Prenatal Screening and Diagnosis of ß-Thalassemia in India: Is ARMS-PCR Enough?

Accurate and timely prenatal diagnosis of thalassemia is cornerstone to the success of thalassemia control; currently parents are screened for ß-thalassemia mutations by ARMS-PCR and subsequently chorionic villus sampling is done. We did an audit to ascertain whether the present design is adequate and determined the role of sequencing for pre-natal diagnosis of beta-thalassemia. This was a retrospective analysis of prenatal testing data collected over 10 years, (2010-2019). ARMS-PCR was done to identify the beta-globin mutation followed by CVS wherever indicated. Data was classified into 3 groups:-5 most commonly occurring mutations (group 1), less common mutations (group 2) and mutations not detected (group 3). Total number of cases studied were 2128. Mean age of the cohort was 29.30 years (range 18-48 years). Approximately 90% individuals had one of the 5 common mutations in decreasing order of frequency: IVS 1-5 G>C (1297/2128); Codon 26G>A/HbE (451/2128); codon 30G>C (69/2128); codon 15G>A (61/2128); FS 41-42-CTTT (48/2128). Undetected mutations amounted to 7.3% (156/2128). Mean haemoglobin was highest in the group 2 (12.46 g/dl) followed by the group 1 (11.20 g/dl) and least in group 3 (10.99 g/dl). MCV, MCH and MCHC showed similar trends. ANOVA on all these parameters, except RDW, within groups and for individual mutations, were statistically significant (p < 0.001). The hemogram-HPLC-ARMS-PCR-CVS approach is a cost-effective and established method but tends to miss out a considerable number of thalassemia mutations (~7%), emphasizing the role of sequencing in difficult cases. This needs to be addressed while formulating guidelines for thalassemia screening in future.

Investigation on the Trace Elemental Profile of Sewage Workers in Kolkata, an Indian Megacity.

BACKGROUND: Environmental pollution has become a global health risk. Exposure to pollutants at the work place, i.e. occupational exposure, is one of the areas that need immediate attention. The civic drainage workers are exposed to pollutants present in the wastewater they handle and most of them are toxic heavy metals. Exposure to such pollutants may be a health hazard, since it can lead to the imbalance in nutrient elements status. DESIGN AND METHODS: In the present study, profiling of trace elements in the blood of drainage worker population from an Indian megacity, Kolkata, was carried out by energy dispersive x-ray fluorescence (EDXRF) and compared with the control group population of the same area. RESULTS: The elements detected by EDXRF spectrometry include P, S, Cl, K, Fe, Cu, Zn, Se, Br, and Rb. By using ANOVA with 5% significance level, we observed significant alterations in the trace elements status, iron over loading, selenium deficiency, and in Cu-Zn ratio. Gender specific variations within the same population were also observed. CONCLUSIONS: The results indicate that the drainage workers have altered elemental profile in comparison to that of control population. Significance for public healthEnvironmental pollution is a global health risk and awareness among sewage workers is growing very slowly in many developing countries. Due to this fact, workers are often exposed to different pollutants which are responsible for several health complications. Imbalances in the presence of trace elements in blood are a symptom of different health status and could also indicate new health perspectives for the future. In the present scenario, this paper is essential since this kind of analysis has not been done yet, especially regarding the health status of sewage workers. We hope this initial study will be a starting point for future investigations.

Polymorphism of ACE gene as the genetic predisposition of coronary artery disease in Eastern India.

AIM: A case control study was designed to assess whether the prevalence of ACE gene polymorphism has any role in the development of CAD. METHODS: The study included unrelated 217 cases with CAD and 255 healthy controls. PCR was done using primers followed by agarose gel electrophoresis for study of different ACE gene polymorphisms. Multiple logistic regression analysis was carried out to find association between studied genotypes and lifestyle as well as biochemical risk factors. RESULTS: Both DD [OR: 2.16; 95%CI: (60.60-67.40)] and ID [OR: 1.48; 95%CI: (93.28-97.72)] genotypes of the ACE gene showed significant associations in the development of CAD. Coexistence of diabetes and hypertension found to be risk modifier of the disease. Tobacco intake in various forms elevates the risk of the disease among the cases with risk genotypes. CONCLUSION: ID and DD genotypes of ACE gene came out to be predisposing factors for the CAD cases in our study population.

Incidence of anaemia among the female tea garden workers in a tea plantation in upper Assam, India.

Nutritional anaemia is the most common type and cause of anaemia. Tea garden workers constitute approximately 1/5th of population in the state of Assam. Reports indicate higher prevalence of undernutrition in this population. The present study was designed to focus on the incidence of anaemia among the female tea garden workers and to examine the effect of iron with vitamin B12 and folic acid supplementation in them. Two hundred and forty-four female tea garden workers who could be persuaded to participate in the study were sampled for haemoglobin typing by agarose gel electrophoresis (pH 8.9). Their haematological parameters were measured on automated cell counter to read Hb, RBC, MCV, MCH and MCHC. BMI was calculated from weight/height in metre2. The study revealed 182 anaemic (<11.0 g/dl) subjects which included 32 cases of hereditary anaemia and 150 cases of nutritional anaemia. Iron supplementation for duration of 3 months was given to 150 workers out of total 182 subjects who had haemoglobin <11.0 g/dl. Thirteen subjects with haemoglobin >11.0 g/dl were studied as controls. Level of haemoglobin was rechecked after 3 months by Sahli's method. Based on the size of red blood cells it was observed that the workers suffered from all three forms of anaemia viz, microcytic, normocytic and macrocytic. There was a significant rise in haemoglobin concentration when the anaemic individuals took iron with vitamin B12 and folic acid supplement for three consecutive months supported by two doses of anthelminthic drug. The present study showed that nutritional anaemia is a major problem in the female tea garden workers and arrangement of supplementation therapy is utmost necessary to improve their general condition and work efficiency.

Polymorphisms of CYP1A1, GSTM1 and GSTT1 Loci as the Genetic Predispositions of Oral Cancers and Other Oral Pathologies: Tobacco and Alcohol as Risk Modifiers.

Polycyclic aromatic hydrocarbons of tobacco require activation by phase I enzymes, such as cytochrome-P4501A1 (CYP1A1) to become an ultimate carcinogen, which are subjected to detoxification by phase II enzymes, especially glutathione S-transferases (GSTs). A study was designed to find whether genetic predisposition are risk modifiers of oral pathologies. The study included 102 cases with Oral Cancers (OCs), 68 cases with nonmalignant pathologies, 100 cases as control group. GSTM1 null genotype was associated with increased risk of OCs but not with benign pathologies. Deleted GSTT1 was associated with all pathologies. Both m1m2 and m2m2 polymorphisms of CYP1A1 were associated with oral pathologies.

Cytogenetic monitoring in human oral cancers and other oral pathology: the micronucleus test in exfoliated buccal cells.

Oral cancer is a lifestyle-related cancer, with tobacco as a primary factor. Progression of oral cancer develops over several years from the stage of leukoplakia, erythroplakia, etc. A micronucleus test was applied to oral mucosal cells, considering them as the target site for carcinogens and cytogenetic damage. The test has been established as a reliable biomarker for differential prevalence of MN indices among oral cancers, pre-cancers, non-malignant oral pathologies, and healthy controls for the first time. Buccal scrapings were collected from 63 patients with cancer and pre-cancerous lesions, 42 with non-malignant oral problems, and 100 healthy controls. The analysis revealed that MN frequencies in cancer and pre-cancerous cases were 4-fold elevated (p < 0.001) and 3.87-fold (p < 0.002) elevated for other non-malignant pathologies. Significant associations between use of tobacco in various forms and development of oral pathologies are also established. The relative cancer risk for smoking healthy controls with a definite MN frequency was also found to be significant. The results indicate the validity of the MN test as a cytogenetic marker for the development of several oral pathologies.

Prevalence of CYP1A1 and GST polymorphisms in the population of northeastern India and susceptibility of oral cancer.

Individual cancer susceptibility is the result of several host factors, including differences in lifestyle habits and genetic susceptibility. There is a correlation between CYP1A1 polymorphism (MspI) and oral cancer susceptibility. Individuals carrying the deletions of GSTM1 and GSTT1 are at high risk of developing oral cancers. In the present study on healthy tribal and nontribal individuals of Assam, we found that the genetic variation of GSST polymorphisms is evident (p = 0.20) with differential dose of toxic exposure. Prevalence of different polymorphic alleles of CYP1A1 also proves the same result. A mini-case-control study with very small sample size showed no marked increase in the risk of developing oral cancer as the frequencies of the studied GST genotypes did not show any statistical significance. But GSTT1-null genotypes were found to have higher risk of developing leukoplakia (OR 1.94, 95% CI 2.61-18.54). CYP1A1 genotype m2 allele was also not found to be associated with the risk of developing leukoplakias in the population.

Alpha-thalassemia among tribal populations of Eastern India.

Five hundred and thirteen unrelated subjects belonging to various tribes of West Bengal, Arunachal Pradesh and Assam in Eastern India, were screened for the presence of alpha-thalassemia (thal) gene deletion(s) as a possible cause of unexplained anemia (Hb < 11 g/dL and/or MCH <28 pg, MCV < 78 fL). As reported earlier, beta-globin gene mutant alleles were found with a frequency of up to 20% in some tribes. In the present study, alpha-globin gene deletion alleles were found in 18% of subjects from West Bengal, 3.9% from Arunachal Pradesh and 3.84% from Assam tribesmen. Coexistence of alpha- and beta-globin gene abnormalities was observed in up to 18% of some tribal groups. The high inbreeding rate and lack of appropriate medical care make these populations particularly vulnerable.