (FACS): IL-1 alone did not cause destruction (Hoorens et al., 2001) contradictory to the increased 51 Cr release following IL-1 exposure demonstrated by (Rabinovitch et al., 1990), and contrasting the destructive effect of cytokine mixture exposure (Delaney et al., 1997; Hoorens et al., 1999; Hoorens et al., 2001).

Hence, evaluating beta-cell functional data after cytokine exposure, the experimental setting needs to be taken into account, e.g. (i) when comparing single beta-cell preparations from rats and humans the purity in rats is reported as more than 92% (Hoorens et al., 1999) contrasting 69-82% for the human enriched beta-cells single cell preparations (Delaney et al., 1997; Hoorens et al., 1999), and (ii) when comparing whole islets to single cell preparation as the yield of single cells only represents a small fraction of the total number of beta-cells (Pipeleers et al., 1985), and it should be considered that these pure beta-cells after FACS purification, might represent a selected, resistant "survivor population" of beta-cells. Furthermore, the Ca ++ concentration of the culture media influences the effect of IL-1 in mouse islets (Helqvist et al., 1989). Moreover, islet isolation procedures vary slightly from laboratory to laboratory and from isolation of rat and human islets (accepting cold-preservation hours) (Keymeulen et al., 1998). Obviously, the use of single cell preparations has the advantage of studying beta-cell specific effects, however the experimental set-up might be too simple to illustrate the pathology of the cytokine mediated beta-cell destruction in vivo.

In conclusion: In both rat and human islets, exposure to cytokine mixture has been shown to impair beta-cell function demonstrated as e.g. inhibited insulin release, destruction of DNA and induced cytotoxicity. Within islets, beta-cells added bystander cells (MØ and endothelial cells) or in single beta-cell preparations grown in high density, the IL-1 mediated beta-cell destruction is suggested to be due to high local production of toxic substances e.g. NO, contrasting the failure of IL-1 induced beta-cell destruction in single cell preparations grown in low density. Furthermore, different experimental settings and conditions as well as islet/beta-cell handling should be kept in mind when comparing data. An altered proteome profile within these settings may be demonstrated and associated to the outcome.

3.2.3. Intracellular cytokine induced pathways

Cytokine receptors on beta-cells provide the basis of cytokine induced signalling in beta-cells (Dinarello, 1997). A review of the signal transduction pathways for IL-1, IFN γ and TNF has been given by Eizirik and Mandrup-Poulsen in (Eizirik et al., 2001b) and recently in (Donath et al., 2003) (see Figure 4 ).

In short:

(i) IL-1: Binding of IL-1 to the IL-1R1 allowing docking of the IL-1 receptor accessory protein mediates signal transduction through three major pathways: activation of (a) nuclear factor kB (NF κ B), (b) mitogen activated protein kinases (MAPK) and (c) protein kinase C (PKC). However, involvement of G-proteins in IL-1 induced NO release and subsequent demise of the pancreatic beta-cell has been suggested (Tannous et al., 2002).

(ii) IFN γ : Interaction between IFN γ and IFN γ receptor 1 leads to the activation of the Janus tyrosine kinases 1 and 2 (JAK1 and JAK2), followed by activation of the signal transducer and activator of transcription 1 molecules (STAT1). STAT1 being a transcription factor translocating to the nucleus and through binding to gamma-activated sites (GAS) initiates transcription of many (hundreds) genes. Interferon regulatory factor 1 (IRF-1) a transcription factor as well, being one of the STAT1 activated genes is subsequently expressed and binds to interferon-stimulated response elements (ISRE) in other genes e.g. iNOS. Further, STAT1 regulates caspase expression and thereby influence the cellular response to pro-apoptotic stimuli.

(iii) TNF: signals through the TNF receptor. TNF belongs to a large family, also containing e.g. FasL. Two TNF receptors exists, p60 and p80, the first containing the death domain (DD), that when activated subsequently leads to activation of e.g. NF κ B, MAPKs and/or caspase mediated apoptosis. The effect of TNF will not be discussed further as the selected signalling transduction candidate gene (IRF-1) mainly operates in the IFN signalling cascade.

3.2.3.1. Nuclear Factor kappa Beta (NF κ B) and
Interferon Regulating Factor-1 (IRF-1)

In rat islets, activation of NF κ B is required for IL-1 induced iNOS expression (Saldeen et al., 1994; Bedoya et al., 1995; Flodstrom et al., 1996a; Darville et al., 1998). In unstimulated cells, NF κ B is located inactively in the cytoplasma due to binding to the inhibitor IkB. IL-1 mediated signalling leads to phosphorylation and degrading of IkB allowing NF κ B to translocate to the nucleus (Gilmore, 1999), bind and initiate or adjust the promoter activity of promoters containing a NF κ B binding sites (Pahl, 1999). Expression profiling studies also detect an increased expression of NF κ B after cytokine exposure (Rieneck et al., 2000; Mose-Larsen et al., 2001; Cardozo et al., 2001a; Cardozo et al., 2001b). Besides being implicated in the transcription of iNOS, NF κ B has been associated to other inflammatory response genes (Tak et al., 2001) like: MnSOD (Darville et al., 2000), Fas (Darville et al.), A20 (Grey et al., 1999) and IkB (Cardozo et al., 2001a). Hence, NF κ B regulates the expression of several response genes which have been suggested to be stimulus and cell-type specific (Karin et al., 2000). A substantiation of the role of NF κ B in cytokine mediated beta-cell destruction came from a study, blocking the NF κ B translocation into the nucleus by infecting rat beta-cells with a non-degradable mutated form of IkB. Cytokine induced iNOS and Fas expression was inhibited and beta-cell survival was significantly improved (Heimberg et al., 2001). Furthermore, inhibition of NF κ B in insulin producing MIN6 cells provided partial protection of IL-1 β /IFN γ /TNF α induced apoptosis, indicating a role of NF κ B in apoptosis signalling (Baker et al., 2001). Finally, NF κ B1 (p50) deficient mice are not susceptible to multiple low-dose streptozotocin-induced diabetes (Mabley et al., 2002).

Inhibition of NF κ B by pyrrolidine dithiocarbamate (PDTC) in human islets treated with IL-1 β /IFN γ /TNF α lead to inhibition of nitrite production (Flodstrom et al., 1996a), suggesting a role of NF κ B in iNOS expression in human islets as well. However, IL-1 alone also enhanced NF κ B activation, but failed to induce iNOS expression in human islets (Flodstrom et al., 1996a) indicating that NF κ B is a necessary but not sufficient factor in inducing iNOS expression. As blockage of general protein synthesis by cyclohexamide has shown to inhibit iNOS mRNA transcription in insulin-producing RINm5F and HIT-cells (Eizirik et al., 1993b) activation of protein synthesis of another transcription factor seems necessary, since activation of NF κ B is not dependent upon active protein synthesis (Grimm et al., 1993). As (i) IL-1 alone induces NF κ B (Flodstrom et al., 1996a) and IRF-1 (Johannesen et al., 2001b) in rat islets, (ii) IRF-1 requires de novo protein synthesis and has been suggested to play a role in IL-1 mediated NO production in rat islets (Akabane et al., 1995), and (iii) macrophages from IRF-1 deficient mice did not produce NO at immuno-stimulation (Kamijo et al., 1994), IRF-1 expression was suggested as the "missing link" in IL-1 β exposed human islets for iNOS expression and NO production (Eizirik et al., 1996c). On the other hand, recent studies using islets from IRF-1-/- mice in vitro and in vivo have provided conflicting results. In vitro studies using both FACS purified beta-cells and whole islets from IRF-1-/- mice suggested that IRF-1 expression, probably within the non-beta-cells present in whole islets, was involved in cytokine induced islets cell damage (Pavlovic et al., 1999b). Contradictory, when islets from IRF-1-/- mice were allografted into alloxan induced diabetic recipient mice, reduced graft survival time was observed compared to IRF1+/+ control islets, suggesting a possible protective role of IRF-1 in this in vivo model (Gysemans et al., 2001). However, recently, Baker et al have demonstrated that cytokine stimulated IRF-1 deficient islets express a T-cell chemotaxin (inducible protein (IP)-10) in higher concentrations - possibly leading to homing of T-cells and higher local cytokine concentrations - than wild type animals (Baker et al., 2003). This finding suggests an explanation of the paradox between the ability of IRF-1-/- islets to resist cytokine induced destruction in vitro and the observed accelerated graft failure in vivo.

Such experiments using single gene knock-out animals in very exact experimental designs asking specific questions give equally specific answers, illustrating that only very specific conclusions can be drawn and that caution regarding extrapolation to more general conclusions should be taken. However, IRF-1 seems to play a role in the overall outcome of cytokine exposed beta-cells, and this possible dual effect of IRF-1 is in line with data for NF κ B, as this transcription factor also has been shown to possess a role in both deleterious and protective mechanisms mediated by cytokines (Heimberg et al., 2001; Cardozo et al., 2001b). Such dual effect could limit the use of knock-ins or knock-outs regarding any putative dynamic function or critical time-window of action for the gene or protein in question.

3.2.3.2. Mitogen Activated Protein Kinase (MAPK)

As suggested above, cytokine mediated NO-independent beta-cell destruction exists in which MAPK signalling has been involved and suggested to lead to apoptosis mediated beta-cell destruction (Mandrup-Poulsen, 2001). The IL-1 β induced MAPK activity in rat islets has shown to be synergistically increased by TNF α and IFN γ (Andersen et al., 2000). The following members of the MAPK family have been identified in beta-cells (Welsh, 1996; Larsen et al., 1998): (i) extracellular regulated signal-kinase (ERK) mainly activated by mitogens, growth factors and cellular stress, (ii) p38, and (iii) c-jun N-terminal kinase (JNK) both activated by cellular stress: e.g. cytokines and irradiation (Widmann et al., 1999).

JNK has been identified as a MAPK mediator to induce apoptosis, as inhibition of JNK reduces IL-1 mediated apoptosis in beta-cell lines (Ammendrup et al., 2000; Bonny et al., 2001). Activation of JNK may be regulated by Ca ++ influx through high voltage-activated (HVA) Ca ++ channels, as blockage of HVA channels has been found to significantly reduce IL-1 stimulated JNK activity in beta-cells (Størling et al., 2001). Activation of the caspase cascade has been suggested to be MAPK mediated in beta-cells (Eizirik et al., 2001b) and by IFN γ induced ICE expression (caspase 1) in rat and human islets (Karlsen et al., 2000). Caspases are present as inactive precursors, which when activated leads to cleavage of many proteins resulting in dismantling of the cell (Thornberry et al., 1998).

Cross-talk between MAPK signalling and NO mediated beta-cell destruction has been described as the induction of iNOS - mainly being dependent upon NF κ B - also was shown to be regulated by p38 and ERK (Larsen et al., 1998; Bellmann et al., 2000). Furthermore, cytokine produced NO may positively feed-back the MAPK signal (Binzer et al., 2001), possibly explaining the protracted activation of MAPK in cytokine exposed beta-cells (Larsen et al., 1998).

In conclusion: In beta-cells cytokines elicit a variety of different signalling pathways, some leading to NO production resulting in necrotic as well as apoptotic cell death. Elucidating the functional relevance of a single factor e.g. NF κ B or IRF-1, caution needs to be taken regarding (i) the efficacy and specificity of chemical blocking of the factor, and (ii) the read-out (usually being a parameter further downstream in the pathway) due to redundancy in such complex pathways.

3.2.4. Protective mechanisms

That the beta-cell is not a passive bystander cell to its own destruction is illustrated by the various defence mechanisms activated by the beta-cell when exposed to toxic stimuli - a race between the deleterious and protective mechanisms is induced. However, a reduced stress-induced defence capacity in the beta-cell has been demonstrated (Welsh et al., 1995b; Lenzen et al., 1996; Burkart et al., 2000) and this may lead to higher susceptibility for destruction of the beta-cell compared to other cell-types (Andersen, 1999) - hence, reduced protective capacity may add to the fact, that in beta-cells the deleterious mechanisms prevail. Concordant results have been provided from a clinical setting, as the total antioxidant status was lower in ICA-positive compared to ICA-negative first degree relatives to T1DM patients (Rocic et al., 1997), and the total antioxidant activity was lower in T1DM patients compared to healthy controls (Maxwell et al., 1997).

From the expression profiling studies various cytokine or NO protective gene-transcript and/or proteins have been demonstrated, e.g. catalase, ceruloplasmine, GADD-153, Gas 5, -6, gluthathione S-transferase, gluthathione peroxidase, glutamine - γ glytanyl transferase, heme oxygenase, HSP 27, -40, -70, metallothionein, MnSOD, MX 1, SOD-B (for references see Chapter 3.1.1. "Expressing profiling"). Only a selection - those that specifically have been evaluated in insulin producing cells or in islets - will be further described here.

Heat shock proteins have been demonstrated as being one of the protective molecules in islets exposed to cytokines. The function of HSP70 in general has been associated to chaperoning (Bukau et al., 1998; Nollen et al., 1999) and in cellular defence HSP70 has been proposed to participate in repair of damaged nuclei as most HSP70 is found in the nuclei after heat shock (Welch et al., 1991). HSP70 has also been suggested to protect mitochondrial function against oxidative injury, as heat shock induced HSP70 prevented H 2 O 2 induced mitochondrial damage (Polla et al., 1996). Finally, HSP70 has been shown to provide cellular protection by interfering with apoptosis induction (Buzzard et al., 1998; Jaattela et al., 1998) possibly by inhibition of JNK and p38 (Gabai et al., 1997; Mosser et al., 1997).

In rat islets expression of a cytokine induced protein with a molecular weight of approximately 72 kDa was initially demonstrated by Helqvist et al. (Helqvist et al., 1989), and subsequently the identity of HSP72 was confirmed in IL-1 exposed mouse and rat islets (Eizirik et al., 1990; Helqvist et al., 1991a; Welsh et al., 1991b). The HSP72 expression was exclusively found in FACS sorted beta-cells and not in alfa-cells (Strandell et al., 1995). A protective role of HSP72 against the deleterious effect of IL-1 in islets was shown by liposomal delivery of HSP72 into rat islets (Margulis et al., 1991). Heat shock treatment induced increased resistance in rat islets against NO, oxygen radicals and STZ toxicity in vitro (Bellmann et al., 1995), and over-expression of HSP70 conferred resistance against NO induced (NO-donor) cell lysis (approximately 50% reduction) in RINm5f cells (Bellmann et al., 1996). In contrast, when HSP70 over-expressing RIN cells were stimulated with cytokines an enhanced p38 MAP kinase dependent increase in nitrite production was seen (Bellmann et al., 2000). This apparent paradox of HSP70 possessing chaperone as well as cytokine properties has been suggested to be due to intracellular vs. extracellular actions of HSP70, as extracellular acting HSP70 has been shown to stimulate cytokine production (Asea et al., 2000).

Another stress protein, heme oxygenase (HSP32), has also been found to be upregulated selectively in IL-1 stimulated rat islets beta-cells (Helqvist et al., 1991a; Strandell et al., 1995). Heme oxygenase has been shown to be induced and cause cytoprotection of beta-cells and other cells exposed to NO (Motterlini et al., 1996).

Besides induction of stress proteins, low expression of the antioxidant enzymes MnSOD, catalase and gluthathione peroxidase (GSH) in rodent islets (Lenzen et al., 1996; Tiedge et al., 1997) have been associated with increased susceptibility to free radicals, as antioxidant administration or over-expression of antioxidant enzymes reduced cytokine induced beta-cell function and/or destruction in rodent and human islets (Sumoski et al., 1989; Welsh et al., 1994; Tiedge et al., 1997; Tiedge et al., 1998; Tiedge et al., 1999; Lortz et al., 2000; Moriscot et al., 2000). Selective increase in MnSOD was found in FACS purified beta-cells and not in alfa-cells contrasting identical levels in both cell types when unstimulated (Strandell et al., 1995). In RIN cells, IL-1 elicited a parallel time-dependent iNOS and MnSOD mRNA expression (Bigdeli et al., 1994). Blockage of IL-1 induced iNOS expression by aminoguanidin (AG) did not inhibit MnSOD mRNA expression, and the NO-donor SNP did not induce MnSOD mRNA expression (Bigdeli et al., 1994) indicating MnSOD being upregulated by IL-1 independently of NO. Inhibited gene transcription by actinomycin D blocked the expression of both iNOS and MnSOD in contrast to inhibition of protein synthesis by cyclohexamide blocking only iNOS expression (Bigdeli et al., 1994) - indicating different mechanisms or pathways controlling the expression of iNOS and MnSOD. This was confirmed by the observation that MnSOD expression was independent of NF κ B activation (Bedoya et al., 1995). However, over-expression of MnSOD reduced cytokine-induced activation of NF κ B by more than 80% associated with iNOS activity at basal/unstimulated level and a significantly reduced iNOS protein expression compared to control (Azevedo-Martins, 2003). Generally, this low antioxidant defence capacity of beta-cells has been considered to be an important aspect of oxygen free-radical induced damage leading to beta-cell death (Ho et al., 1999).

Differences in the protective capacity between human and rodent islets have been demonstrated. Human islets have been shown to be more resistant than rodent islets to damage from NO (Eizirik et al., 1994c) alloxan, hydrogen peroxide and stretozotocin (Eizirik, 1996b). Moreover, the basal content of HSP70 and the activity of catalase and SOD have been demonstrated to be higher in human islets compared to rat and mouse islets (Welsh et al., 1995b; Burkart et al., 2000). Other protective factors may influence the observed species differences as (i) the degree of resistance to NO, alloxan and streptozotocin has been described as highest in human, less in mouse and lowest in rat, (ii) the expression of HSP70 demonstrated to be highest in human, less in rat and mouse, and finally (iii) the activity of catalase and SOD observed to be highest in human, less in rat and lowest in mouse (Eizirik, 1996b).

In cytokine stimulated FACS purified beta-cells and RINm5F cells, HSP70 has been demonstrated to up- and down-regulate by use of 2D-gel protein and mRNA array analyses, respectively (Rieneck et al., 2000; Mose-Larsen et al., 2001; Cardozo et al., 2001a). Increased expression of MnSOD was identified in cytokine treated RINm5F cells, whereas blockage of NF κ B reduced both the expression of HSP70 and MnSOD as well as iNOS in FACS purified rat beta-cells replicating the finding of NF κ B being involved in destructive as well as protective signalling pathways (Cardozo et al., 2001b). Moreover, beside the up-regulation of MnSOD and HSP70 in cytokine exposed primary rat beta-cells, downregulation of gas6 and glutathione peroxidase both representing defence/repair genes was demonstrated in beta-cells (Cardozo et al., 2001a).

Taken together, the beta-cell possess a variety of different protective capacities. These can be activated (i) directly by cytokine signalling pathways and (ii) indirectly by cytokine mediated formation of free radicals. Species and possibly strain dependent protective capacities seem to exist, although beta-cells in general seem to have low basal antioxidant levels.

Conclusions from cytokine beta-cell destruction: Since the first version of "The Copenhagen Model" of cytokine mediated beta-cell destruction was proposed, the effects of cytokine exposure to beta-cell have been extensively studied. New technologies have been used and have generated much new information. However, although the understanding of the complexity of the involved processes and their interactions has expanded significantly but has not yet been fully depicted, the initial proposed idea of a cytokine mediated race between deleterious and protective mechanisms within the beta-cell still stands.

3.3. Selection of proteins/genes
using "The combined candidate gene approach"

On the basis of the above described "Selected candidate gene approach": (i) "The Copenhagen Model", (ii) the functional derived expressional data thereof and (iii) various genome scans, three proteins have been selected for evaluation in this thesis: inducible nitric oxide synthase (iNOS), interferon regulating factor 1 (IRF1), and mortalin.

The iNOS was chosen since:
- NO was the first major cytokine mediated effector molecule identified in rat islets leading to selective beta-cell destruction
- the possibility to further characterise the role of iNOS in two rat strains previously identified as being differently sensitive to IL-1 β , and
- the un-revealed impact of iNOS/NO in human beta-cell destruction/T1DM.
As IRF1 has been shown to be involved in the cytokine mediated activation of iNOS lead to genetic characterisation of IRF1 and a descriptive role of IRF1 in the rat strains, and finally,
Mortalin being newly identified at the 2D protein gels and hypothesized to play a role in "The Copenhagen Model" of T1DM due its role in senescence and mitochondrial function. Furthermore, the gene encoding for mortalin was located to a genetic region shown to be linked to other autoimmune diseases.

Much information regarding the role of iNOS and NO in cytokine mediated beta-cell destruction was awailable at the time of selection - less information was available for IRF-1 and none for mortalin. This differentiated knowledge prior to initiation of the studies for each of the selected proteins was also considered in order to demonstrate all parts of the genetic and functional characterisation - although not necessarily for the same protein ( Table 4 ).

In order to evaluate candidate genes for association to T1DM, it is necessary to obtain information of the DNA sequence of the selected gene in order to identify testable genetic variations within the collected population. Secondly, to examine and understand a functional relevance of these genetic variants in-depth knowledge of regulatory mechanisms becomes mandatory.

Hence, the selected proteins were characterised in the following way:

The gene encoding the protein was evaluated for genetic polymorphisms and these were tested for linkage using (E)TDT analyses in a nationwide Danish T1DM family collection. Re- garding IRF-1, a previously described polymorphism was examined within the family collection.
In an attempt to compare the protein and mRNA response in two genetically different rats - expression of selected transcripts in cytokine exposed isolated islets from two rat strains (not spontaneously developing diabetes) was characterised. After genetic characterisation of the rat iNOS promoter from both strains, the iNOS promoters were functionally tested and associated to the expressed iNOS level.
Ideally, over-expressing of the selected protein in a beta-cell line followed by cytokine exposure is warranted to further characterise the effect of the protein in cytokine mediated beta-cell destruction. However, the rat mortalin over-expression studies performed in a mouse-fibroblast cell line (NIH-3T3) illustrated the effect of high mortalin expressional levels per se and allowed for comparison to over-expression studies of human and mouse mortalin.

3.4. Methods

Within the genetic orientated papers: cloning, screening for and verification of polymorphisms as well as establishing typing assays - generally accepted techniques and methodology were used and are not discussed further. The genetic analyses have concentrated upon association and linkage to T1DM in a Danish nationwide collection by methods previously described. Papers focusing on islet responsiveness to cytokines and functional evaluation of expression of selected proteins were also based upon classical and accepted methodology. However, promoter activity assays and real-time PCR have been introduced in the laboratory and used according to the manufacture's description and guidelines. Naturally, the use of promoter assay in cell lines comprises confounders: in cell lines - not fully resembling the relevant naïve cell type - possibly employment of different signalling transduction pathways might affect the regulation of the promoter in question. The systematic confounding factors in reporter assays being inter- and intra-assay transfection efficacy, inter-assay differences in reporter signal due to e.g. variation in plasmid DNA constructs and quantification, have been controlled to the extent possible in the studies.

Conclusion from Chapter 3

"The combined approach to select candidate genes" has the strength of employing multi-string identification of susceptibility genes - one being the response of the target organ to cytokine exposure. Three candidate genes have been selected. The following chapter presents the evaluation in terms of (i) association to T1DM in a nationwide Danish T1DM family collection preceeded by a seach for genetic variants within the genes, (ii) expression of these genes in cytokine exposed islets from two rat strains in order to study inter-strain target organ responses of cytokine mediated beta-cell destruction. Finally, (iii) an over-expression study of rat mortalin demonstrates the effect of mortalin per se and allowed for comparison to over-expression studies for human and mouse mortalin, although over-expressing of the selected protein in a beta-cell line followed by cytokine exposure is warrented to further characterise the effect of the protein in cytokine mediated beta-cell destruction.

4. Genetic and functional analysis
of the selected candidate genes

In this chapter the selected candidate genes - iNOS, IRF-1 and mortalin - will be reviewed. As most of the papers contributing to this thesis describe aspects of rat and human iNOS in cytokine mediated beta-cell destruction as a model for T1DM, the review of the iNOS gene and protein comprises the majority of this chapter. Hence, the section "Modes of cytokine mediated beta-cell destruction" has been included here as the chapter has special focus on iNOS/NO in beta-cell destruction. Genetic characterisation of iNOS, IRF-1 and mortalin within the Danish nationwide T1DM collection will be demonstrated, descriptive evaluation of the expression in cytokine exposed islets from two rat strains, as well as functional evaluations of the candidate genes expressed in the rat.

4.1. The inducible nitrite oxide synthase (iNOS)

Nitric oxide (NO) formation is generated by the enzymes of nitric oxide synthase (NOS) family converting L-arginine to citruline and NO. It is a potent biologic mediator of diverse physiologic and pathophysiologic effects. It has been implicated in blood pressure regulation, neurotransmission, antimicrobial defence mechanisms, modulation of inflammatory response (Moncada et al., 1991) and autoimmunity (Bogdan, 1998; Singh et al., 2000) in part by modulating the Th1/Th2 response (Taylor-Robinson et al., 1994; Nukaya et al., 1995; Wei et al., 1995; Kolb et al., 1998; Niedbala et al., 1999). The NOS family comprises: neuronal NOS (nNOS or NOS1), the inducible NOS (iNOS or NOS2), and endothelial NOS (eNOS or NOS3). The nNOS and eNOS are constitutively expressed (cNOS), named after the cells in which they were originally discovered, synthesise NO dependent on calmodulin (CaM) and Ca ++ (Nathan et al., 1994), function in signal transduction cascades by linking temporal changes in Ca ++ level to NO production, and serves as activators of soluble guanylate cyclase (Ignarro et al., 1995). Both cNOS isoforms participate in homeostatic cell to cell signalling and are regulated independently of the inflammatory responses (Bredt et al., 1991; Sessa et al., 1992). Induced NO production was initially identified in LPS stimulated MØ (Stuehr et al., 1985), and iNOS expression requires de novo protein synthesis following cellular stimulation by LPS or cytokines (Hughes et al., 1990; Eizirik et al., 1991). The enzyme is predominantly soluble (Hevel et al., 1991), and binds CaM tightly even in absence of Ca ++ , hence being Ca ++ independent (Cho et al., 1992). Furthermore, the enzyme produces much larger amounts of NO when stimulated than the cNOS's (Cho et al., 1992). Once produced, NO quickly (T 1/2 are seconds) undergoes spontaneous oxidation to the inactive metabolites nitrite and nitrate (NO 2 - and NO 3 - ). iNOS has been implicated in numerous human diseases, including neurodegenerative, autoimmune, cardiovascular, inflammatory and a number of human cancers, for review see (Kröncke et al., 1998). Even though iNOS expression appears to have many beneficial roles in the acute septic response (e.g. hepatoprotection (Taylor et al., 1998)), over-expression can be detrimental (Szabo et al., 1994; Cobb et al., 1996).

4.1.1. Modes of cytokine mediated beta-cell destruction

In general, cytokine mediated beta-cell destruction involves the toxicity of generated free radicals (FR) e.g. reactive oxygen species (ROS), which have the capacity to oxidize and thereby damage cellular components (Chapple, 1997). Besides the toxic effect of ROS, oxidation of proteins may turn them into autoantigens thereby initiating/continuating the immune reaction toward the beta-cell (Karlsen et al., 1998). In rat islets, the majority of the inhibitory effects and toxicity of cytokines are believed to be mediated by expression of free radicals generated in the beta-cells - e.g. the nitrous oxide radical formation - by expression of the inducible form of nitric oxide synthesis (iNOS) and consequently the synthesis of nitric oxide (NO) (Southern et al., 1990), reviewed by (Nerup et al., 1994; Mandrup-Poulsen, 1996; Eizirik et al., 1996c; Eizirik et al., 1997).

The toxic effects of cytokines exposed to beta-cells have been shown to lead to necrosis as well as apoptosis. This chapter will demonstrate that NO can lead to both forms of cell deaths. Besides the induction of ROS - mainly leading to necrosis, the transcription factors JNK and NF κ B have been demonstrated to be essential regulators of cytokine signalling leading mainly to apoptotic cell death (See 3.2.3 "Intracellular cytokine induced pathways"). Furthermore, cytokines have been shown to activate Ca ++ channels and caspases inducing apoptotic cell death (for review see also (Bergholdt et al., 2003)).

4.1.2. Nitric Oxide - necrosis and apoptosis -
in cytokine mediated beta-cell destruction

Necrosis is usually the result of acute cellular dysfunction in response to massive cell injury caused by sudden severe ischemia, chemical, physical or thermal injury, leading to loss of the selective permeability of the cell membrane (Majno et al., 1995). Due to disruption of membranes, cellular content and material are released to the exterior triggering an acute inflammatory response by attracting proinflammatory cells (Gores et al., 1990; Haslett, 1992). Necrosis is generally considered a passive, non-energy dependent process associated to rapid cellular ATP depletion. In islets exposed to IL-1 a down-regulation of an ATP- syntheses subunit has been observed (Mose-Larsen et al., 2001; Sparre et al., 2002), however, whether this is a primary or secondary effect is unknown.

As previously described, NO was the first effector molecule identified to mediate the deleterious effects of cytokine mediated beta-cell destruction as a pathogenetic model for T1DM (Southern et al., 1990). This has further been substantiated by:

exposure of NO donors to rat islets (Kroncke et al., 1993; Cunningham, 1994; Sjöholm, 1996; Eizirik et al., 1996a) and beta-cells (Dimatteo et al., 1997) leading to beta-cell destruction
over-expression of the iNOS gene under the RIP (Takamura et al., 1998) mimicking IL-1 mediated beta-cell destruction, and
blockage of NO production (Andersen et al., 1996) protecting IL-1 exposed beta-cells. Moreover,
using the mRNA display technique in cytokine exposed insulin producing cells up-regulation of iNOS and AS (see below) was observed (Rieneck et al., 2000; Cardozo et al., 2001a; Cardozo et al., 2001b; Nielsen et al., 2004).

As iNOS catalyses the reaction arginine to citrulline and NO, the amount of generated NO depends upon availability of arginine. Transportation of arginine has been demonstrated to cross the cell membrane into the cell by use of the transport system y+CAT and being synthesised intracellularly from citrulline by argininosuccinate synthase (AS) (citrulline-NO cycle) or by protein degradation (Morris et al., 1994). IL-1 has been shown to increase the expression of AS (Flodström et al., 1995), to inhibit the enzyme arginase converting arginine to arnithine and urea (Cunningham et al., 1997), and to increase the transport of arginine into rat beta-cells (Flodstrom et al., 1999a). All these IL-1 mediated effects increase iNOS substrate availability securing the NO production.

NO has been identified to:

nitrosylate the Fe-S center of the aconitase enzyme in the Krebs cycle thereby inactivating its function (Welsh et al., 1991a) and
induce DNA strand breaks in mitochondrial DNA (Wilson et al., 1997), probably contributing to the initially described IL-1 mediated reduction of glucose oxidation and mitochondrial organelle dysfunction. Furthermore, NO has been shown to
induce nuclear DNA strand breaks in rat islets (Delaney et al., 1993), a process activating the enzyme poly(ADP-ribose) polymerase (PARP), participating in DNA repair process but consuming nicotinamide adenine nucleotide (NAD) thereby further depleting the cell of energy. In rat and man, nicotinamide, an inhibitor of PARP activation, has been shown to reduce cell loss in islets exposed to NO donors (Radons et al., 1994; Eizirik et al., 1996a). Finally, PARP deficient mice have been shown to be less sensitive to NO and FOR mediated cell death (Heller et al., 1995).

Besides the functionally inhibitory and deleterious effects of NO itself, reaction between NO and superoxide leads to the toxic radical peroxynitrite (ONOO - ) reported to be an even more potent oxidant and cytotoxic mediator than superoxide or nitric oxide (Szabó, 1996). Peroxynitrite has been identified in NOD mice infiltrated by mononuclear cells (Suarez-Pinzon et al., 1997) and inhibition of iNOS and scavenging peroxynitrite prevented diabetes development in NOD mice (Suarez-Pinzon et al., 2001) as well as in the multiple low-dose streptozotocin induced diabetes model (Mabley et al., 2004). Peroxynitrite has also been identified in human islets exposed to cytokine mixture (Lakey et al., 2001). Human islets exposed to peroxynitrite display acute DNA strand break and decreased glucose metabolism leading to cell death (Delaney et al., 1996).

In addition to the direct tissue damage mediated by NO, a report has suggested that NO may enhance and/or preserve the Th1 cytokine profile in the NOD mouse. The Th1 response is activated by IL-12, a MØ cytokine, and suppression of the IL-12 production in NOD may inhibit the progress of the initial benign Th2 insulitis to the destructive Th1 insulitis process, reduce iNOS mRNA expression and decrease diabetes incidence, for review see (Rothe et al., 1999). Thus it seems that NO production facilitates and maintains the destructive Th1 insulitis process. Finally, it has been shown that activated MØ facilitates islet destruction by CD8 + T-cells through a NO synthesis-dependent pathway (Gurlo et al., 1999).

Hence, NO or derivatives thereof lead to acute cellular dysfunction depleting the cell of energy. Results from iNOS -/- transgenic mice have suggested that NO is predominantly involved in necrosis and not apoptotic cell-death (Liu et al., 2000). However, the same study also recognised the existence of NO-independent effector mechanisms as beta-cell destruction was only partly protected in the iNOS -/- beta-cells following cytokine exposure.

Apoptosis or programmed cell death is an energy requiring process (Cummings et al., 1997), naturally occurring during embryogenesis and in normal tissue turnover and constitutes a common mechanism of cell replacement, tissue remodelling and removal of damaged cells (DeLong, 1998). Morphologically, apoptosis is characterised by condensation and margination of the chromatin towards the nuclear membrane, cellular shrinkage, detachment from neighbouring cells, inter-nucleosomal DNA fragmentation and formation of "apoptotic bodies". These apoptotic bodies are almost immediately phagocytosed, preventing exposure of cellular content to the exterior and thereby inflammatory response.

In line with the original concept proposed in "The Copenhagen Model" suggesting that cytokine exposure induces a race between deleterious and protective mechanisms, (Nerup et al., 1994) it follows that cytokine exposure initiates many different responses within the beta-cell. Besides, as indicated from the different signalling pathways activated by IL-1 β , INF γ and TNF α it would seem unlikely that only one cytokine mediated effector arm should exist.

Indeed, apoptosis can be demonstrated in beta-cells: in the post-partum pancreas (Scaglia et al., 1995), in the neonatal pancreas (Scaglia et al., 1997), in response to hyperinsulinemia induced by transplantation of an insulinoma (Blume et al., 1995) and in islets where glucose promotes survival of rat pancreatic beta-cells by activating synthesis of proteins which suppress a constitutive apoptotic program (Hoorens et al., 1996).

In the literature, reports suggesting NO independent apoptosis exist in islets, as:

blocking NO-production does not fully inhibit cytokine mediated apoptosis (Eizirik et al., 1994a; Rabinovitch et al., 1994a; Delaney et al., 1997; Hoorens et al., 2001), and
apoptosis has been detected in cytokine exposed islets from iNOS-/- mice (Liu et al., 2000; Zumsteg et al., 2000)

It has been suggested that cytokines can induce Fas expression upon the cell surface of the beta-cell (Stassi et al., 1997). Consequently, Fas/FasL interaction between the beta-cell and T-cell (CD4 + and CD8 + ) present in the insulitis infiltrate may activate caspases leading to apoptosis of the beta-cell. This initial finding has subsequently been challenged, and the involvement of Fas/FasL as effector molecules for beta-cell destruction in T1DM remains controversial, for review see (Eizirik et al., 2001b). Furthermore, cytokines have (i) shown to induce MAPK (Larsen et al., 1998) and caspase 1 (ICE) (Karlsen et al., 2000) in beta-cells, and (ii) been suggested to mediate beta-cell apoptosis, which lead to focusing at related signalling pathways and apoptotic effector mechanisms as putative mediators of beta-cells death.

However, apoptosis in islets/beta-cells has been identified following exposure to NO, peroxynitrite or cytokine mediated NO-effects (Mabley et al., 1997; Hadjivassiliou et al., 1998; Saldeen, 2000).

Moreover, other reports suggest NO influence on the apoptotic process, as:

blocking NO synthesis leads to reduced PARP-cleavage (indicator of apoptosis) after 24h exposure of islets to IFN/TNF/IL-1, and reduced number of necrotic and apoptotic cells in the islets significantly (Saldeen, 2000)
NO/oxidative stress decreased redox function modifying the cytokine-induced apoptotic pathway (Stamler, 1994; Dimatteo et al., 1997; Hampton et al., 1998)
NO induced DNA strand breaks may induce apoptosis per se (Ankarcrona et al., 1994; Kaneto et al., 1995) or through activation of the tumour suppressor protein p53 (Messmer et al., 1994)
Endoplasmatic reticulum stress (perturbations leading to accumulation of malfolded proteins in that compartment) has been suggested to activate JNK in non-beta-cells, however this coupling is not understood (Urano et al., 2000), and NO induced ER stress could be coupled to the pro-apoptotic JNK pathway,
NO is needed to induce apoptosis in FACS purified rat beta-cells by combinations of viral products and cytokines (Liu et al., 2001), and finally
NO induces ER depletion of Ca ++ leading to ER stress and subsequent induction of apoptosis by the CHOP apoptosis (a C/EBP homologous protein, induced by ER stress and plays a role in growth arrest and cell death) (Oyadomari et al., 2001),

Hence, it seems that beta-cell destruction involves NO as well as non-NO-dependent effector arms leading to both necrosis as well as apoptosis. As both forms of beta-cell destruction have been demonstrated, factors influencing the outcome are of interest. Different study designs may help identify such factors - however, the mode of destruction may be influenced by the study design, e.g. beta-cell single cell suspensions versus islet studies. In rat single cells, both apoptosis and necrosis occur, and full apoptotic effect and necrotic index were observed at relatively low cytokine mix concentrations, possible due to limited numbers of cytokine receptors (Eizirik et al., 2001b). In contrast, in rat islets exposed to cytokine mixture a higher increment in necrosis than in apoptosis was seen (Saldeen, 2000), possibly due to higher intra-islet concentrations of NO produced by both beta-cells and non-beta-cells. A study combining exposure of IL-1 and doubled stranded RNA (dsRNA), imitating virus exposure/infection induced NO-dependent apoptosis in contrast to IFN γ + dsRNA exposure leading to NO-independent death of unknown pathways (Liu et al., 2001), illustrating that different agents can trigger various destructive pathways. Further, the potency of the "hit" versus the defence properties may influence the destructive pathway taken as more severe attack tends to lead to necrosis, as the cell depletes from energy to fulfil the apoptotic program (Lemasters et al., 1999). This may partly explain the finding of apoptosis being more pronounced in human than in rodent islets (Delaney et al., 1997; Hoorens et al., 1999; Hoorens et al., 2001) as (i) human islets are better protected against oxidative stress (see elsewhere) partly due to higher cellular amounts of HSP70 (Welsh et al., 1995b; Burkart et al., 2000) serving an anti-apoptotic effect (Jaattela, 1999), and (ii) a better capacity to continue glucose oxidation resulting in higher ATP-production (regarding human islets, see (Eizirik et al., 1994a) and mouse islets, see (Cetkovic-Cvrlje et al., 1994)), necessary to fulfil the apoptotic program, despite similar amounts of NO are produced (Eizirik et al., 1997). Heterogeneity within beta-cells may also contribute to the outcome. Heterogeneity has been described in FACS purified rat beta-cells (Pipeleers, 1992) - (i) diverse sub-population of beta-cells differently responding in insulin secretion at identical glucose concentration, (ii) high glucose sensitivity associated to high general protein synthesis - could serve as an example of the impact of beta-cell phenotype or functional state. High glucose sensitivity has been proposed to provide a better anti-apoptotic protein response due to the general induced protein synthesis of glucose in beta-cells (Hoorens et al., 1996). On the other hand, islets exposed to high glucose concentrations and high insulin secretion were more sensitive to the deleterious effects of IL-1 exposure compared to low glucose concentrations - suggesting the beta-cell to be "a moving target" (Helqvist, 1994) - a phenomenon not only related to high insulin secretion, as the IL-1 sensitivity were dependent upon the stimulus leading to insulin secretion (Johannesen et al., 1990). This is in line with higher iNOS expression in "high glucose-responsive FACS-purified beta-cells" indicating intercellular differences of beta-cell responsiveness to IL-1 related to the beta-cell glucose-responsiveness (Ling et al., 1998) supporting the existence of variation in beta-cell phenotype. Finally, IL-1 has been demonstrated to induce beta-cell adaptation shown as a reduced cellular sensitivity to conditions that cause necrosis but not to cytokine induced apoptosis (Ling et al., 2000). This adaptation seemed to be independent of NO production as these findings were confirmed in arginine free conditions as well as independent of heme oxygenase and HSP70 as these proteins were not elevated in arginine free condition (Ling et al., 2000).

Lessons from the BB-rat have demonstrated necrosis being the predominant type of islet cell death during development of insulin-dependent diabetes (Fehsel et al., 2003).

Taken together: In beta-cells, NO and NO-independent induced necrotic and apoptotic destruction takes place following cytokine exposure. Whether the necrotic or the apoptotic process - or both - are effectuated may be influenced by e.g. the potency of the "hit" versus the defence properties. Hence, the beta-cell destructive process is dependent upon e.g. (i) the stimuli exposed to the beta-cell and (ii) the functional state of the beta-cell and (iii) possibly influenced by beta-cell heterogeneity. The mode of beta-cell death is still controversial and hence, further studies into these areas e.g. illumination of intercellular, -species or -individual differences - are needed to develop testable preventional actions to diminish cytokine mediated beta-cell destruction.

4.1.3. Intercellular, -species and -individual differences
affecting cytokine cytotoxicity

Differences in cytokine sensitivity have been described between single cells and isolated islets. Moreover, heterogeneity among FACS purified rat beta-cells regarding glucose sensitivity has been suggested to be associated with pro-apoptotic protein response (Hoorens et al., 1996) - suggesting that variation in sensitivity to cytokines in beta-cells may depend upon the beta-cell itself and other cell types in the islet. Only very few phenotypic characteristics are available regarding beta-cell age. In 2001, Bonner-Weir speculated whether the secretory and biosynthetic heterogeneity of FACS purified beta-cells was influenced by the age of the beta-cells (Bonner-Weir, 2001) - hence, the age of the beta-cells could influence cytokine susceptibility. Furthermore, differences in cytokine susceptibility have been demonstrated comparing a beta-cell line to a pre-beta-cell line, the former being the most sensitive (Nielsen et al., 1999). Finally, induction of cardiac iNOS expression increases with age in rats (Rosas et al., 2001). However, no difference in cytokine sensitivity was demonstrated in vitro using neonatal versus adult islets (Mandrup-Poulsen et al., 1987).

As previously described, inter-species differences exist between human and rat islets regarding:

the ability of IL-1 vs cytokine mix capable of leading to beta-cell destruction, initially associated to IL-1 not being able to induce iNOS and NO in human islets, in contrast to human hepatocytes (Geller et al., 1995) where IL-1 can induce iNOS expression indicating tissue differences as well,
different defence capacities, and
different capacity to continue glucose oxidation leading to higher ATP concentrations possibly favouring apoptosis being the leading destructive process in human beta-cells in contrast to necrosis in rat beta-cells.

Moreover, different sensitivity towards NO has been proposed as it was previously shown that inhibition of iNOS expression failed to protect human islets against the deleterious effects of cytokine mixture exposure (Eizirik et al., 1994a; Delaney et al., 1997). However, the evidence of the toxic effect of peroxynitrite in human beta-cells argues for a role of NO in human beta-cell destruction, as inhibition of iNOS did not abolish peroxynitrite formation, possible due to an unchanged basal NO production, and did not prevent beta-cell destruction (Lakey et al., 2001). Besides, NO donors are able to destroy beta-cells in human islets (Eizirik et al., 1996a), and as iNOS induction is detected in human beta-cells (Arnush et al., 1998) and by neighboring non-beta-cells (Pavlovic et al., 1999), it might be speculated that cytokines can lead to deleterious local intra-islet concentrations of NO. Furthermore, equal susceptibility towards NO-donors in human islets and rat beta-cells has been demonstrated (Delaney et al., 1996; Hoorens et al., 2001).

Comparing human and bovine islets - bovine islets being less susceptible to damage by human cytokines compared to human islets (Piro et al., 2001) - demonstrates inter-species differences.

Besides inter-species differences in islet and beta-cells, alveolar macrophages from rat, hamster, monkey and man have been examined under identical experimental conditions. Clear differences between rodent and the primate species were demonstrated in iNOS expression and nitrite production after LPS and IFN γ stimulation (Jesch et al., 1997).

The different phenotypic characteristics within species may be influenced by genetic variation, as exemplified by the different iNOS regulation in macrophages from chicken of different genetic backgrounds (Hussain et al., 1998). Finally, the islets from diabetes-resistant BB rats have been shown to mount a HSP70 response after heat stress in contrast to the diabetes-prone BB rat. (Bellmann et al., 1997). The lack of a protective stress response in islet cells from diabetes-prone BB rats could be important for initiation or propagation of the disease process.

In vivo as well as in vitro, strain-dependent differences in cytokine responsiveness have been demonstrated between two rat strains, Brown Norway and Wistar Kyoto (Reimers et al., 1996). This difference has been found to be associated to different IRF-1, iNOS and HSP70 expression levels, whereas no difference in IL-1R1 expression could be demonstrated (Johannesen et al., 2001b). The study design, however, did not allow the conclusion that a causal relation between IRF-1 and iNOS exists, but it was speculated that polymorphisms in the IRF-1 gene, as well as quantitative differences in the transcriptional regulation could be involved.

In conclusion: Inter- and intra-individual differences and heterogeneity among beta-cells may potentially influence the cytokine sensitivity and might correlate to defence capacities and the level of the induced oxidative stress. These differences could e.g. be influenced by genetic factors hence, a genetic evaluation of involved proteins could increase our understanding of some of the inter- and intra-individual differences.

4.1.4. Genetic structure of the rat iNOS gene

The remaining part of this chapter will describe the structure and functional regulation of the rat and human iNOS gene. The iNOS gene regulation in insulin producing cells and different iNOS promoter sequences within the BN and WKY rat strains will be in focus. Furthermore, in order to understand the genetic impact of iNOS in T1DM an evaluation of the human iNOS gene sequence becomes mandatory as identification of sequence variations are needed to test for genetic association - here illustrated by transmission disequilibrium within the Danish T1DM family collection.

In 1993, Nunokawa was the first to clone the coding sequence of rat iNOS gene (Nunokawa et al., 1993) and in 1996 the first part of the rat iNOS promoter was cloned first by Eberhardt (Eberhardt et al., 1996). The structure of the rat iNOS gene is outlined in Figure 5 .

4.1.4.1. The rat iNOS: promoter region

In the rat iNOS promoter, more than 20 transcription-binding factor (TFB) sites are known and represent (i) LPS-related response elements (NF-IL6 and NF κ B (Lowenstein et al., 1993)), (ii) IFN γ -related response elements (IRF-1 and STAT1 (Lowenstein et al., 1993) (Teng et al., 2002)) and (iii) IL-1 β -related response elements (NF κ B and C/EBP (Teng et al., 2002)). Homology of the rat iNOS promoter from different strains is high: > 95% (Johannesen et al., 2003), see Figure 6 , but decreasing when comparing the rat iNOS promoter to the iNOS promoter of mouse and human (73% and 55% homology, respectively) (Zhang et al., 1998).

Whether the observed sequence differences represent tissue-dependent or intra-strain-dependent differences or "simple sequence inconsistency" is unknown. Two studies using different tissue sources have compared the iNOS promoter from different rat strains, and independently identified a GT-repeat polymorphism in position -1685 to -1634 from the transcription start site (Deng, 1998; Johannesen et al., 2003) - suggesting the difference to be strain and not tissue-dependent. Comparing the BN-rat to WKY-rat, a polymorphism in position +222 within exon 1 was identified (Johannesen et al., 2003).

In Table 5 " Rat iNOS promoter cloning and function ", the structural and functional findings of different rat iNOS promoters cloned from different tissues and tested in various cell-types are listed.

Expressional control of the iNOS promoter has been shown to be tissue and/or cell as well as species specific. The mouse MØ iNOS promoter organisation are characterised by two distinct regions of importance, Region I and Region II, in the initial 1.2 kb of the promoter conferring full promoter activity (Lowenstein et al., 1993). Within the rat and the human iNOS promoter, 3.2 kb and 16 kb, respectively, are needed for the highest promoter activation (Vera et

al., 1996a; Zhang et al., 1998). Further, the JAK/STAT pathway mediates the LPS/IFN γ induced iNOS expression in mouse RAW294.7 cells (Gao et al., 1997), whereas inhibition of the JAK/STAT pathway in RASMC enhance iNOS induction by LPS / IFN γ (Marrero et al., 1998). The latter could also be explained by cell-specificity, as IL-1 β alone is able to induce iNOS expression in cultured RASMC (Kanno et al., 1993; Koide et al., 1994), but neither in rat pulmonary SMC (Nakayama et al., 1992) nor in the SMC cell-line A7r5 (Spink et al., 1995). Further, testing the same mouse iNOS promoter clone in three different cell-types - RAW264.7, VSMC and RASMC, the two first systems by LPS and the latter by cytokine mix - demonstrated that different TBF sites were of importance. In RAW264.7 and RASMC, the lower NF κ B site was indispensable whereas it was the upper NF κ B site in VSMC (Xie et al., 1994; Spink et al., 1995; Perrella et al., 1996). Finally, a 1.7 kb rat iNOS promoter in RASMC gave a 13 fold induction by cytokines (Zhang et al., 1998), whereas in Swiss3T3 cells only 3-4 fold induction was observed for the same promoter sequence and stimulation (Eberhardt et al., 1996). In 2001, Zhang demonstrated that the iNOS promoter activity of the initial 3.2 kb rat iNOS promoter was only negative feed-back regulated by NO within the MØ, in contrast to findings within the RASMC (Zhang et al., 2001).

Much work has focused upon cytokine induced iNOS expression and modulation hereof in insulin producing cells (see previous chapters). Only few studies have explored iNOS promoter gene regulation in insulin producing cells using promoter activity assays. Early studies using deletional constructs of the rat iNOS promoter to identify significant TFB sites revealed that NF- κ B, GAS, ISRE binding-sites were crucial in the cytokine mediated NO-dependent pathway (Darville et al., 1998). Again, cell specificity was observed, as IFN γ (inducing the transcription factors STAT1 α and IRF-1, binding to GAS and ISRE, respectively,) enhanced IL-1 β mediated iNOS promoter activity in RINm5F cells whereas in primary rat beta-cells, IFN γ neither increased the iNOS promoter activity nor iNOS mRNA expression, but did induce a two-fold increase in NO (Darville et al., 1998). Bellmann showed that over-expression of HSP70 led to enhanced IL-1 β induced rat iNOS promoter activity (testing 1 kb of the promoter) in RIN cells through an increased activity of MAPK p38 (Bellmann et al., 2000). Further, based upon the model of viral induced beta-cell destruction, a synthetic dsRNA (PIC) in combination with IFN γ was able to induce NF κ B dependent iNOS promoter activity in primary rat beta-cells (Liu et al., 2001). Finally, two studies describe abrogation of cytokine induced iNOS promoter activity: (i) the suppressor of cytokine signalling-3 (SOCS-3) abrogated the rat iNOS promoter activity (1.8 kb) in rat INS-beta-cells (Karlsen et al., 2001) and (ii) the pituitary adenylate cyclase-activating polypeptide (PACAP) abrogated the mouse iNOS promoter activity (1.6 kb) in the mouse beta-cell line, β TC cells (Sekiya et al., 2000).

As depicted above, structural features within the iNOS promoter as well as the cellular environment in which the iNOS promoter operates influence iNOS promoter activity. In an attempt to by-pass the influence of cellular environment, Johannesen et al. tested the rat iNOS promoters from two different rat strains (BN and WKY), in the same test-cell system (Johannesen et al., 2003). Previously, isolated islets from these rat strains were identified as having IL-1 mediated strain-dependent nitrite, iNOS mRNA and protein expression profiles (Johannesen et al., 2001b). Cloning and sequencing of the iNOS promoters identified two polymorphisms within the promoter region spanning -1744 bp to +333 bp. The upper GT-repeat polymorphism gave rise to a WT-1 (-KTS) TFB site (Bickmore et al., 1992) in the BN rat strain approximately 1650 bp upstream the promoter. For details regarding Wilm's Tumor, please see Table 6 ).

Strain-dependent and IL-1 dose-response of the tested iNOS promoter sequence spanning -1744 to +267 was demonstrated in a luciferase assay co-expressing the transcription factor WT-1. The promoter activity assay revealed higher iNOS promoter activity of the BN than of the WKY iNOS promoter (Johannesen et al., 2003), whereas data generated in vitro from rat islets culture IL-1 dose- and time dependently revealed higher iNOS mRNA and protein expression levels and nitrite production from WKY islets (Johannesen et al., 2001b). This apparent controversy needs to be evaluated in the light of: (i) not full length rat iNOS promoter was tested and hence, additional promoter differences between these two rat strains may exist - indirectly evidenced by the fact that probably due to sequence variations it was not possible to construct a common upper cloning primer further 5' upstream than the one used. (ii) Information regarding the role of the 3' UTR in mRNA stability should be explored, and finally (iii) indeed, the differences in BN and WKY intracellular milieu may influence the respective promoters as illustrated by the findings of Darville (Darville et al., 1998).

4.1.4.2. The rat iNOS: cDNA/gDNA region

In Table 7 " Rat cDNA/gDNA iNOS cloning " studies characterizing the iNOS cDNA are listed.

As seen, the homology between the various iNOS clones from different cell-types is very high ( > 99%), as is the homology of iNOS between rat and mouse MØ, (approximately 92%). The cytokine induced iNOS gene sequence in rat islets is identical to other rat iNOS sequences from other tissues (Karlsen et al., 1995). The translation initiation codon has been located in exon two (Keinanen et al., 1999), as in the mouse and in the human gene (Chartrain et al., 1994). The stop codon is placed in e27 leaving a 3'UTR of 495 bp in length (Keinanen et al., 1999). Sporadic base-pair mutations/mismatches have been identified between the separate clonings when compared to each other, some leading to amino acid changes. None so far have involved known co-factor binding sites. Whether any functional relevance exists for these variations is unknown at present. Co-factor binding sites as depicted in the figure "Rat iNOS gene structure" include CaM, FMN, FAD, NADPH, Heme. Each binding site appears to lie in separate exons, except for CaM, spanning e13 and e14 (Keinanen et al., 1999).

In conclusion: The studies regarding the rat and mouse iNOS promoters have revealed (i) intra- and interspecies differences in genomic sequence, (ii) a complex regulatory mechanism controlling the promoter activity involving various transcription binding sites, and (iii) an intra- and interspecies dependent functional regulation. These differences might influence the different levels of iNOS expression demonstrated within the islets from BN and WKY rats, hence being of importance to the different response of cytokine exposure between these two rat strains.

4.1.5. Genetic structure of the human iNOS gene

The human iNOS cDNA was initially cloned from LPS and cytokine stimulated hepatocytes by Geller at al. (Charles et al., 1993; Geller et al., 1993; Sherman et al., 1993; Hokari et al., 1994). Subsequently, a variety of human cell-lines and cell-types have been shown to express iNOS, including human pancreatic islet cells (Flodstrom et al., 1996a; Corbett et al., 1996b; Flodstroem et al., 1997; Arnush et al., 1998; Scarim et al., 1998; Pavlovic et al., 1999; Karlsen et al., 2000; Chen et al., 2001; Heitmeier et al., 2001) - each of these cDNAs shows > 99% homology to the human hepatocyte sequence, for review see (Taylor et al., 2000).

The genetic structure, chromosomal localisation including the promoter region are outlined in Figure 7 .

4.1.5.1. The human iNOS: promoter region

A number of different groups have cloned and functionally tested various parts of the human promoter region. Promoter activity has been identified as far as 16kb upstream of the transcription start site (Vera et al., 1996a).

The existence of several transcription-binding factor (TBF) sites (cytokine-response elements (CRE)) have been shown (Vera et al., 1996a). More than 30 putative TBF sites are identified within the first 1.5 kb, although this region does not exhibit any significant activity in promoter activity analysis probably due to nucleotide exchanges within the LPS/IFN γ -responsive region leading to hypo-responsiveness of LPS/IFN γ (Zhang et al., 1996; Spitsin et al., 1997). However, putative cytokine response elements are identified within the interval -3.8 kb to -16 kb: AP-1, NF κ B, γ IRE, NF-IL6, GAS, IRF-E, ISRE, TNF-RE Oct-1 and STAT1 (Vera et al., 1996a; Linn et al., 1997; Chu et al., 1998).

Functional analysis of the promoter region: The initial studies elucidating the transcriptional control of iNOS used murine MØ. Two regions of importance were identified within the first 1.0 kb upstream from transcription start site (Lowenstein et al., 1993; Xie et al., 1993): an NF-KB site at -85 to -76 (Xie et al., 1994a) and an IRF-E/ISRE at -923 to -913 (Martin et al., 1994), the latter serving as an enhancer site, as IFN γ or LPS exposure to this site alone could not induce promoter activity (Lowenstein et al., 1993). In contrast, the proximal 16kb from transcription start site of human NOS2 cloned from hepatocytes was needed to show maximal activity when tested in the human hepatocyte cell line AKN-1 (Vera et al., 1996a). Based on several different promoter activity assays (luciferase and CAT assays) various central areas have been identified:

-3.8 kb to -16 kb, especially -7.0 kb to -16 kb: no identification of specific elements (Vera et al., 1996a)
NF κ B site at position -115 to -106 (Nunokawa et al., 1996)
-10.7 to -8.7 kb, potential transcription factor binding sites identified: 2xOct-1, 3xIRF-1, 4xSTAT1, 3xAP-1 and 2NF- κ B (Linn et al., 1997)
5xNF-KB sites important in -7.2 to -4.7 kb, especially at -5.8kb verified by site-directed mutagenesis (Taylor et al., 1998)
an area of inhibition at -7.3 to -6.8 kb (Chu et al., 1998), and between -351 and -632 (Pance et al., 2002)
importance of the AP-1 sites (-5301 and -5115) and the NF-KB site (-115 and -8283) (Marks-Konczalik et al., 1998), and
synergistic regulation of promoter activity of NF κ B (-115 to -106) and of the A-activator-binding site (AABS: an CCAAT/ enhancer binding protein (C/EBP)-binding site)(-192 to -184) (Sakitani et al., 1998). C/EBP is also found to be important in murine iNOS promoter regulation (-153 to -142bp upstream: NF-IL6 binding site) (Dlaska et al., 1999) as well as in human iNOS promoter regulation (-205/+88 bp region) along with the NF κ B site (Kolyada et al., 2001).
Finally, two repeat polymorphisms (CCTTT) n and (TAAA) n have been demonstrated to influence the human iNOS promoter activity (Warpeha et al., 1999; Morris et al., 2002). The functional mechanism of these findings is presently unknown.

The latter two repeat polymorphisms influencing the human promoter activity have been tested in diseases, where iNOS have been proposed to influence the pathogenetic process, see Table 8 . Additionally, an iNOS promoter sequence variation search might identify nucleotide substitutions involving known or unknown TFB sites of functional relevance.

4.1.5.2. The human iNOS: exon organisation

cDNA has been cloned from hepatocytes (Geller et al., 1993), chrondrocytes (Charles et al., 1993; Maier et al., 1994), DLD-1 cells (Sherman et al., 1993), fibroblast (Chartrain et al., 1994), a human glioblastome cell line (A-172) (Hokari et al., 1994) two human cosmid DNA libraries (Xu et al., 1996), heart and skeletal muscle (Adams et al., 1998). The cDNA sequence obtained from hepatocytes (Geller et al., 1993) reveals 4145 bp, an open reading frame of 3459 bp encoding 1153 amino acids and has an estimated mass of 131kDa. The gene spans approximately 38kb and comprises 27 exons. Unprocessed pseudogenes (Park et al., 1997) have been described. iNOS is distinct from the other human NOS genes nNOS (Kishimoto et al., 1992) and eNOS (Marsden et al., 1993) located at chromosomes 12q24.2 and 7q35-36, respectively.

Taken together: The human iNOS gene possesses a long promoter region comprising several transcription binding factor sites and promoter activity has been identified as long as 16kb upstream of the transcription start site. Repeat sequences within the human iNOS promoter influencing the promoter activity have been demonstrated. Furthermore, the coding region spans approximately 38 kb and comprises 27 exons.

4.1.6. Expressional control of iNOS

Initially, it was believed that the iNOS activity was regulated predominantly at the transcriptional level (Cho et al., 1992), which partly were based upon the high number of transcription binding sites within the promoter region (Vera et al., 1996a). Subsequently, increasing evidence points towards the importance of both 5' and 3' UTRs being implicated in the regulation of gene expression (Kozak, 1992; Altmann et al., 1993). Vodovotz was the first to show that post-transcriptional mechanisms such as decreasing mRNA stability, reducing mRNA translation and increasing degradation of iNOS protein influenced NO production in TGF β exposed mouse peritoneal MØ (Vodovotz et al., 1993; Vodovotz, 1997).

In summary:

Various stimuli increase promoter activity by different TBF (see Figures in promoter sections). The intensifying effect by using cytokine mix compared to single cytokines suggests interaction of signal transduction pathways (Taylor et al., 1998). HSR attenuated the iNOS promoter activity (De-Vera et al., 1996; Vera et al., 1996)
Structural diversity in the 5'UTR in mRNA isolated from stimulated cells (freshly isolated alveolar MØ, bronchial epithelial cells and several types of cultured cells) has suggested alternative splicing as an additional way of regulating the expression of the gene (Chu et al., 1995)
Tissue specificity exists: expressed transcription binding factors vary in different cells types (Chu et al., 1995; Kolyada et al., 1996), and the transcription of the human iNOS gene has shown tissue specific regulation using human cell-lines from pulmonary and hepatic biliary epithel (Mellott et al., 2001).
Alternative splicing at the mRNA level (Eissa et al., 1996; Park et al., 2000), in exon 8 and exon 9 (Park et al., 1996; Eissa, 1998). This could explain the finding of Adams and co-workers (Adams et al., 1998) only showing 79% homology both at the protein and nucleotide level when cloning iNOS cDNA and protein from cardiac and skeletal muscle.
Effect of 3'UTR region at expression: 1.1 kb of the iNOS promoter and approximately 1.5 kb of the 3'UTR inserted in luciferase constructs showed lower basal activity and hence relatively higher stimulated activity compared to the construct without 3'UTR (Nunokawa et al., 1997), suggesting that the 3'UTR region may alter the mRNA stability (Geng et al., 1995; Belin et al., 2000).

Several promoter activity studies have shown low levels of promoter activity in the absence of cytokine stimulation (Vera et al., 1996a), and in vivo , Kobzik et al. have demonstrated iNOS expression without cytokine stimulation in epithelial cells and alveolar MØ lining the larger airways of humans by immunochemistry (Kobzik et al., 1993). It is speculated that low grade basal expression of iNOS mRNA takes place in many tissues, but these transcrips are highly unstable in the absence of cytokines - a putative effect of iNOS 3'UTR. Cytokines may stabilize iNOS mRNA, hence transcription increases (Nunokawa et al., 1996). Finally:

Post-translatory events: Cytokine stimulation of DLD-1 cells indicates a > 20 fold steady-state of iNOS mRNA (Salzman et al., 1996), which is in contrast to iNOS promoter activity (luciferase activity of 13.1 kb) where only 2-4 fold increase was observed (Linn et al., 1997) - could be due to 3'UTR effects
The activity of the iNOS enzyme requires binding of many co-factors (FAD, FMN, NADPH, tetrahydrobiopterin and calmodulin) (Marletta, 1993; Fossetta et al., 1996)

In conclusion: iNOS seems to be under tight expressional control at various levels which seems adequate as iNOS possess many different beneficial functions in various cellular systems in normal physiology, however leads to detrimental effects when expressed inadequately. Hence, an understanding of the various ways the expression of iNOS is controlled becomes essential, when searching for and evaluating genetic variation within the gene that might influence its expressional control.

4.1.7. Genetic variations in the human iNOS gene

Within the human iNOS promoter region, four polymorphisms have been described (i) G/C (position -969, subsequently corrected to position -954) (Kun et al., 1998; Kun et al., 2001), (ii) (TAAA) n (position -754 to -739) (Bellamy et al., 1997), (iii) (CCTTT) n (position -2662 to -2608) (Xu et al., 1997) and (iv) C/T (position -1173) (Hobbs et al., 2002). The (CCTTT) n repeat polymorphism has been functionally tested in vitro, associating the 14 repeat allele to high promoter activity (Warpeha et al., 1999).

Different allelic frequency of the (CCTTT) n repeat polymorphism has been observed between ethnically diverse populations (Africa, Europe, Asia and Caribbean) (Xu et al., 2000) and China (Lu et al., 2002). The G/C (position -966) (Kun et al., 1998) has not been identified in any Caucasians tested so far (Kun et al., 1998) (Johannesen et al., 2000b).

In Table 8 " iNOS promoter polymorphisms in human diseases " publications are listed examining the above polymorphisms within various diseases, in which iNOS mediated NO production has been suggested to have a possible pathogenical role.

It appears from the table that in most diseases tested no association has been found, though some inconsistent findings within hypertension (Glenn et al., 1999) (Rutherford et al., 2001) and malaria (Levesque et al., 1999; Kun et al., 2001; Ohashi et al., 2002) are seen. Regarding T1DM, no genetic association/linkage was identified (Johannesen et al., 2000b). However, association to the iNOS promoter has been reported in a subset of T1DM patients suffering from nephropathy/retinopathy (Warpeha et al., 1999; Johannesen et al., 2000a; Kumaramanickavel et al., 2002; Morris et al., 2002).

The coding region of the human iNOS gene has been characterised by (Xu et al., 1996) identifying intron/exon splice sites. Three papers have identified polymorphisms within the exons (Johannesen et al., 2001a; Shen et al., 2002; Levecque et al., 2003). The paper of Johannesen et al. tested the identified polymorphisms for linkage to T1DM. In total, 10 polymorphisms were identified from a complete iNOS gene scan of all exons. The four most common polymorphisms (in exon 1, 8, 16 and 20) were tested for linkage using the TDT analysis. Linkage was identified for T1DM among HLA DR3/4 positive individuals having a T at the C/T polymorphisms in exon 16. Furthermore, haplotypes were constructed and tested by ETDT although no increase in genetic information of disease susceptibility could be demonstrated. However, the C/T polymorphism in exon 16 gave rise to an amino acid shift Ser 608 Leu only six amino acids from a region identified as being of importance to the Ca ++ independency of iNOS (Daff et al., 1999; Johannesen et al., 2001a). As this polymorphism may have functional implications it would be interesting to test this polymorphism in other autoimmune diseases, in which iNOS mediated NO-production has been proposed in the pathogenesis (Singh et al., 2000).

In the genome scans of T1DM, the region in which human iNOS is located (17q11) has not been demonstrated to be in linkage with T1DM (see Chapter 2) with the exception of Vaessen demonstrating linkage of 17q24 to T1DM in a small genetically isolated Dutch population (Vaessen et al., 2002). Furthermore, a genome scan of Crohns Disease in a Jewish population demonstrated linkage to the chromosomal regions 17q21-23 (Ma et al., 1999). The distances between 17q11 (NOS2, position 50.6 cM at http://research.marshfieldclinic.org) and 17q24 (D17S2059, position 93.3 cM at http://research.marshfieldclinic.org) and 17q21 (D17S787, position 75.0 cM at http://research.marshfieldclinic.org) are 42.7 cM and 24.4 cM, respectively. Hence, these distances do not support the iNOS gene being an obvious candidate gene within these regions in the respective populations.

In conclusion: Polymorphisms within the iNOS gene promoter region have been tested for association to several different diseases. However, only the studies within diabetic retinopathy/nephropathy, arterial hypertension and malaria have been replicated, and the association has only been reproduced for diabetic retinopathy/nephropathy. No association to T1DM of the iNOS promoter polymorphisms has been shown; however linkage for the exon 16 polymorphism was demonstrated in high risk HLA T1DM individuals. Despite recent findings from a genome scan of possible T1DM linkage to the iNOS gene region in a genetic isolate, testing of association and linkage of the iNOS gene to T1DM should be replicated in other populations to confirm or reject the present findings.

4.1.8. Critical transcription factors for iNOS transcription

Only the genetics of IRF-1 and NF κ B in T1DM will be briefly reviewed, as these genes have been examined in relation to T1DM. Furthermore, interaction of IRF-1 and NF κ B during activation of iNOS transcription has been illustrated (Saura et al., 1999), as well as a NF κ B binding motif in the IRF-1 gene has been demonstrated (Miyamoto et al., 1988)

4.1.8.1. IRF-1

Interferons involved in antiviral defence, cell growth regulation and immune activation, elicit their effects through transcriptional activation of the target genes, e.g. iNOS which possesses specific consensus DNA-binding recognition sites for IRF-1 in their promoters. These interferon-regulated genes are regulated through the JAK-STAT pathway and the interferon regulatory factors (IRFs). Additionally, the IRFs also act as transcription factors for the IFNs. The IRF family is rapidly expanding in number and covers a broad range of activities, for review see (Mamane et al., 1999).

In IRF-1-/- mice, the gene has been shown to be involved in T-cell selection and maturation, as these mice are 90% deficient of mature CD8 + T-cells (Matsuyama et al., 1993). In disease models of autoimmunity in mice lacking IRF-1-/- was shown to be protected against the mortality mediated by TNF and IFN- γ , possibly due to the impaired production of TNF and IFN γ , as IRF-1-/- mice have similar mortality to coinjections of TNF and IFN γ as wild type mice (Senaldi et al., 1999). Furthermore, mice lacking IRF-1 in a model of EAE demonstrate higher Th2-type cytokine responses thereby protected from severe autoimmune brain inflammation (Buch et al., 2003). This observation is in line with the previous finding of IRF-1 deficient mice having an impaired Th1 and enhanced Th2 response (Lohoff et al., 1997). Finally, IRF-1 along with TGF- β and STAT-1 have been implicated in refining the regulation of class II MHC genes through differential control of class II transactivator (CIITA) promoters (Piskurich et al., 1999).

Indeed, IRF-1 may possess an important regulatory role regarding cytokine mediated iNOS expression: IFN- γ induced binding of IRF-1 to the ISRE sequence of the RAW264.7 iNOS promoter - this binding activity was reduced in cells pre-treated with IL-4. Moreover, IL-4 down-regulated the IFN- γ induced IRF-1 mRNA expression (Coccia et al., 2000). Finally, IL-4 has also been shown to suppress IFN- γ stimulated iNOS transcription by elevating the level of IRF-2 which, through competition, prevents IRF-1 from binding to ISRE in the iNOS promoter (Paludan et al., 1999).

The role of IRF-1 and NF κ B in IL-1 mediated beta-cell destruction has been discussed in a previous chapter.

The IRF-1 gene has been assigned to chromosome 5q31.1 by fluorescent in situ hybridisation (Willman et al., 1993). The gene is 7.72 kb in length and comprises 10 exons (Cha et al., 1992). Several genetic polymorphisms within the gene have been identified:

promoter -300G/T, 4396 A/G, 6355 G/A (Noguchi et al., 2000), were identified by SSCP in order to test for association to asthma using TDT. The 6355G/A polymorphism was very rare. The -300G/T polymorphism was in nearly complete linkage disequilibrium with the 4396A/G which by TDT did not show significant transmission to atopy- or asthma-affected children. Recent studies from patients with chronic hepatitis C have identified association to the -300A allele (Promrat et al., 2002), and Saito and colleagues demonstrated that in chronic hepatitis C patients being -300A/A the Th1-type CD4 + cell population was significantly increased by IFN β administration (Saito et al., 2002). Promoter assay studies of the IRF-1 promoter (Saito et al., 2001) suggest that the single nucleotide polymorphisms identified contribute to determining responses to interferons.
GT-repeat in intron 7 (Kroef et al., 1993). This polymorphism has been tested by Johannesen et al. without finding any association to T1DM (Johannesen et al., 1997), but has been demonstrated to associate to childhood atopic asthma in a Japanese population (Nakao et al., 2001).
A C/T polymorphism in intron 6 of the IRF-1 gene has in a gene-gene (to p21 and p53) and gene-environmental testing been associated to cervical cancer susceptibility in Korean women (Park et al., 2003).
HinfI polymorphism in the 3'UTR, position 1688 with reference to EMBL sequence HSIRF1 (Donn et al., 2001) showed association to juvenile idiopathic arthritis. Seegers et al studied this polymorphism in Celiac Disease by use of TDT without finding any distorted transmission from parents to affected offspring (Seegers et al., 2003).

Finally, genetic variations within the IRF-2 gene have been examined in atopic dermatitis with contradictory results (Nishio et al., 2001) and (Hosomi et al., 2002).

4.1.8.2. NF κ B

Only two studies have tested a polymorphism within the NF κ B in different T1DM populations. Hegazy et al demonstrated association of alleles to T1DM (Hegazy et al., 2001) which could not be confirmed in a Danish T1DM collection (Gylvin et al., 2002).

Recently, a new gene (SUMO4, a I κ B α modifier) has been identified in the IDDM5 region at chromosome 6q25 being associated to T1DM (Guo et al., 2004). This study demonstrates that fine mapping of a chromosomal region linked to T1DM can successfully lead to identification of new genes possibly modifying the genetic risk of T1DM.

In conclusion: Obviously, genes encoding transcription factors being of importance to a candidate gene may themselves be candidate genes. However, only a limited number of studies have been performed testing association of iNOS related genes in T1DM - hence, no genetic predisposition to T1DM for IRF-1 and NF κ B can be confirmed or rejected and further studies are needed.

4.2. Mortalin

Mortalin was initially identified as a 66-kDa protein of pI 5.9 in mouse embryonic fibroblasts (MEF) (Wadhwa et al., 1991), later shown to be a member of the mouse HSP70 family (Wadhwa et al., 1993a). Its presence in the cytosol was correlated to the normal mortal phenotype, in contrast to its absence in the cytosolic fraction of immortal cells (Wadhwa et al., 1993a). Microinjection of anti-mortalin antibodies into senescent mouse cells led to transient stimulation of cell division, suggesting an anti-proliferative function of the protein (Wadhwa et al., 1993a), hence the name mortalin. Subsequently, an isoform of mortalin in mouse was identified in immortalized cells as well as in the perinuclear space (Wadhwa et al., 1993b). The isoform associated to the normal mortal phenotype has a uniform pancytosolic distribution (mot-1) and the immortal phenotype located perinuclarly (mot-2) only differs at two amino acids. It was shown later that in the mouse the mot-1 and mot-2 genes segregated in two mouse generations (Kaul et al., 2000a), which illustrates that the mot-1 and mot-2 genes are allelic in mice, and were assigned to mouse chromosome 18 (Kaul et al., 1995; Ohashi et al., 1995). Transfection of mouse mot-1 cDNA (pancytosolic form) induced cellular senescence in NIH 3T3 cells, whereas mot-2 cDNA (perinuclear form) did not impart any equivalent effect (Wadhwa et al., 1993c).

In the rat, a homologue protein named Grp75 (glucose regulated protein, 75kDa) was identified (Mizzen et al., 1989; Massa et al., 1995), a resident mitochondrial matrix protein, mediating the import of translocation-competent proteins into the mitochondria and subsequent assembly of proteins within this organelle (Mizzen et al., 1991). In normal rat tissue, expression studies on mortalin have revealed functional in vivo characteristics: non-dividing tissues and cells are observed to have higher levels of expression than the ones with division potential, supporting an anti-proliferative function of mortalin in normal tissue. However, in samples of brain tumour tissue the expression was dysregulated and non-pancytosolic distributed, suggesting its involvement in pathways leading to malignant transformation (Kaul et al., 1997). Hence: mouse mot-1 cDNA and pancytosolic distribution of mortalin are associated to mortality in normal cells ("mot-1 effect") - contrasting perinuclear localisation associated to immortality/malignancy ("mot-2 effect").

In 1993, the human counterpart to the mouse mortalin gene was cloned from B-lymphoblastomas under the name PBP74, a new member of the HSP70 family, suggested to be involved in antigen processing, however not inducible by heat (Domanico et al., 1993). In 1995 it was cloned under the name mitochondrial-HSP75 (mthsp75) due to its subcellular fraction (Bhattacharyya et al., 1995). Mortalin cDNA isolates from normal and immortalized human cells showed differential localisation patterns by staining (Wadhwa et al., 1995a) but identical sequences, implying that (i) cellular distribution rather than the presence or absence of the protein marks cellular mortal and immortal phenotypes, and (ii) the differential distribution of the protein in human cells is due to e.g. protein modifications and does not originate from distinct cDNA's as in mouse cells. Similar to the mouse mot-2 cDNA, human mortalin induced malignant transformation of NIH 3T3 cells (Kaul et al., 1998a), and stable transfected human lung fibroblast with human mortalin underwent extended population doublings in vitro (Kaul et al., 2003). Further, it has been shown that differentiation of HL-60 promyelocytic leukemia cells was accompanied by a decreased level of human mortalin expression (Xu et al., 1999), whereas over-expression of mortalin impaired the growth advantage of the leukemia cells and attenuated their differentiation (Xu et al., 1999). Recently, targeting mortalin using RNA-helicase-linked hybrid ribozymes successfully suppressed the expression of mortalin in transformed human cells, which resulted in growth arrest (Wadhwa et al., 2003). Transient transfection of cells with human mortalin cDNA led to a delay in the development of apoptosis after serum deprivation (Taurin et al., 2002), and finally, over-expression of mot-2 resulted in reduced level of Ras and phosphorylated ERK2, involved in the apoptotic pathway (Wadhwa et al., 2003). All these studies support a "mot-2" effect of human mortalin in various experimental settings.

4.2.1. Mortalin expression

Mortalin has been shown to be expressed in all cell types and tissues studied so far, including pancreas and islets of Langerhans (Wadhwa et al., 1995a; Kaul et al., 1997; John et al., 2000; Mose-Larsen et al., 2001; Johannesen et al., 2004). Expression levels of mortalin have been correlated to muscle activity (Ornatsky et al., 1995), mitochondrial activity (Ibi et al., 1996) and biogenesis (the accepted theory that life can originate only from pre-existing life and never from non-living material) (Takahashi et al., 1998). Various stimuli can induce mortalin expression:

glucose deprivation (Mizzen et al., 1989)
calcium ionophores (Resendez-E. et al., 1985)
ischemia (Massa et al., 1995)
hyperthyroidism (Craig et al., 1998)
ozone (Wu et al., 1999)
IL-1/nitric oxide (John et al., 2000; Mose-Larsen et al., 2001; Johannesen et al., 2004)

Furthermore, mortalin has been demonstrated to interact with and inhibit the function of the tumour suppressor p53 (Wadhwa et al., 1998; Wadhwa et al., 1999; Kaul et al., 2001; Wadhwa et al., 2002d), which partly can explain why mortalin is able to induce immortality. The mortalin - p53 interaction can be abrogated by MKT-077 (a lipophilic cationic dye possessing anti-tumour effect) which binds to mot-2 and lead p53 translocate to the nucleus, followed by growth arrest of the tumour cells (Wadhwa et al., 2000a). Mortalin-p53 complexes have also been detected in mitochondria during p53-induced apoptosis, implicating a role of mortalin in apoptosis (Marchenko et al., 2000). This indicates that mortalin may possess a role in cell fate determination (Rivolta et al., 2002). Besides binding to p53, mortalin has also been shown to bind (i) fibroblast growth factor-1 and aiding in its intracellular trafficking (Mizukoshi et al., 1999; Mizukoshi et al., 2001), (ii) and the IL-1RI and mortalin have been suggested to take part in IL-1RI internalisation (Sacht et al., 1999).

Increasing evidence supports a role of mortalin in mitochondrial function. Previously, it has been shown in yeast that the distribution of mitochondria changes in response to heat shock treatment (Collier et al., 1993) and recently, mortalin has been proposed to be essential for optimizing the functions of as-yet-unidentified heat-labile proteins in the mitochondrial matrix in controlling the mitochondrial morphology (Kawai et al., 2001). These studies suggest that mortalin may play an important role regarding mitochondrial function and that the differentiated distributions of mortalin in immortal versus mortal cells, at least in part, may be related to altered mitochondria morphology and function. That mortalin is involved in mitochondrial function can also explain the associations of mortalin to cellular energy supply, regulation of calcium levels, apoptosis, cellular localisation and cellular immortality (Wadhwa et al., 2002a).

In line with these observations are the findings of Johannesen et al of inter-individual expression of mortalin in isolated islets of Langerhans from two rat strains, the strain being most susceptible to the cytotoxic effect of IL-1 having the highest expressing of mortalin (Johannesen et al., 2004).

In summary: Being a protein involved in cell fate determination possibly by its involvement in mitochondrial functioning mortalin has been demonstrated to be upregulated in cytokine exposed islets of Langerhans. Hence, mortalin is a relevant protein/gene to study further in cytokine mediated beta-cell destruction.

4.2.2. Human mortalin gene

The human mortalin is encoded as a large protein containing a 46 residue pre-sequence which is not present in the mature protein purified from cells (Domanico et al., 1993). This pre-sequence shares features common to other mitochondrial targeting sequences (Bhattacharyya et al., 1995). Mitochondrial targeting proteins serve as facilitators for mitochondrial proteins to target and enter the mitochondria (Hartl et al., 1989). In accordance, the mitochondria are a central localisation of mortalin - but not unique - in human immortalised cell lines (Ran et al., 2000).

As described above, mortalin can be induced by various forms of cellular stress and is associated to the determination of cell fate. Furthermore, rat mortalin expression was:

identified in IL-1 exposed/NO-treated islets of Langerhans (John et al., 2000; Mose-Larsen et al., 2001), and
associated to different IL-1 sensitivity in two rat strains (Johannesen et al., 2004). Moreover,
NIH3T3 cells over-expressing rat mortalin induced decreased cellular survival (Johannesen et al., 2004), and finally
human mortalin has been localised to chromosome 5q31 (Kaul et al., 1995) - see Table 9

- thus, the human mortalin gene qualify as a T1DM candidate gene.

Table 9: " Diseases associated to chromosome 5q31 ", reflects papers reporting genome scan data that positively identifies 5q31 as a genomic region of interest. It is not the aim of this review to specifically compare to other studies within each of the diseases that either can or cannot confirm the findings listed in the table. The list simply illustrates that this genomic region possibly may enhance susceptibility of several immune mediated diseases.

The study of Johannesen et al (Johannesen et al., 2004) is the first paper to identify polymorphisms in the human mortalin gene. Three nucleotide polymorphisms were identified within the coding region, however none of them led to amino acid substitutions. Neither the tested polymorphisms, the D5S500 dinucleotide marker located close to the gene nor constructed haplotypes were identified to be linked to T1DM in this Danish Caucasoid collection (Johannesen et al., 2004). These identified polymorphisms are obvious SNPs to be tested within other of the diseases listed in the Table 9, Diseases associated to chromosome 5q31 .

In the study of Johannesen et al (Johannesen et al., 2004) overlapping PCR-products based on cDNA sequence were used to screen the coding sequence for polymorphisms. Our cDNA sequencing data were 100% identical to the published mRNA-based sequences except for the identified SNP's. When initially establishing typing assays for the identified polymorphisms, the use of the cDNA designed primer pairs in genomic DNA revealed only 97 to 99 percent identity between the cDNA and genomic DNA sequences within the same individual, the variation depending upon the primer set used. This inconsistency led us to initiate a NCBI BLAST search for genomic sequences that could possibly explain the deviating sequence results using cDNA versus genomic DNA material (pseudogenes?). At www.ncbi.nlm.nih.gov/LocusLink/ mortalin has been given the Locus ID 3313 and symbol HSPA9B. Two loci links are given: 5q31.1 and 2q36.1 - corresponding to the mRNA sequences L11066 and L15189, respectively. These mRNA sequences show 99% identity. A BLAST search in NCBI of L15189 (chromosome 2) identified a BAC clone, RP11-71J24 (GenBank accession number AC009302) located at the human chromosome 2 (227M, GenBank) where a part of the BAC clone showed 94% homology to the full length published L15189 mortalin cDNA sequence. The nucleotide sequence we obtained in genomic DNA material using the cDNA based primers showed 100% similarity to this specific BAC clone and hence, was not identical to the published cDNA sequences or the sequences we obtained in cDNA material. Subsequently, the gDNA material based typing assays of the polymorphisms were based upon the human sequence of chromosome 5 (AC011385) at the time it was available to the public. Using this sequence to design primers, a 100% sequence homology was obtained between cDNA and gDNA sequence for each tested individual, see Figure 8 .

However, should the mortalin gene be located at chromosome 2, then the marker D2S339 is located less than 1.3 cM (230.1-228.8 cM) from the putative localisation of HSPA9B at chromosome 2. We have previously tested the D2S126 marker (a marker of the IDDM13 locus at 2q33) (Larsen et al., 1999) without finding any evidence of linkage or association to T1DM in the Danish population, and since no recombination between D2S339 and D2S126 has been demonstrated the lack of linkage of mortalin to T1DM in the Danish population seemed substantiated.

In 2000, Xie et al published (Xie et al., 2000) the exon/intron organisation of the HSPA9 (human Mortalin) gene as part of their search for variations in the human mortalin gene. This was performed using a BAC-clone (15L17) as template, not available to the public. They used intron-based primer sequences to amplify the 17 identified exons followed by direct sequencing of amplified PCR products. They identified a C to T substitution in the BAC-clone 15L17 corresponding to position 1933 in the human mortalin gene. As Xie et al used a BAC clone as template in contrast to full genomic DNA in our design, they were not in a position to identify putative pseudogenes. In future studies it should be of no difference whether to use the primer pairs designed by Xie et al or by Johannesen et al.

In conclusion: Mortalin (i) is a protein induced by various forms of cellular stress, associated to determination of cell fate, and functionally involved in e.g. mitochondrial function; (ii) has been located to chromosome 5q31, a region of putative interest in immune mediated diseases and, (iii) has been identified and demonstrated to be up-regulated in cytokine exposed rat islets of Langerhans - used as a model for beta-cell destruction in T1DM. Hence, mortalin was considered a candidate gene in the pathogenesis of T1DM. Furthermore, the inter-individual expression of mortalin was associated to different IL-1 sensitivity in two rat strains suggesting inter-individual expressional control of this candidate gene being of importance in cytokine mediated beta-cell destruction. However, the precise pathogenitical involvement of mortalin needs further exploration. In a Danish national wide collected T1DM family collection, the mortalin gene could not be demonstrated to be in linkage to T1DM. In order to finally exclude the mortalin gene as a susceptibility gene in T1DM, additional screenings for polymorphisms in the 5' UTR and the 3'UTRs are requested, as well as the identified polymorphisms in the gene should be tested in other T1DM collections.

Conclusion from Chapter 4

By means of a combined candidate gene approach based upon an experimentally testable pathogenetic model of cytokine mediated beta-cell destruction, three genes were selected: the iNOS, the IRF-1 and the mortalin genes. These genes were examined for sequence variation and tested for association to diabetes in a population based nationwide Danish T1DM collection. The expression patterns of the selected genes were examined in a rat model using two different rat strains showing different sensitivity to cytokine exposure as defined by different insulin release from cytokine exposed islets in vitro.

In beta-cells, NO and NO-independent induced necrotic and apoptotic destruction takes place following cytokine exposure. Whether the necrotic or the apoptotic processes - or both - are effectuated may be influenced by e.g. the potency of the "cytokine hit" versus the defence properties of the beta-cell. iNOS is expressed in cytokine exposed human beta-cells/islets and the evidence of the toxic effect of peroxynitrite and NO donors argues in favour of a role of NO in human beta-cell destruction. Furthermore, inter-individual expression levels of iNOS in the rat model and its association to riNOS promoter polymorphism, and the genetic association of the human iNOS gene to T1DM is a further substantiation of a role of NO in T1DM pathogenesis. However, a polymorphism in the IRF-1 gene (a transcription factor of the iNOS gene) was not demonstrated to be associated to T1DM in a Danish collection.

Mortalin expression - associated to determination of cell fate as demonstrated by decreased cellular survival when over-expressed in NIH-3T3 cells - was differentially expressed in the rat model suggesting an inter-individual expressional control of this candidate gene. However, in a Danish nation-wide T1DM family collection the mortalin gene could not be demonstrated to be in linkage to T1DM.

References

Adams, V., S. Krabbes, H. Jiang, J. Yu, A. Rahmel, S. Gielen, G. Schuler and R. Hambrecht (1998). Complete coding sequence of inducible nitric oxide synthase from human heart and skeletal muscle of patients with chronic heart failure. Nitric oxide 2(4): 242-249.
Adorini, L., S. Gregori and L. C. Harrison (2002). Understanding autoimmune diabetes: insights from mouse models. Trends in Molecular Medicine 8(1): 31-38.
Akabane, A., I. Kato, S. Takasawa, M. Unno, H. Yonekura, T. Yoshimoto and H. Okamoto (1995). Nicotinamide inhibits IRF-1 mRNA induction and prevents IL-1 beta induced nitric oxide synthase expression in pancreatic beta-cells. Biochem Biophys Res Com 215: 524-530.
Akerblom, H. K., O. Vaarala, H. Hyoty, J. Ilonen and M. Knip (2002). Environmental factors in the etiology of type 1 diabetes. American Journal of Medical Genetics 115(1): 18-29.
Algar, E. M., T. Khromykh, S. I. Smith, D. M. Blackburn, G. J. Bryson and P. J. Smith (1996). A WT1 antisense oligonucleotide inhibits proliferation and induces apoptosis in myeloid leukaemia cell lines. Oncogene 12(5): 1005-1014.
Altmann, D. (1993). Practical statitics for medical research. Capman and hall: 456.
Altmann, M. and H. Trachsel (1993). Regulation of translation initiation and modulation of cellular physiology. Trends in biochemical sciences 18(11): 429-432.
Altmuller, J., L. J. Palmer, G. Fischer, H. Scherb and M. Wjst (2001). Genomewide scans of complex human diseases: True linkage is hard to find. American Journal of Human Genetics 69(5): 936-950.
Ammendrup, A., A. Maillard, K. Nielsen, N. A. Andersen, P. Serup, O. D. Madsen, T. Mandrup-Poulsen and C. Bonny (2000). The c-Jun amino-terminal kinase pathway is preferentially activated by interleukin-1 and controls apoptosis in differentiating pancreatic beta-cells. Diabetes 49(9): 1468-1476.
Andersen, H. U. (1999). The role of the target organ in cytokine-mediated beta-cell destruction: Possible relevance for the pathogenesis of insulin dependent diabetes mellitus. Danish Medical Bulletin 46(2): 127-164.
Andersen, H. U., S. J. Fey, P. M. Larsen, A. Nawrocki, K. R. Hejnæs, T. Mandrup-Poulsen and J. Nerup (1997). Interleukin -1 beta induced changes in the protein expression of rat islets: A computerized database. Electrophoresis 18: 2091-2103.
Andersen, H. U., P. M. Larsen, S. J. Fey, A. E. Karlsen, T. Mandrup-Poulsen and J. Nerup (1995). Two-dimentionel gel electrophoresis of rat islets proteins: Interleukin-1 beta induced changes in protein expression are reduced by L-argenine depletion and nicotinamide. Diabetes 44: 400-407.
Andersen, H. U., D. Mauricio, A. E. Karlsen, T. Mandrup-Poulsen, J. H. Nielsen and J. Nerup (1996). Interleukin-1 β -induced nitric oxide production from isolated rat islets is modulated by D-glucose and 3-isobutyl-1-methyl xanthine. Eur J Endocrinol 134: 251-259.
Andersen, N. A., C. M. Larsen and T. Mandrup-Poulsen (2000). TNFalpha and IFNgamma potentiate IL-1beta induced mitogen activated protein kinase activity in rat pancreatic islets of Langerhans. Diabetologia 43(11): 1389-1396.
Ankarcrona, M., J. M. Dypbukt, B. Brüne and P. Nicotera (1994). Interleukin-1 beta-induced nitric oxide production activates apoptosis in pancreatic RINm5F cells. Experimental cell research 213(1): 172-177.
Aparicio, J. M., A. Wakisaka, A. Takada, N. Matsuura and T. Yoshiki (1990). Non-HLA genetic factors and insulin dependent diabetes mellitus in the Japanese: TCRA, TCRB and TCRG, INS, THY1, CD3D and ETS1. Disease markers 8(5): 283-294.
Argentaro, A., B. Wapelhorst, P. Concannon and V. R. Harley (2001). Linkage studies of SOX13, the ICA12 autoantigen gene, in families with type 1 diabetes. Molecular Genetics and Metabolism 72(4): 356-359.
Armstrong, J. F., K. Pritchard-Jones, W. A. Bickmore, N. D. Hastie and J. B. L. Bard (1992). The expression of the Wilms' tumour gene, WT1, in the developing mammalian embryo. Mechanisms of Development 40(1-2): 85-97.
Arnush, M., M. R. Heitmeier, A. L. Scarim, M. H. Marino, P. T. Manning and J. A. Corbett (1998). IL-1 produced and released endogenously within human islets inhibits beta cell function. Journal of Clinical Investigation 102(3): 516-526.
Asea, A., S. Kraeft, E. Kurt-Jones, M. Stevenson, L. Chen, R. Finberg, G. Koo and S. Calderwood (2000). HSP70 stimulates cytokine production through a CD14-dependant pathway, demonstrating its dual role as a chaperone and cytokine. Nature Medicine 6: 435-442.
Avoustin, P., L. Briant, C. De-Préval and A. Cambon-Thomsen (1992). Polymorphism study of TCR alpha and gamma genes in insulin dependent diabetes mellitus (IDDM) multiplex families. Autoimmunity 14(2): 97-100.
Awata, T., K. Inoue, I. Inoue, T. Abe, H. Takino, Y. Kanazawa and S. Katayama (2000). Lack of association of the Ala45Thr variant in the BETA2/ NEUROD1 with type 1 diabetes in Japanese. Diabetes Research and Clinical Practice 49(1): 61-63.
Awata, T., C. Matsumoto, T. Urakami, R. Hagura, S. Amemiya and Y. Kanazawa (1994). Association of polymorphism in the interferon gamma gene with IDDM. Diabetologia 37(11): 1159-1162.
Azevedo-Martins, A. (2003). Improvement of the mitochondrial antioxidant defense status prevents cytokine-induced nuclear factor-kappaB activation in insulin-producing cells. Diabetes. 52: 93-101.
Bach, J. F. (1994). Insulin-dependent diabetes mellitus as an autoimmune disease. Endocrine reviews 15(4): 516-542.
Bach, J. F., H. J. Garchon and P. van-Endert (2001). Genetics of human type 1 diabetes mellitus. 4: 1-30.
Bain, S. C., A. H. Barnett and J. A. Todd (1992). Lack of association between type 1 diabetes and the glucokinase gene. Lancet 340(8810): 54-55.
Baker, M. S., X. Chen, A. R. Rotramel, J. J. Nelson and D. B. Kaufman (2003). Interferon regulatory factor-1 down-regulates cytokine- induced IP-10 expression in pancreatic islets. Surgery (St Louis) 134(2): 134-141.
Baker, M. S., X. J. Chen, X. C. Cao and D. B. Kaufman (2001). Expression of a dominant negative inhibitor of NF-kappa B protects MIN6 beta-cells from cytokine-induced apoptosis. Journal of Surgical Research 97(2): 117-122.
Barbaux, S., P. Niaudet, M. C. Gubler, J. P. Grünfeld, F. Jaubert, F. Kuttenn, C. N. Fékété, N. Souleyreau-Therville, E. Thibaud, M. Fellous and K. McElreavey (1997). Donor splice-site mutations in WT1 are responsible for Frasier syndrome. Nature genetics 17(4): 467-470.
Barbosa, J., S. Rich, T. Dunsworth and J. Swanson (1982). Linkage disequilibrium between insulin-dependent diabetes and the Kidd blood group Jkb allele. Journal of clinical endocrinology and metabolism 55(1): 193-195.
Bassuny, W. M., K. Ihara, N. Matsuura, S. Ahmed, H. Kohno, R. Kuromaru, K. Miyako and T. Hara (2002). Association study of the NRAMP1 gene promoter polymorphism and early-onset type 1 diabetes. Immunogenetics 54(4): 282-285.
Beck, K. F., W. Eberhardt, S. Walpen, M. Apel and J. Pfeilschifter (1998). Potentiation of nitric oxide synthase expression by superoxide in interleukin 1 beta-stimulated rat mesangial cells. FEBS letters 435(1): 35-38.
Beck, K. F. and R. B. Sterzel (1996). Cloning and sequencing of the proximal promoter of the rat iNOS gene: activation of NFkappaB is not sufficient for transcription of the iNOS gene in rat mesangial cells. FEBS letters 394(3): 263-267.
Becker, K. G. (1999). Comparative genetics of type 1 diabetes and autoimmune disease: Common loci, common pathways? Diabetes 48(7): 1353-1358.
Becker, K. G., R. M. Simon, J. E. Bailey-Wilson, B. Freidlin, W. E. Biddison, H. F. McFarland and J. M. Trent (1998). Clustering of non-major histocompatibility complex susceptibility candidate loci in human autoimmune diseases. Proceedings of the National Academy of Sciences of the United States of America 95(17): 9979-9984.
Bedoya, F. J., M. Flodström and D. L. Eizirik (1995). Pyrrolidine dithiocarbamate prevents IL-1-induced nitric oxide synthase mRNA, but not superoxide dismutase mRNA, in insulin producing cells. Biochemical and biophysical research communications 210(3): 816-822.
Belin, V. D., C. Southern and I. C. Green (2000). Influence of the 3'UTR on IL-1beta stimulation of an iNOS reporter system. Diabetologia 43(Suppl 1): A122.
Bellamy, R. and A. V. Hill (1997). A bi-allelic tetranucleotide repeat in the promoter of the human inducible nitric oxide synthase gene. Clinical genetics 52(3): 192-193.
Bellmann, K., V. Burkart, J. Bruckhoff, H. Kolb and J. Landry (2000). p38-dependent enhancement of cytokine-induced nitric-oxide synthase gene expression by heat shock protein 70. J Biol Chem 275(24): 18172-18179.
Bellmann, K., L. Hui, J. Radons, V. Burkart and H. Kolb (1997). Low stress response enhances vulnerability of islet cells in diabetes-prone BB rats. Diabetes 46(2): 232-236.
Bellmann, K., M. Jäättelä, D. Wissing, V. Burkart and H. Kolb (1996). Heat shock protein hsp70 overexpression confers resistance against nitric oxide. FEBS letters 391(1-2): 185-188.
Bellmann, K., A. Wenz, J. Radons, V. Burkart, R. Kleemann and H. Kolb (1995). Heat shock induces resistance in rat pancreatic islet cells against nitric oxide, oxygen radicals and streptozotocin toxicity in vitro. Journal of clinical investigation 95(6): 2840-2845.
Bendtzen, K., T. Mandrup-Poulsen, J. Nerup, J. H. Nielsen, C. A. Dinarello and M. Svenson (1986). Cytotoxicity of human pI 7 interleukin-1 for pancreatic islets of Langerhans. Science 232(4757): 1545-1547.
Ben-Salem, M., N. Ghanem, T. Kneissi, M. Abbal, J. M. Dugoujon, J. Ben-Chibani, F. Ellouze and G. Lefranc (1991). Immunogenetic markers (BF, C2, C4, 21-OH, TNF alpha, TCR beta, Ig) and insulin-dependent diabetes in the Tunisian population: serological and molecular study. Annales de biologie clinique 49(7): 389-396.
Bergholdt, R., P. Ghandil, J. Johannesen, O. P. Kristiansen, I. Kockum, H. Luthman, K. S. Ronningen, J. Nerup, C. Julier and F. Pociot (2004). Genetic and functional evaluation of an interleukin-12 polymorphism (IDDM18) in families with type 1 diabetes - art. no. e39. Journal of Medical Genetics 41(4): 31-37.
Bergholdt, R., P. E. Heding, K. Nielsen, R. L. Nolsoe, T. Sparre, J. Størling, J. Nerup, F. Pociot and T. Mandrup-Poulsen (2003). Type 1 diabetes mellitus: an inflammatory disease of the islet. In: Eisenbarth GS (ed) Type 1 Diabetes: molecular, cellular and clinical immunology. http://www. uchsedu/misc/diabetes/bdc.html
Bergholdt, R., A. E. Karlsen, J. Johannesen, P. M. Hansen, C. A. Dinarello, J. Nerup and F. Pociot (1995). Characterization of polymorphisms of an interleukin 1 receptor type 1 gene (IL1RI) promotor region (P2) and their relation to insulin-dependent diabetes mellitus (IDDM). The Danish Study Group of Diabetes in Childhood. Cytokine 7(7): 727-733.
Bergholdt, R., Z. Larsen, N. Andersen, J. Johannesen, O. Kristensen, T. Mandrup-Poulsen, J. Nerup, F. Pociot, DSGD and DIEGG (2000). Characterization of new polymorphisms in the 5'UTR of the human interleukin-1 receptor type 1 (IL1R1) gene: linkage to type 1 diabetes and correlation to IL-1RI plasma level. GAI 1: 495-500.
Bhatia, E., J. B. Buse and R. A. Jackson (1988). T-cell antigen receptor alpha chain polymorphisms in insulin-dependent diabetes. Journal of autoimmunity 1(5): 389-397.
Bhattacharyya, T., A. Karnezis, S. Murphy, T. Hoang, B. Freeman, B. Phillips and R. Morimoto (1995). Cloning and subcellular localization of human mitochondrial hsp70. J of Biol Chem 270: 1705-1710.
Bickmore, W. A., K. Oghene, M. H. Little, A. Seawright, V. van-Heyningen and N. D. Hastie (1992). Modulation of DNA binding specificity by alternative splicing of the Wilms tumor wt1 gene transcript. Science 257 (5067): 235-237.
Bieda, K., M. A. Pani, A. Van-der, B., C. Seidl, R. R. Tonjes, F. Gorus, K. H. Usadel and K. Badenhoop (2002). A retroviral long terminal repeat adjacent to the HLA DQB1 gene (DQ-LTR13) modifies Type I diabetes susceptibility on high risk DQ haplotypes. Diabetologia 45(3): 443-447.
Bigdeli, N., A. Niemann, S. Sandler and D. L. Eizirik (1994). Dissociation between interleukin-1 beta-induced expression of mRNA for superoxide dismutase and nitric oxide synthase in insulin-producing cells. Biochemical and biophysical research communications 203(3): 1542-1547.
Bilbao, J. R., A. Martín-Pagola, B. Calvo, N. Perez-de, G., Gepv-N. and L. Castaño (2002). Contribution of MIC-A polymorphism to type 1 diabetes mellitus in Basques. Annals of the New York Academy of Sciences 958: 321-324.
Binzer, J., N. A. Andersen, J. Størling and T. Mandrup-Poulsen (2001). Interleukin-1 β induced p38 and C-jun N-terminal kinase activities in isolated rat pancreatic islets is nitric oxide dependent. Diabetes Metab 17: A39.
Blume, N., J. Skouv, L. I. Larsson, J. J. Holst and O. D. Madsen (1995). Potent inhibitory effects of transplantable rat glucagonomas and insulinomas on the respective endogenous islet cells are associated with pancreatic apoptosis. Journal of clinical investigation 96(5): 2227-2235.
Bogdan, C. (1998). The multiplex function of nitric oxide in (auto)immunity. Journal of Experimental Medicine 187(9): 1361-1365.
Boitard, C., S. Caillat-Zucman and J. Timsit (1997). Insulin-dependent diabetes and human leucocyte antigens. Diabetes & metabolism 23 Suppl 2: 22-28.
Bonfanti, R., E. Bognetti, F. Meschi, A. Brunelli, M. C. Riva, M. R. Pastore, G. Calori and G. Chiumello (1998). Residual beta-cell function and spontaneous clinical remission in type 1 diabetes mellitus: the role of puberty. Acta diabetologica 35(2): 91-95.
Bonner-Weir, S. (2001). beta-Cell turnover: Its assessment and implications. Diabetes 50(Suppl 1): S20-S24.
Bonny, C., A. Oberson, S. Negri, C. Sauser and D. F. Schorderet (2001). Cell-permeable peptide inhibitors of JNK novel blockers of beta-cell death. Diabetes 50(1): 77-82.
Borch-Johnsen, K. (1989). The prognosis of insulin-dependent diabetes mellitus. An epidemiological approach. Dan Med Bull 36: 336-349.
Bottini, N. (2004). A functional variant of lymphoid tyrosine phosphatase is associated with type I diabetes. Nature genetics. 36: 337-338.
Bredt, D. S., P. M. Hwang, C. E. Glatt, C. Lowenstein, R. R. Reed and S. H. Snyder (1991). Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature 351(6329): 714-718.
Breslow and Day (1987). The design and analysis of cohort studies in statistical methods in cancer research. Science Publications, Lyon 2.
Buch, T., C. Uthoff-Hachenberg and A. Waisman (2003). Protection from autoimmune brain inflammation in mice lacking IFN-regulatory factor-1 is associated with Th2-type cytokines. International Immunology 15(7): 855-859.
Bugawan, T. L., M. Alejandrino, D. B. Mirel, A. Panelo, C. M. Solfelix, P. Pozzilli, R. Buzzetti, R. L. Reynolds, A. M. Valdes and H. A. Erlich (2001). Association of specific IL4 receptor SNPs with type I diabetes in Filipinos. American Journal of Human Genetics 69(4 Supplement): 403.
Bukau, B. and A. L. Horwich (1998). The Hsp70 and Hsp60 chaperone machines. Cell 92(3): 351-366.
Burkart, V., H. Liu, K. Bellmann, D. Wissing, M. Jaattela, M. G. Cavallo, P. Pozzilli, K. Briviba and H. Kolb (2000). Natural resistance of human beta cells toward nitric oxide is mediated by heat shock protein 70. Journal of Biological Chemistry 275(26): 19521-19528.
Buzzard, K. A., A. J. Giaccia, M. Killender and R. L. Anderson (1998). Heat shock protein 72 modulates pathways of stress-induced apoptosis. Journal of Biological Chemistry 273(27): 17147-17153.
Caillat-Zucman, S., E. Bertin, J. Timsit, C. Boitard, R. Assan and J. Bach (1993). Protection from insulin-dependent diabetes mellitus is linked to a peptide transporter gene. European Journal of Immunology 23(8): 1784-1788.
Caillat-Zucman, S., S. Daniel, I. Djilali-Saiah, J. Timsit, H. J. Garchon, C. Boitard and J. F. Bach (1995). Family study of linkage disequilibrium between TAP2 transporter and HLA class II genes. Absence of TAP2 contribution to association with insulin-dependent diabetes mellitus. Human immunology 44(2): 80-87.
Call, K. M., T. Glaser, C. Y. Ito, A. J. Buckler, J. Pelletier, D. A. Haber, E. A. Rose, A. Kral, H. Yeger, W. H. Lewis and a. et (1990). Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms' tumor locus. Cell 60(3): 509-520.
Calzada, J. E., M. A. Lopez-Nevot, Y. Beraun and J. Martin (2002). No evidence for association of the inducible nitric oxide synthase promoter polymorphism with Trypanosoma cruzi infection. Tissue Antigens 59(4): 316-319.
Camacho, A., S. Balladares, C. Alaez, E. Infante, H. Flores, C. Robles and C. Gorodezky (2002). Analysis of single nucleotide polymorphisms (SNPs) of eleven cytokines in Mexican patients with type 1 diabetes. Secondary association with TNF-alpha. Tissue Antigens 59(2 Supplement): 113.
Cantagrel, A., F. Navaux, P. Loubet-Lescoulie, F. Nourhashemi, G. Enault, M. Abbal, A. Constantin, M. Laroche and B. Mazieres (1999). Interleukin-1 beta, interleukin-1 receptor antagonist, interleukin-4, and interleukin-10 gene polymorphisms: relationship to occurrence and severity of rheumatoid arthritis. Arthritis and Rheumatism 42(6): 1093-1100.
Caplen, N. J., A. Patel, A. Millward, R. D. Campbell, S. Ratanachaiyavong, F. S. Wong and A. G. Demaine (1990). Complement C4 and heat shock protein 70 HSP70 genotypes and Type I diabetes mellitus. Immunogenetics 32(6): 427-430.
Cardozo, A. K., H. Heimberg, Y. Heremans, R. Leeman, B. Kutlu, M. Kruhoffer, T. Orntoft and D. L. Eizirik (2001b). A comprehensive analysis of cytokine-induced and nuclear factor-kappa B-dependent genes in primary rat pancreatic beta-cells. Journal of Biological Chemistry 276(52): 48879-48886.
Cardozo, A. K., M. Kruhoffer, R. Leeman, T. Orntoft and D. L. Eizirik (2001a). Identification of novel cytokine-induced genes in pancreatic & beta-cells by high-density oligonucleotide arrays. Diabetes 50(5): 909-920.
Cartegni, L., S. L. Chew and A. R. Krainer (2002). Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nature Reviews Genetics 3(4): 285-298.
Celis, J. E., M. Kruhøffer, I. Gromova, C. Frederiksen, M. Ostergaard, T. Thykjaer, P. Gromov, J. Yu, H. Pálsdóttir, N. Magnusson and T. F. Orntoft (2000). Gene expression profiling: monitoring transcription and translation products using DNA microarrays and proteomics. FEBS letters 480(1): 2-16.
Cetkovic-Cvrlje, M. and D. L. Eizirik (1994). TNF-alpha and IFN-gamma potentiate the deleterious effects of IL-1 beta on mouse pancreatic islets mainly via generation of nitric oxide. Cytokine 6(4): 399-406.
Cha, Y., S. H. Sims, M. F. Romine, M. Kaufmann and A. B. Deisseroth (1992). Human interferon regulatory factor 1: intron-exon organization. DNA and cell biology 11(8): 605-611.
Chang, T. J., H. H. Lei, J. I. Yeh, K. C. Chiu, K. C. Lee, M. C. Chen, T. Y. Tai and L. M. Chuang (2000). Vitamin D receptor gene polymorphisms influence susceptibility to type 1 diabetes mellitus in the Taiwanese population. Clinical Endocrinology 52(5): 575-580.
Chapple, I. L. (1997). Reactive oxygen species and antioxidants in inflammatory diseases. Journal of clinical periodontology 24(5): 287-296.
Charles, I. G., R. M. Palmer, M. S. Hickery, M. T. Bayliss, A. P. Chubb, V. S. Hall, D. W. Moss and S. Moncada (1993). Cloning, characterization, and expression of a cDNA encoding an inducible nitric oxide synthase from the human chondrocyte. Proceedings of the National Academy of Sciences of the United States of America 90(23): 11419-11423.
Charlton, B. and K. J. Lafferty (1995). The Th1/Th2 balance in autoimmunity. Current opinion in immunology 7(6): 793-798.
Chartrain, N. A., D. A. Geller, P. P. Koty, N. F. Sitrin, A. K. Nussler, E. P. Hoffman, T. R. Billiar, N. I. Hutchinson and J. S. Mudgett (1994). Molecular cloning, structure, and chromosomal localization of the human inducible nitric oxide synthase gene. Journal of biological chemistry 269(9): 6765-6772.
Chauffert, M., A. Cissé, D. Chevenne, J. F. You, S. Michel and F. Trivin (1997). Susceptibility to type 1 diabetes in the Senegalese population is linked to HLA-DQ and not TAP and LMP genes. Diabetes care 20(8): 1299-1303.
Chen, M., P. Proost, C. Gysemans, C. Mathieu and D. L. Eizirik (2001). Monocyte chemoattractant protein-1 is expressed in pancreatic islets from prediabetic NOD mice and in interleukin-1beta-exposed human and rat islet cells. Diabetologia 44(3): 325-332.
Cho, H. J., Q. W. Xie, J. Calaycay, R. A. Mumford, K. M. Swiderek, T. D. Lee and C. Nathan (1992). Calmodulin is a subunit of nitric oxide synthase from macrophages. Journal of experimental medicine 176(2): 599-604.
Chong, M. M. W., Y. Chen, R. Darwiche, N. L. Dudek, W. Irawaty, P. Santamaria, J. Allison, T. W. H. Kay and H. E. Thomas (2004). Suppressor of cytokine signaling-1 overexpression protects pancreatic beta cells from CD8(+) T cell-mediated autoimmune destruction. Journal of Immunology 172(9): 5714-5721.
Christensen, U. B., P. M. Larsen, S. J. Fey, H. U. Andersen, A. Nawrocki, T. Sparre, T. Mandrup-Poulsen and J. Nerup (2000). Islet protein expression changes during diabetes development in islet syngrafts in BB-DP rats and during rejection of BB- DP islet allografts. Autoimmunity 32(1): 1-15.
Christy, M., A. Green, B. Christau, H. Kromann and J. Nerup (1979). Epidemiologic studies of insulin-dependent diabetes mellitus. Diabetes Care 2(2): 127-130.
Chu, S. C., J. Marks-Konczalik, H. P. Wu, T. C. Banks and J. Moss (1998). Analysis of the cytokine-stimulated human inducible nitric oxide synthase (iNOS) gene: characterization of differences between human and mouse iNOS promoters. Biochemical and biophysical research communications 248(3): 871-878.
Chu, S. C., H. P. Wu, T. C. Banks, N. T. Eissa and J. Moss (1995). Structural diversity in the 5'-untranslated region of cytokine-stimulated human inducible nitric oxide synthase mRNA. Journal of biological chemistry 270(18): 10625-10630.
Chuang, L., T. Jou, H. Wu, T. Tai and B. J. Lin (1996). A rapid method to study heat shock protein 70-2 gene polymorphism in insulin-dependent diabetes mellitus. Pancreas 13(3): 268-272.
Cinek, O., P. Dreviacutenek, Z. Sumniacutek, B. Bendlova, P. Sedlakovaacute, S. Kolouskovaacute, M. Snajderovaacute and J. Vavrinec (2003). NEUROD polymorphism Ala45Thr is associated with Type 1 diabetes mellitus in Czech children. Diabetes Research and Clinical Practice 60(1): 49-56.
Cobb, J. P. and R. L. Danner (1996). Nitric oxide and septic shock. Journal of the American Medical Association 275(15): 1192-1196.
Coccia, E. M., E. Stellacci, G. Marziali, G. Weiss and A. Battistini (2000). IFN-gamma and IL-4 differently regulate inducible NO synthase gene expression through IRF-1 modulation. International immunology 12(7): 977-985.
Coffeen, C., C. McKenna, A. Koeppen, N. Plaster, N. Maragakis, J. Mihalopoulos, J. Schwankhaus, K. Flanigang, R. Gregg, L. Ptacek and Y. Fu (2000). Genetic localization of an autosomal dominant leukodystrophy mimicking chronic progressive multiple sclerosis to chromosome 5q31. Hum Mol Genet 9: 787-793.
Collier, N. C., M. P. Sheetz and M. J. Schlesinger (1993). Concomitant changes in mitochondria and intermediate filaments during heat shock and recovery of chicken embryo fibroblasts. Journal of cellular biochemistry 52(3): 297-307.
Concannon, P., H. A. Erlich, C. Julier, G. Morahan, J. Nerup, F. Pociot, J. Todd and S. Rich (Submitted). Linkage analysis of type 1 diabetes combining four genome-wide scans in 1435 multiplex families.
Concannon, P., K. Gogolinewens, D. Hinds, B. Wapelhorst, V. Morrison, B. Stirling, M. Mitra, J. Farmer, S. Williams, N. Cox, G. Bell, N. Risch and R. Spielman (1998). A second-generation screen of the human genome for susceptibility to insulin-dependent diabetes-mellitus. Nature Genet 19: 292-296.
Concannon, P., J. A. Wright, L. G. Wright, D. R. Sylvester and R. S. Spielman (1990). T-cell receptor genes and insulin-dependent diabetes mellitus (IDDM): no evidence for linkage from affected sib pairs. American journal of human genetics 47(1): 45-52.
Corbett, J. A., G. Kwon, M. H. Marino, C. P. Rodi, P. M. Sullivan, J. Turk and M. L. McDaniel (1996b). Tyrosine kinase inhibitors prevent cytokine-induced expression of iNOS and COX-2 by human islets. American journal of physiology 270(6 Pt 1): C1581-C1587.
Cox, N. J., B. Wapelhorst, V. A. Morrison, L. Johnson, L. Pinchuk, R. S. Spielman, J. A. Todd and P. Concannon (2001). Seven regions of the genome show evidence of linkage to type 1 diabetes in a consensus analysis of 767 multiplex families. Am J Hum Genet 69(4): 820-830.
Craig, E. E., A. Chesley and D. A. Hood (1998). Thyroid hormone modifies mitochondrial phenotype by increasing protein import without altering degradation. American journal of physiology 275(6 Pt 1): C1508-C1515.
Crowe, R. R., V. Vieland, S. D. Detera-Wadleigh, D. L. Garver, P. V. Gejman, I. Hovatta and E. Shink (1999). Report of the Chromosome 5 Workshop of the Sixth World Congress on Psychiatric Genetics. American Journal of Medical Genetics - Neuropsychiatric Genetics 88(3): 229-232.
Cucca, F., M. Congia, J. Trowsdale and S. H. Powis (1994). Insulin-dependent diabetes mellitus and the major histocompatibility complex peptide transporters TAP1 and TAP2: no association in a population with a high disease incidence. Tissue antigens 44(4): 234-240.
Cummings, M. C., C. M. Winterford and N. I. Walker (1997). Apoptosis. American journal of surgical pathology 21(1): 88-101.
Cunningham, J. M., JG; Delaney, CA; Green, IC (1994). The effect of nitric-oxide donors on insulin-secretion, cyclic-gmp and cyclic-amp in rat islets of langerhans and the insulin-secreting cell-lines hit-t15 and rinm5f. Molecular and Cellular Endocrinology 102: 23-29.
Cunningham, J. M., J. G. Mabley and I. C. Green (1997). Interleukin 1beta-mediated inhibition of arginase in RINm5F cells. Cytokine 9(8): 570-576.
Daff, S., I. Sagami and T. Shimizu (1999). The 42-amino acid insert in the FMN domain of neuronal nitric-oxide synthase exerts control over Ca(2+)/calmodulin-dependent electron transfer. Journal of Biological Chemistry 274: 30589-30595.
Darville, M. I. and D. L. Eizirik (1998). Regulation by cytokines of the inducible nitric oxide synthase promoter in insulin producing cells. Diabetologia 41: 1101-1108.
Darville, M. I., Y. S. Ho and D. L. Eizirik (2000). NF-kappaB is required for cytokine-induced manganese superoxide dismutase expression in insulin-producing cells. Endocrinology 141(1): 153-162.
Darville, M. I., D. Liu, M. Chen and D. L. Eizirik (2001). Molecular regulation of Fas expression in beta-cells. Diabetes 50(Suppl 1): S83.
Davies, J. L., Y. Kawaguchi, S. T. Bennett, J. B. Copeman, H. J. Cordell, L. E. Pritchard, P. W. Reed, S. C. L. Gough, S. C. Jenkins, S. M. Palmer, K. M. Balfour, B. R. Rowe, M. Farrall, A. H. Barnett, S. C. Bain, et al. (1994). A genome-wide search for human type 1 diabetes susceptibility genes. Nature 371: 130-136.
Davies, R., A. Moore, A. Schedl, E. Bratt, K. Miyahawa, M. Ladomery, C. Miles, A. Menke, V. van-Heyningen and N. Hastie (1999). Multiple roles for the Wilms' tumor suppressor, WT1. Cancer Res 59(7): 1747S-1750S.
Davoodi-Semiromi, A., J. J. Yang and J. X. She (2002). IL-12p40 is associated with type 1 diabetes in Caucasian-American families. Diabetes 51(7): 2334-2336.
DCCT (1993). The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. The Diabetes Control and Complications Trial Research Group. New England Journal of Medicine 329: 977-986.
DCCT, Epidemiology of Diabetes Interventions and Complications Research Group (2003). Sustained effect of intensive treatment of type 1 diabetes mellitus on development and progression of diabetic nephropathy: the Epidemiology of Diabetes Interventions and Complications (EDIC) study. JAMA 290(16): 2159-2167.
Delaney, C., D. Pavlovic, A. Hoorens, D. Pipeleers and D. Eizirik (1997). Cytokines induce deoxyribonucleic acid strand breaks and apoptosis in human pancreatic islet cells. Endocrinology 138: 2610-2614.
Delaney, C., B. Tyrberg, L. Bouwens, H. Vaghef, B. Hellman and D. Eizirik (1996). Sensitivity of human pancreatic islets to peroxynitrite-induced cell dysfunction and death. Febs Letters 394: 300-306.
Delaney, C. A., M. H. Green, J. E. Lowe and I. C. Green (1993). Endogenous nitric oxide induced by interleukin-1 beta in rat islets of Langerhans and HIT-T15 cells causes significant DNA damage as measured by the "comet" assay. FEBS letters 333(3): 291-295.
DeLong, M. J. (1998). Apoptosis: A modulator of cellular homeostasis and disease states. Annals of the New York Academy of Sciences 842: 82-90.
Delrieu, O., D. Dubois-Laforgue, J. Timsit, E. Tournier-Lasserve and S. Caillat-Zucman (2001). A dinucleotide repeat polymorphism at the poly(ADP-ribose) polymerase gene is not associated with predisposition to type 1 diabetes in French Caucasians. Journal of autoimmunity 17(2): 137-140.
Deng, A. (1998). Molecular analysis of the rat inducible nitric oxide synthase genes. Clin Exp Hypertension 20: 53-65.
Deng, A. Y., L. Gu, J. P. Rapp, C. Szpirer and J. Szpirer (1994). Chromosomal assignment of 11 loci in the rat by mouse-rat somatic hybrids and linkage. Mammalian genome 5(11): 712-716.
Deng, A. Y. and J. P. Rapp (1995). Locus for the inducible, but not a constitutive, nitric oxide synthase cosegregates with blood pressure in the Dahl salt-sensitive rat. Journal of clinical investigation 95(5): 2170-2177.
Deng, G., A. Muir, N. Maclaren and J. She (1995). Association of LMP2 and LMP7 genes within the major histocompatibility complex with insulin-dependent diabetes mellitus: population and family studies. American journal of human genetics 56(2): 528-534.
Deng, H. W. and W. M. Chen (2001). The power of the transmission disequilibrium test (TDT) with both case-parent and control-parent trios. Genetical Research 78(3): 289-302.
Denys, P., P. Malvaux, B. Van-Den, H., W. Tanghe and W. Proesmans (1967). Association of an anatomo-pathological syndrome of male pseudohermaphroditism, Wilms' tumor, parenchymatous nephropathy and XX/ XY mosaicism. Archives francaises de pediatrie 24(7): 729-739.
De-Vera, M. E., J. M. Wong, J. Y. Zhou, E. Tzeng, H. R. Wong, T. R. Billiar and D. A. Geller (1996). Cytokine-induced nitric oxide synthase gene transcription is blocked by the heat shock response in human liver cells. Surgery 120(2): 144-149.
DiLorenzo, T. P., R. T. Graser, T. Ono, G. J. Christianson, H. D. Chapman, D. C. Roopenian, S. G. Nathenson and D. V. Serreze (1998). Major histocompatibility complex class I-restricted T cells are required for all but the end stages of diabetes development in nonobese diabetic mice and use a prevalent T cell receptor alpha chain gene rearrangement. Proceedings of the National Academy of Sciences of the United States of America 95(21): 12538-12543.
Dimatteo, M. A., A. C. Loweth, S. Thomas, J. G. Mabley, N. G. Morgan, J. R. Thorpe and I. C. Green (1997). Superoxide, nitric-oxide, peroxynitrite and cytokine combinations all-cause functional impairment and morphological-changes in rat islets of langerhans and insulin-secreting cell-lines, but dictate cell-death by different mechanisms. Apoptosis 2(2): 164-177.
Dinarello, C. A. (1997). Interleukin-1. Cytokine & growth factor reviews 8(4): 253-265.
Ding, H., H. Cheng, Z. Fu, L. Yan and G. Yang (2001). Relationship of large multifunctional proteasome 7 gene polymorphism with susceptibility to type 1 diabetes mellitus and DR3 gene. Chinese medical journal 114(12): 1263-1266.
Dlaska, M. and G. Weiss (1999). Central role of transcription factor NF-IL6 for cytokine and iron-mediated regulation of murine inducible nitric oxide synthase expression. Journal of immunology 162(10): 6171-6177.
Domanico, S., D. DeNagel, J. Dahlseid, J. Green and S. Pierce (1993). Cloning of the gene encoding peptide-binding protein 74 shows that it is a new member of the heat shock protein 70 family. Mol and Cell Biol 13: 3598-3610.
Donath, M. Y., J. Størling, K. Maedler and T. Mandrup-Poulsen (2003). Inflammatory mediators and islet beta-cell failure: a link between type 1 and type 2 diabetes. Journal of molecular medicine (Berlin, Germany) 81(8): 455-470.
Donn, R. P., J. H. Barrett, A. Farhan, A. Stopford, L. Pepper, E. Shelley, N. Davies, W. E. R. Ollier and W. Thomson (2001). Cytokine gene polymorphisms and susceptibility to juvenile idiopathic arthritis. Arthritis & Rheumatism 44(4): 802-810.
Drash, A., F. Sherman, W. H. Hartmann and R. M. Blizzard (1970). A syndrome of pseudohermaphroditism, Wilms' tumor, hypertension, and degenerative renal disease. Journal of pediatrics 76(4): 585-593.
Dubouix, A., I. Gennero, M. Nieto, N. Ser, H. Hannaire-Broutin, J. P. Tauber, J. Pourrat, J. Fauvel, P. Barthe, H. Chap and J. P. Salles (2000). Polymorphism of the 5' untranslated region of NHE1 gene associated with type-I diabetes. Molecular Cell Biology Research Communications 3(3): 141-144.
Dupont, S., C. Dina, E. H. Hani and P. Froguel (1999). Absence of replication in the french population of the association between BETA2/NEUROD-A45T polymorphism and Type 1 diabetes. Diabetes and Metabolism 25(6): 516-517.
Eberhardt, W., D. Kunz, R. Hummel and J. Pfeilschifter (1996). Molecular cloning of the rat inducible nitric oxide synthase gene promoter. Biochem Biophys Res Commun 223(3): 752-756.
Eberhardt, W., C. Plüss, R. Hummel and J. Pfeilschifter (1998). Molecular mechanisms of inducible nitric oxide synthase gene expression by IL-1beta and cAMP in rat mesangial cells. Journal of immunology 160(10): 4961-4969.
Eckenrode, S., M. P. Marron, R. Nicholls, M. C. K. Yang, J. J. Yang, L. C. G. Fonseca and J. X. She (2000). Fine-mapping of the type 1 diabetes locus (IDDM4) on chromosome 11q and evaluation of two candidate genes (FADD and GALN) by affected sibpair and linkage-disequilibrium analyses. Human Genetics 106(1): 14-18.
Eerligh, P., G. Valdigem, B. P. C. Koeleman, B. O. Roep and M. J. Giphart (2002). Association of vitamin D receptor with type 1 diabetes mellitus using FokI in the Dutch population. European Journal of Immunogenetics 29(2): 146-PY - 2002 Conferences, Congresses, Review Annuals.
Einarsdottir, E., I. Soderstrodiem, A. Lofgren-Burstrodiem, S. Haraldsson, S. Nilsson-Ardnor, C. Penha-Goncalves, L. Lind, G. Holmgren, M. Holmberg, K. Asplund and D. Holmberg (2003). The CTLA4 region as a general autoimmunity factor: An extended pedigree provides evidence for synergy with the HLA locus in the etiology of type 1 diabetes mellitus, Hashimoto's thyroiditis and Graves' disease. European Journal of Human Genetics 11(1): 81-84.
Eisenbarth, G. S. and J. M. Jasinski (2004). Disease prevention with islet autoantigens. Endocrinology and metabolism clinics of North America 33(1): 59-73, viii.
Eissa, N. T., A. J. Strauss, C. M. Haggerty, E. K. Choo, S. C. Chu and J. Moss (1996). Alternative splicing of human inducible nitric-oxide synthase mRNA. tissue-specific regulation and induction by cytokines. Journal of biological chemistry 271(43): 27184-27187.
Eissa, N. Y., JW; Haggerty, CM; Choo, EK; Palmer, CD; Moss, J (1998). Cloning and characterization of human inducible nitric-oxidesynthase splice variants - a domain, encoded by exon-8 and exon-9, is critical for dimerization. Proceedings of the National Academy of Science of the United States of America 95: 7625-7630.
Eizirik, D. (1996b). Beta-cell defence and repair mechanisms in human pancreatic islets. Hormone and Metabolic Research 28: 302-305.
Eizirik, D., C. Delaney, M. Green, J. Cunningham, J. Thorpe, D. Pipeleers, C. Hellerstrom and I. Green (1996a). Nitric oxide donors decrease the function and survival of human pancreatic islets. Molecular and Cellular Endocrinology 118: 71-83.
Eizirik, D. L. (1988). Interleukin-1 induced impairment in pancreatic islet oxidative metabolism of glucose is potentiated by tumor necrosis factor. Acta endocrinologica 119(3): 321-325.
Eizirik, D. L., K. Bendtzen and S. Sandler (1991). Short exposure of rat pancreatic islets to interleukin-1 beta induces a sustained but reversible impairment in beta-cell function: influence of protease activation, gene transcription, and protein synthesis. Endocrinology 128(3): 1611-1616.
Eizirik, D. L., A. Björklund and N. Welsh (1993b). Interleukin-1-induced expression of nitric oxide synthase in insulin-producing cells is preceded by c-fos induction and depends on gene transcription and protein synthesis. FEBS letters 317(1-2): 62-66.
Eizirik, D. L., M. Flodstroem, A. E. Karlsen and N.Welch (1996c). The harmony of the spheres: inducible nitric oxide and related genes in the pancreatic beta cells. Diabetologia 39: 875-890.
Eizirik, D. L. and T. Mandrup-Poulsen (2001b). A choice of death - the signal-transduction of immune- mediated beta-cell apoptosis. Diabetologia 44(12): 2115-2133.
Eizirik, D. L. and D. Pavlovic (1997). Is there a role for nitric oxide in beta-cell dysfunction and damage in IDDM? Diabetes/Metabolism Reviews 13(4): 293-307.
Eizirik, D. L., D. G. Pipeleers, Z. Ling, N. Welsh, C. Hellerström and A. Andersson (1994c). Major species differences between humans and rodents in the susceptibility to pancreatic beta-cell injury. Proceedings of the National Academy of Sciences of the United States of America 91(20): 9253-9256.
Eizirik, D. L., S. Sandler, N. Welsh, M. Cetkovic-Cvrlje, A. Nieman, D. A. Geller, D. G. Pipeleers, K. Bendtzen and C. Hellerström (1994a). Cytokine suppress human islet function irrespective of their effects on nitric oxide generation. J Clin Invest 93: 1968-1974.
Eizirik, D. L., M. Welsh, E. Strandell, N. Welsh and S. Sandler (1990). Interleukin-1 beta depletes insulin messenger ribonucleic acid and increases the heat shock protein hsp70 in mouse pancreatic islets without impairing the glucose metabolism. Endocrinology 127(5): 2290-2297.
Eizirik, D. L., N. Welsh and C. Hellerström (1993c). Predominance of stimulatory effects of interleukin-1 beta on isolated human pancreatic islets. Journal of clinical endocrinology and metabolism 76(2): 399-403.
Esposito, L., N. J. Hill, L. E. Pritchard, F. Cucca, C. Muxworthy, M. E. Merriman, A. Wilson, C. Julier, M. Delepine, J. Tuomilehto, E. Tuomilehto-Wolf, C. Ionesco-Tirgoviste, L. Nistico, N. Visalli, M. Baroni, et al. (1998). Genetic analysis of chromosome 2 in type 1 diabetes: Analysis of putative loci IDDM7, IDDM12, and IDDM13 and candidate genes NRAMP1 and IA-2 and the interleukin-1 gene cluster. Diabetes 47(11): 1797-1799.
Esposito, L., V. Lampasona, E. Bonifacio, E. Bosi and M. Ferrari (1997). Lack of association of DMB polymorphism with insulin-dependent diabetes. Journal of autoimmunity 10(4): 395-400.
Fassbender, W. J., B. Goertz, K. Weismüller, B. Steinhauer, H. Stracke, D. Auch, T. Linn and R. G. Bretzel (2002). VDR gene polymorphisms are overrepresented in german patients with type 1 diabetes compared to healthy controls without effect on biochemical parameters of bone metabolism. Hormone and metabolic research Hormon- und Stoffwechselforschung Hormones et metabolisme 34(6): 330-337.
Fava, D., S. Gardner, D. Pyke and R. D. Leslie (1998). Evidence that the age at diagnosis of IDDM is genetically determined. Diabetes care 21(6): 925-929.
Federici, M., A. Petrone, O. Porzio, C. Bizzarri, D. Lauro, R. D'Alfonso, I. Patera, M. Cappa, L. Nistico, M. Baroni, G. Sesti, U. di-Mario, R. Lauro and R. Buzzetti (2003). The Gly(972)- > Arg IRS-1 variant is associated with type 1 diabetes in continental Italy. Diabetes 52(3): 887-890.
Fehsel, K., V. Kolb-Bachofen and K. D. Kroncke (2003). Necrosis is the predominant type of islet cell death during development of insulin-dependent diabetes mellitus in BB rats. Laboratory Investigation 83(4): 549-559.
Feugeas, J. P., H. Caillens, J. C. Poirier, D. Charron, A. Marcelli-Barge and J. L. Wautier (1997). Influence of metabolic and genetic factors on tumour necrosis factor-alpha and lymphotoxin-alpha production in insulin-dependent diabetes mellitus. Diabetes & metabolism 23(4): 295-301.
Feutren, G., L. Papoz, R. Assan, B. Vialettes, G. Karsenty, P. Vexiau, H. Du-Rostu, M. Rodier, J. Sirmai, A. Lallemand and a. et (1986). Cyclosporin increases the rate and length of remissions in insulin-dependent diabetes of recent onset. Results of a multicentre double-blind trial. Lancet 2 (8499): 119-124.
Field, L. L. (2002). Genetic linkage and association studies of Type I diabetes: challenges and rewards. Diabetologia 45(1): 21-35.
Field, L. L. and V. Bonnevie-Nielsen (1999). Locus for antiviral enzyme 2',5'oligoadenylate synthetase (OAS) on chromosome 12q24 influences predisposition to Type 1 diabetes. American Journal of Human Genetics 65(4): A59-PY - 1999.
Field, L. L., Z. Larsen, F. Pociot, J. Nerup, R. Tobias and V. Bonnevie-Nielsen (2002). Evidence for a locus (IDDM16) in the immunoglobulin heavy chain region on chromosome 14q32.3 producing susceptibility to type 1 diabetes. 3(6): 338-344.
Field, L. L., F. Pociot, J. Nerup and V. Bonnevie-Nielsen (1999). Significant association between interferon regulatory factor 2 (IRF2) gene and Type 1 diabetes. Diabetologia 42(SUPPL. 1): A73.
Field, L. L., D. K. Stephure and R. G. McArthur (1991). Interaction between T cell receptor beta chain and immunoglobulin heavy chain region genes in susceptibility to insulin-dependent diabetes mellitus. American journal of human genetics 49(3): 627-634.
Flodstroem, M. and D. L. Eizirik (1997). Interferon-gamma-induced interferon regulatory factor-1 (IRF-1) expression in rodent and human islet cells precedes nitric oxide production. Endocrinology 138: 2747-2753.
Flodstrom, M., M. Chen, A. Smismans, F. Schuit, D. Pipeleers and D. Eizirik (1999a). Interleukin 1beta increases arginine accumulation and activates the citrulline-NO cycle in rat pancreatic beta cells. Cytokine 11: 400-407.
Flodstrom, M., N. Welsh and D. L. Eizirik (1996a). Cytokines activate the nuclear factor kappa-B (NF-kappa-B) and induce nitric oxide production in human pancreatic islets. FEBS Letters 385(1-2): 4-6.
Flodström, M., A. Niemann, F. J. Bedoya, S. M. Morris and D. L. Eizirik (1995). Expression of the citrulline-nitric oxide cycle in rodent and human pancreatic beta-cells: induction of argininosuccinate synthetase by cytokines. Endocrinology 136(8): 3200-3206.
Fossetta, J. D., X. D. Niu, C. A. Lunn, P. J. Zavodny, S. K. Narula and D. Lundell (1996). Expression of human inducible nitric oxide synthase in Escherichia coli. FEBS letters 379(2): 135-138.
Furuta, H., M. Ishigame, M. Nishikimi, M. Furuta, H. Wakasaki, T. Hanabusa, M. Nishi, T. Sanke and K. Nanjo (2001). Missense mutations in the superoxide dismutase 2 (SOD2) gene associated with type I diabetes. Diabetologia 44(Suppl 1): A 83.
Gabai, V. L., A. B. Meriin, D. D. Mosser, A. W. Caron, S. Rits, V. I. Shifrin and M. Y. Sherman (1997). Hsp70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance. Journal of biological chemistry 272(29): 18033-18037.
Gale, E. A. M. (2002). The rise of childhood type 1 diabetes in the 20th century. Diabetes 51(12): 3353-3361.
Galea, E., D. Reis and D. Feinstein (1994). Cloning and expression of inducible nitric oxide synthase from rat astrocytes. J of Neuroscience Research 37: 406-414.
Gambelunghe, G., M. Ghaderi, A. Cosentino, A. Falorni, P. Brunetti, A. Falorni and C. B. Sanjeevi (2000). Association of MHC Class I chain-related A (MIC-A) gene polymorphism with Type I diabetes. Diabetologia 43(4): 507-514.
Gao, J., D. C. Morrison, T. J. Parmely, S. W. Russell and W. J. Murphy (1997). An interferon-gamma-activated site (GAS) is necessary for full expression of the mouse iNOS gene in response to interferon-gamma and lipopolysaccharide. Journal of biological chemistry 272(2): 1226-1230.
Gao, P. S., H. Kawada, T. Kasamatsu, X. Q. Mao, M. H. Roberts, Y. Miyamoto, M. Yoshimura, Y. Saitoh, H. Yasue, K. Nakao, C. N. Adra, J. F. Kun, S. Moro-oka, H. Inoko, L. P. Ho, et al. (2000). Variants of NOS1, NOS2, and NOS3 genes in asthmatics. Biochemical and Biophysical Research Communications 267(3): 761-763.
Garban, H., D. Marquez, T. Magee, J. Moody, T. Rajavashisth, J. Rodriguez, A. Hung, D. Vernet, J. Rajfer and N. Gonzalez-Cadavid (1997). Cloning of rat and human inducible penile nitric oxide synthase. Application for gene therapy of erectile dysfunction. Biol Reproduc 56: 954-963.
Geller, D., C. Lowenstein, R. Shapiro, A. Nussler, M. D. Silvio, S. Wang, D. Nakayama, R. Simmons, S. Snyder and T. Billiar (1993). Molecular cloning and expression of inducible nitric oxide synthase from human hepatocytes. Proceedings of the National Academy of Sciences of the United States of America 90: 3491-3495.
Geller, D. A., M. E. de-Vera, D. A. Russell, R. A. Shapiro, A. K. Nussler, R. L. Simmons and T. R. Billiar (1995). A central role for IL-1 beta in the in vitro and in vivo regulation of hepatic inducible nitric oxide synthase. IL-1 beta induces hepatic nitric oxide synthesis. Journal of immunology 155(10): 4890-4898.
Geng, Y. and M. Lotz (1995). Increased intracellular Ca2+ selectively suppresses IL-1-induced NO production by reducing iNOS mRNA stability. Journal of cell biology 129(6): 1651-1657.
Geng, Y. J., M. Almqvist and G. K. Hansson (1994). cDNA cloning and expression of inducible nitric oxide synthase from rat vascular smooth muscle cells. Biochimica et biophysica acta 1218(3): 421-424.
Gepts, W. (1965). Pathologic anatomy of the pancreas in juvenile diabetes mellitus. Diabetes 14: 619-633.
Gessler, M., A. Poustka, W. Cavenee, R. L. Neve, S. H. Orkin and G. A. Bruns (1990). Homozygous deletion in Wilms tumours of a zinc-finger gene identified by chromosome jumping. Nature 343(6260): 774-778.
Giannoukakis, N. and P. D. Robbins (2002). Gene and cell therapies for diabetes mellitus: strategies and clinical potential. BioDrugs 16(3): 149-173.
Giannoukakis, N., W. A. Rudert, M. Trucco and P. D. Robbins (2000). Protection of human islets from the effects of interleukin- 1β by adenoviral gene transfer of an IκB repressor. Journal of Biological Chemistry 275(47): 36509-36513.
Gilmore, T. D. (1999). The Rel/NF- kappaB signal transduction pathway: introduction. Oncogene 18(49): 6842-6844.
Glenn, C. L., W. Y. S. Wang and B. J. Morris (1999). Different frequencies of inducible nitric oxide synthase genotypes in older hypertensives. Hypertension 33(4): 927-932.
Gores, G. J., B. Herman and J. J. Lemasters (1990). Plasma membrane bleb formation and rupture: a common feature of hepatocellular injury. Hepatology 11(4): 690-698.
Gough, S. C., P. J. Saker, L. E. Pritchard, T. R. Merriman, M. E. Merriman, B. R. Rowe, S. Kumar, T. Aitman, A. H. Barnett and R. C. Turner (1995). Mutation of the glucagon receptor gene and diabetes mellitus in the UK: association or founder effect? Human molecular genetics 4(9): 1609-1612.
Green, A., P. K. Andersen, A. J. Svendsen and K. Mortensen (1992). Increasing incidence of early onset type 1 (insulin-dependent) diabetes mellitus: a study of Danish male birth cohorts. Diabetologia 35(2): 178-182.
Green, A., G. Brutti, C. Patterson, G. Dahlquist, G. Soltesz, A. Green, E. Schober, I. Weets, C. Vandevalle, F. Gorus, M. Coeckelberghs, M. D. Caju, V. Christov, V. Tzaneva, V. Iotova, et al. (2000). Variation and trends in incidence of childhood diabetes in Europe. Lancet 355: 873-876.
Green, A. and C. C. Patterson (2001). Trends in the incidence of childhood-onset diabetes in Europe 1989-1998. Diabetologia 44: B3-B8.
Grey, S. T., M. B. Arvelo, W. Hasenkamp, F. H. Bach and C. Ferran (1999). A20 inhibits cytokine-induced apoptosis and nuclear factor kappaB-dependent gene activation in islets. Journal of Experimental Medicine 190(8): 1135-1145.
Grimm, S. and P. A. Baeuerle (1993). The inducible transcription factor NF-kappaB: structure-function relationship of its protein subunits. Biochemical journal 290 ( Pt 2): 297-308.
Guja, C., S. Marshall, K. Welsh, M. Merriman, A. Smith, J. A. Todd and C. Ionescu-Tîrgoviste (2002). The study of CTLA-4 and vitamin D receptor polymorphisms in the Romanian type 1 diabetes population. 6(1): 75-81.
Guja, C. P., J. A. Todd, K. Welsh, S. Marshall and C. Ionescu-Tirgoviste (2002). Interleukin 10 gene polymorphisms in Romanian type 1 diabetic families. Diabetologia 45(Suppl 2): A 111.
Guo, D., M. Li, Y. Zhang, P. Yang, S. Eckenrode, D. Hopkins, W. Zheng, S. Purohit, R. H. Podolsky, A. Muir, J. Wang, Z. Dong, T. Brusko, M. Atkinson, P. Pozzilli, et al. (2004). A functional variant of SUMO4, a new I kappa B alpha modifier, is associated with type 1 diabetes. Nature genetics 36(8): 837-841.
Guo, H., C. Q. Cai and P. C. Kuo (2002). Hepatocyte nuclear factor-4alpha mediates redox sensitivity of inducible nitric-oxide synthase gene transcription. Journal of biological chemistry 277(7): 5054-5060.
Gupta, M., L. Nikitina-Zake, M. Zarghami, M. Landin-Olsson, I. Kockum, A. Lernmark and C. B. Sanjeevi (2003). Association between the transmembrane region polymorphism of MHC class I chain related gene-A and type 1 diabetes mellitus in Sweden. Human Immunology 64(5): 553-561.
Gurlo, T., K. Kawamura and H. von-Grafenstein (1999). Role of inflammatory infiltrate in activation and effector function of cloned islet reactive nonobese diabetic CD8+ T cells: involvement of a nitric oxide-dependent pathway. Journal of immunology (Baltimore, Md. 163(11): 5770-5780.
Gygi, S. P., Y. Rochon, B. R. Franza and R. Aebersold (1999). Correlation between protein and mRNA abundance in yeast. Molecular and cellular biology 19(3): 1720-1730.
Gylvin, T., R. Bergholdt, J. Nerup and F. Pociot (2002). Characterization of a nuclear-factor-kappa B (NFkappaB) genetic marker in type 1 diabetes (T1DM) families. 3(7): 430-432.
Gysemans, C. A., D. Pavlovic, R. Bouillon, D. L. Eizirik and C. Mathieu (2001). Dual role of interferon-γ signalling pathway in sensitivity of pancreatic beta cells to immune destruction. Diabetologia 44(5): 567-574.
Györffy, B., B. Vásárhelyi, D. Krikovszky, L. Madácsy, A. Tordai, T. Tulassay and A. Szabó (2002). Gender-specific association of vitamin D receptor polymorphism combinations with type 1 diabetes mellitus. European journal of endocrinology 147(6): 803-808.
Haber, D. A., R. L. Sohn, A. J. Buckler, J. Pelletier, K. M. Call and D. E. Housman (1991). Alternative splicing and genomic structure of the Wilms tumor gene WT1. Proceedings of the National Academy of Sciences of the United States of America 88(21): 9618-9622.
Hadjivassiliou, V., M. H. L. Green, R. F. L. James, S. M. Swift, H. A. Clayton and I. C. Green (1998). Insulin secretion, DNA damage, and apoptosis in human and rat islets of Langerhans following exposure to nitric oxide, peroxynitrite, and cytokines. Nitric Oxide 2(6): 429-441.
Hamilton-Williams, E. E., S. E. Palmer, B. Charlton and R. M. Slattery (2003). Beta cell MHC class I is a late requirement for diabetes. Proceedings of the National Academy of Sciences of the United States of America 100(11): 6688-6693.
Hammes, A., J. K. Guo, G. Lutsch, J. R. Leheste, D. Landrock, U. Ziegler, M. C. Gubler and A. Schedl (2001). Two splice variants of the Wilms' tumor 1 gene have distinct functions during sex determination and nephron formation. Cell 106(3): 319-329.
Hampton, M. B. and S. Orrenius (1998). Redox regulation of apoptotic cell death. BioFactors 8(1-2): 1-5.
Hanenberg, H., V. Kolb-Bachofen, G. Kantwerk-Funke and H. Kolb (1989). Macrophage infiltration precedes and is a prerequisite for lymphocytic insulitis in pancreatic islets of pre-diabetic BB rats. Diabetologia 32(2): 126-134.
Hansen, L., J. N. Jensen, S. Urioste, H. V. Petersen, F. Pociot, H. Eiberg, O. P. Kristiansen, T. Hansen, P. Serup, J. Nerup and O. Pedersen (2000). NeuroD/BETA2 gene variability and diabetes - No associations to late-onset type 2 diabetes but an A45 allele may represent a susceptibility marker for type 1 diabetes among Danes. Diabetes 49(5): 876-878.
Hartl, F. U., N. Pfanner, D. W. Nicholson and W. Neupert (1989). Mitochondrial protein import. Biochimica et biophysica acta 988(1): 1-45.
Hashimoto, L., C. Habita, J. Beressi, M. Delepine, C. Besse, A. Cambon-Thomsen, I. Deschapms, J. Rotter, S. Djoulah, M. James, P. Froguel, J. Weissenbach, G. M. Lathrop and C. Julier (1994). Genetic mapping of a susceptibility locus for insulin-dependent mellitus on chromosome 11q. Nature 371: 161-164.
Hashimoto, M., N. Nakamura, H. Obayashi, F. Kimura, A. Moriwaki, G. Hasegawa, H. Shigeta, Y. Kitagawa, K. Nakano, M. Kondo, M. Ohta and M. Nishimura (1999). Genetic contribution of the BAT2 gene microsatellite polymorphism to the age-at-onset of insulin-dependent diabetes mellitus. Human Genetics 105(3): 197-199.
Haslett, C. (1992). Resolution of acute inflammation and the role of apoptosis in the tissue fate of granulocytes. Clinical science 83(6): 639-648.
Hastie, N. D. (2001). Life, sex, and WT1 isoforms - Three amino acids can make all the difference. Cell 106(4): 391-394.
Heding, P. E., A. E. Karlsen, R. Veijola, J. Nerup and F. Pociot (2001). No evidence of a functionally significant polymorphism of the BCL2 gene in Danish, Finnish and Basque type 1 diabetes families. 2(7): 398-400.
Hegazy, D. M., D. A. O'Reilly, B. M. Yang, A. D. Hodgkinson, B. A. Millward and A. G. Demaine (2001). NFkappaB polymorphisms and susceptibility to type 1 diabetes. 2(6): 304-308.
Heimberg, H., Y. Heremans, C. Jobin, R. Leemans, A. K. Cardozo, M. Darville and D. L. Eizirik (2001). Inhibition of cytokine-induced NF-kappa B activation by adenovirus-mediated expression of a NF-kappaB super-repressor prevents beta-cell apoptosis. Diabetes 50(10): 2219-2224.
Heinzmann, A., X. Q. Mao, M. Akaiwa, R. T. Kreomer, P. S. Gao, K. Ohshima, R. Umeshita, Y. Abe, S. Braun, T. Yamashita, M. H. Roberts, R. Sugimoto, K. Arima, Y. Arinobu, B. Yu, et al. (2000). Genetic variants of IL-13 signalling and human asthma and atopy. Human Molecular Genetics 9(4): 549-559.
Heitmeier, M. R., M. Arnush, A. L. Scarim and J. A. Corbett (2001). Pancreatic beta-cell damage mediated by beta-cell production of interleukin-1 - A novel mechanism for virus-induced diabetes. Journal of Biological Chemistry 276(14): 11151-11158.
Heller, B., Z. Q. Wang, E. F. Wagner, J. Radons, A. Bürkle, K. Fehsel, V. Burkart and H. Kolb (1995). Inactivation of the poly(ADP-ribose) polymerase gene affects oxygen radical and nitric oxide toxicity in islet cells. Journal of biological chemistry 270(19): 11176-11180.
Helqvist, S. (1994). Interleukin 1-beta-mediated destruction of pancreatic beta- cells in-vitro - a model of beta-cell destruction in insulin- dependent diabetes-mellitus. Danish Medical Bulletin 41(2): 151-166.
Helqvist, S., P. Bouchelouche, H. U. Andersen and J. Nerup (1989). Modulation of calcium flux influences interleukin 1 beta effects on insulin release from isolated islets of Langerhans. Acta endocrinologica 121(3): 447-455.
Helqvist, S., B. S. Polla, J. Johannesen and J. Nerup (1991a). Heat shock protein induction in rat pancreatic islets by recombinant human interleukin 1 beta. Diabetologia 34: 150-156.
Helqvist, S., B. Sehested-Hansen, J. Johannesen, H. Ullits-Andersen, J. Høiriis-Nielsen and J. Nerup (1989). Interleukin 1 induces new protein formation in isolated rat islets of Langerhans. Acta endocrinologica 121(1): 136-140.
Helqvist, S., U. W. Zumsteg, G. A. Spinas, J. P. Palmer, T. Mandrup-Poulsen, J. Egeberg and J. Nerup (1991b). Repetitive exposure of pancreatic islets to interleukin-1 beta. An in vitro model of pre-diabetes? Autoimmunity 10 (4): 311-318.
Hermann, R., M. Knip, R. Veijola, O. Simell, A. P. Laine, H. K. Akerblom, P. H. Groop, C. Forsblom, K. Pettersson-Fernholm and J. Ilonen (2003). Temporal changes in the frequencies of HLA genotypes in patients with Type 1 diabetes-indication of an increased environmental pressure? Diabetologia 46(3): 420-425.
Herold, K. C., J. B. Burton, F. Francois, E. Poumian-Ruiz, M. Glandt and J. A. Bluestone (2003). Activation of human T cells by FcR nonbinding anti-CD3 mAb, hOKT3gamma1(Ala-Ala). Journal of clinical investigation 111(3): 409-418.
Hevel, J. M., K. A. White and M. A. Marletta (1991). Purification of the inducible murine macrophage nitric oxide synthase. Identification as a flavoprotein. Journal of biological chemistry 266(34): 22789-22791.
Hibberd, M. L., B. A. Millward, F. S. Wong and A. G. Demaine (1992). T-cell receptor constant beta chain polymorphisms and susceptibility to type 1 diabetes. Diabetic medicine 9(10): 929-933.
Hirschhorn, J. N. (2003). Genetic epidemiology of type 1 diabetes. Pediatric diabetes 4(2): 87-100.
Hirschhorn, J. N., K. Lohmueller, E. Byrne and K. Hirschhorn (2002). A comprehensive review of genetic association studies. Genetics in Medicine 4(2): 45-61.
Hitman, G. A., G. C. Toms, B. J. Boucher, L. Garde, P. Baker, J. Awad and H. Festenstein (1989). 2'-5' oligoadenylate synthetase and its relationship to HLA and genetic markers of insulin-dependent diabetes mellitus. Immunogenetics 30(6): 427-431.
Ho, E. and T. M. Bray (1999). Antioxidants, NFkappaB activation, and diabetogenesis. Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N. Y.) 222(3): 205-213.
Hobbs, M. R., V. Udhayakumar, M. C. Levesque, J. Booth, J. M. Roberts, A. N. Tkachuk, A. Pole, H. Coon, S. Kariuki, B. L. Nahlen, E. D. Mwaikambo, A. L. Lal, D. L. Granger, N. M. Anstey and J. B. Weinberg (2002). A new NOS2 promoter polymorphism associated with increased nitric oxide production and protection from severe malaria in Tanzanian and Kenyan children. Lancet 360(9344): 1468-1475.
Hodge, S. E., C. E. Anderson, K. Neiswanger, R. Rubin, R. S. Sparkes, M. C. Sparkes, M. Crist, M. A. Spence, P. I. Terasaki and D. L. Rimoin (1983). Association studies between Type 1 (insulin-dependent) diabetes and 27 genetic markers: lack of association between Type 1 diabetes and Kidd blood group. Diabetologia 25(4): 343-347.
Hofmann, W., H. D. Royer, M. Drechsler, S. Schneider and B. Royer-Pokora (1993). Characterization of the transcriptional regulatory region of the human WT1 gene. Oncogene 8(11): 3123-3132.
Hokari, A., M. Zeniya and H. Esumi (1994). Cloning and functional expression of human inducible nitric oxide synthase (NOS) cDNA from a glioblastoma cell line A- 172. Journal of Biochemistry (Tokyo) 116(3): 575-581.
Hoorens, A. and D. Pipeleers (1999). Nicotinamide protects human beta cells against chemically-induced necrosis, but not against cytokine-induced apoptosis. Diabetologia 42: 55-59.
Hoorens, A., G. Stange, D. Pavlovic and D. Pipeleers (2001). Distinction between interleukin-1-induced necrosis and apoptosis of islet cells. Diabetes 50(3): 551-557.
Hoorens, A., C. Van-de, M., G. Klöppel and D. Pipeleers (1996). Glucose promotes survival of rat pancreatic beta cells by activating synthesis of proteins which suppress a constitutive apoptotic program. Journal of clinical investigation 98(7): 1568-1574.
Hosomi, N., K. Fukai, N. Oiso, A. Kato, T. Murakami, M. Ishii, K. Nakajima and S. Shibahara (2002). Lack of association of a promoter polymorphism of interferon regulatory factor 2 (IRF-2) gene with psoriasis in Japan. American Journal of Human Genetics 71(4 Supplement): 462-PY.
Hughes, J. H., J. R. Colca, R. A. Easom, J. Turk and M. L. McDaniel (1990). Interleukin 1 inhibits insulin secretion from isolated rat pancreatic islets by a process that requires gene transcription and mRNA translation. Journal of Clinical Investigation 86(3): 856-863.
Hussain, I. and M. Qureshi (1998). The expression and regulation of inducible nitric oxide synthase gene differ in macrophages from chickens of different genetic background. Vet Immun Immunopath 61: 317-329.
Hyttinen, V., J. Kaprio, L. Kinnunen, M. Koskenvuo and J. Tuomilehto (2003). Genetic liability of type 1 diabetes and the onset age among 22,650 young Finnish twin pairs: A nationwide follow-up study. Diabetes 52(4): 1052-1055.
Ibi, T., K. Sahashi, J. Ling, K. Marui and T. Mitsuma (1996). Immunostaining of mitochondrial heat shock proteins (mtHSPs) in skeletal muscle fibers of mitochondrial cytopathy. Rinsho shinkeigaku. Clinical neurology 36(1): 61-64.
Ide, A., E. Kawasaki, N. Abiru, F. Sun, R. Takahashi, H. Kuwahara, N. Fujita, A. Kita, K. Oshima, H. Sakamaki, S. Uotani, H. Yamasaki, Y. Yamaguchi and K. Eguchi (2002). Genetic association between interleukin-10 gene promoter region polymorphisms and type 1 diabetes age-at-onset. Human immunology 63(8): 690-695.
Ignarro, L. and F. E. Murad (1995). Advances in Pharmacology, Vol. 34. Nitric oxide: biochemistry, molecular biology, and therapeutic implications. Advances in Pharmacology 34: xxiv+530.
Ihara, K., S. Ahmed, F. Nakao, N. Kinukawa, R. Kuromaru, N. Matsuura, I. Iwata, S. Nagafuchi, H. Kohno, K. Miyako and T. Hara (2001). Association studies of CTLA-4, CD28, and ICOS gene polymorphisms with type 1 diabetes in the Japanese population. Immunogenetics 53(6): 447-454.
Imberti, L., A. Sottini, S. Signorini, S. Pirovano, A. Albertini and R. Gorla (1999). CCR5 delta32 deletion in autoimmune diseases. Clinical Chemistry and Laboratory Medicine 37: S486.
Inoue, T., A. H. Kwon, M. Oda, M. Kaibori, Y. Kamiyama, M. Nishizawa, S. Ito and T. Okumura (2000). Hypoxia and heat inhibit inducible nitric oxide synthase gene expression by different mechanisms in rat hepatocytes. Hepatology 32(5): 1037-1044.
Ito, M., M. Tanimoto, H. Kamura, M. Yoneda, Y. Morishima, K. Takatsuki, T. Itatsu and H. Saito (1988). Association of HLA-DR phenotypes and T-lymphocyte-receptor beta-chain-region RFLP with IDDM in Japanese. Diabetes 37(12): 1633-1636.
Iwashina, M., Y. Hirata, T. Imai, K. Sato and F. Marumo (1996). Molecular cloning of endothelial, inducible nitric oxide synthase gene from rat aortic endothelial cell. European journal of biochemistry 237(3): 668-673.
Iwata, I., S. Nagafuchi, H. Nakashima, S. Kondo, T. Koga, Y. Yokogawa, T. Akashi, T. Shibuya, Y. Umeno, T. Okeda, S. Shibata, S. Kono, M. Yasunami, H. Ohkubo and Y. Niho (1999). Association of polymorphism in the neuroD/BETA2 gene with type 1 diabetes in the Japanese. Diabetes 48(2): 416-419.
Jackson, D. G. and J. D. Capra (1993). TAP1 alleles in insulin-dependent diabetes mellitus: A newly defined centromeric boundary of disease susceptibility. Proceedings of the National Academy of Sciences of the United States of America 90(23): 11079-11083.
Jackson, D. G. and J. D. Capra (1995). TAP2 association with insulin-dependent diabetes mellitus is secondary to HLA-DQB1. Human immunology 43(1): 57-65.
Jahromi, M., A. Millward and A. Demaine (2000). A CA repeat polymorphism of the IFN-gamma gene is associated with susceptibility to type 1 diabetes. Journal of interferon & cytokine research 20(2): 187-190.
Jahromi, M. M., B. A. Millward and A. G. Demaine (2000). A polymorphism in the promoter region of the gene for interleukin-6 is associated with susceptibility to type 1 diabetes mellitus. Journal of interferon & cytokine research 20(10): 885-888.
Janssens, S. P., A. Shimouchi, T. Quertermous, D. B. Bloch and K. D. Bloch (1992). Cloning and expression of a cDNA encoding human endothelium-derived relaxing factor/nitric oxide synthase. Journal of biological chemistry 267(21): 14519-14522.
Jenhani, F., R. Bardi, Y. Gorgi, K. Ayed and M. Jeddi (1992). C4 polymorphism in multiplex families with insulin dependent diabetes in the Tunisian population: standard C4 typing methods and RFLP analysis. Journal of autoimmunity 5(2): 149-160.
Jesch, N. K., M. Dörger, G. Enders, G. Rieder, C. Vogelmeier, K. Messmer and F. Krombach (1997). Expression of inducible nitric oxide synthase and formation of nitric oxide by alveolar macrophages: an interspecies comparison. Environmental health perspectives 105 Suppl 5: 1297-1300.
Johannesen, J., S. Helqvist and J. Nerup (1990). Not all insulin secretagogues sensitize pancreatic islets to recombinant human interleukin 1 beta. Acta endocrinologica 123(4): 445-452.
Johannesen, J., A. E. Karlsen, J. Nerup, F. Pociot and the Danish Study Group for Diabetes in Childhood (1997). No association or linkage to IDDM of an interferon regulatory factor 1 (IRF-1) gene polymorphism in the Danish population. Eur J Immunogenetics 24: 377-383.
Johannesen, J., A. E. Karlsen, F. Pociot, S. G. Roenn and J. Nerup (2003). Strain dependent rat iNOS promoter activity - Correlation to identified WT1 transcription factor binding site. Autoimmunity 36(3): 167-175.
Johannesen, J., A. Pie, A. E. Karlsen, C. M. Larsen, A. Jensen, H. Vissing, O. P. Kristiansen, J. Nerup and DSBD (2004). Is mortalin a candidate gene for T1DM. Autoimmunity, In Press.
Johannesen, J., A. Pie, F. Pociot, O. P. Kristiansen, A. E. Karlsen and J. Nerup (2001a). Linkage of the human inducible nitric oxide synthase gene to type 1 diabetes. J. Clin. Endocrinol. Metab. 86(6): 2792-2796.
Johannesen, J., F. Pociot, A. E. Karlsen, T. Mandrup-Poulsen and J. Nerup (2001b). Strain-dependent difference in inducible nitric oxide synthesis (NOS) expression in rat pancreatic islets correlates with interferon regulating factor 1 (IRF-1) and heat shock protein 70 (HSP70) expression. Eur. Cytokine Netw. 12(3): 501-509.
Johannesen, J., F. Pociot, O. Kristiansen, A. Karlsen, J. Nerup, DIEGG and DSGD (2000b). No evidence for linkage in the promoter region of the inducible nitric oxide synthase gene (NOS2) in a Danish type 1 diabetes population. Genes and Immunity 1: 362-366.
Johannesen, J., L. Tarnow, H. H. Parving, J. Nerup and F. Pociot (2000a). CCTTT-repeat polymorphism in the human NOS2-promoter confers low risk of diabetic nephropathy in type 1 diabetic patients. Diabetes Care 23(4): 560-562.
Johansson, S., B. A. Lie, E. Thorsby and D. E. Undlien (2001). The polymorphism in the 3-prime; untranslated region of ILI2b has a negligible effect on the susceptibility to develop type I diabetes in Norway. Immunogenetics 53(7): 603-605.
John, N., H. Andersen, S. Fey, P. Larsen, P. Roepstroff, M. Larsen, F. Pociot, A. Karlsen, J. Nerup, I. Green and T. Mandrup-Poulsen (2000). Cytokine- or chemically -derived nitric oxide alters the expression of proteins detected by two-dimensional gel electrophoresis in neonatal rat islets of Langerhans. Diabetes 49: 1819-1829.
Johnson, G. C., F. Payne, S. Nutland, H. Stevens, E. Tuomilehto-Wolf, J. Tuomilehto and J. A. Todd (2002). A comprehensive, statistically powered analysis of GAD2 in type 1 diabetes. Diabetes 51(9): 2866-2870.
Jun, H. and J. Yoon (2003). A new look at viruses in type 1 diabetes. Diabetes-Metabolism Research and Reviews 19(1): 8-31.
Jungblut, P. R., U. Zimny-Arndt, E. Zeindl-Eberhart, J. Stulik, K. Koupilova, K. P. Pleissner, A. Otto, E. C. Muller, W. Sokolowska-Kohler, G. Grabher and G. Stoffler (1999). Proteomics in human disease: Cancer, heart and infectious diseases. Electrophoresis 20(10): 2100-2110.
Jaattela, M. (1999). Heat shock proteins as cellular lifeguards. Annals of Medicine 31(4): 261-271.
Jaattela, M., D. Wissing, K. Kokholm, T. Kallunki and M. Egeblad (1998). Hsp7O exerts its anti-apoptotic function downstream of caspase-3-like proteases. EMBO Journal 17(21): 6124-6134.
Kamijo, R., H. Harada, T. Matsuyama, M. Bosland, J. Gerecitano, D. Shapiro, J. Le, S. I. Koh, T. Kimura and S. J. Green (1994). Requirement for transcription factor IRF-1 in NO synthase induction in macrophages. Science 263(5153): 1612-1615.
Kaneto, H., J. Fujii, H. G. Seo, K. Suzuki, T. Matsuoka, M. Nakamura, H. Tatsumi, Y. Yamasaki, T. Kamada and N. Taniguchi (1995). Apoptotic cell death triggered by nitric oxide in pancreatic beta-cells. Diabetes 44(7): 733-738.
Kanno, K., Y. Hirata, T. Imai and F. Marumo (1993). Induction of nitric oxide synthase gene by interleukin in vascular smooth muscle cells. Hypertension 22(1): 34-39.
Karin, M. and Y. Ben-Neriah (2000). Phosphorylation meets ubiquitination: The control of NF- kappaB activity. Annual Review of Immunology (18): 621-663.
Karlsen, A., D. Pavlovic, K. Nielsen, J. Jensen, H. Andersen, F. Pociot, T. Mandrup-Poulsen, D. Eizirik and J. Nerup (2000). Interferon-gamma induces interleukin-1 converting enzyme expression in pancreatic islets by an interferon regulatory factor-1-dependent mechanism. J Clin Endocrin Metab 85: 830-836.
Karlsen, A. E., H. U. Andersen, H. Vissing, P. M. Larsen, S. J. Fey, B. G. Cuartero, O. D. Madsen, J. S. Petersen, S. B. Mortensen, T. Mandrup-Poulsen, E. Boel and J. Nerup (1995). Cloning and expression of cytokine inducible nitric oxide cDNA from rat islets of Langerhans. Diabetes 44: 753-758.
Karlsen, A. E. and T. Dyrberg (1998). Molecular mimicry between non-self, modified self and self in autoimmunity. Seminars in immunology 10(1): 25-34.
Karlsen, A. E., S. G. Rønn, K. Lindberg, J. Johannesen, E. D. Galsgaard, F. Pociot, J. H. Nielsen, T. Mandrup-Poulsen, J. Nerup and N. Billestrup (2001). Suppressor of cytokine signaling 3 (SOCS-3) protects beta-cells against interleukin-1beta - and interferon-gamma-mediated toxicity. Proc Natl Acad Sci U S A 98(21): 12191-12196.
Karlsen, A. E., T. Sparre, K. Nielsen, J. Nerup and F. Pociot (2001). Proteome analysis - A novel approach to understand the pathogenesis of Type 1 diabetes mellitus. Disease Markers 17(4): 205-216.
Karvonen, M., M. Viik-Kajander, E. Moltchanova, I. Libman, R. LaPorte and J. Tuomilehto (2000). Incidence of childhood type 1 diabetes worldwide. Diabetes Care 23(10): 1516-1526.
Kaul, S., R. Wadhwa, Y. Matsuda, P. Hensler, O. Pereira-Smith, Y. Komatsu and Y. Mitsui (1995). Mouse and human chromosomal assignments of mortalin, a novel member of the murine hsp70 family of proteins. Febs Letters 361: 269-272.
Kaul, S. C., E. Duncan, T. Sugihara, R. R. Reddel, Y. Mitsui and R. Wadhwa (2000a). Structurally and functionally distinct mouse hsp70 family members Mot-1 and Mot-2 proteins are encoded by two alleles. DNA research 7(3): 229-231.
Kaul, S. C., E. L. Duncan, A. Englezou, S. Takano, R. R. Reddel, Y. Mitsui and R. Wadhwa (1998a). Malignant transformation of NIH3T3 cells by overexpressed of mot-2 protein. Oncogene 17(7): 907-911.
Kaul, S. C., M. Matsui, S. Takano, T. Sugihara, Y. Mitsui and R. Wadhwa (1997). Expression analysis of mortalin, a unique member of the Hsp70 family of proteins, in rat tissues. Experimental Cell Research 232(1): 56-63.
Kaul, S. C., R. R. Reddel, Y. Mitsui and R. Wadhwa (2001). An N-terminal region of mot-2 binds to p53 in vitro. 3(2): 110-114.
Kaul, S. C., T. Yaguchi, K. Taira, R. R. Reddel and R. Wadhwa (2003). Overexpressed mortalin (mot-2)/mthsp70/GRP75 and hTERT cooperate to extend the in vitro lifespan of human fibroblasts. Experimental Cell Research 286(1): 96-101.
Kawabata, Y., H. Ikegami, Y. Kawaguchi, T. Fujisawa, M. Hotta, H. Ueda, M. Shintani, K. Nojima, M. Ono, M. Nishino, H. Taniguchi, S. Noso, K. Yamada, N. Babaya and T. Ogihara (2000). Age-related association of MHC class I chain-related gene A (MICA) with type 1 (insulin-dependent) diabetes mellitus. Human Immunology 61(6): 624-629.
Kawaguchi, Y., H. Ikegami, M. Fukuda, Y. Fujioka, K. Shima and T. Ogihara (1993). Polymorphism of HSP70 gene is not associated with type 1 (insulin-dependent) diabetes mellitus in Japanese. Diabetes research and clinical practice 21(2-3): 103-107.
Kawaguchi, Y., H. Ikegami, M. Fukuda, K. Takekawa, Y. Fujioka, T. Fujisawa, H. Ueda and T. Ogihara (1994). Absence of association of TAP and LMP genes with type 1 (insulin-dependent) diabetes mellitus. Life Sciences 54(26): 2049-2053.
Kawahara, D. J. and J. S. Kenney (1991). Species differences in human and rat islet sensitivity to human cytokines. Monoclonal anti-interleukin-1 (IL-1) influences on direct and indirect IL-1-mediated islet effects. Cytokine 3(2): 117-124.
Kawai, A., S. Nishikawa, A. Hirata and T. Endo (2001). Loss of the mitochondrial Hsp70 functions causes aggregation of mitochondria in yeast cells. Journal of cell science 114(Pt 19): 3565-3574.
Kay, T. W., I. L. Campbell and L. C. Harrison (1991). Characterization of pancreatic T lymphocytes associated with beta cell destruction in the non-obese diabetic (NOD) mouse. Journal of autoimmunity 4(2): 263-276.
Keinanen, R., N. Vartiainen and J. Koistinaho (1999). Molecular cloning and characterization of the rat inducible nitric oxide synthase (iNOS) gene. Gene 234: 297-305.
Kelly, H. and M. J. Garlepp (1993). T cell receptor haplotypes in families of patients with insulin-dependent diabetes mellitus. Clinical and experimental immunology 91(2): 226-231.
Kennedy, D., T. Ramsdale, J. Mattick and M. Little (1996). An RNA recognition motif in Wilms' tumour protein (WT1) revealed by structural modelling. Nature genetics 12(3): 329-331.
Kent, J., M. Lee, A. Schedl, S. Boyle, J. Fantes, M. Powell, N. Rushmere, C. Abbott, V. van-Heyningen and W. A. Bickmore (1997). The reticulocalbin gene maps to the WAGR region in human and to the Small eye Harwell deletion in mouse. Genomics 42(2): 260-267.
Keymeulen, B., Z. Ling, F. K. Gorus, G. Delvaux, L. Bouwens, A. Grupping, C. Hendrieckx, M. Pipeleers-Marichal, C. Van-Schravendijk, K. Salmela and D. G. Pipeleers (1998). Implantation of standardized beta-cell grafts in a liver segment of IDDM patients: Graft and recipient characteristics in two cases of insulin- independence under maintenance immunosuppression for prior kidney graft. Diabetologia 41(4): 452-459.
Kinjo, Y., N. Matsuura, Y. Yokota, S. Ohtsu, K. Nomoto, I. Komiya, J. Sugimoto, Y. Jinno and N. Takasu (2001). Identification of nonsynonymous polymorphisms in the superantigen-coding region of IDDMK(1,2)22 and a pilot study on the association between IDDMK(1,2)22 and type 1 diabetes. Journal of Human Genetics 46(12): 712-716.
Kinugawa, K., T. Shimizu, A. Yao, O. Kohmoto, T. Serizawa and T. Takahashi (1997). Transcriptional regulation of inducible nitric oxide synthase in cultured neonatal rat cardiac myocytes. Circulation research 81(6): 911-921.
Kirk, R. L., P. R. Ranford, S. W. Serjeantson, A. R. Thompson, S. M. Munirathnam-Chetty, L. John, V. Mohan, A. Ramachandran, C. Snehalatha and M. Viswanathan (1985). HLA, complement C2, C4, properdin factor B and glyoxalase types in South Indian diabetics. Diabetes research and clinical practice 1(1): 41-47.
Kirk, R. L., P. R. Ranford, J. Theophilus, H. M. Ahuja, N. K. Mehra and M. C. Vaidya (1982). The rare factor BfSl of the properdin system strongly associated with insulin-dependent diabetes in north India. Tissue antigens 20(4): 303-304.
Kishimoto, J., N. Spurr, M. Liao, L. Lizhi, P. Emson and W. Xu (1992). Localization of brain nitric oxide synthase (NOS) to human chromosome 12. Genomics 14(3): 802-804.
Kloting, I., B. Van-Den, J., N. Kloting and B. Radovic (2003). Alleles of diabetes-resistant BN rats contribute to insulin- dependent type 1 diabetes mellitus. Journal of Autoimmunity 20(2): 119-123.
Klupa, T., M. Malecki, L. Hanna, J. Sieradzka, J. Frey, J. H. Warram, J. Sieradzki and A. S. Krolewski (1999). Amino acid variants of the vitamin D-binding protein and risk of diabetes in white Americans of European origin. European Journal of Endocrinology 141(5): 490-493.
Kobzik, L., D. S. Bredt, C. J. Lowenstein, J. Drazen, B. Gaston, D. Sugarbaker and J. S. Stamler (1993). Nitric oxide synthase in human and rat lung: immunocytochemical and histochemical localization. American journal of respiratory cell and molecular biology 9(4): 371-377.
Koide, M., Y. Kawahara, T. Tsuda, I. Nakayama and M. Yokoyama (1994). Expression of nitric oxide synthase by cytokines in vascular smooth muscle cells. Hypertension 23(1 Suppl): I45-I48.
Kolb, H. (1997). Benign versus destructive insulitis. Diabetes/metabolism Reviews 13(3): 139-146.
Kolb, H. and V. Kolb-Bachofen (1998). Nitric oxide in autoimmune disease: cytotoxic or regulatory mediator? Immunology Today 19: 556-561.
Kolb-Bachofen, V., S. Epstein, U. Kiesel and H. Kolb (1988). Low-dose streptozocin-induced diabetes in mice. Electron microscopy reveals single-cell insulitis before diabetes onset. Diabetes 37(1): 21-27.
Kolyada, A. Y. and N. E. Madias (2001). Transcriptional regulation of the human iNOS gene by IL-1beta in endothelial cells. Molecular medicine 7(5): 329-343.
Kolyada, A. Y., N. Savikovsky and N. E. Madias (1996). Transcriptional regulation of the human iNOS gene in vascular-smooth-muscle cells and macrophages: evidence for tissue specificity. Biochemical and biophysical research communications 220(3): 600-605.
Komaki, S., M. Kohno, N. Matsuura, M. Shimadzu, N. Adachi, R. Hoshide, S. Nishiyama and I. Matsuda (1998). The polymorphic 43Thr bcl-2 protein confers relative resistance to autoimmunity: An analytical evaluation. Human Genetics 103(4): 435-440.
Konno, S., N. Hizawa, E. Yamaguchi, E. Jinushi and M. Nishimura (2001). (CCTTT)(n) repeat polymorphism in the NOS2 gene promoter is associated with atopy. Journal of Allergy and Clinical Immunology 108(5): 810-814.
Korpinen, E., P. H. Groop, A. Rautio, L. Madácsy, A. Reunanen, O. Vaarala and H. K. Akerblom (1999). N-acetyltransferase-2 polymorphism, smoking and type 1 diabetic nephropathy. Pharmacogenetics 9(5): 627-633.
Korsgren, O. and L. Jansson (1994). Characterization of mixed syngeneic-allogeneic and syngeneic-xenogeneic islet-graft rejections in mice. Evidence of functional impairment of the remaining syngeneic islets in xenograft rejections. Journal of clinical investigation 93(3): 1113-1119.
Kozak, M. (1992). Regulation of translation in eukaryotic systems. Annual review of cell biology 8: 197-225.
Kretowski, A., K. Mironczuk, A. Karpinska, U. Bojaryn, M. Kinalski, Z. Puchalski and I. Kinalska (2002). Interleukin-18 promoter polymorphisms in type 1 diabetes. Diabetes 51(11): 3347-3349.
Krikovsky, D., B. Vásárhelyi, A. Treszl, A. Körner, A. Tordai, T. Tulassay and L. Madácsy (2002). Genetic polymorphism of interleukin-1beta is associated with risk of type 1 diabetes mellitus in children. European journal of pediatrics 161(9): 507-508.
Kristiansen, O. P., A. E. Karlsen, Z. M. Larsen, J. Johannesen, F. Pociot, T. Mandrup-Poulsen, DIEGG and DSGD (2004). Identification of a type 1 diabetes-associated CD4 promoter haplotype with high constitutive activity. Scandinavian journal of immunology 59(6): 582-591.
Kristiansen, O. P., Z. M. Larsen, J. Johannesen, J. Nerup, T. Mandrup-Poulsen, F. Pociot, D.I.E.G.G. and D.S.G.D. (1999). No linkage of P187S polymorphism in NAD(P)H: Quinone oxidoreductase (NQO1/DIA4) and type 1 diabetes in the Danish population. Human Mutation 14(1): 67-70.
Kristiansen, O. P., Z. M. Larsen and F. Pociot (2000). CTLA-4 in autoimmune diseases--a general susceptibility gene to autoimmunity? 1(3): 170-184.
Kristiansen, O. P., R. L. Nolsoe, H. Holst, S. Reker, Z. M. Larsen, J. Johannesen, J. Nerup, F. Pociot and T. Mandrup-Poulsen (2000). The intercellular adhesion molecule-1 K469E polymorphism in type 1 diabetes. Immunogenetics 52(1-2): 107-111.
Kristiansen, O. P., F. Pociot, E. P. Bennett, H. Clausen, J. Johannesen, J. Nerup and T. Mandrup-Poulsen (2000). IDDM7 links to insulin-dependent diabetes mellitus in Danish multiplex families but linkage is not explained by novel polymorphisms in the candidate gene GALNT3. The Danish Study Group of Diabetes in Childhood and The Danish IDDM Epidemiology and Genetics Group. Human mutation 15(3): 295-296.
Kristiansen, O. P., F. Pociot, J. Johannesen, R. Bergholdt, C. A. Dinarello, J. Nerup and T. Mandrup-Poulsen (2000). Linkage disequilibrium testing of four interleukin-1 gene-cluster polymorphisms in Danish multiplex families with insulin-dependent diabetes mellitus. Cytokine 12(2): 171-175.
Kristiansen, O. P., M. Zamani, J. Johannesen, T. Mandrup-Poulsen, J. Cassiman-J., J. Nerup and F. Pociot (1998). Linkage and association between a CD4 gene polymorphism and IDDM in Danish IDDM patients. Diabetes 47(2): 281-283.
Kroef, M. J., R. Willemze and J. E. Landegent (1993). Dinucleotide repeat polymorphism in the interferon regulating factor 1 (IRF1) gene. Human molecular genetics 2(10): 1748-PY - 1993.
Kroncke, K., M. Rodriguez, H. Kolb and V. Kolb-Bachofen (1993). Cytotoxicity of activated rat macrophages against syngeneic islet cells is arginine-dependent, correlates with citrulline and nitrite concentrations and is identical to lysis by the nitric oxide donor nitroprusside. Diabetologia 36: 14-24.
Kroncke, K. D., V. Kolb-Bachofen, B. Berschick, V. Burkart and H. Kolb (1991). Activated macrophages kill pancreatic syngeneic islet cells via arginine-dependent nitric oxide generation. Biochemical and Biophysical Research Communications 175(3): 752-758.
Kröncke, K. D., K. Fehsel and V. Kolb-Bachofen (1998). Inducible nitric oxide synthase in human diseases. Clinical and experimental immunology 113(2): 147-156.
Kumaramanickavel, G., S. Sripriya, R. N. Vellanki, N. K. Upadyay, S. S. Badrinath, V. Rajendran, B. Sukumar, V. L. Ramprasad and T. Sharma (2002). Inducible nitric oxide synthase gene and diabetic retinopathy in Asian Indian patients. Clinical Genetics 61(5): 344-348.
Kun, J. F., B. Mordmueller, D. J. Perkins, J. May, O. Mercereau-Puijalon, M. Alpers, J. B. Weinberg and P. G. Kremsner (2001). Nitric oxide synthase 2Lambarene (G-954C), increased nitric oxide production, and protection against malaria. Journal of Infectious Diseases 184(3): 330-336.
Kun, J. F. J., B. Mordmuller, B. Lell, L. G. Lehman, D. Luckner and P. G. Kremsner (1998). Polymorphism in promoter region of inducible nitric oxide synthase gene and protection against malaria. Lancet 351(9098): 265-266.
Kuo, P. C., K. Abe and R. A. Schroeder (2000). Superoxide enhances interleukin 1beta-mediated transcription of the hepatocyte-inducible nitric oxide synthase gene. Gastroenterology 118(3): 608-618.
Kuo, P. C., K. Y. Abe and R. A. Schroeder (1997). Oxidative stress increases hepatocyte iNOS gene transcription and promoter activity. Biochemical and biophysical research communications 234(2): 289-292.
Kutlu, B., A. K. Cardozo, M. I. Darville, M. Kruhoffer, N. Magnusson, T. Orntoft and D. L. Eizirik (2003). Discovery of gene networks regulating cytokine-induced dysfunction and apoptosis in insulin-producing INS-1 cells. Diabetes 52(11): 2701-2719.
Kyvik, K. O., A. Green and H. Beck-Nielsen (1995). Concordance rates of insulin dependent diabetes mellitus: a population based study of young Danish twins. BMJ 311(7010): 913-917.
Laitinen, T. (2004). Characterization of a common susceptibility locus for asthma- related traits. Science. 304: 300-304.
Lakey, J. R., W. L. Suarez-Pinzon, K. Strynadka, G. S. Korbutt, R. V. Rajotte, J. G. Mabley, C. Szabó and A. Rabinovitch (2001). Peroxynitrite is a mediator of cytokine-induced destruction of human pancreatic islet beta cells. Laboratory investigation; a journal of technical methods and pathology 81(12): 1683-1692.
Larsen, C., K. Wadt, L. Juhl, H. Andersen, A. Karlsen, M. Su, K. Seedorf, L. Shapiro, C. Dinarello and T. Mandrup-Poulsen (1998). Interleukin-1beta-induced rat pancreatic islet nitric oxide synthesis requires both the p38 and extracellular signal- regulated kinase 1/2 mitogen-activated protein kinases. J Biol Chem 273: 15294-15300.
Larsen, Z., O. Kristiansen, E. Mato, J. Johannesen, M. Puig-Domingo, A. d. Leiva, J. Nerup and F. Pociot (1999). IDDM12 (CTLA4) on 2q33 and IDDM13 on 2q34 in genetic susceptibility to type 1 diabetes (insulin-dependent). Autoimmunity 31: 35-42.
Larsen, Z. M., A. D. Angelo, M. Cattaneo, J. Nerup, I. Biunno, M. Zollo and F. Pociot (2001). Complete mutation scanning of the human SEL 1L gene: a candidate gene for type 1 diabetes. Acta Diabetologica 38(4): 191-192.
Larsen, Z. M., J. Johannesen, O. P. Kristiansen, J. Nerup, F. Pociot and F. Pociot (2004). Evidence for linkage on chromosome 4p16.1 in Type 1 diabetes Danish families and complete mutation scanning of the WFS1 (Wolframin) gene. Diabetic Medicine 21(3): 218-222.
Lea, R. A., R. P. Curtain, A. G. Shepherd, P. J. Brimage and L. R. Griffiths (2001). No evidence for involvement of the human inducible nitric oxide synthase (iNOS) gene in susceptibility to typical migraine. American Journal of Medical Genetics 105(1): 110-113.
Lee, K. H., K. W. Wucherpfennig and D. C. Wiley (2001). Structure of a human insulin peptide-HLA-DQ8 complex and susceptibility to type I diabetes. Nature Immunology 2(6): 501-507.
Lee, K. U., M. K. Kim, K. Amano, C. Y. Pak, M. A. Jaworski, J. G. Mehta and J. W. Yoon (1988). Preferential infiltration of macrophages during early stages of insulitis in diabetes-prone BB rats. Diabetes 37(8): 1053-1058.
Lee, Y., F. Huang, C. Wang, F. Lo, K. Tsan, C. Hsu, C. Huang, S. Chang and J. Chang (2000). Polymorphism in the transmembrane region of the MICA gene and type 1 diabetes. Journal of Pediatric Endocrinology & Metabolism 13(5): 489-496.
Lemasters, J. J., T. Qian, C. A. Bradham, D. A. Brenner, W. E. Cascio, L. C. Trost, Y. Nishimura, A. L. Nieminen and B. Herman (1999). Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death. Journal of Bioenergetics and Biomembranes 31(4): 305-319.
Lenzen, S., J. Drinkgern and M. Tiedge (1996). Low antioxidant enzyme gene expression in pancreatic islets compared with various other mouse tissues. Free radical biology & medicine 20(3): 463-466.
Lernmark, A. and J. Ott (1998). Sometimes it's hot, sometimes it's not. Nature genetics 19(3): 213-214.
Levecque, C., A. Elbaz, J. Clavel, F. Richard, J. Vidal, P. Amouyel, C. Tzourio, A. Alperovitch and M. Chartier-Harlin (2003). Association between Parkinson's disease and polymorphisms in the nNOS and iNOS genes in a community-based case-control study. Human Molecular Genetics 12(1): 79-86.
Levesque, M. C., M. R. Hobbs, N. M. Anstey, T. N. Vaughn, J. A. Chancellor, A. Pole, D. J. Perkins, M. A. Misukonis, S. J. Chanock, D. L. Granger and J. B. Weinberg (1999). Nitric oxide synthase type 2 promoter polymorphisms, nitric oxide production, and disease severity in Tanzanian children with malaria. Journal of Infectious Diseases 180(6): 1994-2002.
Lhotta, K., M. Auinger, F. Kronenberg, K. Irsigler and P. König (1996). Polymorphism of complement C4 and susceptibility to IDDM and microvascular complications. Diabetes care 19(1): 53-55.
Liblau, R. S., S. M. Singer and H. O. McDevitt (1995). Th1 and Th2 CD4+ T cells in the pathogenesis of organ-specific autoimmune diseases. Immunology today 16(1): 34-38.
Lie, B. A., J. A. Todd, F. Pociot, J. Nerup, H. E. Akselsen, G. Joner, K. Dahl-Jorgensen, K. S. Ronningen, E. Thorsby and D. E. Undlien (1999). The predisposition to type 1 diabetes linked to the human leukocyte antigen complex includes at least one non-class II gene. American Journal of Human Genetics 64(3): 793-800.
Ling, Z., M. C. Chen, A. Smismans, D. Pavlovic, F. Schuit, D. L. Eizirik and D. G. Pipeleers (1998). Intercellular differences in interleukin 1beta-induced suppression of insulin synthesis and stimulation of noninsulin protein synthesis by rat pancreatic beta-cells. Endocrinology 139(4): 1540-1545.
Ling, Z., P. A. In't-Veld and D. G. Pipeleers (1993). Interaction of interleukin-1 with islet beta-cells. Distinction between indirect, aspecific cytotoxicity and direct, specific functional suppression. Diabetes 42(1): 56-65.
Ling, Z., C. Van-de, M., D. L. Eizirik and D. G. Pipeleers (2000). Interleukin-1beta-induced alteration in a beta-cell phenotype can reduce cellular sensitivity to conditions that cause necrosis but not to cytokine-induced apoptosis. Diabetes 49(3): 340-345.
Linn, S. C., P. J. Morelli, I. Edry, S. E. Cottongim, C. Szabó and A. L. Salzman (1997). Transcriptional regulation of human inducible nitric oxide synthase gene in an intestinal epithelial cell line. American journal of physiology 272(6 Pt 1): G1499-G1508.
Little, M., G. Holmes, W. Bickmore, V. van-Heyningen, N. Hastie and B. Wainwright (1995). DNA binding capacity of the WT1 protein is abolished by Denys-Drash syndrome WT1 point mutations. Human molecular genetics 4(3): 351-358.
Little, M., G. Holmes and P. Walsh (1999). WT1: what has the last decade told us? BioEssays 21(3): 191-202.
Little, M. and C. Wells (1997). A clinical overview of WT1 gene mutations. Human mutation 9(3): 209-225.
Liu, D., M. Darville and D. L. Eizirik (2001). Double-Stranded Ribonucleic Acid (RNA) Induces beta-Cell Fas Messenger RNA Expression and Increases Cytokine-Induced beta-Cell Apoptosis. Endocrinology 142(6): 2593-2599.
Liu, D., D. Pavlovic, M. Chen, M. Flodstrom, S. Sandler and D. Eizirik (2000). Cytokines induce apoptosis in beta-cells isolated from mice lacking the inducible isoform of nitric oxide synthase (iNOS-/-). Diabetes 49: 1116-1122.
Lockhart, D. and E. Winzeler (2000). Genomics, gene expression and DNA arrays. Nature 405: 827-836.
Lohmueller, K. (2003). Meta-analysis of genetic association studies supports a contribution of common variants to susceptibility to common disease. Nature genetics. 33: 177-182.
Lohoff, M., D. Ferrick, H. W. Mittrucker, G. S. Duncan, S. Bischof, M. Rollinghoff and T. W. Mak (1997). Interferon regulatory factor-1 is required for a T helper 1 immune response in vivo. Immunity 6(6): 681-689.
Lorenzen, T., F. Pociot, P. Hougaard and J. Nerup (1994). Long-term risk of iddm in first-degree relatives of patients with iddm. Diabetologia 37(3): 321-327.
Lorenzen, T., F. Pociot, L. Stilgren, O. Kristiansen, J. Johannesen, P. Olsen, A. Walmar, A. Larsen, N. Albrechtsen, P. Eskildsen, O. Andersen and J. N. J (1998). Predictors of IDDM recurrence risk in offspring of Danish IDDM patients. Danish IDDM Epidemiology and Genetics Group. Diabetologia 41: 666-673.
Lortz, S., M. Tiedge, T. Nachtwey, A. Karlsen, J. Nerup and S. Lenzen (2000). Protection of insulin-producing RINm5F cells against cytokine-mediated toxicity through overexpression of antioxidant enzymes. Diabetes 49: 1123-1130.
Los, H., G. Koppelman and D. Postma (1999). The importance of genetic influences in asthma. Eur Resp J 14: 1210-1227.
Lotfi, K., G. Sund, R. Lowe, J. Graham, M. Landin-Olsson, I. Kockum, S. Deeb and A. Lernmark (1997). The beta cell glucokinase promoter variant is an unlikely risk factor for diabetes mellitus. Diabetes Incidence Study in Sweden (DISS). Diabetologia 40(8): 959-962.
Lowe, R. M., J. Graham, G. Sund, I. Kockum, M. Landin-Olsson, J. B. Schaefer, C. T.örn, A. Lernmark and G. Dahlquist (2000). The length of the CTLA-4 microsatellite (AT)N-repeat affects the risk for type 1 diabetes. Diabetes Incidence in Sweden Study Group. Autoimmunity 32(3): 173-180.
Lowenstein, C. J., E. W. Alley, P. Raval, A. M. Snowman, S. H. Snyder, S. W. Russell and W. J. Murphy (1993). Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc Natl Acad Sci U S A 90(20): 9730-9734.
Lu, X., P. Lu, R.-Y. Xing, Q.-Y. Sun, Z.-W. Qiu, L. Han, X.-b. Zhou and X.-F. Zheng (2002). A short tandem repeat polymorphism in the inducible nitric oxide synthase gene in Chinese population. Acta Genetica Sinica 29(4): 290-293.
Ma, L., A. Penfornis, X. Wang, D. Schoenfeld, E. Tuomilehto-Wolf, K. Metcalfe, G. Hitman and D. Faustman (1997). Evaluation of TAP1 polymorphisms with insulin dependent diabetes mellitus in Finnish diabetic patients. The Childhood Diabetes in Finland (DiMe) Study Group. Human immunology 53(2): 159-166.
Ma, Y., J. D. Ohmen, Z. Li, L. G. Bentley, C. McElree, S. Pressman, S. R. Targan, N. Fischel-Ghodsian, J. I. Rotter and H. Yang (1999). A genome-wide search identifies potential new susceptibility loci for Crohn's disease. Inflammatory bowel diseases 5(4): 271-278.
Mabley, J. G., V. Belin, N. John and I. C. Green (1997). Insulin-like growth factor I reverses interleukin-1beta inhibition of insulin secretion, induction of nitric oxide synthase and cytokine-mediated apoptosis in rat islets of Langerhans. FEBS letters 417(2): 235-238.
Mabley, J. G., G. Haskó, L. Liaudet, F. Soriano, G. J. Southan, A. L. Salzman and C. Szabó (2002). NFkappaB1 (p50)-deficient mice are not susceptible to multiple low-dose streptozotocin-induced diabetes. Journal of endocrinology 173(3): 457-464.
Mabley, J. G., G. J. Southan, A. L. Salzman and C. Szabo (2004). The combined inducible nitric oxide synthase inhibitor and free radical scavenger guanidinoethyldisulfide prevents multiple low-dose streptozotocin-induced diabetes in vivo and interleukin-1 beta-induced suppression of islet insulin secretion in vitro. Pancreas 28(2): E39-E44.
Madden, S. L., D. M. Cook, J. F. Morris, A. Gashler, V. P. Sukhatme and I. I. I. Rauscher-F.J. (1991). Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science 253(5027): 1550-1553.
Maedler, K., P. Sergeev, F. Ris, J. Oberholzer, H. I. Joller-Jemelka, G. A. Spinas, N. Kaiser, P. A. Halban and M. Y. Donath (2002). Glucose-induced beta cell production of IL-1 beta contributes to glucotoxicity in human pancreatic islets. Journal of Clinical Investigation 110(6): 851-860.
Maheswaran, S., S. Park, A. Bernard, J. F. Morris, F. J. Rauscher, D. E. Hill and D. A. Haber (1993). Physical and functional interaction between WT1 and p53 proteins. Proc Natl Acad Sci U S A 90(11): 5100-5104.
Maier, R., G. Bilbe, J. Rediske and M. Lotz (1994). Inducible nitric oxide synthase from human articular chondrocytes: cDNA cloning and analysis of mRNA expression. Biochimica et biophysica acta 1208(1): 145-150.
Majno, G. and I. Joris (1995). Apoptosis, oncosis, and necrosis. An overview of cell death. American journal of pathology 146(1): 3-15.
Malecki, M. T., T. Klupa, D. Moczulski, J. H. Warram and A. S. Krolewski (2000). No evidence of association between vitamin D receptor (VDR) gene polymorphisms and type 1 diabetes in Caucasians. Diabetologia 43(Suppl 1): A7-PY - 2000.
Mamane, Y., C. Heylbroeck, P. Génin, M. Algarté, M. J. Servant, C. LePage, C. DeLuca, H. Kwon, R. Lin and J. Hiscott (1999). Interferon regulatory factors: the next generation. Gene 237(1): 1-14.
Mandrup-Poulsen, T. (1996). The role of interleukin-1 in the pathogenesis of IDDM. Diabetologia 39: 1005-1029.
Mandrup-Poulsen, T. (2001). beta-Cell apoptosis: Stimuli and signaling. Diabetes 50(Suppl 1): S58-S63.
Mandrup-Poulsen, T., K. Bendtzen, C. A. Dinarello and J. Nerup (1987b). Human tumor necrosis factor potentiates human interleukin 1-mediated rat pancreatic beta-cell cytotoxicity. Journal of immunology 139(12): 4077-4082.
Mandrup-Poulsen, T., K. Bendtzen, J. Nerup, C. A. Dinarello, M. Svenson and J. H. Nielsen (1986a). Affinity-purified human interleukin I is cytotoxic to isolated islets of Langerhans. Diabetologia 29(1): 63-67.
Mandrup-Poulsen, T., K. Bendtzen, J. Nerup, J. Egeberg and J. H. Nielsen (1986b). Mechanisms of pancreatic islet cell destruction. Dose-dependent cytotoxic effect of soluble blood mononuclear cell mediators on isolated islets of Langerhans. Allergy 41(4): 250-259.
Mandrup-Poulsen, T., K. Bendtzen, J. H. Nielsen, G. Bendixen and J. Nerup (1985). Cytokines cause functional and structural damage to isolated islets of Langerhans. Allergy 40(6): 424-429.
Mandrup-Poulsen, T., J. Egeberg, J. Nerup, K. Bendtzen, J. H. Nielsen and C. A. Dinarello (1987a). Ultrastructural studies of time-course and cellular specificity of interleukin-1 mediated islet cytotoxicity. Acta pathologica, microbiologica, et immunologica Scandinavica Section C, Immunology 95(2): 55-63.
Mandrup-Poulsen, T., G. A. Spinas, S. J. Prowse, B. S. Hansen, D. W. Jørgensen, K. Bendtzen, J. H. Nielsen and J. Nerup (1987). Islet cytotoxicity of interleukin 1. Influence of culture conditions and islet donor characteristics. Diabetes 36(5): 641-647.
Mansfield, M. W., M. H. Stickland, A. M. Carter and P. J. Grant (1994). Polymorphisms of the plasminogen activator inhibitor-1 gene in type 1 and type 2 diabetes, and in patients with diabetic retinopathy. Thrombosis and haemostasis 71(6): 731-736.
Marchenko, N. D., A. Zaika and U. M. Moll (2000). Death signal-induced localization of p53 protein to mitochondria - A potential role in apoptotic signaling. Journal of Biological Chemistry 275(21): 16202-16212.
Margulis, B. A., S. Sandler, D. L. Eizirik, N. Welsh and M. Welsh (1991). Liposomal delivery of purified heat shock protein hsp70 into rat pancreatic islets as protection against interleukin 1 beta-induced impaired beta-cell function. Diabetes 40(11): 1418-1422.
Marks-Konczalik, J., S. C. Chu and J. Moss (1998). Cytokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor kappaB-binding sites. Journal of biological chemistry 273(35): 22201-22208.
Marletta, M. A. (1993). Nitric oxide synthase structure and mechanism. Journal of biological chemistry 268(17): 12231-12234.
Marquet, S., L. Abel, D. Hillaire and A. Dessein (1999). Full results of the genome-wide scan which localises a locus controlling the intensity of infection by Schistosoma mansoni on chromosome 5q31-q33. European Journal of Human Genetics 7(1): 88-97.
Marrero, M. B., V. J. Venema, H. He, R. B. Caldwell and R. C. Venema (1998). Inhibition by the JAK/STAT pathway of IFNgamma- and LPS-stimulated nitric oxide synthase induction in vascular smooth muscle cells. Biochemical and biophysical research communications 252(2): 508-512.
Marron, M. P., D. I. Hopkins, P. Yong-Soo and J. X. She (1999). NeuroD/Beta2 polymorphism is not associated with type 1 diabetes in Chinese, Korean, or Caucasian populations. Journal of Endocrine Genetics 1(2): 73-77.
Marron, M. P., L. J. Raffel, H. J. Garchon, C. O. Jacob, M. Serrano-Rios, M. T. Martinez-Larrad, W. P. Teng, Y. Park, Z. X. Zhang, D. R. Goldstein, Y. W. Tao, G. Beaurain, J. F. Bach, H. S. Huang, D. F. Luo, et al. (1997). Insulin-dependent diabetes mellitus (IDDM) is associated with CTLA4 polymorphisms in multiple ethnic groups. Human molecular genetics 6(8): 1275-1282.
Marsden, P., H. Heng, C. Duff, X.-M. Shi, L.-C. Tsui and A. Hall (1994). Localization of the human gene for inducible nitric oxide synthase (NOS2) to chromosome 17q11.2-q12. Genomics 19: 183-185.
Marsden, P. A., H. H. Heng, S. W. Scherer, R. J. Stewart, A. V. Hall, X. M. Shi, L. C. Tsui and K. T. Schappert (1993). Structure and chromosomal localization of the human constitutive endothelial nitric oxide synthase gene. Journal of biological chemistry 268(23): 17478-17488.
Marsden, P. A., K. T. Schappert, H. S. Chen, M. Flowers, C. L. Sundell, J. N. Wilcox, S. Lamas and T. Michel (1992). Molecular cloning and characterization of human endothelial nitric oxide synthase. FEBS letters 307(3): 287-293.
Marth, G., R. Yeh, M. Minton, R. Donaldson, Q. Li, S. G. Duan, R. Davenport, R. D. Miller and P. Y. Kwok (2001). Single-nucleotide polymorphisms in the public domain: how useful are they? Nature Genetics 27(4): 371-372.
Martin, E., C. Nathan and Q. W. Xie (1994). Role of interferon regulatory factor 1 in induction of nitric oxide synthase. Journal of Experimental Medicine 180(3): 977-984.
Martin, J., J. E. Calzada and A. Nieto (1999). Inducible nitric oxide synthase (NOS2) gene polymorphism and parasitic diseases. Lancet 353(9146): 72.
Martínez-Naves, E., M. Peña and C. López-Larrea (1993). T-cell receptor alpha, delta, and gamma chain genes in insulin-dependent diabetes mellitus. European journal of immunogenetics 20(5): 317-325.
Massa, S., F. Longo, J. Zuo, S. Wang, J. Chen and F. Sharp (1995). Cloning of rat grp75, an hsp70-family member, and its expression in normal and ischemic brain. J of Neurosci Res 40: 807-819.
Matsuyama, T., T. Kimura, M. Kitagawa, K. Pfeffer, T. Kawakami, N. Watanabe, T. M. K.ündig, R. Amakawa, K. Kishihara, A. Wakeham and a. et (1993). Targeted disruption of IRF-1 or IRF-2 results in abnormal type I IFN gene induction and aberrant lymphocyte development. Cell 75(1): 83-97.
Maugendre, D., M. Alizadeh, A. Gauthier, I. Guilhem, C. Pouillaud, B. Genetet, H. Allannic and G. Semana (1996). Genetic heterogeneity between type 1a and type 1b insulin-dependent diabetes mellitus: HLA Class II and TAP gene analysis. Tissue Antigens 48(5): 540-548.
Maxwell, S. R., H. Thomason, D. Sandler, C. Leguen, M. A. Baxter, G. H. Thorpe, A. F. Jones and A. H. Barnett (1997). Antioxidant status in patients with uncomplicated insulin-dependent and non-insulin-dependent diabetes mellitus. European journal of clinical investigation 27(6): 484-490.
McCormack, R. M., A. P. Maxwell, D. Carson, C. C. Patterson and D. A. Savage (2001). No association between IL-10 and TGFB1 polymorphisms and type 1 diabetes mellitus. Diabetes 50(Suppl 2): A492-PY - 2001 Conferences, Congresses, Review Annuals.
McCormack, R. M., A. P. Maxwell, D. J. Carson, C. C. Patterson, D. Middleton and D. A. Savage (2002). The IL12B 3' untranslated region DNA polymorphism is not associated with early-onset type 1 diabetes. 3(7): 433-435.
McDermott, M. F., A. Ramachandran, B. W. Ogunkolade, E. Aganna, D. Curtis, B. J. Boucher, C. Snehalatha and G. A. Hitman (1997). Allelic variation in the vitamin D receptor influences susceptibility to IDDM in Indian Asians. Diabetologia 40(8): 971-975.
McDermott, M. F., G. Schmidt-Wolf, A. A. Sinha, M. Koo, M. A. Porter, L. Briant, A. Cambon-Thomsen, N. K. Maclaren, D. Fiske, S. Bertera, M. Trucco, C. I. Amos, H. O. McDevitt and D. L. Kastner (1996). No linkage or association of telomeric and centromeric T-cell receptor beta-chain markers with susceptibility to type 1 insulin-dependent diabetes in HLA-DR4 multiplex families. European journal of immunogenetics 23(5): 361-370.
McGinnis, R. (2000). General equations for Pt, Ps, and the power of the TDT and the affected-sib-pair test. American Journal of Human Genetics 67(5): 1340-1347.
McMillan, S. A., C. A. Graham, P. J. Hart, D. R. Hadden and T. A. McNeill (1990). A T cell receptor beta chain polymorphism is associated with patients developing insulin-dependent diabetes after the age of 20 years. Clinical and experimental immunology 82(3): 538-541.
McTernan, C. L., L. C. Stewart, C. H. Mijovic and A. H. Barnett (2000). Assessment of the non-HLA-DR-DQ contribution to IDDM1 in British Caucasian families: analysis of LMP7 polymorphisms. Diabetic Medicine 17(9): 661-666.
Mein, C., L. Esposito, M. Dunn, G. Johnson, A. Timms, J. Goy, A. Smith, L. Sebag-Montefiore, M. Merriman, A. Wilson, L. Pritchard, F. Cucca, A. Barnett, S. B. SC and J. Todd (1998). A search for type 1 diabetes susceptibility genes in families from the United Kingdom. Nature Genet 19: 297-300.
Mellott, J. K., H. S. Nick, M. F. Waters, T. R. Billiar, D. A. Geller and S. E. Chesrown (2001). Cytokine-induced changes in chromatin structure and in vivo footprints in the inducible NOS promoter. American journal of physiology Lung cellular and molecular physiology 280(3): L390-L399.
Menke, A., L. McInnes, N. D. Hastie and A. Schedl (1998). The Wilms' tumor suppressor WT1: approaches to gene function. Kidney international 53(6): 1512-1518.
Menke, A. L., A. Shvarts, N. Riteco, R. C. van-Ham, E. van-der, A.J. and A. G. Jochemsen (1997). Wilms' tumor 1-KTS isoforms induce p53-independent apoptosis that can be partially rescued by expression of the epidermal growth factor receptor or the insulin receptor. Cancer Res 57(7): 1353-1363.
Messmer, U. K., M. Ankarcrona, P. Nicotera and B. Bruene (1994). P53 expression in nitric oxide-induced apoptosis. FEBS Letters 355(1): 23-26.
Metcalfe, K. A., G. A. Hitman, F. Pociot, R. Bergholdt, E. Tuomilehto-Wolf, J. Tuomilehto, M. Viswanathan, A. Ramachandran and J. Nerup (1996). An association between type 1 diabetes and the interleukin-1 receptor type 1 gene. The DiMe Study Group. Childhood Diabetes in Finland. Human immunology 51(1): 41-48.
Meyer, G., H. Donner, J. Herwig, H. Boehles, K. H. Usadel and K. Badenhoop (2001). Screening for an AIRE-1 mutation in patients with Addison's disease, type 1 diabetes, Graves' disease and Hashimoto's thyroiditis as well as in APECED syndrome. Clinical Endocrinology 54(3): 335-338.
Millward, B. A., K. I. Welsh, R. D. Leslie, D. A. Pyke and A. G. Demaine (1987). T cell receptor beta chain polymorphisms are associated with insulin-dependent diabetes. Clinical and experimental immunology 70(1): 152-157.
Mirel, D. B., A. M. Valdes, L. C. Lazzeroni, R. L. Reynolds, H. A. Erlich and J. A. Noble (2002). Association of IL4R haplotypes with type 1 diabetes. Diabetes 51(11): 3336-3341.
Miyamoto, M., T. Fujita, Y. Kimura, M. Maruyama, H. Harada, Y. Sudo, T. Miyata and T. Taniguchi (1988). Regulated expression of a gene encoding a nuclear factor, IRF-1, that specifically binds to IFN-beta gene regulatory elements. Cell 54(6): 903-913.
Mizukoshi, E., M. Suzuki, A. Loupatov, T. Uruno, H. Hayashi, T. Misono, S. Kaul, R. Wadhwa and T. Imamura (1999). Fibroblast growth factor-1 interacts with the glucose-regulated protein GRP75/mortalin. Biochem J 343: 461-466.
Mizukoshi, E., M. Suzuki, T. Misono, A. Loupatov, E. Munekata, S. C. Kaul, R. Wadhwa and T. Imamura (2001). Cell-cycle dependent tyrosine phosphorylation on mortalin regulates its interaction with fibroblast growth factor-1. Biochemical and biophysical research communications 280(4): 1203-1209.
Mizzen, L. A., C. Chang, J. I. Garrels and W. J. Welch (1989). Identification, characterization, and purification of two mammalian stress proteins present in mitochondria, grp 75, a member of the hsp 70 family and hsp 58, a homolog of the bacterial groEL protein. Journal of biological chemistry 264(34): 20664-20675.
Mizzen, L. A., A. N. Kabiling and W. J. Welch (1991). The two mammalian mitochondrial stress proteins, grp 75 and hsp 58, transiently interact with newly synthesized mitochondrial proteins. Cell regulation 2(2): 165-179.
Mochizuki, M., S. Amemiya, K. Kobayashi, K. Kobayashi, T. Ishihara, M. Aya, K. Kato, A. Kasuga and S. Nakazawa (2002). The association of Ala45Thr polymorphism in NeuroD with child-onset Type 1a diabetes in Japanese. Diabetes research and clinical practice 55(1): 11-17.
Moghaddam, P. H., P. De-Knijf, B. O. Roep, A. Van-Der, B., A. Naipal, F. Gorus, F. Schuit, M. J. Giphart and B. D. Registry (1998). Genetic structure of IDDM1: Two separate regions in the major histocompatibility complex contribute to susceptibility or protection. Diabetes 47(2): 263-269.
Moghaddam, P. H., A. H. Zwinderman, P. De-Knijff, B. O. Roep, R. F. Schipper, A. Van-Der, B., A. Naipal, F. Gorus, F. Schuit, M. J. Giphart and B. D. Registry (1997). TNFa microsatellite polymorphism modulates the risk of IDDM in Caucasians with the high-risk genotype HLA DQA1*0501- DQB1*0201/DQA1*0301-DQB1*0302. Diabetes 46(9): 1514-1515.
Moncada, S. and E. A. Higgs (1991). Endogenous nitric oxide: physiology, pathology and clinical relevance. European journal of clinical investigation 21(4): 361-374.
Monos, D. S., M. Kamoun, I. A. Udalova, E. Csanky, B. Cizman, R. L. Turetskaya, J. B. Smirnova, V. G. Zharkov, D. Gasser, C. M. Zmijewski, R. S. Spielman and S. A. Nedospasov (1995). Genetic polymorphism of the human tumor necrosis factor region in insulin-dependent diabetes mellitus linkage disequilibrium of TNFab microsatellite alleles with HLA haplotypes. Human Immunology 44(2): 70-79.
Morahan, G., D. Huang, S. I. Ymer, M. R. Cancilla, K. Stephen, P. Dabadghao, G. Werther, B. D. Tait, L. C. Harrison and P. G. Colman (2001). Linkage disequilibrium of a type 1 diabetes susceptibility locus with a regulatory IL12B allele. Nature genetics 27(2): 218-221.
Moriscot, C., F. Pattou, J. Kerr-Conte, M. Richard, P. Lemarchand and P. Benhamou (2000). Contribution of adenoviral-mediated superoxide dismutase gene transfer to the reduction in nitric oxide-induced cytotoxicity on human islets and INS-1 insulin-secreting cells. Diabetologia 43: 625-631.
Morris, B. J., C. L. Glenn, D. E. L. Wilcken and X. L. Wang (2001). Influence of an inducible nitric oxide synthase promoter variant on clinical variables in patients with coronary artery disease. Clinical Science 100(5): 551-556.
Morris, B. J., M. A. Markus, C. L. Glenn, D. J. Adams, S. Colagiuri and X. L. Wang (2002). Association of a functional inducible nitric oxide synthase promoter variant with complications in type 2 diabetes. Journal of Molecular Medicine-Jmm 80(2): 96-104.
Morris, S. M. and T. R. Billiar (1994). New insights into the regulation of inducible nitric oxide synthesis. American journal of physiology 266(6 Pt 1): E829-E839.
Morrison, V. A., S. Onengut-Gumuscu and P. Concannon (2004). A functional variant of IRS1 is associated with type 1 diabetes in families from the US and UK. Molecular genetics and metabolism 81(4): 291-294.
Mose-Larsen, P., S. Fey, M. Larsen, A. Nawrocki, H. Andersen, H. Kähler, C. Heilmann, V. MC., P. Roepstorff, F. Pociot, A. Karlsen and J. Nerup (2001). Proteome analysis of IL-1b induced changes in protein expression in rat islets of Langerhans. Diabetes 50: 1056-1063.
Mosmann, T. R., H. Cherwinski, M. W. Bond, M. A. Giedlin and R. L. Coffman (1986). Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. Journal of immunology 136(7): 2348-2357.
Mosser, D. D., A. W. Caron, L. Bourget, C. Denis-Larose and B. Massie (1997). Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Molecular and cellular biology 17(9): 5317-5327.
Motterlini, R., R. Foresti, M. Intaglietta and R. M. Winslow (1996). NO-mediated activation of heme oxygenase: endogenous cytoprotection against oxidative stress to endothelium. American journal of physiology 270(1 Pt 2): H107-H114.
Mrozikiewicz, P. M., N. Drakoulis and I. Roots (1994). Polymorphic arylamine N-acetyltransferase (NAT2) genes in children with insulin-dependent diabetes mellitus. Clinical pharmacology and therapeutics 56(6 Pt 1): 626-634.
Muhammad, B. J., P. G. Swift, N. T. Raymond and J. L. Botha (1999). Partial remission phase of diabetes in children younger than age 10 years. Archives of disease in childhood 80(4): 367-369.
Müller, A., P. Schott-Ohly, C. Dohle and H. Gleichmann (2002). Differential regulation of Th1-type and Th2-type cytokine profiles in pancreatic islets of C57BL/6 and BALB/c mice by multiple low doses of streptozotocin. Immunobiology 205(1): 35-50.
Nakane, M., H. H. Schmidt, J. S. Pollock, U. Förstermann and F. Murad (1993). Cloned human brain nitric oxide synthase is highly expressed in skeletal muscle. FEBS letters 316(2): 175-180.
Nakanishi, K., T. Kobayashi, T. Murase and K. Kosaka (1994). Lack of association of the transporter associated with antigen processing with Japanese insulin-dependent diabetes mellitus. Metabolism Clinical and Experimental 43(8): 1013-1017.
Nakao, F., K. Ihara, K. Kusuhara, Y. Sasaki, N. Kinukawa, A. Takabayashi, S. Nishima and T. Hara (2001). Association of IFN-gamma and IFN regulatory factor 1 polymorphisms with childhood atopic asthma. Journal of Allergy and Clinical Immunology 107(3): 499-504.
Nakayama, D. K., D. A. Geller, C. J. Lowenstein, H. D. Chern, P. Davies, B. R. Pitt, R. L. Simmons and T. R. Billiar (1992). Cytokines and lipopolysaccharide induce nitric oxide synthase in cultured rat pulmonary artery smooth muscle. American journal of respiratory cell and molecular biology 7(5): 471-476.
Naluai, A. T., S. Nilsson, A. H. Gudjónsdóttir, A. S. Louka, H. Ascher, J. Ek, B. Hallberg, L. Samuelsson, B. Kristiansson, T. Martinsson, O. Nerman, L. M. Sollid and J. Wahlström (2001). Genome-wide linkage analysis of Scandinavian affected sib-pairs supports presence of susceptibility loci for celiac disease on chromosomes 5 and 11. European journal of human genetics 9(12): 938-944.
Nathan, C. and Q. W. Xie (1994). Nitric oxide synthases: roles, tolls, and controls. Cell 78(6): 915-918.
Nejentsev, S., Z. Gombos, A. P. Laine, R. Veijola, M. Knip, O. Simell, O. Vaarala, H. K. Akerblom and J. Ilonen (2000). Non-class IIHLA gene associated with type 1 diabetes maps to the 240-kb region near HLA-B. Diabetes 49(12): 2217-2221.
Nejentsev, S., A. Laine, O. Simell and J. Ilonen (2000). Intercellular adhesion molecule-1 (ICAM-1) K469E polymorphism: No association with type 1 diabetes among Finns. Tissue Antigens 55(6): 568-570.
Nepom, G. T. and W. W. Kwok (1998). Molecular basis for HLA-DQ associations with IDDM. Diabetes 47(8): 1177-1184.
Nerup, J., O. O. Andersen, G. Bendixen, J. Egeberg and J. E. Poulsen (1971). Anti-pancreatic cellular hypersensitivity in diabetes mellitus. Diabetes 20(6): 424-427.
Nerup, J., T. Mandrup-Poulsen, S. Helqvist, H. U. Andersen, F. Pociot, J. I. Reimers, B. G. Cuartero, A. E. Karlsen, U. Bjerre and T. Lorenzen (1994). On the pathogenesis of IDDM. Diabetologia 37 (suppl 2): S82-S89.
Nerup, J., P. Platz and O. O. Andersen (1974). HL-A antigens and diabetes mellitus. Lancet I: 864-866.
Nerup, J., F. Pociot and European Consortium for IDDM Studies (2001). A genomewide scan for type 1-diabetes susceptibility in Scandinavian families: identification of new loci with evidence of interactions. Am J Hum Genet 69(6): 1301-1313.
Nervi, S., S. Nicodeme, C. Gartioux, C. Atlan, M. Lathrop, D. Reviron, P. Naquet, F. Matsuda, J. Imbert and B. Vialettes (2002). No association between lck gene polymorphisms and protein level in type 1 diabetes. Diabetes 51(11): 3326-3330.
Ng, Y. C., P. Jacobs and J. A. Johnson (2001). Productivity losses associated with diabetes in the US. Diabetes care 24(2): 257-261.
Niedbala, W., X. Q. Wei, D. Piedrafita, D. Xu and F. Y. Liew (1999). Effects of nitric oxide on the induction and differentiation of Th1 cells. European Journal of Immunology 29(8): 2498-2505.
Nielsen, K. (2004). Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1ß. Diabetologia, 47: 62-74.
Nielsen, K., A. E. Karlsen, M. Deckert, O. D. Madsen, P. Serup, T. Mandrup-Poulsen and J. Nerup (1999). Beta-cell maturation leads to in vitro sensitivity to cytotoxins. Diabetes 48(12): 2324-2332.
Nielsen, K., M. Kruhøffer, T. Ørntoft, T. Sparre, H. Wang, C. Wollheim, M. C. Jørgensen, J. Nerup and A. E. Karlsen (2004). Gene expression profiles during beta-cell maturation and after IL-1beta exposure reveal important roles of Pdx-1 and Nkx6.1 or IL-1beta sensitivity. In press.
Nishimura, M., H. Obayashi, E. Maruya, M. Ohta, H. Tegoshi, M. Fukui, G. Hasegawa, H. Shigeta, Y. Kitagawa, K. Nakano, H. Saji and N. Nakamura (2000). Association between type 1 diabetes age-at-onset and intercellular adhesion molecule-1 (ICAM-1) gene polymorphism. Human Immunology 61(5): 507-510.
Nishino, M., H. Ikegami, Y. Kawaguchi, T. Fujisawa, Y. Kawabata, M. Shintani, M. Ono, M. Horiki, E. Kawasaki and T. Ogihara (2001). Polymorphism in gene for islet autoantigen, IA-2, and type 1 diabetes in Japanese subjects. Human immunology 62(5): 518-522.
Nishio, Y., E. Noguchi, S. Ito, E. Ichikawa, Y. Umebayashi, F. Otsuka and T. Arinami (2001). Mutation and association analysis of the interferon regulatory factor 2 gene (IRF2) with atopic dermatitis. Journal of Human Genetics 46(11): 664-667.
Nistico, L., R. Buzzetti, L. E. Pritchard, A. Van-der, D., C. Giovannini, E. Bosi, M. T. Martinez-Larrad, M. S. Rios, C. C. Chow, C. S. Cockram, K. Jacobs, C. Mijovic, S. C. Bain, A. H. Barnett, C. L. Vandewalle, et al. (1996). The CTLA-4 gene region of chromosome 2q33 is linked to, and associated with, type 1 diabetes. Human Molecular Genetics 5(7): 1075-1080.
Nisticò, L., G. Giorgi, M. Giordano, A. Galgani, A. Petrone, S. D'Alfonso, M. Federici, U. Di-Mario, P. Pozzilli, R. Buzzetti and I. Cascino (2002). IL12B polymorphism and type 1 diabetes in the Italian population: a case-control study. Diabetes 51(5): 1649-1650.
Nithiyananthan, R., J. M. Heward, A. Allahabadia, A. H. Barnett, J. A. Franklyn and S. C. L. Gough (2000). A heterozygous deletion of the autoimmune regulator (AIRE1) gene, autoimmune thyroid disease, and type 1 diabetes: No evidence for association. Journal of Clinical Endocrinology & Metabolism 85(3): 1320-1322.
Niven, M. J., C. Caffrey, R. H. Moore, J. A. Sachs, V. Mohan, H. Festenstein, M. L. Hoover and G. A. Hitman (1990). T-cell receptor beta-subunit gene polymorphism and autoimmune disease. Human immunology 27(4): 360-367.
Niwa, M., Y. Kawai, N. Nakamura and S. Futaki (1997). The structure of the promoter region for rat inducible nitric oxide synthase gene. Life sciences 61(5): PL 45-PL 49.
Noble, J. A., A. M. Valdes, T. L. Bugawan, R. J. Apple, G. Thomson and H. A. Erlich (2002). The HLA class I A locus affects susceptibility to type 1 diabetes. Human Immunology 63(8): 657-664.
Noble, J. A., A. M. Valdes, M. Cook, W. Klitz, G. Thomson and H. A. Erlich (1996). The role of HLA class II genes in insulin-dependent diabetes mellitus: molecular analysis of 180 Caucasian, multiplex families. American journal of human genetics 59(5): 1134-1148.
Noble, J. A., A. White, D. B. Mirel, A. M. Valdes, R. Reynolds, G. Zangenberg, L. Lazzeroni, A. Grupe, G. Peltz and H. A. Erlich (2001). A polymorphism in the TCF7 locus is associated with type 1 diabetes in Caucasians. American Journal of Human Genetics 69(4 Supplement): 226.
Noguchi, E., M. Shibasaki, T. Arinami, K. Yamakawa-Kobayashi, Y. Yokouchi, K. Takeda, A. Matsui and H. Hamaguchi (2000). Mutation screening of interferon regulatory factor 1 gene (IRF-1) as a candidate gene for atopy/asthma. Clinical and Experimental Allergy 30(11): 1562-1567.
Nollen, E. A. A., J. F. Brunsting, H. Roelofsen, L. A. Weber and H. H. Kampinga (1999). In vivo chaperone activity of heat shock protein 70 and thermotolerance. Molecular and Cellular Biology 19(3): 2069-2079.
Nolsoe, R., O. Kristiansen, K. Sanghongpitag, Z. Larsen, J. Johannesen, A. Karlsen, F. Pociot, J. Nerup, C. Verge and T. Mandrup-Poulsen (2000). Complete molecular scanning of the human Fas gene: mutational analysis and linkage studies in families with Type 1 diabetes mellitus. Diabetologia 43: 800-808.
Nolsoe, R. L., O. P. Kristiansen, Z. M. Larsen, J. Johannesen, F. Pociot and T. Mandrup-Poulsen (2002). Complete mutation scan of the human Fas ligand gene: Linkage studies in Type I diabetes mellitus families. Diabetologia 45(1): 134-139.
Notkins, A. L. and A. Lernmark (2001). Autoimmune type 1 diabetes: Resolved and unresolved issues. Journal of Clinical Investigation 108(9): 1247-1252.
Nukaya, I., K. Takagi, T. Kawabe and Y. Suketa (1995). Suppression of cytokine production in T helper type 2 cells by nitric oxide in comparison with T helper type 1 cells. Microbiology and immunology 39(9): 709-714.
Nunokawa, Y., N. Ishida and S. Tanaka (1993). Cloning of inducible nitric oxide synthase in rat vascular smooth muscle cells. Biochemical and biophysical research communications 191(1): 89-94.
Nunokawa, Y., S. Oikawa and S. Tanaka (1996). Human inducible nitric oxide synthase gene is transcriptionally regulated by nuclear factor-kappaB dependent mechanism. Biochemical and biophysical research communications 223(2): 347-352.
Nunokawa, Y., S. Oikawa and S. Tanaka (1997). Expression of human inducible nitric oxide synthase is regulated by both promoter and 3'-regions. Biochemical and biophysical research communications 233(2): 523-526.
Obayashi, H., N. Nakamura, M. Fukui, H. Tegoshi, M. Fujii, M. Ogata, G. Hasegawa, H. Shigeta, Y. Kitagawa, K. Nakano, M. Kondo, I. Fukui, E. Maruya, H. Saji, M. Ohta, et al. (1999). Influence of TNF microsatellite polymorphisms (TNFa) on age-at-onset of insulin-dependent diabetes mellitus. Human Immunology 60(10): 974-978.
Oda, M., K. Sakitani, M. Kaibori, T. Inoue, Y. Kamiyama and T. Okumura (2000). Vicinal dithiol-binding agent, phenylarsine oxide, inhibits inducible nitric-oxide synthase gene expression at a step of nuclear factor-kappaB DNA binding in hepatocytes. Journal of biological chemistry 275(6): 4369-4373.
Ohashi, J., I. Naka, J. Patarapotikul, H. Hananantachai, S. Looareesuwan and K. Tokunaga (2002). Significant association of longer forms of CCTTT microsatellite repeat in the inducible nitric oxide synthase promoter with severe malaria in Thailand. Journal of Infectious Diseases 186(4): 578-581.
Ohashi, M., M. Oyanagi, K. Hatakeyama, M. Inoue and R. Kominami (1995). The gene encoding PBP74/CSA/motalin-1, a novel mouse hsp70, maps to mouse chromosome 18. Genomics 30(2): 406-407.
Ohkubo, T., T. Awata, K. Inoue, S. Kurihara, M. Watanabe, K. Inukai, I. Inoue and S. Katayama (2001). A novel polymorphism in the promoter region of the IL-4 gene is associated with type 1 diabetes in Japanese. Diabetologia 44(Suppl 1): A 80.
Olivès, B., M. Merriman, P. Bailly, S. Bain, A. Barnett, J. Todd, J. P. Cartron and T. Merriman (1997). The molecular basis of the Kidd blood group polymorphism and its lack of association with type 1 diabetes susceptibility. Human molecular genetics 6(7): 1017-1020.
Onengut-Gumuscu, S., K. G. Ewens, R. S. Spielman and P. Concannon (2004). A functional polymorphism (1858C/T) in the PTPN22 gene is linked and associated with type I diabetes in multiplex families. Genes and immunity 5(8): 678-680.
Ongagna, J. C., M. C. Kaltenbacher, R. Sapin, M. Pinget and A. Belcourt (2001). The HLA-DQB alleles and amino acid variants of the vitamin D-binding protein in diabetic patients in Alsace. Clinical biochemistry 34(1): 59-63.
O'Reilly, L. A., P. R. Hutchings, P. R. Crocker, E. Simpson, T. Lund, D. Kioussis, F. Takei, J. Baird and A. Cooke (1991). Characterization of pancreatic islet cell infiltrates in NOD mice: effect of cell transfer and transgene expression. European journal of immunology 21(5): 1171-1180.
Ornatsky, O. I., M. K. Connor and D. A. Hood (1995). Expression of stress proteins and mitochondrial chaperonins in chronically stimulated skeletal muscle. Biochemical journal 311 ( Pt 1): 119-123.
Owerbach, D., F. J. Naya, M. J. Tsai, S. V. Allander, D. R. Powell and K. H. Gabbay (1997). Analysis of candidate genes for susceptibility to type I diabetes: a case-control and family-association study of genes on chromosome 2q31-35. Diabetes 46(6): 1069-1074.
Oyadomari, S., K. Takeda, M. Takiguchi, T. Gotoh, M. Matsumoto, I. Wada, S. Akira, E. Araki and M. Mori (2001). Nitric oxide-induced apoptosis in pancreatic beta cells is mediated by the endoplasmic reticulum stress pathway. Proc. Natl. Acad. Sci. U. S. A. 98(19): 10845-10850.
Pahan, K., X. J. Liu, M. J. McKinney, C. Wood, F. G. Sheikh and J. R. Raymond (2000). Expression of a dominant-negative mutant of p21(ras) inhibits induction of nitric oxide synthase and activation of nuclear factor-kappa B in primary astrocytes. Journal of Neurochemistry 74(6): 2288-2295.
Pahan, K., J. R. Raymond and I. Singh (1999). Inhibition of phosphatidylinositol 3-kinase induces nitric-oxide synthase in lipopolysaccharide- or cytokine-stimulated C6 glial cells. Journal of biological chemistry 274 (11): 7528-7536.
Pahan, K., F. G. Sheikh, A. M. Namboodiri and I. Singh (1998). Inhibitors of protein phosphatase 1 and 2A differentially regulate the expression of inducible nitric-oxide synthase in rat astrocytes and macrophages. Journal of biological chemistry 273(20): 12219-12226.
Pahl, H. L. (1999). Activators and target genes of Rel/NF- kappaB transcription factors. Oncogene 18(49): 6853-6866.
Paludan, S. R., S. Ellermann-Eriksen, J. Lovmand and S. C. Mogensen (1999). Interleukin-4-mediated inhibition of nitric oxide production in interferon-gamma-treated and virus-infected macrophages. Scandinavian journal of immunology 49(2): 169-176.
Pance, A., A. Chantome, S. Reveneau, F. Bentrari and J. F. Jeannin (2002). A repressor in the proximal human inducible nitric oxide synthase promoter modulates transcriptional activation. 16(6): 631-633.
Pani, M. A., K. Bieda, J. Wood, K. H. Usadel and K. Badenhoop (2001). A novel vitamin D receptor gene polymorphism confers susceptibility to type 1 diabetes mellitus. Diabetologia 44(Suppl 1): A 53.
Pani, M. A., H. Donner, J. Herwig, K. H. Usadel and K. Badenhoop (1999). Vitamin D binding protein alleles and susceptibility for type 1 diabetes in Germans. Autoimmunity 31(1): 67-72.
Pani, M. A., M. Knapp, H. Donner, J. Braun, M. P. Baur, K. H. Usadel and K. Badenhoop (2000). Vitamin D receptor allele combinations influence genetic susceptibility to type 1 diabetes in Germans. Diabetes 49(3): 504-507.
Pani, M. A., J. P. Wood, K. Bieda, R. R. Toenjes, K. H. Usadel and K. Badenhoop (2002). The variable endogenous retroviral insertion in the human complement C4 gene: a transmission study in type I diabetes mellitus. Human immunology 63(6): 481-484.
Park, C. S., G. Krishna, M. S. Ahn, J. H. Kang, W. G. Chung, D. J. Kim, H. K. Hwang, J. N. Lee, S. G. Paik and Y. N. Cha (2000). Differential and constitutive expression of neuronal, inducible, and endothelial nitric oxide synthase mRNAs and proteins in pathologically normal human tissues. Nitric Oxide-Biology and Chemistry 4(5): 459-471.
Park, C. S., H. S. Lee, H. Y. Lee and G. Krishna (1997). An unprocessed pseudogene of inducible nitric oxide synthase gene in human. Nitric oxide 1(4): 294-300.
Park, C. S., R. Park and G. Krishna (1996). Constitutive expression and structural diversity of inducible isoform of nitric oxide synthase in human tissues. Life sciences 59(3): 219-225.
Park, J. S., S. J. Lee, J. Myeong, S. E. Namkoong, S. J. Um and S. H. Jee (2003). Polymorphisms of p53, p21 and IRF-1 and cervical cancer susceptibility in Korean women. Proceedings of the American Association for Cancer Research Annual Meeting 44: 1081.
Park, Y., H. Lee, C. B. Sanjeevi and G. S. Eisenbarth (2001). MICA polymorphism is associated with type 1 diabetes in the Korean population. Diabetes care 24(1): 33-38.
Pascual, M., M. A. Lopez-Nevot, R. Caliz, B. P. C. Koeleman, A. Balsa, D. Pascual-Salcedo and J. Martin (2002). Genetic determinants of rheumatoid arthritis: The inducible nitric oxide synthase (NOS2) gene promoter polymorphism. Genes and Immunity 3(5): 299-301.
Patrick, S. L., C. S. Moy and R. E. LaPorte (1989). The world of insulin-dependent diabetes mellitus: what international epidemiologic studies reveal about the etiology and natural history of IDDM. Diabetes/metabolism reviews 5(7): 571-578.
Pavlovic, D., M. Chen, L. Bouwens, D. Eizirik and D. Pipeleers (1999). Contribution of ductal cells to cytokine responses by human pancreatic islets. Diabetes 48: 29-33.
Pavlovic, D., M. Chen, C. Gysemans, C. Mathieu and D. Eizirik (1999b). The role of interferon regulatory factor-1 in cytokine-induced mRNA expression and cell death in murine pancreatic beta-cells. Eur Cyt Net 10: 403-412.
Payami, H., G. Thomson, U. Motro and a. et (1985). The affected sib method. IV. Sib trios. Annals of Human Genetics 49(4): 303-314.
Pelletier, J., W. Bruening, C. E. Kashtan, S. M. Mauer, J. C. Manivel, J. E. Striegel, D. C. Houghton, C. Junien, R. Habib, L. Fouser and a. et (1991). Germline mutations in the Wilms' tumor suppressor gene are associated with abnormal urogenital development in Denys-Drash syndrome. Cell 67(2): 437-447.
Penfornis, A., E. Tuomilehto-Wolf, D. L. Faustman and G. A. Hitman (2002). Analysis of TAP2 polymorphisms in Finnish individuals with type I diabetes. Human Immunology 63(1): 61-70.
Perrella, M. A., C. Patterson, L. Tan, S. F. Yet, C. M. Hsieh, M. Yoshizumi and M. E. Lee (1996). Suppression of interleukin-1beta-induced nitric-oxide synthase promoter/enhancer activity by transforming growth factor-beta1 in vascular smooth muscle cells. Evidence for mechanisms other than NF-kappaB. Journal of biological chemistry 271(23): 13776-13780.
Pipeleers, D. G. (1992). Heterogeneity in pancreatic beta-cell population. Diabetes 41(7): 777-781.
Pipeleers, D. G., P. A. In't-Veld, M. Van-de Winkel, E. Maes, F. C. Schuit and W. Gepts (1985). A new in vitro model for the study of pancreatic A and B cells. Endocrinology 117(3): 806-816.
Piro, S., R. Lupi, F. Dotta, G. Patanè, M. A. Rabuazzo, L. Marselli, C. Santangelo, M. Realacci, S. Del-Guerra, F. Purrello and P. Marchetti (2001). Bovine islets are less susceptible than human islets to damage by human cytokines. Transplantation 71(1): 21-26.
Piskurich, J. F., M. W. Linhoff, Y. Wang and J. P. Ting (1999). Two distinct gamma interferon-inducible promoters of the major histocompatibility complex class II transactivator gene are differentially regulated by STAT1, interferon regulatory factor 1, and transforming growth factor beta. Molecular and cellular biology 19(1): 431-440.
Pociot, F. (1996). Insulin-dependent diabetes mellitus - a polygenic disorder? Dan Med Bull (Thesis) 43: 216-248.
Pociot, F., L. Briant, C. V. Jongeneel, J. Mölvig, H. Worsaae, M. Abbal, M. Thomsen, J. Nerup and A. Cambon-Thomsen (1993). Association of tumor necrosis factor (TNF) and class II major histocompatibility complex alleles with the secretion of TNF-alpha and TNF-beta by human mononuclear cells: a possible link to insulin-dependent diabetes mellitus. Eur J Immunol 23(1): 224-231.
Pociot, F. and A. E. Karlsen (2002). Combined genome and proteome approach to identify new susceptibility genes. Am. J. Med. Genet. 115(1): 55-60.
Pociot, F., A. E. Karlsen, C. B. Pedersen, M. Aalund and J. Nerup (2004). Novel analytical methods applied to type 1 diabetes genome-scan data. American Journal of Human Genetics 74(4): 647-660.
Pociot, F., Z. M. Larsen, P. Zavattari, E. Deidda, J. Nerup, M. Cattaneo, R. Chiaramonte, P. Comi, M. Sabbadini, M. Zollo, I. Biunno and F. Cucca (2001). No evidence for SEL1L as a candidate gene for IDDM11-conferred susceptibility. Diabetes-Metabolism Research and Reviews 17(4): 292-295.
Pociot, F., T. Lorenzen and J. Nerup (1993). A manganese superoxide dismutase (SOD2) gene polymorphism in insulin-dependent diabetes mellitus. Disease markers 11(5-6): 267-274.
Pociot, F. and M. F. McDermott (2002). Genetics of type 1 diabetes mellitus. Genes and Immunity 3(5): 235-249.
Pociot, F., J. Mølvig, L. Wogensen, H. Worsaae and J. Nerup (1992). A TaqI polymorphism in the human interleukin-1 beta (IL-1 beta) gene correlates with IL-1 beta secretion in vitro. Eur J Clin Invest 22(6): 396-402.
Pociot, F., K. Nørgaard, N. Hobolth, O. Andersen, J. Nerup and the Danish Study Group of Diabetes in Childhood (1993). A nationwide population-based study of the familial aggregation of insulin-dependent diabetes in Denmark. Diabetologia 36: 870-875.
Pociot, F., J. S. Ronningen, R. Bergholdt, T. Lorenzen, J. Johannesen, K. Ye, C. A. Dinarello and J. Nerup (1994). Genetic susceptibility markers in Danish patients with type 1 (insulin-dependent) diabetes evidence for polygenecity in man. Autoimmunity 19(3): 169-178.
Pociot, F., K. S. Ronningen and K. Nerup (1993). Polymorphic analysis of the human MHC-linked heat shock protein 70 (HSP70-2) and HSP70-Hom genes in insulin-dependent diabetes mellitus (IDDM). Scandinavian Journal of Immunology 38(5): 491-495.
Pociot, F., R. Veijola, J. Johannesen, P. M. Hansen, T. Lorenzen, A. E. Karlsen, H. Reijonen, M. Knip and J. Nerup (1997). Analysis of an interferon-gamma gene (IFNG) polymorphism in Danish and Finnish insulin-dependent diabetes mellitus (IDDM) patients and control subjects. Danish Study Group of Diabetes in Childhood. Journal of interferon & cytokine research 17(2): 87-93.
Polla, B. S., S. Kantengwa, D. François, S. Salvioli, C. Franceschi, C. Marsac and A. Cossarizza (1996). Mitochondria are selective targets for the protective effects of heat shock against oxidative injury. Proceedings of the National Academy of Sciences of the United States of America 93(13): 6458-6463.
Polymeropoulos, M. H., R. G. Swift and M. Swift (1994). Linkage of the gene for Wolfram syndrome to markers on the short arm of chromosome 4. Nature genetics 8(1): 95-97.
Prevost, G., I. Fajardy, P. Fontaine, P. M. Danze and C. Besmond (1999). Human RAGE GLY82SER dimorphism and HLA class II DRB1-DQA1- DQB1 haplotypes in type 1 diabetes. European Journal of Immunogenetics 26(5): 343-348.
Pritchard, L. E., Y. Kawaguchi, P. W. Reed, J. B. Copeman, J. L. Davies, A. H. Barnett, S. C. Bain and J. A. Todd (1995). Analysis of the CD3 gene region and type 1 diabetes: application of fluorescence-based technology to linkage disequilibrium mapping. Human molecular genetics 4(2): 197-202.
Pritchard-Jones, K., S. Fleming, D. Davidson, W. Bickmore, D. Porteous, C. Gosden, J. Bard, A. Buckler, J. Pelletier, D. Housman and et al. (1990). The candidate Wilms' tumour gene is involved in genitourinary development. Nature 346(6280): 194-197.
Promrat, K., G. Tang, D. Wang, C. Gonzalez, X. Tong, L. Zu, M. Ghany, Y. Park, H. J. Alter, J. I. Rotter, J. H. Hoofnagle, H. Yang and T. J. Liang (2002). The association of interferon regulatory factor-1 (IRF-1) promoter polymorphism with HCV infection and interferon treatment response. Hepatology 36(4 Part 2): 282A.
Prud'homme, G. J., B. R. Lawson and A. N. Theofilopoulos (2001). Anticytokine gene therapy of autoimmune diseases. Expert Opin Biol Ther 1(3): 359-373.
Pugliese, A., Z. L. Awdeh, A. Galluzzo, E. J. Yunis, C. A. Alper and G. S. Eisenbarth (1992). No independent association between HSP70 gene polymorphisms and IDDM. Diabetes 41(7): 788-791.
Pugliese, A. and G. Eisenbarth (2003). Type I diabetes mellitus of man: Genetic susceptibility and resistance. In: Eisenbarth GS (ed) Type 1 Diabetes: molecular, cellular and clinical immunology. http://www.uchsedu/ misc/diabetes/bdc.html
Pugliese, A. and D. Miceli (2002). The insulin gene in diabetes. Diabetes-Metabolism Research and Reviews 18(1): 13-25.
Punzalan, C., C. Cai, R. A. Schroeder and P. C. Kuo (1999). Redox regulation of the rat hepatocyte iNOS promoter. Surgery 126(2): 450-455.
Rabinovitch, A. (1994c). Immunoregulatory and cytokine imbalances in the pathogenesis of IDDM. Therapeutic intervention by immunostimulation? Diabetes 43(5): 613-621.
Rabinovitch, A. and W. L. Suarez-Pinzon (1998b). Cytokines and their roles in pancreatic islet beta-cell destruction and insulin-dependent diabetes mellitus. Biochemical pharmacology 55(8): 1139-1149.
Rabinovitch, A., W. L. Suarez-Pinzon, K. Strynadka, R. Schulz, J. R. T. Lakey, G. L. Warnock and R. V. Rajotte (1994a). Human pancreatic islet beta-cell destruction by cytokines is independent of nitric oxide production. Journal of Clinical Endocrinology & Metabolism 79(4): 1058-1062.
Rabinovitch, A., W. Sumoski, R. V. Rajotte and G. L. Warnock (1990). Cytotoxic effects of cytokines on human pancreatic islet cells in monolayer culture. Journal of clinical endocrinology and metabolism 71(1): 152-156.
Radons, J., B. Heller, A. Bürkle, B. Hartmann, M. L. Rodriguez, K. D. Kröncke, V. Burkart and H. Kolb (1994). Nitric oxide toxicity in islet cells involves poly(ADP-ribose) polymerase activation and concomitant NAD+ depletion. Biochemical and biophysical research communications 199(3): 1270-1277.
Rambrand, T., F. Pociot, K. Rønningen, J. Nerup and B. K. Michelsen (1997). Genetic markers for glutamic acid decarboxylase do not predict insulin-dependent diabetes mellitus in pairs of affected siblings. The Danish Study Group of Diabetes in Childhood. Human genetics 99(2): 177-185.
Ran, Q., R. Wadhwa, R. Kawai, S. C. Kaul, R. N. Sifers, R. J. Bick, J. R. Smith and O. M. Pereira-Smith (2000). Extramitochondrial localization of mortalin/mthsp70/PBP74/GRP75. Biochemical and Biophysical Research Communications 275(1): 174-179.
Rau, H., H. Donner, K. H. Usadel and K. Badenhoop (1997). Polymorphisms of tumor necrosis factor receptor 2 are not associated with insulin-dependent diabetes mellitus or Graves' disease. Tissue Antigens 49(5): 535-536.
Rau, H., A. Nicolay, H. Donner, K. H. Usadel and K. Badenhoop (1997). Polymorphisms of TAP1 and TAP2 genes in German patients with type 1 diabetes mellitus. European Journal of Immunogenetics 24(3): 229-236.
Rau, H., K. H. Usadel, S. Ommert and K. Badenhoop (1995). PVU II polymorphism of LST-1 (leucocyte specific transcript-1) in type I diabetes mellitus, Graves' disease and healthy controls. European journal of immunogenetics 22(3): 277-282.
Reddy, J. C. and J. D. Licht (1996). The WT1 Wilms' tumor suppressor gene: how much do we really know? Biochim Biophys Acta 1287(1): 1-28.
Redondo, M. J., L. Yu, M. Hawa, T. Mackenzie, D. A. Pyke, G. S. Eisenbarth and R. D. G. Leslie (2001). Heterogeneity of Type I diabetes: analysis of monozygotic twins in Great Britain and the United States. Diabetologia 44(3): 354-362.
Reijonen, H., S. Silvennoinen-Kassinen, J. Ilonen and M. Knip (1990). Lack of association of T cell receptor beta-chain constant region polymorphism with insulin-dependent diabetes mellitus in Finland. Clinical and experimental immunology 81(3): 396-399.
Reimers, J. I., H. U. Andersen, D. Mauricio, F. Pociot, A. E. Karlsen, J. S. Petersen, T. Mandrup-Poulsen and J.Nerup (1996). Strain-dependent differences in sensitivity of rat beta-cells to IL-1 β in vitro and in vivo. Diabetes 45: 771-778.
Reimsnider, S. K., S. E. Eckenrode, M. P. Marron, A. Muir and J. X. She (2000). IL4 and IL4R alpha genes are not linked or associated with type 1 diabetes. Pediatric Research 47(2): 246-249.
Resendez-E., J., J. W. Attenello, A. Grafsky, C. S. Chang and A. S. Lee (1985). Calcium ionophore A23187 induces expression of glucose-regulated genes and their heterologous fusion genes. Molecular and cellular biology 5(6): 1212-1219.
Rieneck, K., L. F. Bovin, K. Josefsen, K. Buschard, M. Svenson and K. Bendtzen (2000). Massive parallel gene expression profiling of RINm5F pancreatic islet beta-cells stimulated with interleukin-1 beta. APMIS 108(12): 855-872.
Ringel, J., S. Engeli, A. Distler and A. M. Sharma (1999). Pro12Ala missense mutation of the peroxisome proliferator activated receptor and diabetes mellitus. Biochemical and Biophysical Research Communications 254(2): 450-453.
Rioux, J., M. Silverberg, M. Daly, A. Steinhart, R. McLeod, A. Griffiths, T. Green, T. Brettin, V. Stone, S. Bull, A. Bitton, C. Williams, G. Greenberg, Z. Cohen, E. Lander, et al. (2000). Genomewide search in Canadian families with inflammatory bowel disease reveals two novel susceptibility loci. Am J Hum Genet 66: 1863-1870.
Risch, N. (1987). Assessing the role of HLA-linked and unlinked determinants of disease. American journal of human genetics 40(1): 1-14.
Risch, N., E. Burchard, E. Ziv and H. Tang (2002). Categorization of humans in biomedical research: Genes, race and disease. Genome Biology 3(7): 2007.1-2007.12.
Risch, N. and K. Merikangas (1996). The future of genetic studies of complex human diseases. Science 273(5281): 1516-1517.
Risch, N. and J. Teng (1998). The relative power of family-based and case-control designs for linkage disequilibrium studies of complex human diseases. I. DNA pooling. Genome Research 8(12): 1273-1288.
Rivolta, M. N. and M. C. Holley (2002). Asymmetric segregation of mitochondria and mortalin correlates with the multi-lineage potential of inner ear sensory cell progenitors in vitro. Brain research Developmental brain research 133(1): 49-56.
Robinson, W. P., J. Barbosa, S. S. Rich and G. Thomson (1993). Homozygous parent affected sib pair method for detecting disease predisposing variants - application to insulin- dependent diabetes-mellitus. Genetic Epidemiology 10(5): 273-288.
Rocic, B., M. Vucic, J. Knezevic-Cuca, A. Radica, I. Pavlic-Renar, V. Profozic and Z. Metelko (1997). Total plasma antioxidants in first-degree relatives of patients with insulin-dependent diabetes. Experimental and clinical endocrinology and diabetes 105(4): 213-217.
Roep, B. O. (2003). The role of T-cells in the pathogenesis of Type 1 diabetes: From cause to cure. Diabetologia 46(3): 305-321.
Rosas, G. O., S. J. Zieman, M. Donabedian, K. Vandegaer and J. M. Hare (2001). Augmented age-associated innate immune responses contribute to negative inotropic and lusitropic effects of lipopolysaccharide and interferon & gamma. Journal of Molecular and Cellular Cardiology 33(10): 1849-1859.
Rothe, H. and H. Kolb (1999). Strategies of protection from nitric oxide toxicity in islet inflammation. Journal of Molecular Medicine 77(1): 40-44.
Rowe, R. E., B. Wapelhorst, G. I. Bell, N. Risch, R. S. Spielman and P. Concannon (1995). Linkage and association between insulin-dependent diabetes mellitus (IDDM) susceptibility and markers near the glucokinase gene on chromosome 7. Nature genetics 10(2): 240-242.
Rutherford, S., M. P. Johnson, R. P. Curtain and L. R. Griffiths (2001). Chromosome 17 and the inducible nitric oxide synthase gene in human essential hypertension. Human Genetics 109(4): 408-415.
Rønningen, K. S., D. E. Undlien, R. Ploski, N. Maouni, R. J. Konrad, E. Jensen, E. Hornes, H. Reijonen, M. Colonna and D. S. Monos (1993). Linkage disequilibrium between TAP2 variants and HLA class II alleles; no primary association between TAP2 variants and insulin-dependent diabetes mellitus. European journal of immunology 23(5): 1050-1056.
Sacht, G., R. Brigelius-Flohe, M. Kiess, H. Sztajer and L. Flohe (1999). ATP-sensitive association of mortalin with the IL-1 receptor type I. Biofactors 9: 49-60.
Saito, H., S. Tada, H. Ebinuma, K. Wakabayashi, T. Takagi, Y. Saito, N. Nakamoto, S. Kurita and H. Ishii (2001). Interferon regulatory factor 1 promoter polymorphism and response to type 1 interferon. Journal of Cellular Biochemistry: 191-200.
Saito, H., S. Tada, K. Wakabayashi, N. Nakamoto, M. Takahashi, M. Nakamura, H. Ebinuma and H. Ishii (2002). The detection of IRF-1 promoter polymorphisms and their possible contribution to T helper 1 response in chronic hepatitis C. Journal of Interferon and Cytokine Research 22(6): 693-700.
Sakitani, K., M. Nishizawa, K. Inoue, Y. Masu, T. Okumura and S. Ito (1998). Synergistic regulation of inducible nitric oxide synthase gene by CCAAT/ enhancer-binding protein beta and nuclear factor-kappaB in hepatocytes. Genes to cells 3(5): 321-330.
Saldeen, J. (2000). Cytokines induce both necrosis and apoptosis via a common Bcl-2-inhibitable pathway in rat insulin-producing cells. Endocrinology 141(6): 2003-2010.
Saldeen, J. and N. Welsh (1994). Interleukin-1 beta induced activation of NF-kappa B in insulin producing RINm5F cells is prevented by the protease inhibitor N alpha-p-tosyl-L-lysine chloromethylketone. Biochemical and biophysical research communications 203(1): 149-155.
Salzman, A. L., A. G. Denenberg, I. Ueta, M. Oconnor, S. C. Linn and C. Szabo (1996). Induction and activity of nitric-oxide synthase in cultured human intestinal epithelial monolayers. American Journal of Physiology-Gastrointestinal and Liver Physiology 33(4): G565-G573.
Sandler, S., A. Andersson and C. Hellerström (1987). Inhibitory effects of interleukin 1 on insulin secretion, insulin biosynthesis, and oxidative metabolism of isolated rat pancreatic islets. Endocrinology 121(4): 1424-1431.
Sandler, S., K. Bendtzen, L. A. Borg, D. L. Eizirik, E. Strandell and N. Welsh (1989). Studies on the mechanisms causing inhibition of insulin secretion in rat pancreatic islets exposed to human interleukin-1 beta indicate a perturbation in the mitochondrial function. Endocrinology 124(3): 1492-1501.
Sandler, S., D. L. Eizirik, C. Svensson, E. Strandell, M. Welsh and N. Welsh (1991). Biochemical and molecular actions of interleukin-1 on pancreatic beta-cells. Autoimmunity 10(3): 241-253.
Sanjeevi, C. B., A. Kanungo, L. Berzina, A. Shtauvere-Brameus, M. Ghaderi and K. C. Samal (2002). MHC class I chain-related gene a alleles distinguish malnutrition-modulated diabetes, insulin-dependent diabetes, and non-insulin- dependent diabetes mellitus patients from eastern India. Annals of the New York Academy of Sciences 958: 341-344.
Sartoris, S., A. Brendolan, A. Degola, M. G. Testi, R. Chignola, A. Scarpa, M. Scardoni, G. Contreas, L. Pinelli, C. Lunardi, R. Beri, C. Pera, G. B. Ferrara, A. P. Riviera, G. Tridente, et al. (2000). Analysis of CIITA encoding AIR-1 gene promoters in insulin- dependent diabetes mellitus and rheumatoid arthritis patients from the Northeast of Italy: Absence of sequence variability. Human Immunology 61(6): 599-604.
Saura, M., C. Zaragoza, C. Bao, A. McMillan and C. Lowenstein (1999). Interaction of interferon regulatory factor-1 and nuclear factor kappaB during activation of inducible nitric oxide synthase transcription. J Mol Biol 289: 459-471.
Saura, M., C. Zaragoza, M. Díaz-Cazorla, O. Hernández-Perera, E. Eng, C. J. Lowenstein, D. P.érez-Sala and S. Lamas (1998). Involvement of transcriptional mechanisms in the inhibition of NOS2 expression by dexamethasone in rat mesangial cells. Kidney international 53(1): 38-49.
Savostianov, K. V., D. A. Chistiakov, L. N. Shcherbacheva, G. G. Mamaeva, M. I. Balabolkin and V. V. Nosikov (2002). Polymorphic locus D6S392 near the Mn-dependent superoxide dismutase gene is associated with diabetes mellitus in the Moscow population. Terapevticheskii arkhiv 74(10): 25-27.
Scaglia, L., C. J. Cahill, D. T. Finegood and S. Bonner-Weir (1997). Apoptosis participates in the remodeling of the endocrine pancreas in the neonatal rat. Endocrinology 138(4): 1736-1741.
Scaglia, L., F. E. Smith and S. Bonner-Weir (1995). Apoptosis contributes to the involution of beta cell mass in the post partum rat pancreas. Endocrinology 136(12): 5461-5468.
Scarim, A., M. Heitmeier and J. Corbett (1998). Heat shock inhibits cytokine-induced nitric oxide synthase expression by rat and human islets. Endocrinology 139: 5050-5057.
Scharnhorst, V., O. Kranenburg, E. van-der, A.J. and A. G. Jochemsen (1997). Differential regulation of the Wilms' tumor gene, WT1, during differentiation of embryonal carcinoma and embryonic stem cells. Cell growth and differentiation 8(2): 133-143.
Schroeder, R. A., J. S. Gu and P. C. Kuo (1998). Interleukin 1beta-stimulated production of nitric oxide in rat hepatocytes is mediated through endogenous synthesis of interferon gamma. Hepatology 27(3): 711-719.
Seegers, D., M. E. A. Borm, M. J. Van-Belzen, C. J. J. Mulder, J. Bailing, J. B. A. Crusius, J. W. R. Meijer, C. Wijmenga, A. S. Pena and G. Bouma (2003). IL 12B and IRF1 gene polymorphisms and susceptibility to celiac disease. European Journal of Immunogenetics 30(6): 421-425.
Segurado, O. G., C. M. Giles, P. Iglesias-Casarrubios, A. Corell, J. Martinez-Laso, J. L. Vicario and A. Arnaiz-Villena (1991). C4 Chido 3 and 6 distinguish two diabetogenic haplotypes: HLA-B49, SC01,DR4,DQw8 and B8,SC01,DR3,DQw2. Immunobiology 183(1-2): 12-22.
Sekiya, K., H. Nagasaki, N. Ozaki, A. Suzuki, Y. Miura and Y. Oiso (2000). Pituitary adenylate cyclase-activating polypeptide prevents cytokine-induced cytotoxicity via inhibition of inducible nitric oxide synthase expression in betaTC cells. Biochem Biophys Res Commun 278(1): 211-216.
Senaldi, G., C. Shaklee, J. Guo, L. Martin, T. Boone, T. W. Mak and T. Ulich (1999). Protection against the mortality associated with disease models mediated by TNF and IFN-gamma in mice lacking IFN regulatory factor-1. Journal of Immunology 163: 6820-6826.
Serreze, D. V., H. D. Chapman, D. S. Varnum, I. Gerling, E. H. Leiter and L. D. Shultz (1997). Initiation of autoimmune diabetes in NOD/Lt mice is MHC class I-dependent. Journal of immunology 158(8): 3978-3986.
Serreze, D. V. and E. H. Leiter (2001). Genes and cellular requirements for autoimmune diabetes susceptibility in nonobese diabetic mice. 4: 31-67.
Sessa, W. C., J. K. Harrison, C. M. Barber, D. Zeng, M. E. Durieux, D. D. D'Angelo, K. R. Lynch and M. J. Peach (1992). Molecular cloning and expression of a cDNA encoding endothelial cell nitric oxide synthase. Journal of biological chemistry 267(22): 15274-15276.
Sham, P. and D. Curtis (1995). An extended transmission/disequilibrium test (TDT) for multi-allele marker loci. Ann Hum Genet 59: 323-336.
Sharma, A., P. Prakash and M. Singh (2003). Autoimmune perspective of insulin-dependent diabetes mellitus: cytokines as therapeutic targets. Drugs of the Future 28(1): 31-42.
She, J. (1998). Genetic susceptibility factors in type 1 diabetes: linkage, disequilibrium and functional analyses. Current opinion in immunology, 10: 682-689.
She, J. X. (1996). Susceptibility to type I diabetes: HLA-DQ and DR revisited. Immunology today 17(7): 323-329.
Sheehy, M. J., L. M. Meske, C. A. Emler, J. R. Rowe, G. Neme-De, M.H., C. A. Ingle, A. Chan, M. Trucco and T. W. Mak (1989). Allelic T-cell receptor alpha complexes have little or no influence on susceptibility to type 1 diabetes. Human Immunology 26(4): 261-271.
Shen, J., R. T. Wang and X. P. Xu (2002). Newfound Tsp polymorphism of iNOS gene in Chinese and its correlation with the susceptibility of stomach and cardiac cancer. Proceedings of the American Association for Cancer Research Annual Meeting 43: 571.
Sherman, P. A., V. E. Laubach, B. R. Reep and E. R. Wood (1993). Purification and cDNA sequence of an inducible nitric oxide synthase from a human tumor cell line. Biochemistry 32(43): 11600-11605.
Sherry, S. T., M. Ward and K. Sirotkin (2000). Use of molecular variation in the NCBI dbSNP database. Human Mutation 15(1): 68-75.
Shtauvere-Brameus, A., P. Dabadghao, I. Rumba and C. B. Sanjeevi (2002). Tumor necrosis factor-alpha allele 2 shows an association with insulin-dependent diabetes mellitus in Latvians. Annals of the New York Academy of Sciences 958: 357-361.
Shtauvere-Brameus, A., M. Ghaderi, I. Rumba and C. B. Sanjeevi (2002). Microsatellite allele 5 of MHC class I chain-related gene a increases the risk for insulin-dependent diabetes mellitus in latvians. Annals of the New York Academy of Sciences 958: 349-352.
Sieradzki, J., M. T. Malecki, T. Klupa, L. Hanna, J. Sieradzka, J. Frey and A. S. Krolewski (1999). Amino acid variants of the vitamin D-binding protein are not associated with type 1 diabetes in Caucasians. Diabetologia 42(SUPPL. 1): A91.
Simeonovic, C. J., R. Ceredig and J. D. Wilson (1990). Effect of GK1.5 monoclonal antibody dosage on survival of pig proislet xenografts in CD4+ T cell-depleted mice. Transplantation 49(5): 849-856.
Singh, V. K., S. Mehrotra, P. Narayan, C. M. Pandey and S. S. Agarwal (2000). Modulation of autoimmune diseases by nitric oxide. Immunologic Research 22(1): 1-19.
Sjöholm, A. (1996). Nitric oxide donor SIN-1 inhibits insulin release. American journal of physiology 271(4 Pt 1): C1098-C1102.
Skrabic, V., T. Zemunik, M. Situm and J. Terzic (2003). Vitamin D receptor polymorphism and susceptibility to type 1 diabetes in the Dalmatian population. Diabetes Research and Clinical Practice 59(1): 31-35.
Smyth, D., J. D. Cooper, J. E. Collins, J. M. Heward, J. A. Franklyn, J. M. Howson, A. Vella, S. Nutland, H. E. Rance, L. Maier, B. J. Barratt, C. Guja, C. Ionescu-T.îrgoviste, D. A. Savage, D. B. Dunger, et al. (2004). Replication of an association between the lymphoid tyrosine phosphatase locus (LYP/PTPN22) with type 1 diabetes, and evidence for its role as a general autoimmunity locus. Diabetes 53(11): 3020-3023.
Southern, C., D. Schulster and I. Green (1990). Inhibition of insulin secretion by interleukin-1-beta and tumor necrosis factor-alpha via an L-argenine-dependent nitric oxide generating mechanism. FEBS Lett. 276: 42-44.
Sparre, T., U. B. Christensen, P. M. Larsen, S. J. Fey, K. Wrzesinski, P. Roepstorff, T. Mandrup-Poulsen, F. Pociot, A. E. Karlsen and J. Nerup (2002). IL-1beta induced protein changes in diabetes prone BB rat islets of langerhans identified by proteome analysis. Diabetologia 45(11): 1550-1561.
Spielman, R. and W. Ewens (1996). The TDT and other family-based tests for linkage disequilibrium and association. Am Hum J Gen 59: 983-989.
Spielman, R. S., R. E. McGinnis and W. J. Ewens (1993). Transmission test for linkage disequilibrium: the insulin gene region and insulin-dependent diabetes mellitus (IDDM). Am J Hum Genet 52: 506-516.
Spinas, G. A., B. S. Hansen, S. Linde, W. Kastern, J. Mølvig, T. Mandrup-Poulsen, C. A. Dinarello, J. H. Nielsen and J. Nerup (1987). Interleukin 1 dose-dependently affects the biosynthesis of (pro)insulin in isolated rat islets of Langerhans. Diabetologia 30(7): 474-480.
Spink, J., J. Cohen and T. J. Evans (1995). The cytokine responsive vascular smooth muscle cell enhancer of inducible nitric oxide synthase. Activation by nuclear factor-kappa B. Journal of biological chemistry 270(49): 29541-29547.
Spitsin, S. V., J. L. Farber, M. Bertovich, G. Moehren, H. Koprowski and F. H. Michaels (1997). Human- and mouse-inducible nitric oxide synthase promoters require activation of phosphatidylcholine-specific phospholipase C and NF-kappa B. Molecular medicine (Cambridge, Mass.) 3(5): 315-326.
Spitsin, S. V., H. Koprowski and F. H. Michaels (1996). Characterization and functional analysis of the human inducible nitric oxide synthase gene promoter. Molecular medicine 2(2): 226-235.
Stamler, J. S. (1994). Redox signaling: nitrosylation and related target interactions of nitric oxide. Cell 78(6): 931-936.
Staneková, D., M. Niks, M. Buc, Z. Starsia and Michalková (1993). Genetic polymorphism of factor B (Bf) and C3 component of complement in type 1 (insulin-dependent) diabetes mellitus: BFQO allele observed in a diabetic child. Folia biologica 39(3): 117-123.
Stassi, G., R. De-Maria, G. Trucco, W. Rudert, R. Testi, A. Galluzzo, C. Giordano and M. Trucco (1997). Nitric oxide primes pancreatic beta cells for Fas-mediated destruction in insulin-dependent diabetes mellitus. Journal of experimental medicine 186(8): 1193-1200.
Steiner, L., K. Kroncke, K. Fehsel and V. K.-B. V. (1997). Endothelial cells as cytotoxic effector cells: cytokine-activated rat islet endothelial cells lyse syngeneic islet cells via nitric oxide. Diabetologia 40: 150-155.
Strandell, E., K. Buschard, J. Saldeen and N. Welsh (1995). Interleukin-1-beta induces the expression of HSP70, heme oxygenase and Mn-SOD in FACS-purified rat islet beta-cells, but not in alpha-cells. Immunology Letters 48(2): 145-148.
Stuehr, D. J. and M. A. Marletta (1985). Mammalian nitrate biosynthesis: mouse macrophages produce nitrite and nitrate in response to Escherichia coli lipopolysaccharide. Proceedings of the National Academy of Sciences of the United States of America 82(22): 7738-7742.
Størling, J., S. V. Zaitsev, N. A. Andersen, I. L. Kapelioukh, A. E. Karlsen, P. O. Berggren and T. Mandrup-Poulsen (2001). Calcium signalling via L-type calcium channel mediates IL-1-induced activation of C-jun N-terminal kinase and p38 in pancreatic beta-cells. Diabetologia 44: A129.
Suarez-Pinzon, W., C. Szabo and A. Rabinovitch (1997). Development of autoimmune diabetes in NOD mice is associated with the formation of peroxynitrite in pancreatic islet beta-cells. Diabetes 46: 907-911.
Suarez-Pinzon, W. L., J. G. Mabley, K. Strynadka, R. F. Power, C. Szabo and A. Rabinovitch (2001). An inhibitor of inducible nitric oxide synthase and scavenger of peroxynitrite prevents diabetes development in NOD mice. Journal of Autoimmunity 16(4): 449-455.
Sumoski, W., H. Baquerizo and A. Rabinovitch (1989). Oxygen free radical scavengers protect rat islet cells from damage by cytokines. Diabetologia 32(11): 792-796.
Svejgaard, A. and L. P. Ryder (1994). HLA and disease associations: detecting the strongest association. Tissue antigens 43(1): 18-27.
Svensson, J., B. Carstensen, A. Molbak, B. Christau, H. B. Mortensen, J. Nerup and K. Borch-Johnsen (2002). Increased, risk of childhood type I diabetes in children born after 1985. Diabetes Care 25(12): 2197-2201.
Swift, R. G., M. H. Polymeropoulos, R. Torres and M. Swift (1998). Predisposition of Wolfram syndrome heterozygotes to psychiatric illness. Molecular psychiatry 3(1): 86-91.
Syapin, P. J., J. D. Militante, D. K. Garrett and L. Ren (2001). Cytokine-induced iNOS expression in C6 glial cells: transcriptional inhibition by ethanol. Journal of Pharmacology and Experimental Therapeutics 298(2): 744-752.
Szabó, C. (1996). The pathophysiological role of peroxynitrite in shock, inflammation, and ischemia-reperfusion injury. Shock 6(2): 79-88.
Szabo, C. and C. Thiemermann (1994). Invited opinion - role of nitric-oxide in hemorrhagic, traumatic, and anaphylactic shock and thermal-injury. Shock 2(2): 145-155.
Szalai, C., A. Csaszar, A. Czinner, T. Szabo, P. Panczel, L. Madacsy and A. Falus (1999). Chemokine receptor CCR2 and CCR5 polymorphisms in children with insulin-dependent diabetes mellitus. Pediatric Research 46 (1): 82-84.
Tabiin, M. T., B. E. Tuch, L. Bai, X. Han and A. M. Simpson (2001). Susceptibility of insulin-secreting hepatocytes to the toxicity of pro-inflammatory cytokines. Journal of Autoimmunity 17(3): 229-242.
Tabone, T. and G. Morahan (2003). Definition of polymorphisms in the gene encoding the interleukin-12 receptor B1 subunit: testing linkage disequilibrium with Type I diabetes susceptibility. 4(3): 222-227.
Tabor, H. K., N. J. Risch and R. M. Myers (2002). Candidate-gene approaches for studying complex genetic traits: practical considerations. Nature Reviews Genetics 3(5): 391-A396.
Taillon-Miller, P., Z. Gu, Q. Li, L. Hillier and P. Y. Kwok (1998). Overlapping genomic sequences: a treasure trove of single-nucleotide polymorphisms. Genome Research 8(7): 748-754.
Tak, P. P. and G. S. Firestein (2001). NF-kappa B: a key role in inflammatory diseases. Journal of Clinical Investigation 107(1): 7-11.
Takahashi, K., S. Suzuki, N. Matsuura and J. Satoh (2001). A novel NRAMP1 promoter polymorphism associates with type 1 diabetes mellitus in Japanese subjects. Diabetes 50(Suppl 2): A502-A503.
Takahashi, M., A. Chesley, D. Freyssenet and D. A. Hood (1998). Contractile activity-induced adaptations in the mitochondrial protein import system. American journal of physiology 274(5 Pt 1): C1380-C1387.
Takamura, T., I. Kato, N. Kimura, T. Nakazawa, H. Yonekura, S. Takasawa and H. Okamoto (1998). Transgenic mice overexpressing type 2 nitric-oxide synthase in pancreatic gb cells develop insulin-dependent diabetes without insulitis. Journal of Biological Chemistry 273(5): 2493-2496.
Tannous, M., R. Veluthakal, R. Amin and A. Kowluru (2002). IL-1 beta-induced nitric oxide release from insulin-secreting beta-cells: Further evidence for the involvement of GTP- binding proteins. Diabetes and Metabolism 28(6): S78-S84.
Taurin, S., V. Seyrantepe, S. N. Orlov, T. L. Tremblay, P. Thibault, M. R. Bennett, P. Hamet and A. V. Pshezhetsky (2002). Proteome analysis and functional expression identify mortalin as an antiapoptotic gene induced by elevation of [Na + ] i /[K + ]i ratio in cultured vascular smooth muscle cells. Circulation Research 91(10): 915-922.
Taylor, B. S., L. H. Alarcon and T. R. Billiar (1998). Inducible nitric oxide synthase in the liver: regulation and function. Biochemistry Biokhimiia 63(7): 766-781.
Taylor, B. S., M. E. de-Vera, R. W. Ganster, Q. Wang, R. A. Shapiro, S. M. Morris, T. R. Billiar and D. A. Geller (1998). Multiple NF-kappaB enhancer elements regulate cytokine induction of the human inducible nitric oxide synthase gene. Journal of biological chemistry 273(24): 15148-15156.
Taylor, B. S. and D. A. Geller (2000). Molecular regulation of the human inducible nitric oxide synthase (iNOS) gene. Shock 13(6): 413-424.
Taylor-Robinson, A. W., F. Y. Liew, A. Severn, D. Xu, S. J. McSorley, P. Garside, J. Padron and R. S. Phillips (1994). Regulation of the immune response by nitric oxide differentially produced by T helper type 1 and T helper type 2 cells. European journal of immunology 24(4): 980-984.
Tegoshi, H., G. Hasegawa, H. Obayashi, K. Nakano, Y. Kitagawa, M. Fukui, S. Matsuo, M. Deguchi, M. Ohta, M. Nishimura, N. Nakamura and T. Yoshikawa (2002). Polymorphisms of interferon-gamma gene CA-repeat and interleukin-10 promoter region (-592A/C) in Japanese type I diabetes. Human immunology 63(2): 121-128.
Teng, X., H. Zhang, C. Snead and J. D. Catravas (2000). A reverse nuclear factor-kappaB element in the rat type II nitric oxide synthase promoter mediates the induction by interleukin-1beta and interferon-gamma in rat aortic smooth muscle cells. General pharmacology 34(1): 9-16.
Teng, X., H. Zhang, C. Snead and J. D. Catravas (2002). Molecular mechanisms of iNOS induction by IL-1 beta and IFN-gamma in rat aortic smooth muscle cells. American journal of physiology Cell physiology 282(1): C144-C152.
Thivolet, C., A. Bendelac, P. Bedossa, J. F. Bach and C. Carnaud (1991). CD8+ T cell homing to the pancreas in the nonobese diabetic mouse is CD4+ T cell-dependent. Journal of immunology 146(1): 85-88.
Thomas, H. E., R. Darwiche, J. A. Corbett and T. W. H. Kay (2002). Interleukin-1 plus gamma-interferon-induced pancreatic beta- cell dysfunction is mediated by beta-cell nitric oxide production. Diabetes 51(2): 311-316.
Thomas, H. E., W. Irawaty, R. Darwiche, T. C. Brodnicki, P. Santamaria, J. Allison and T. W. H. Kay (2004). IL-1 receptor deficiency slows progression to diabetes in the NOD mouse. Diabetes 53(1): 113-121.
Thomson, G., W. P. Robinson, M. K. Kuhner, S. Joe, M. J. MacDonald, J. L. Gottschall, J. Barbosa, S. S. Rich, J. Bertrams, M. P. Baur and et al. (1988). Genetic heterogeneity, modes of inheritance, and risk estimates for a joint study of Caucasians with insulin-dependent diabetes mellitus. American journal of human genetics 43(6): 799-816.
Thornberry, N. A. and Y. Lazebnik (1998). Caspases: Enemies within. Science 281(5381): 1312-1316.
Thorsby, E. and K. S. Rønningen (1993). Particular HLA-DQ molecules play a dominant role in determining susceptibility or resistance to type 1 (insulin-dependent) diabetes mellitus. Diabetologia 36(5): 371-377.
Tiedge, M., S. Lortz, J. Drinkgern and S. Lenzen (1997). Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin-producing cells. Diabetes 46(11): 1733-1742.
Tiedge, M., S. Lortz, R. Munday and S. Lenzen (1998). Complementary action of antioxidant enzymes in the protection of bioengineered insulin-producing RINm5F cells against the toxicity of reactive oxygen species. Diabetes 47(10): 1578-1585.
Tiedge, M., S. Lortz, R. Munday and S. Lenzen (1999). Protection against the co-operative toxicity of nitric oxide and oxygen free radicals by overexpression of antioxidant enzymes in bioengineered insulin-producing RINm5F cells. Diabetologia 42(7): 849-855.
Timon, M., A. Arnaiz-villena, P. Perez-aciego, P. Morales, D. Benmamar and J. R. Regueiro (1991). A diallelic RFLP of the CD3-epsilon chain of the clonotypic T-lymphocyte receptor is not associated with certain autoimmune diseases. Human Genetics 86(4): 363-364.
Todd, J. A. and M. Farrall (1997). Panning for gold: Genomewide scanning in type 1 diabetes. Diabetes Reviews 5(3): 284-291.
Todd, J. A. and L. S. Wicker (2001). Genetic protection from the inflammatory disease type 1 diabetes in humans and animal models. Immunity 15(3): 387-395.
Ton, C. C., H. Hirvonen, H. Miwa, M. M. Weil, P. Monaghan, T. Jordan, V. van-Heyningen, N. D. Hastie, H. Meijers-Heijboer, M. Drechsler and a. et (1991). Positional cloning and characterization of a paired box- and homeobox-containing gene from the aniridia region. Cell 67(6): 1059-1074.
Torn, C., M. Gupta, L. N. Zake, C. B. Sanjeevi and M. Landin-Olsson (2003). Heterozygosity for MICA5.0/MICA5.1 and HLA-DR3-DQ2/DR4-DQ8 are independent genetic risk factors for latent autoimmune diabetes in adults. Human Immunology 64(9): 902-909.
Ueda, H., J. M. M. Howson, L. Esposito, J. Heward, H. Snook, G. Chamberlain, D. B. Rainbow, K. M. D. Hunter, A. N. Smith, G. Di-Genova, M. H. Herr, I. Dahlman, F. Payne, D. Smyth, C. Lowe, et al. (2003). Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease. Nature (London) 423(6939): 506-511.
Undlien, D. E., H. E. Akselsen, G. Joner, K. Dahl-J.ørgensen, O. S.øvik, K. S. R.ønningen and E. Thorsby (1997). No independent associations of LMP2 and LMP7 polymorphisms with susceptibility to develop IDDM. Diabetes 46(2): 307-312.
Undlien, D. E., I. Kockum, K. S. Rønningen, R. Lowe, C. B. Saanjeevi, J. Graham, B. A. Lie, H. E. Akselsen, A. Lernmark and E. Thorsby (1999). HLA associations in type 1 diabetes among patients not carrying high-risk DR3-DQ2 or DR4-DQ8 haplotypes. Tissue Antigens 54(6): 543-551.
Undlien, D. E., B. A. Lie and E. Thorsby (2001). HLA complex genes in type 1 diabetes and other autoimmune diseases. Which genes are involved? Trends in genetics 17(2): 93-100.
Urano, F., X. Z. Wang, A. Bertolotti, Y. H. Zhang, P. Chung, H. P. Harding and D. Ron (2000). Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1. Science 287(5453): 664-666.
Vaessen, N., P. Heutink, J. J. Houwing-Duistermaat, P. J. Snijders, T. Rademaker, L. Testers, M. R. Batstra, L. A. Sandkuijl, C. M. van-Duijn and B. A. Oostra (2002). A genome-wide search for linkage-disequilibrium with type 1 diabetes in a recent genetically isolated population from the Netherlands. Diabetes 51(3): 856-859.
Vafiadis, P., S. T. Bennett, J. A. Todd, J. Nadeau, R. Grabs, C. G. Goodyer, S. Wickramasinghe, E. Colle and C. Polychronakos (1997). Insulin expression in human thymus is modulated by INS VNTR alleles at the IDDM2 locus. Nature genetics 15(3): 289-292.
Van-Endert, P. M., R. S. Liblau, S. D. Patel, L. Fugger, T. Lopez, F. Pociot, J. Nerup and H. O. McDevitt (1994). Major histocompatibility complex-encoded antigen processing gene polymorphism in IDDM. Diabetes 43(1): 110-117.
Vara, E., J. Arias-Diaz, C. García and J. L. Balibrea (1994). Cytokine-induced inhibition of lipid synthesis and hormone secretion by isolated human islets. Pancreas 9(3): 316-323.
Veijola, R., M. Knip, R. Puukka, H. Reijonen, D. W. Cox and J. Ilonen (1996). The immunoglobulin heavy-chain variable region in insulin-dependent diabetes mellitus: affected-sib-pair analysis and association studies. American journal of human genetics 59(2): 462-470.
Veijola, R., M. Knip, H. Reijonen, P. Vähäsalo, R. Puukka and J. Iionen (1995). Effect of genetic risk load defined by HLA-DQB1 polymorphism on clinical characteristics of IDDM in children. European journal of clinical investigation 25(2): 106-112.
Vella, A., J. M. Howson, B. J. Barratt, R. C. Twells, H. E. Rance, S. Nutland, E. Tuomilehto-Wolf, J. Tuomilehto, D. E. Undlien, K. S. R.ønningen, C. Guja, C. Ionescu-Tîrgoviste, D. A. Savage and J. A. Todd (2004). Lack of association of the Ala(45)Thr polymorphism and other common variants of the NeuroD gene with type 1 diabetes. Diabetes 53(4): 1158-1161.
Vera, d. M., R. Shapiro, A. Nussler, J. Mudgett, R. Simmons, S. Morris, T. J. Billiar and D. Geller (1996a). Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: initial analysis of the human NOS2 promoter. PNAS 93: 1054-1059.
Vera, M. d., Y. Kim, H. Wong, Q. Wang, T. Billiar and D. Geller (1996). Heat shock response inhibits cytokine-inducible nitric oxide synthase expression in rat hepatocytes. Hepatology 24: 1238-1245.
Verge, C. F., P. Vardi, S. Babu, F. Bao, H. A. Erlich, T. Bugawan, D. Tiosano, L. P. Yu, G. S. Eisenbarth and P. R. Fain (1998). Evidence for oligogenic inheritance of type 1 diabetes in a large Bedouin Arab family. J. Clin. Invest. 102(8): 1569-1575.
Vodovotz, Y. (1997). Control of nitric oxide production by transforming growth factor- beta1: mechanistic insights and potential relevance to human disease. Nitric Oxide - Biology and Chemistry 1(1): 3-17.
Vodovotz, Y., C. Bogdan, J. Paik, Q. W. Xie and C. Nathan (1993). Mechanisms of suppression of macrophage nitric oxide release by transforming growth factor beta. Journal of experimental medicine 178(2): 605-613.
Wadhwa, R., H. Ando, H. Kawasaki, K. Taira and S. C. Kaul (2003). Targeting mortalin using conventional and RNA-helicase-coupled hammerhead ribozymes. EMBO Reports 4(6): 595-601.
Wadhwa, R., S. Kaul, Y. Ikawa and Y. Sugimoto (1991). Protein markers for cellular mortality and immortality. Mutat Res 256: 243-254.
Wadhwa, R., S. Kaul, Y. Ikawa and Y. Sugimoto (1993a). Identification of a novel member of mouse hsp70 family. Its association with cellular mortal phenotype. J Biol Chem 268: 6615-6621.
Wadhwa, R., S. Kaul, Y. Mitsui and Y. Sugimoto (1993b). Differential subcellular distribution of mortalin in mortal and immortal mouse and human fibroblasts. Experimental Cell Research 207: 442-448.
Wadhwa, R., S. Kaul, Y. Sugimoto and Y. Mitsui (1993c). Induction of cellular senescence by transfection of cytosolic mortalin cDNA in NIH 3T3 cells. J of Biol Chem 268: 22239-22242.
Wadhwa, R., O. Pereira-Smith, R. Reddel, Y. Sugimoto, Y. Mitsui and S. Kaul (1995a). Correlation between complementation group for immortality and the cellular distribution of mortalin. Experimental Cell Research 216: 101-106.
Wadhwa, R., T. Sugihara, A. Yoshida, H. Nomura, R. R. Reddel, R. Simpson, H. Maruta and S. C. Kaul (2000a). Selective toxicity of MKT-077 to cancer cells is mediated by its binding to the hsp70 family protein mot-2 and reactivation of p53 function. Cancer research 60(24): 6818-6821.
Wadhwa, R., K. Taira and S. C. Kaul (2002a). An Hsp70 family chaperone, mortalin/mthsp70/PBP74/Grp75: What, when, and where? Cell Stress & Chaperones 7(3): 309-316.
Wadhwa, R., S. Takano, Y. Mitsui and S. C. Kaul (1999). NIH 3T3 cells malignantly transformed by mot-2 show inactivation and cytoplasmic sequestration of the p53 protein. Cell research 9(4): 261-269.
Wadhwa, R., S. Takano, M. Robert, A. Yoshida, H. Nomura, R. R. Reddel, Y. Mitsui and S. C. Kaul (1998). Inactivation of tumor suppressor p53 by Mot-2, a hsp70 family member. Journal of Biological Chemistry 273(45): 29586-29591.
Wadhwa, R., T. Yaguchi, M. K. Hasan, Y. Mitsui, R. R. Reddel and S. C. Kaul (2002d). Hsp70 family member, mot-2/mthsp70/GRP75, binds to the cytoplasmic sequestration domain of the p53 protein. Experimental cell research 274(2): 246-253.
Wadhwa, R., T. Yaguchi, M. K. Hasan, K. Taira and S. C. Kaul (2003). Mortalin-MPD (mevalonate pyrophosphate decarboxylase) interactions and their role in control of cellular proliferation. Biochemical and Biophysical Research Communications 302(4): 735-742.
Wang, C., M. L. Rivas, G. A. Burghen, E. C. Hudson and R. J. Wyatt (1989). C4 and Bf phenotypes in black and Caucasian patients with childhood onset insulin dependent diabetes mellitus. Journal of clinical & laboratory immunology 30(4): 183-190.
Wapelhorst, B., G. I. Bell, N. Risch, R. S. Spielman and P. Concannon (1995). Linkage and association studies in insulin-dependent diabetes with a new dinucleotide repeat polymorphism at the GAD65 locus. Autoimmunity 21(2): 127-130.
Warpeha, K. M., W. Xu, L. Liu, I. G. Charles, C. C. Patterson, F. Ah-Fat, S. Harding, P. M. Hart, U. Chakravarthy and A. E. Hughes (1999). Genotyping and functional analysis of a polymorphic (CCTTT)(n) repeat of NOS2A in diabetic retinopathy. FASEB Journal 13(13): 1825-1832.
Watkins, P. J. and P. K. Thomas (1998). Diabetes mellitus and the nervous system. Journal of neurology, neurosurgery, and psychiatry 65(5): 620-632.
Wei, X. Q., I. G. Charles, A. Smith, J. Ure, G. J. Feng, F. P. Huang, D. Xu, W. Muller, S. Moncada and F. Y. Liew (1995). Altered immune responses in mice lacking inducible nitric oxide synthase. Nature 375(6530): 408-411.
Welch, W. J., H. S. Kang, R. P. Beckmann and L. A. Mizzen (1991). Response of mammalian cells to metabolic stress; changes in cell physiology and structure/function of stress proteins. Current topics in microbiology and immunology 167: 31-55.
Welsh, N. (1996). Interleukin-1 beta-induced ceramide and diacylglycerol generation may lead to activation of the c-Jun NH2-terminal kinase and the transcription factor ATF2 in the insulin-producing cell line RINm5F. Journal of biological chemistry 271(14): 8307-8312.
Welsh, N., D. L. Eizirik, K. Bendtzen and S. Sandler (1991a). Interleukin-1 beta-induced nitric oxide production in isolated rat pancreatic islets requires gene transcription and may lead to inhibition of the Krebs cycle enzyme aconitase. Endocrinology 129(6): 3167-3173.
Welsh, N., B. Margulis, K. Bendtzen and S. Sandler (1994). Liposomal delivery of antioxidant enzymes protects against hydrogen peroxide- but not interleukin-1 beta-induced inhibition of glucose metabolism in rat pancreatic islets. Journal of endocrinology 143(1): 151-156.
Welsh, N., B. Margulis, L. A. Borg, H. J. Wiklund, J. Saldeen, M. Flodström, M. A. Mello, A. Andersson, D. G. Pipeleers and C. Hellerström (1995b). Differences in the expression of heat-shock proteins and antioxidant enzymes between human and rodent pancreatic islets: implications for the pathogenesis of insulin-dependent diabetes mellitus. Molecular medicine 1(7): 806-820.
Welsh, N., M. Welsh, S. Lindquist, D. L. Eizirik, K. Bendtzen and S. Sandler (1991b). Interleukin-1 beta increases the biosynthesis of the heat shock protein hsp70 and selectively decreases the biosynthesis of five proteins in rat pancreatic islets. Autoimmunity 9(1): 33-40.
Wen, J., M. Han and D. Liu (2000). Molecular cloning and characterization of the promoter region of the inducible nitric oxide synthase gene of the rat. Biochemistry Biokhimiia 65(11): 1327-1330.
WHO (1999). Definition, Diagnosis and Classification of Diabetes Mellitus and its Complications. WHO, Report of a WHO Consultation Part 1: 1-59.
Wicker, L. S., J. A. Todd and L. B. Peterson (1995). Genetic control of autoimmune diabetes in the NOD mouse. Annual review of immunology 13: 179-200.
Widmann, C., S. Gibson, M. B. Jarpe and G. L. Johnson (1999). Mitogen-activated protein kinase: Conservation of a three- kinase module from yeast to human. Physiological Reviews 79(1): 143-180.
Willman, C. L., C. E. Sever, M. G. Pallavicini, H. Harada, N. Tanaka, M. L. Slovak, H. Yamamoto, K. Harada, T. C. Meeker, A. F. List and et al. (1993). Deletion of IRF-1, mapping to chromosome 5q31.1, in human leukemia and preleukemic myelodysplasia. Science 259(5097): 968-971.
Wilson, G. L., N. J. Patton and S. P. LeDoux (1997). Mitochondrial DNA in beta-cells is a sensitive target for damage by nitric oxide. Diabetes 46(8): 1291-1295.
Wong, S., S. Moore, S. Orisio, A. Millward and A. G. Demaine (1991). Susceptibility to type I diabetes in women is associated with the CD3 epsilon locus on chromosome 11. Clinical and experimental immunology 83(1): 69-73.
Wood, E. R., H. Berger, P. A. Sherman and E. G. Lapetina (1993). Hepatocytes and macrophages express an identical cytokine inducible nitric oxide synthase gene. Biochemical and biophysical research communications 191(3): 767-774.
Wood, J. P., M. A. Pani, K. Bieda, G. Meyer, K. H. Usadel and K. Badenhoop (2002). A recently described polymorphism in the CD28 gene on chromosome 2q33 is not associated with susceptibility to type 1 diabetes. European journal of immunogenetics 29(4): 347-349.
Wu, R., Y. H. Zhao, C. G. Plopper, M. M. Chang, K. Chmiel, J. J. Cross, A. Weir, J. A. Last and B. Tarkington (1999). Differential expression of stress proteins in nonhuman primate lung and conducting airway after ozone exposure. American journal of physiology 277(3 Pt 1): L511-L522.
Xie, H., Z. Hu, B. Chyna, S. K. Horrigan and C. A. Westbrook (2000). Human mortalin (HSPA9): a candidate for the myeloid leukemia tumor suppressor gene on 5q31. Leukemia 14(12): 2128-2134.
Xie, Q., Y. Kashiwabara and C. Nathan (1994a). Role of Transcription Factor NF-kappa-B/Rel in Induction of Nitric Oxide Synthase. Journal of Biological Chemistry 269(7): 4705-4708.
Xie, Q. W., Y. Kashiwabara and C. Nathan (1994). Role of transcription factor NF-kB/Rel in induction of nitric oxide synthase. J Biol Chem 269: 4705-4708.
Xie, Q. W., R. Whisnant and C. Nathan (1993). Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide. J Exp Med 177(6): 1779-1784.
Xu, J., H. H. Xiao and A. C. Sartorelli (1999). Attenuation of the induced differentiation of HL-60 leukemia cells by mitochondrial chaperone HSP70. Oncology Research 11(9): 429-435.
Xu, W., I. Charles, L. Liu, S. Moncada and P. Emson (1996). Molecular cloning and structural organization of the human inducible nitric oxide synthase gene (NOS2). Biochemical and Biophysical Research Communications 219: 784-788.
Xu, W., L. Liu, P. C. Emson, C. R. Harrington and I. G. Charles (1997). Evolution of a homopurine-homopyrimidine pentanucleotide repeat sequence upstream of the human inducible nitric oxide synthase gene. Gene 204(1-2): 165-170.
Xu, W. M., S. Humphries, M. Tomita, T. Okuyama, M. Matsuki, D. Burgner, D. Kwiatkowski, L. Z. Liu and I. G. Charles (2000). Survey of the allelic frequency of a NOS2A promoter microsatellite in human populations: Assessment of the NOS2A gene and predisposition to infectious disease. Nitric Oxide-Biology and Chemistry 4(4): 379-383.
Xu, W. M., L. Z. Liu, P. Emson, C. R. Harrington, I. G. McKeith, R. H. Perry, C. M. Morris and I. G. Charles (2000). The CCTTT polymorphism in the NOS2A gene is associated with dementia with Lewy bodies. Neuroreport 11(2): 297-299.
Yamada, S., Y. Motohashi, T. Yanagawa, T. Maruyama, A. Kasuga, H. Hirose, K. Matsubara, A. Shimada and T. Saruta (2001). NeuroD/BETA2 gene G -> A polymorphism may affect onset pattern of type 1 diabetes in Japanese. Diabetes Care 24(8): 1438-1441.
Yamada, S., Y. Motohashi, T. Yanagawa, T. Maruyama, A. Kasuga, H. Hirose, R. Suzuki, K. Matsubara, A. Shimada and T. Saruta (2001). Vitamin D receptor gene polymorphism in type 1 diabetes. Diabetes 50(Suppl 2): A232.
Yamazaki, Y., N. Miyokawa and M. Katagiri (1994). Polymorphism of the TAP genes Japanese healthy control and type I diabetes mellitus. [Hokkaido igaku zasshi] The Hokkaido journal of medical science 69(2): 337-346.
Yan, G., L. Shi, Y. Fu, X. Wang, D. Schoenfeld, L. Ma, A. Penfornis, H. Gebel and D. L. Faustman (1997). Screening of the TAP1 gene by denaturing gradient gel electrophoresis in insulin-dependent diabetes mellitus: Detection and comparison of new polymorphisms between patients and controls. Tissue Antigens 50(6): 576-585.
Yokota, I., S. Satomura, S. Kitamura, Y. Taki, E. Naito, M. Ito, K. Nisisho and Y. Kuroda (2002). Association between vitamin D receptor genotype and age of onset in juvenile Japanese patients with type 1 diabetes. Diabetes care 25(7): 1244-PY - 2002.
Yokouchi, Y., Y. Nukaga, M. Shibasaki, E. Noguchi, K. Kimura, S. Ito, M. Nishihara, K. Yamakawa-Kobayashi, K. Takeda, N. Imoto, K. Ichikawa, A. Matsui, H. Hamaguchi and T. Arinami (2000). Significant evidence for linkage of mite-sensitive childhood asthma to chromosome 5q31-q33 near the interleukin 12 B locus by a genome-wide search in Japanese families. Genomics 66(2): 152-160.
Yu, H., A. Thai and S. Chan (1999). HLA microsatellite associations with insulin-dependent diabetes mellitus in Singaporean Chinese. Human Immunology 60(9): 894-900.
Zake, L. N., M. Ghaderi, Y. S. Park, S. Babu, G. Eisenbarth and C. B. Sanjeevi (2002). MHC class I chain-related gene alleles 5 and 5.1 are transmitted more frequently to type 1 diabetes offspring in HBDI families. Annals of the New York Academy of Sciences 958: 309-311.
Zamani, M. and J. J. Cassiman (1998). Reevaluation of the importance of polymorphic HLA Class II Alleles and amino acids in the susceptibility of individuals of different populations to type I diabetes. American Journal of Medical Genetics 76(2): 183-194.
Zamani-Ghabanbasani, M., I. Buyse, E. Legius, R. Decorte, P. Marynen, R. Bouillon and J. J. Cassiman (1994). Possible association of CD3 and CD4 polymorphisms with insulin-dependent diabetes mellitus (IDDM). Clinical and Experimental Immunology 97(3): 517-521.
Zhai, G., M. Iskandar, K. Barilla and P. J. Romaniuk (2001). Characterization of RNA aptamer binding by the Wilms' tumor suppressor protein WT1. Biochemistry 40(7): 2032-2040.
Zhang, H., X. Chen, X. Teng, C. Snead and J. D. Catravas (1998). Molecular cloning and analysis of the rat inducible nitric oxide synthase gene promoter in aortic smooth muscle cells. Biochem Pharmacol 55(11): 1873-1880.
Zhang, H., C. Snead and J. D. Catravas (2001). Nitric oxide differentially regulates induction of type II nitric oxide synthase in rat vascular smooth muscle cells versus macrophages. Arteriosclerosis, thrombosis, and vascular biology 21(4): 529-535.
Zhang, H., X. Teng, C. Snead and J. D. Catravas (2000). Non-NF-kappaB elements are required for full induction of the rat type II nitric oxide synthase in vascular smooth muscle cells. British journal of pharmacology 130(2): 270-278.
Zhang, J. (2003). Search for Haplotype Interactions That Influence Susceptibility to Type 1 Diabetes, through Use of Unphased Genotype Data. American journal of human genetics, 73: 1385-1401.
Zhang, X., V. E. Laubach, E. W. Alley, K. A. Edwards, P. A. Sherman, S. W. Russell and W. J. Murphy (1996). Transcriptional basis for hyporesponsiveness of the human inducible nitric oxide synthase gene to lipopolysaccharide/interferon-gamma. Journal of leukocyte biology 59(4): 575-585.
Zollner, S. (2004). Evidence for Extensive Transmission Distortion in the Human Genome. American journal of human genetics, 74: 62-72.
Zumsteg, U., S. Frigerio and G. A. Hollander (2000). Nitric oxide production and Fas surface expression mediate two independent pathways of cytokine-induced murine beta-cell damage. Diabetes 49(1): 39-47.
Zumsteg, U., J. I. Reimers, F. Pociot, L. Mørch, S. Helqvist, M. Brendel, R. Alejandro, T. Mandrup-Poulsen, C. A. Dinarello and J. Nerup (1993). Differential interleukin-1 receptor antagonism on pancreatic beta and alpha cells. Studies in rodent and human islets and in normal rats. Diabetologia 36(8): 759-766.