Multicentric study on prevalence and risk factors for hypertension and diabetes in tribal communities in Western and Northern Maharashtra.

BACKGROUND AND OBJECTIVES: Although a number of studies have been conducted on the prevalence of hypertension (HTN) and diabetes on tribal populations in different parts of India, comparative tribe-specific information is very meager. The main objective of this study is to generate tribe-specific information on the noncommunicable disorders (NCDs) and associated risk factors in scheduled tribes (STs) in Coastal and Western Maharashtra. METHODS: The study was conducted on 1864 (females 960) adults (≥18 years) of both sexes in four dominant tribes in the region, namely, Bhils (748), Katkaris (560), Kokana (352), and Thakars (204), using the protocols approved by the Institutional Review Board. The study areas were geographically separated by large distances (250-500 km apart). Prevalence of overweight, diabetes, HTN, and hypercholesterolemia was measured using standard field-based techniques described in our earlier publication. RESULTS: All STs in this study are grossly underweight; the Katkaris are worst affected. The prevalence of obesity (body mass index ≥30 kg/m2), HTN (blood pressure ≥140 mmHg), diabetes (capillary blood glucose >126 mg/dl), and hypercholesterolemia (cholesterol ≥200 mg/dl) was 0.9%, 11.7%, 6.7%, and 0.6% respectively. There are no statistically significant inter-tribal differences in the prevalence of these parameters. Age and obesity appeared to be the most dominant risk factors for HTN. However, there is no clear-cut picture about the influence of risk factors on diabetes or hypercholesterolemia. INTERPRETATION AND CONCLUSION: The prevalence of NCDs is still very low in STs, probably due to near absence of the risk factors such as obesity, sedentary lifestyle, and hyperlipidemia. STs are highly endogamous, and the study areas are separated by large distances ruling out intermingling of tribes. Yet, there is no consistent inter-tribal prevalence pattern for NCDs. Although the sample size is small, the results support the view that environmental factors are likely to be more important than host genetics in the causation of NCDs.

Prevalence and risk factors of hypertension and diabetes in the Katkari tribe of coastal Maharashtra.

BACKGROUND AND OBJECTIVES: Urban and rural India are both going through health epidemiological transition and will soon face huge burden of noncommunicable diseases (NCDs). Information on the status of NCDs in tribals is limited. Although the prevalence of hypertension in scheduled tribes (STs) has been studied in several states by the National Nutrition Monitoring Bureau, tribe-specific data are very scanty. The objective of this study was to generate data on the status of hypertension and diabetes, the two objectively measurable NCDs in Katkaris, the dominant ST in the Raigad district of coastal Maharashtra. METHODS: The study was conducted in 410 adult Katkaris (women 219) of both sexes of ≥18 years of age in three adjoining tehsils of the district. Using the Institution Review Board approved protocol; information was obtained on sociodemographic parameters, educational level, dietary pattern, and substance abuse. Prevalence of overweight, hypertension, and diabetes was measured using standard field-based procedures and techniques. RESULTS: Katkaris, who are mostly landless manual laborers, subsist on a protein-poor, imbalanced diet. About half of women and one-third of men have body mass index (BMI) <18.5 kg/m2, an indication of undernutrition. On the other hand, about 2% of participants were obese (BMI ≥30 kg/m2). The overall prevalence of hypertension and diabetes was 16.8% and 7.3%, respectively. Hypercholesterolemia was recorded in about 3% of the participants. INTERPRETATION AND CONCLUSIONS: Prevalence of diabetes and hypertension in Katkaris is still lower than that of urban and rural populations, closer to the latter. This may be due to the absence of known risk factors such as obesity, sedentary lifestyle, and hyperlipidemia in this community. Fast acculturation of the STs suggests that NCDs will soon become a major health issue in them too. It is time to launch a multicentric national study to gather baseline information on the status of NCDs in STs.

Restriction fragment length polymorphism of the L-myc oncogene in non-Hodgkin's lymphoma patients from India.

We examined 89 non-Hodgkin's lymphoma (NHL) patients of Indian origin for EcoRI restriction fragment length polymorphism (RFLP) of the L-myc gene with a view to testing the hypothesis that the presence of the L-myc S-allele predisposes towards NHL. We found no significant difference either in the distribution of the LL, LS and SS genotypes or in the allelic frequencies between the patient group and the control group with the frequencies of L-myc alleles, L (10.0 kb) and S (6.6 kb), being 0.56 and 0.44, respectively, in the patient group and 0.54 and 0.46, respectively, in the control group. However, a higher proportion (70%) of the S-allele was observed in our control group of normal healthy volunteers. Thus, the presence of L-myc S-allele did not indicate increased susceptibility or predisposition to the malignancy.

Establishment of a human squamous cell carcinoma cell line of the upper aero-digestive tract.

A human squamous cell carcinoma (SCC) cell line has been established from the surgical specimen of an untreated, upper aero-digestive tract tumour, diagnosed as a squamous carcinoma, grade III, of the pyriform fossa. The tumour tissue was grown as a xenograft in an athymic nude mouse and was designated as NT-8. Histological examination of the surgical specimen and the nude mouse tumour showed that the two were identical. NT-8 was subsequently passed by subcutaneous injections into nude mice. After the 6th passage in nude mouse, the tumour was cultured in vitro where it grew as an epithelial cell line, with a typical cobblestone appearance. This cell line was designated as NT-8e. Both the primary tumour as well as xenograft and the cells in culture have retained several common morphological and biochemical characteristics. Immunological markers for epithelial cells including epithelial membrane antigen and cytokeratins were seen in all three, confirming the epithelial lineage. Characterization of the NT-8e cell line including growth parameters, anchorage-independent growth and tumorigenicity in nude mice, chromosome counts and DNA content by flow cytometry have been carried out.

Loss of p53 gene as a biomarker of high risk oral leukoplakias.

We have investigated loss of heterozygosity of p53 tumor suppressor gene in Indian oral cancer patients, individuals with premalignant leukoplakia lesions, and corresponding normal mucosa, to study the status of p53 alleles in oral cancer pathogenesis. Fifty oral cancers, and 42 oral leukoplakia lesions and corresponding clinically normal oral mucosa from 18 individuals, were analysed. Peripheral blood cells (PBCs) from all the individuals and 47 normal healthy volunteers were also included in the study. Polymerase chain reaction(PCR) of p53 Exon4, followed by restriction enzyme digestion with AccII due to the enzyme polymorphic site at Exon4 codon72, was used to detect homozygosity/heterozygosity of p53 alleles, and compared with the allelic pattern in the corresponding PBC. The PCR product subjected to AccII digestion detected 259 bp, 160/99 bp fragments indicating heterozygosity of p53 alleles in 69% of the 139 individuals. On comparison of the p53 allelic distribution in the lesions or tumour tissues, and corresponding PBC, LOH was observed in 20.5% oral tumors and 22% leukoplakias. However, there was no evidence of LOH in the clinically normal mucosa available from 16 individuals with leukoplakia. Our studies demonstrated LOH of p53 allele in early and advanced stages of oral cancers, as well as leukoplakias, perhaps indicating p53 LOH as one of the early events in oral carcinogenesis. Thus, p53 LOH may be useful as a biomarker in defining a certain population of high risk leukoplakias that may progress to oral cancer.

Ability of lymphokine-activated killer cells to lyse mycobacteria-infected cells.

Lymphokine-activated killer (LAK) cells were generated by interleukin-2 activation of peripheral blood lymphocytes obtained from lepromatous leprosy (LL) patients and healthy individuals. The ability of LAK cells to lyse targets (macrophages and T-24, a bladder carcinoma cell line) infected with mycobacteria (Mycobacterium leprae and mycobacterial strain ICRC) was assessed in a 51 chromium-release assay. It was observed that LAK cells generated from LL patients and healthy individuals could preferentially lyse M. leprae or ICRC-pulsed macrophages and T-24 cells, compared to non-pulsed targets. The ability of LAK cells to kill intracellular mycobacteria was demonstrated in colony forming assays. These results indicate a promising role for LAK cells in immunotherapy of leprosy.

Detection and cloning of potent transforming gene(s) from chewing tobacco-related human oral carcinomas.

High molecular weight DNA isolated from 14 primary tumour tissues of human oral carcinoma patients was analysed for transforming activity by NIH3T3 co-transfection assay using pSV2neo gene as a selectable marker, followed by nude mouse tumorigenicity assay. Ten of the patient tumour tissues demonstrated molecular lesions in myc, ras or/and EGF-R genes, whereas 4 patients did not show tumour associated aberrations in these oncogenes. The G418-resistant transfected cells from 12 of 14 individual patients demonstrated transforming potential by colony formation in soft agar and tumour induction in nude mice within 25-80 days. DNAs from the transfected cells, consequent nude mice tumours and corresponding cell lines, contained human Alu sequences. Southern blot hybridisation with ras, myc, EGF-R oncogenes demonstrated the presence of human H-ras oncogene in one of the 12 sets of nude mice tumours. In contrast, DNA from the other 11 sets of nude mice tumours indicated absence of c-myc, N-myc, L-myc, H-ras, K-ras, N-ras and EGF-R genes on Southern analysis. Further, DNAs from five first cycle tumorigenic transformants were subjected to a second cycle of transfection, and induced tumours in nude mice with a shorter latency period of 21-50 days. The secondary transformants contained discrete human Alu sequences; however, the DNA did not hybridise with myc/ras/EGF-R probes. A genomic library was constructed from a second cycle nude mice tumour, using EMBL-3 as the vector. Four human Alu sequence positive clones were isolated on screening 2 x 10(5) plaques, and one of the recombinant clones subjected to fine restriction mapping using 16 restriction enzymes. The lack of association of the nude mice tumour DNA with myc/ras/EGF-R showing aberrations in the primary human tumour, implies activation of an alternative potent transforming gene(s) in the chewing tobacco-related oral carcinomas in India.

Restriction fragment length polymorphisms of the human N-myc gene in normal healthy individuals and oral cancer patients in India.

Southern blot hybridization with N-myc oncogene probes coding for different regions of the N-myc gene demonstrated three polymorphic restriction sites in the Indian population. The SphI and PvuII polymorphic pattern due to the SphI polymorphic site in the second intron and the PvuII polymorphic site in the 3'-region of the human N-myc oncogene respectively, was similar to that reported in the Japanese population. The allelic frequency distribution for SphI polymorphism did not differ significantly for the S1 and S2 alleles representing presence (allele S1) or absence (allele S2) of a SphI site. However, the allelic frequency distribution was distorted in the case of PvuII polymorphism, as the frequency of P1 allele (0.7) indicating presence of PvuII site, was higher than the P2 allele (0.3) indicating absence of PvuII site, in the Indian population. An additional polymorphic HindIII site localised in the second intron of the N-myc gene was also observed in both the Indian oral cancer patients and the normal healthy individuals, indicating that this RFLP was not tumor associated and may perhaps represent N-myc alteration in the Indian population.

Immunoprecipitation of mycobacterial antigens with sera from patients with leprosy.

Pooled sera from leprosy patients across the clinical spectrum, tuberculosis patients and healthy individuals were tested for their reactivity with antigens of Mycobacterium leprae and a panel of cultivable mycobacteria by immunoprecipitation technique. Sera from lepromatous leprosy patients demonstrated exclusive reactivity with the 26-kDa protein of M. tuberculosis H37Ra, 28-kDa protein of M. kansasii, 45-kDa protein of M. smegmatis, and 158, 40 and 14 kDa proteins of M. phlei. Sera from patients with borderline tuberculoid leprosy, tuberculoid leprosy, tuberculosis and health individuals failed to identify these antigens. Our studies indicate that analysis and characterization of immunodominant antigenic epitopes present on proteins of cultivable mycobacteria, sharing cross-reactive epitopes with M. leprae may prove to be important in the serodiagnosis of multibacillary leprosy as well as for developing vaccines for immunotherapy of leprosy.

Immunoreactivity of mycobacterial strain ICRC and Mycobacterium leprae antigens with polyclonal and monoclonal antibodies.

ICRC, a cultivable mycobacterium, is undergoing clinical trials as an antileprosy vaccine in India. In the present study, we have investigated the crossreactivity between antigens of the mycobacterial strains of ICRC and Mycobacterium leprae using polyclonal and monoclonal antibodies in a radioimmunoprecipitation assay. It was observed that polyclonal anti-ICRC and anti-M. leprae antibodies showed predominant reactivity to a 21-kDa protein of the mycobacterial strain ICRC and the 21- and 14-kDa proteins of M. leprae. Crossreactivity between the antigens of the mycobacterial strains ICRC and M. leprae was established further by using M. leprae-specific monoclonal antibody WML06 (reacting with the 14-kDa protein of M. leprae), which identified the 21- and 14-kDa proteins of the mycobacterial strain ICRC. Thus, our studies demonstrate that the 14-kDa protein of M. leprae, which is known to harbor T- and B-cell epitopes, shares crossreactive antigenic determinants with the 21- and 14-kDa proteins of the mycobacterial strain ICRC. We believe that such proteins may provide important reagents for designing subunit vaccines and for determining skin-test reagents.

Enhancing factor, a Paneth cell specific protein from mouse small intestines: predicted amino acid sequence from RT-PCR amplified cDNA and its expression.

Enhancing factor (EF), a Paneth cell specific growth factor modulator, has been identified in our laboratory from mouse small intestines. In this paper we describe generation of an EF specific cDNA by RT-PCR and its sequence. The predicted amino acid sequence was found to be similar to, and hence confirms, the partial amino acid sequence obtained earlier by protein sequencing. In Northern blot analysis, a 1 kb transcript was observed in intestinal RNA alone. In situ hybridization indicated that the EF gene is expressed exclusively in the Paneth cells. The present study indicates that EF is an isoform of PLA2 type II, with a unique tissue distribution, found predominantly in the Paneth cells of the small intestines. Further, based on the properties of EF, we propose that isoforms of PLA2, belonging to type II, may be involved in regulation of cell proliferation via EGF binding.

H-ras-1 restriction fragment length polymorphism in normal individuals and oral cancer patients in India.

Restriction fragment length polymorphism (RFLP) of the human H-ras-1 gene has been indicated as a marker for detection of individuals at high risk of cancer. We have investigated the association of RFLP at the H-ras-1 locus and susceptibility to oral cancer by Southern hybridization analysis in 77 primary oral tumors and 99 healthy donors. The frequency distribution of the BamHI fragments of H-ras-1 revealed homozygous or heterozygous alleles in the two subpopulations. The heterozygous genotype occurred more frequently in the normal subjects (53%) as compared to the cancer patients (36%). Four common alleles-C1 to C4, were noted predominantly in both groups, with rare alleles detected at a lower frequency. The common allele with 7.6 kb BamHI fragment was significantly higher in normals (10%) than in the tumor population (4%) (P < 0.05). However, a similar distribution of rare alleles in both groups indicated that the presence of rare alleles is not indicative of predisposition to oral cancer.