Iodine and selenium: Dietary sources and nutritional status of the population of the Kurdistan Region in Northern Iraq.

AIM: The primary aim of this study was to determine the selenium (Se) and iodine (I) food concentrations and dietary intake of the population living in the Kurdish controlled region of northern Iraq. We also assessed the extent to which iodised salt contributes to dietary iodine intake. METHODOLOGY: Foods and samples of salt and drinking water were analysed, including 300 crops samples from 40 local farms. The results, supplemented by food composition data, were used to assess dietary Se and I intake for 410 volunteers using a semi-quantitative food questionnaire. To directly investigate the nutritional status of individuals, urine samples were also collected from participants. RESULTS: Selenium intake was mainly supplied by protein and cereal sources. Calculated median dietary intake of Se was 62.7 µg d-1 (mean = 66.3 µg d-1) with c. 72 % of participants meeting or exceeding dietary reference intake recommendations for age. Median dietary intake of I, excluding salt consumption, was 94.6 µg d-1 (mean 100.2 µg d-1), increasing to 607.2 µg d-1 when salt (of which >90 % was iodized) was included. Salt intake was estimated to be c.13.5 g d-1 (5400 mg Na d-1) which greatly exceeds WHO recommended intake (< 2000 mg d-1 of Na). Urine iodine concentrations indicated that 98 % of school aged children had excessive iodine intake (≥300 µg L-1) and 80-90 % of all study participants had above average or excessive iodine intake (≥200 µg L-1). CONCLUSIONS: Poultry and rice are the main sources of dietary Se to this population but around a third of children receive an inadequate Se intake. Fresh fruit and vegetables are the main sources of dietary I, but consumption of local foods cannot supply adequate I without iodised salt supplementation. Consumption of iodized salt well above recommended amounts is supplying this population with substantial iodine intake. Interventions to reduce salt intake would help to limit excessive iodine intake whilst also reducing cardio-vascular risks from Na consumption.

Influenza A and respiratory syncytial virus hospital burden in young children in East London.

Epidemiological studies have demonstrated high hospitalization rates attributable to influenza and RSV in children aged 6 months and those aged <12 months, respectively (43 and 92.5/10 000 person-months, respectively). In conclusion, these high paediatric RSV and influenza incidence rates can be used to inform UK policy on childhood influenza immunization and subsequent RSV immunization in the future.

Investigation of varicella-zoster virus variation by heteroduplex mobility assay.

Heteroduplex mobility assays of 37 regions were performed on ten UK isolates of varicella zoster virus, four from cases of zoster, and six from cases of chickenpox. The variation between isolates was found to be 0.061%, which is at least five times lower than any other member of the human herpesvirus family. Fifteen of the 37 regions tested had 29 single nucleotide polymorphisms, and over half the polymorphisms were located in four gene fragments. Of the 29 SNPs, eleven were non-synonymous and these were clustered in six genes. Isolates from a child and her mother to whom she had transmitted the virus, were identical at every locus tested. All other viruses could be distinguished by a combination of SNPs and length polymorphisms of the repeat regions R1, R2 and R5.

Simultaneous treatment of cytomegalovirus and varicella zoster infections in a renal transplant recipient with ganciclovir: use of viral load to monitor response to treatment.

Disseminated zoster occurring simultaneously with cytomegalovirus (CMV) disease in a renal transplant recipient is potentially life threatening. We describe the use of intravenous ganciclovir to treat both infections. The efficacy of treatment was assessed clinically and by the measurement of CMV viral load using the hybrid capture (Murex version 2) and varicella zoster (VZV) viral load using an in-house assay. Results from this case suggest that clinical resolution in severe viral infections such as described below may be related to early control of viraemia.

Development of a fluorogenic polymerase chain reaction assay (TaqMan) for the detection and quantitation of varicella zoster virus.

A TaqMan based polymerase chain reaction (PCR) assay was developed for the detection and quantitation of varicella zoster virus (VZV). This method enables simple, reproducible, sensitive and specific detection and quantification of VZV. The TaqMan assay was able to detect four copies of VZV and did not cross-react with other herpesviruses DNA. The assay has several advantages over conventional PCR. First, in the TaqMan assay there is no need for gel electrophoresis and contact with hazardous chemicals. Second, the method is rapid allowing the analysis of 92 samples within minutes after completion of PCR. Finally, the incorporation of a specific probe into the PCR reaction enhances the sensitivity and specificity of the method compared with conventional PCR. The TaqMan system could, therefore, be a useful tool for the epidemiological and diagnostic investigation of VZV.

Molecular epidemiology of varicella-zoster virus in East London, England, between 1971 and 1995.

The molecular epidemiology of varicella-zoster virus in London, England, between 1971 and 1995 was examined by using two informative polymorphic markers, variable repeat region R5 and a BglI restriction site in gene 54. Viruses from 105 cases of chickenpox and 144 of zoster were typed. Two alleles of R5, A and B, were found at prevalences of 89 and 6%, respectively. No difference in allele frequency between the zoster and chickenpox cases was found, and no change in the frequencies of these alleles was observed to occur over time. By contrast, a BglI restriction site (BglI+) was found with increasing frequency over time among cases of varicella (P < 0.005) and, to a lesser extent, cases of zoster. The BglI+ polymorphism was strongly associated (P < 0.0005) with zoster in subjects who had immigrated to the United Kingdom from countries with low adult immunity to varicella (LAIV). Sixty-three percent of the subjects with zoster who had emigrated from countries with LAIV carried the BglI+ virus, in contrast to 10% of adults who had grown up in countries with high adult immunity to varicella. The significance of these data, in view of the changing epidemiology of chickenpox, is discussed.

Analysis of United Kingdom wild-type strains of varicella-zoster virus: differentiation from the Oka vaccine strain.

In Japan and the United States, where vaccination against varicella-zoster virus (VZV) infection with the live attenuated Oka strain of varicella is routine, cases of chickenpox or shingles occurring in vaccinees can be caused by either wild-type or vaccine virus. Differentiating such cases is important epidemiologically and can be achieved only using molecular typing methods. In the United Kingdom, the Oka vaccine is being considered for use in groups at risk of severe primary varicella, such as seronegative immunocompromised patients and women who may be considering pregnancy. In addition, seronegative health workers who may be occupationally exposed to VZV infection might also be offered vaccination. We analysed 249 U.K. wild-type VZV strains, 105 from cases of chickenpox and 144 from shingles cases, to determine whether they could be distinguished from Oka by the genotyping systems used in Japan and the United States. Four polymorphic loci were examined, a Pst 1 restriction site in gene 38, a Bgl 1 restriction site in gene 54, the R5 repeat region, and the R2 repeat region. The results suggest that U.K. strains of VZV are more similar to U.S. strains than to Japanese strains. All the U.K. wild-type viruses were positive for the Pst 1-1 restriction site, unlike Oka, which is negative. However, one of thirty strains was indistinguishable from Oka at all other loci.

Analysis of islet regenerating (reg) gene polymorphisms in fibrocalculous pancreatic diabetes.

Fibrocalculous pancreatic diabetes (FCPD) is a form of diabetes associated with tropical chronic calcific pancreatitis, seen mostly in developing countries. FCPD is likely to be a multifactorial disease with both environmental and genetic components. The reg 1A gene encodes a protein associated with regeneration of pancreatic islets and has a sequence identical to that of pancreatic stone protein. Since FCPD is associated with both diabetes and pancreatitis, we tested the hypothesis that FCPD may be the result of mutations in the coding regions of the reg 1A gene. Restriction length polymorphisms (RFLPs) and possible sequence variants of the reg 1A gene were studied by RFLP analysis, looking for single-stranded conformational polymorphisms (SSCPs) and direct nucleotide sequencing. In 20 patients with FCPD and 20 control subjects, no RFLPs were detected using 10 restriction enzymes. In 50 patients with FCPD and 50 control subjects, no SSCP variants were detected. Finally, direct nucleotide sequencing of the reg 1A gene from 30 patients with FCPD did not show any differences from the published human reg 1A gene sequence. In conclusion, it seems unlikely that mutations in the coding region of the reg 1A gene are a common cause of FCPD.

Typing of varicella zoster virus by amplification of DNA polymorphisms.

The polymerase chain reaction was used to amplify five variable regions of varicella zoster virus DNA from 20 samples of vesicle fluid. Two of the regions, R1 and R5, were found to be polymorphic, with the former having three alleles (A, B and C) and the latter, two (A and B). The R1 and R5 polymorphisms were stable up to passage five in tissue culture. The sensitivity of the PCR (down to six copies) enabled detection of virus from vesicle fluid dried on glass slides and overall the method was five times more sensitive than conventional tissue culture. The method described is simple, sensitive and informative and provides a means by which questions about the epidemiology and clinical biology of VZV infection may begin to be addressed.

An association in non-insulin-dependent diabetes mellitus subjects between susceptibility to retinopathy and tumor necrosis factor polymorphism.

In IDDM an association between diabetic retinopathy and polymorphic markers of MHC has been described. However, these associations are complicated by a primary association between the MHC and IDDM. Because the pathogenesis of retinopathy is likely to be the same in IDDM and NIDDM, NIDDM subjects with retinopathy would be the ideal population to study for an association with MHC markers. The following South Indian subjects were therefore studied: unselected NIDDM (n = 76), unselected IDDM (n = 99), non-diabetic controls (n = 96), NIDDM subjects with maculopathy (MAC), n = 55, NIDDM subjects with proliferative retinopathy (PR), n = 53, and without retinopathy (LTD), n = 46. DNA was amplified and studied using a microsatellite polymorphism located 3.5 kb upstream of TNF-beta within the MHC class III region on the short arm of chromosome 6. No differences in allelic distribution were observed between the random NIDDM subjects and controls (p = 0.17). Differences in allelic distribution were found between unselected IDDM and controls (P = 0.016) and between the NIDDM subjects with maculopathy and/or proliferative retinopathy and no retinopathy (P = 0.006). This association could be accounted for by those patients with proliferative retinopathy (MAC vs LTD, p = 0.23; MAC vs PR, p = 0.07; and PR vs LTD, p = 0.002).

Insulin receptor substrate-1 gene mutations in NIDDM; implications for the study of polygenic disease.

Variations in the coding regions of the insulin receptor substrate-1 (IRS-1) gene have recently been suggested to contribute to the susceptibility of non-insulin-dependent diabetes mellitus (NIDDM). The purpose of this study was to examine the role of the IRS-1 missense mutations at codons 972 (glycine to arginine) and 513 (alanine to proline) in two diverse populations from South India and Finland at high risk for NIDDM. DNA was amplified and digested with restriction enzymes BstN1 to detect the codon 972 mutation and Dra III to detect the codon 513 mutation. The codon 513 mutation was not found in the study subjects. The codon 972 mutation was present in 10.3% of 126 middle-aged NIDDM subjects and 5.3% of 95 matched control subjects in the South Indians (p = 0.17). In elderly Finnish subjects the frequency of the mutation was 7.5% in 40 NIDDM subjects and 7% in 42 matched control subjects. The frequency of codon 972 mutation in the South Indian NIDDM subjects was very similar to the two previously published studies in Danish and French subjects although each study individually fails to reach conventional levels of significance. The data from all four ethnic groups were analysed together after ascertaining that significant heterogeneity did not exist between the studies. Overall, the frequency of the codon 972 mutation is found in 10.7% NIDDM subjects and 5.8% control subjects (p = 0.02). These studies suggest that the codon 972 mutation of the IRS-1 gene might act as a susceptibility gene predisposing to NIDDM in certain ethnic groups.

Apolipoprotein D gene polymorphism: a new genetic marker for type 2 diabetic subjects in Nauru and south India.

Type 2 diabetes is characterized by abnormalities in both glucose and lipoprotein metabolism and genes involved in lipid metabolism are legitimate candidates for involvement in Type 2 diabetes. We have previously reported an association in Nauruans between a Taq 1 polymorphism of the apolipoprotein D gene (apo D) and Type 2 diabetes. In this study these findings were investigated further in the Nauruan population as well as two other ethnic groups. In South Indian subjects, there was a significant difference in genotype distribution of apo D genotypes between diabetic subjects (n = 110) and controls (n = 88; p = 0.004) which was similar to that previously found in the Nauruan subjects. No such association was seen in elderly Finnish subjects (diabetic n = 69; impaired glucose tolerance n = 26 and normal glucose tolerance n = 31). Linkage between the apo D polymorphism and diabetes in 12 Nauruan families was only excluded under a highly penetrant dominant model and was unlikely under other single gene models. Since the beta cell glucose transporter gene (Glut 2) is found in a similar chromosomal location to apo D, South Indian subjects (diabetic n = 95 and controls n = 56) were typed at this locus. No association between diabetes and the Glut 2 Taq I polymorphism was found in the South Indian subjects. Furthermore, there was no evidence of linkage disequilibrium between the apo D and Glut 2 genes. In conclusion, apo D might act as a modifying gene for Type 2 diabetes in some ethnic groups.

Positive association in the absence of linkage suggests a minor role for the glucokinase gene in the pathogenesis of type 2 (non-insulin-dependent) diabetes mellitus amongst south Indians.

Mutations of the glucokinase gene have been implicated in the development of glucose intolerance in pedigrees with maturity-onset diabetes of the young. However, the contribution of the glucokinase gene to the aetiology of common Type 2 (non-insulin-dependent) diabetes mellitus is uncertain. We have studied the role of the glucokinase gene in the pathogenesis of Type 2 diabetes in South Indians, using both population-association and linkage methodology. A pair of CA-repeat sequences (GCK(3') and GCK(5')) straddling the glucokinase gene were employed as markers, each subject being typed using the polymerase chain reaction and polyacrylamide gel electrophoresis. Comparisons of allele frequencies at these markers were made between 168 Type 2 diabetic subjects and 70 racially-matched control subjects. No differences in allele frequencies were apparent at the GCK(5') marker; however, there were significant differences in allele frequencies at the GCK(3') marker between the Type 2 diabetic subjects and control subjects (chi 2 = 11.6, df = 3, p = 0.009) with an increase of the z allele (78.0% vs 66.4%) and a decrease of the z + 2 allele (13.7% vs 25.0%) amongst the diabetic subjects. Linkage between glucose intolerance and the glucokinase gene was studied in 53 nuclear pedigrees under a variety of genetic models. Linkage was excluded (lod score < -2) at a recombination fraction of zero under five of the ten models used and highly unlikely (-2 < lod score < -1) under the others. The combination of positive association and negative linkage suggests that glucokinase acts as a minor gene influencing the development of Type 2 diabetes within this population.

A genetic study of retinopathy in south Indian type 2 (non-insulin-dependent) diabetic patients.

Genetic marker studies in diabetic retinopathy are controversial and frequently complicated by possible independent associations of Type 1 (insulin-dependent) diabetes mellitus with the markers so far analysed. We have looked for associations of candidate genes with retinopathy in South Indian Type 2 (non-insulin-dependent) diabetic patients; patients were subdivided into those with exudative maculopathy (n = 53), proliferative retinopathy (n = 40) and patients free from diabetic retinopathy with a minimum disease duration of 15 years (n = 45). DNA was extracted from blood samples and studied by Southern blot hybridisation techniques and the following probe enzyme combinations: HLA-DQB1; Taq 1, HLA-DQA1; Taq 1, HLA-DRA; Bgl II, insulin gene hypervariable region; Pvu II and the switch region of the immunoglobulin IgM heavy chain gene (S mu); Sac I. Differences in genotype distributions between the study groups were only detected with the S mu probe which detects polymorphism of both S mu and S alpha 1 (the switch region of IgA). Two alleles of S alpha 1 were detected sized 7.4 kilobase and 6.9 kilobase. The frequency of 6.9 kilobase homozygotes was lower in proliferative retinopathy (19%) compared to patients free from diabetic retinopathy (54%, p = 0.005) and exudative maculopathy (46%, p = 0.03). This data suggests that there is a genetic predisposition to proliferative retinopathy in Type 2 (non-insulin-dependent) diabetes of South Indian origin and that this is determined by polymorphism of the heavy chain immunoglobulin genes located on chromosome 14.