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D Owerbach

Publications and source records attributed to D Owerbach.

At least 37 records · Page 2Linked to original sources
PubMed1988

Tissue-specific expression of transfected human insulin genes in pluripotent clonal rat insulinoma lines induced during passage in vivo.

The pluripotent rat islet tumor cell line MSL-G2 expresses primarily glucagon or cholecystokinin and not insulin in vitro but changes phenotype completely after prolonged in vivo cultivation to yield small-sized hypoglycemic tumors composed almost entirely of insulin-producing beta cells. When a genomic DNA fragment containing the coding and upstream regulatory regions of the human insulin gene was stably transfected into MSL-G2 cells no measurable amounts of insulin or insulin mRNA were detected in vitro. However, successive transplantation of two transfected clones resulted in hypoglycemic tumors that efficiently coexpressed human and rat insulin as determined by human C-peptide-specific immunoreagents. These results demonstrate that cis-acting tissue-specific insulin gene enhancer elements are conserved between rat and human insulin genes. We propose that the in vivo differentiation of MSL-G2 cells and transfected subclones into insulin-producing cells reflects processes of natural beta-cell ontogeny leading to insulin gene expression.

Adenoma, Islet Cell↗
PubMed1987

Molecular biology of the HLA system in insulin-dependent diabetes mellitus.

Genetic studies indicate that the IDDM susceptibility genes in the HLA region are closely linked to the DR3 and DR4 specificities; however, these specificities do not define the actual susceptibility genes. Molecular studies confirm this hypothesis by demonstrating restriction fragment length polymorphism between DNA's of identical DR specificities and thereby separating the DR haplotypes into those strongly or weakly associated with IDDM. Further studies at the nucleotide sequence level demonstrate further heterogeneity, with DR4 being associated with at least three different DQ beta genes and five different genes of the DR beta-1 locus. However, the majority of these subtypes are now recognized either serologically or by T-cell responses in mixed lymphocyte cultures. Furthermore, the sequences associated with IDDM are those most commonly found in DR4 individuals, ie, Dw4 and DQw3.2. Clearly, these and other class II genes must be studied for additional DNA polymorphism and their relevance for IDDM. For example, the DX alpha, 2.1-kb Taql polymorphism shows a stronger correlation with IDDM than DR3. However, it is not even known if the DX alpha genes are expressed. In addition, little is known of the DQ beta and DR beta genes associated with different DR3-associated haplotypes. Furthermore, an IDDM susceptibility gene may contain important differences in flanking or intron sequences controlling expression of these genes. The methods of recombinant DNA technology are enabling these unanswered questions to be addressed.

Base Sequence↗
PubMed1987

Gestational diabetes mellitus is associated with HLA-DQ beta-chain DNA endonuclease fragments.

DNA from Caucasian normal healthy control subjects, non-gravid patients with insulin-dependent diabetes mellitus (IDDM) and gravida with gestational diabetes mellitus (GDM) were analyzed with DNA probes for HLA markers associated with HLA-DR and HLA-DQ to compare the hybridization patterns of their DNA after digestion with restriction endonucleases. We report HLA-DQ beta restriction endonuclease fragments to be presented with increased frequency in Caucasian gravida with GDM as well as in subjects with IDDM. These findings provide further evidence for genetic heterogeneity in GDM and are compatible with the presence of slowly evolving IDDM in some women with "carbohydrate intolerance of variable severity with onset or first recognition during pregnancy".

DNA↗
PubMed1986

Characterization of three HLA-DR beta genes isolated from an HLA-DR 3/4 insulin-dependent diabetic patient.

Three HLA-DR beta genes were isolated from a Swedish HLA-DR3/4 insulin-dependent diabetes mellitus (IDDM) patient and characterized by restriction endonuclease mapping and nucleotide sequence analysis. Two out of the three DNA sequences differed from those of published DR beta-chain sequences. A DR beta-gene probe prepared from exon 4 and flanking sequences was used in a Southern blot analysis of blood donors' DNA and DNA from HLA-DR3/4 IDDM patients and HLA-DR-matched healthy control subjects. This probe differentiated HLA-DR3/4 IDDM patients and HLA-DR-matched controls in the Scandinavian population but not in the North American Caucasoid population.

Amino Acid Sequence↗
PubMed1986

Diabetes mellitus, atherosclerosis, and the 5' flanking polymorphism of the human insulin gene.

On the 5' side of the human insulin gene is a highly polymorphic locus containing 2 major size classes of DNA restriction fragments which segregate in families as stable genetic elements. Fragments with an average size of about 600 base-pairs (bp) (the 'L-allele') seem to be a weak genetic marker for type 1 (insulin-dependent) diabetes mellitus, whereas fragments of an average size of about 2500 bp (the 'U-allele') have hitherto been associated with type 2 (non-insulin-dependent) diabetes mellitus and diabetic hypertriglyceridaemia. Recent evidence does not confirm the association between the U-allele and type 2 diabetes. Our own studies suggest that the U-allele is a fairly strong marker for the development of atherosclerosis with a relative risk for U-carriers of 3.36. The U-allele has not been associated with conventional cardiovascular risk factors such as body weight, blood pressure, or levels of blood glucose, triglycerides or lipoproteins. The putative functions of the polymorphic region in the aetiology of type 1 diabetes and atherosclerosis, and the relation of this region to other genetic markers for these disorders are not known.

Alleles↗
PubMed1985

Insulin-gene flanking sequences, diabetes mellitus and atherosclerosis: a review.

A highly polymorphic locus flanking the human insulin gene contains two major size classes of DNA restriction fragments, which segregate in families as stable genetic elements. The L-allele, i.e. fragments with an average size of about 600 base-pairs seems to be a weak genetic marker for Type 1 (insulin-dependent) diabetes mellitus, whereas the U-allele, i.e. fragments of an average size of about 2500 base-pairs hitherto has been associated with Type 2 (non-insulin-dependent) diabetes mellitus and diabetic hypertriglyceridaemia. The most recent reports on this subject do not confirm an association between the U-allele and Type 2 diabetes. Our own studies indicate that the U-allele is a fairly strong marker for the development of atherosclerosis (relative risk for U-carriers 3.36). The putative functions of the polymorphic region in atherogenesis and the relation of this region to other genetic markers for atherosclerosis are not known.

Alleles↗
PubMed1985

Identification of an HLA-DQ beta-chain related genomic sequence associated with insulin-dependent diabetes.

Restriction fragment length polymorphism detected by a cDNA probe for an HLA-DQ beta-chain gene has revealed a HLA-DR4 linked BamH1 3.7 kb fragment which is rarely found among insulin-dependent diabetic patients. The present analysis demonstrates that the BamH1 3.7 kb fragment present on a HLA-DR4 positive chromosome in a healthy individual contains coding sequences for an HLA-DQ beta-chain gene and that the absence of this fragment among HLA-DR4 positive insulin-dependent patients is due to the loss of a BamH1 restriction within an intervening sequence.

Base Sequence↗
PubMed1984

DNA sequences flanking the insulin gene on chromosome 11 confer risk of atherosclerosis.

The allelic frequency of DNA restriction fragments of a large size class (U alleles) in the polymorphic region flanking the 5'-end of the human insulin gene on chromosome 11 was 2 X 5 times higher in a group of patients with extensive atherosclerosis than in subjects in whom atherosclerosis could not be demonstrated by coronary arteriography and careful clinical examination. The U alleles apparently do not confer risk of atherosclerosis through conventional risk factors such as body weight or blood pressure or levels of blood glucose, triglycerides, cholesterol, or lipoproteins.

Adult↗
PubMed1984

Analysis of a 1963-bp polymorphic region flanking the human insulin gene.

The nucleotide sequence of a long polymorphic region located 365 bp upstream from the human insulin gene is reported. The region is composed of 139 repeating sequences whose consensus structure is related to ACAGGGGTGTGGGG. Expansion in the number of repeating sequences appears to have taken place through duplication and triplication of 112-141-bp regions. However, ancestral polymorphic regions containing additions or deletions of 50 bp or more were not detected in two previous generations.

Base Sequence↗
PubMed1984

Susceptibility to insulin-dependent diabetes defined by restriction enzyme polymorphism of HLA-D region genomic DNA.

DNA fragments complementary to cloned sequences encoding HLA-D region class II antigen alpha- and beta-chains were determined by genomic blotting with DNA from HLA-typed members of 22 complete families, 12 of which had a proband with insulin-dependent diabetes mellitus (IDDM). Analysis of genotypes showed that the DNA sequences were linked to HLA-DR and permitted confirmation of recombinations in two families. Digestion with the restriction enzymes BamHI, EcoRI, and PstI and hybridization with an HLA-D region beta-chain cDNA probe confirmed a BamHI 3.7 kilobase (kb) fragment present at low frequency among diabetic individuals and a BamHI 3.2 kb fragment that was also decreased among the diabetic subjects compared with siblings (P less than 0.05) as well as nonrelated control siblings (P less than 0.02) and their parents (P less than 0.01). BamHI 12.0 kb (P less than 0.05) and 5.8 kb (P less than 0.02, P less than 0.02), EcoRI 20 kb (P less than 0.05, P less than 0.02), and PstI 6.0 kb (P less than 0.05) fragments were more frequent in diabetic individuals compared with nonrelated control siblings or their parents, respectively. Analysis of individual haplotypes revealed that HLA-DR4-containing chromosomes were heterogeneous among controls but that the diabetic individuals showed a similar pattern of restriction fragment length polymorphism. Genomic blotting of blood lymphocyte DNA with a cDNA clone encoding the chain of HLA-D region class II antigens permits detection of fragments that are strongly associated with IDDM.

Adolescent↗
PubMed1983

HLA-D region beta-chain DNA endonuclease fragments differ between HLA-DR identical healthy and insulin-dependent diabetic individuals.

The human HLA-D histocompatibility region encodes class II antigens each of which consists of two polypeptide chains (alpha and beta) inserted in the plasma membrane. These molecules are implicated in the regulation of the immune response but several human diseases are also found to be associated with certain HLA-DR antigens. The occurrence of insulin-dependent (type I) diabetes (IDDM) is strongly associated with HLA-DR3 and/or 4 (ref. 5). The class II antigens, however, show a marked genetic polymorphism associated with the beta-chains which seem, from hybridization studies, to be encoded by several genes. We have therefore used the beta-chain cDNA probe, pDR-beta-1 (refs 8, 10) to test whether there are differences in hybridization pattern between DNA from healthy individuals and diabetic patients, after digestion with restriction endonucleases. Among the HLA-DR 4 and 3/4 individuals, the IDDM patients showed an increased frequency of a PstI 18 kilobase (kb) fragment. A BamHI 3.7 kb fragment, frequent among controls (30-40%), was rarely detected in the IDDM patients (0-2%). These differences may be related to susceptibility to develop the disease.

DNA Restriction Enzymes↗
PubMed1983

Human class II major histocompatibility antigen beta-chains are derived from at least three loci.

Class II antigens of the major histocompatibility complex (MHC) consist of two glycosylated, membrane-integrated polypeptide chains. These cell surface-expressed molecules are involved in several immunobiological events involving cell-cell interactions, most of which seem to require that genetically identical class II antigens, or other molecules controlled by the same region of the MHC, are expressed on the interacting cells. The extensive genetic polymorphism of the class II antigens has rendered analyses in the human system of the number of non-allelic species of class II antigens difficult, although several laboratories have reported the existence of at least two types of human class II antigens. Here we present the results of experiments using restriction enzyme digestions and separation of DNA from individuals homozygous for the MHC followed by hybridization to human class II antigen alpha- and beta-chain cDNA probes. While the alpha-chain probe gave only a single hybridization band, the various beta-chain probes revealed a more complex pattern that is consistent with the existence of at least three separate beta-chain genes or pseudogenes in the human MHC.

Bacterial Proteins↗
PubMed1983

Genetics of diabetes in Nauru: effects of foreign admixture, HLA antigens and the insulin-gene-linked polymorphism.

Genetic factors play a major role in predisposition to diabetes in the Micronesian population of Nauru. In people aged 60 years and older, 83% of full-blooded Nauruans were diabetic compared with 17% of those with ancestral foreign admixture, as detected by HLA typing. HLA distributions also showed a small increased risk for early onset of diabetes (less than 46 years) associated with HLA-Bw22 (Bw56). Variation in the restriction fragment length of DNA near the insulin gene was found, but was not associated with diabetes. The distribution in fragment lengths, previously reported in Caucasoids, was observed in healthy Polynesians, Melanesians and Micronesians.

Adult↗