Ahmad Fazilat ¹, Swarupa R. Didla ², Sashikanth Chitti ², Mrinmayi Joshi ²

Affiliations
1 Department of Biotechnology, Andhra Universitry, Visakhapatnam, IN
2 Translational Research Institute of Molecular Sciences Pvt Ltd, 9-36-1, Visakhapatnam-17, IN

Abstract

Background/Objectives: To understand the genotoxicity and cytotoxicity potentials in tobacco-chewing habitues from coastal Andhra Population. Place and Duration of Study: Translational Research Institute of Molecular Sciences, between October 2013 to October 2014. Methods/Statistical Analysis: Present study was carried out using 35 individuals (28 men, 7 women; age range 25-56 years) who are having a habit of chewing tobacco from 5-20 years were tested along with 35 controls with same age group. Samples were cultured and stained with Hoechst 3325. Scored chromosomal abnormalities, SCE’s, CPK and MI using statistical evaluations. Results/Findings: In this study we have scored the numerical and structural abnormalities, Mitotic Index (MI), Cell Proliferation Kinetics (CPK) and Sister Chromatid Exchanges (SCE’s) of the presented samples have been calculated. Although significant numerical aberration observed, structural abnormalities were notably seen in a few cases. When compared with controls tobacco-chewing habitués does increase induced significant mito depression. Our data reveals that samples CPK values were remarkably decreased when compared to control value. Conclusion/Application: The demonstration of chromosomal damage in lymphocytes from tobacco chewing habitues strongly indicates that tobacco should be categorized as a human carcinogen and major risk factor for oral, throat and pharynx cancers.

Keywords

BrdU-labelling, CA, CPK, MI, SCE

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Ahmad Fazilat ¹, Swarupa R. Didla ², M.S. Chakravarty ³, Saima Wajid ¹

Affiliations
1 Department of Biotechnology, Faculty of Science, Hamdard University (Jamia Hamdard), New Delhi - 110062, IN
2 Department of Human Genetics, Faculty of Science, Andhra University, Vishakhapatnam – 530003, Andhra Pradesh, IN
3 Department of Biotechnology, Faculty of Science, Andhra University, Vishakhapatnam – 530003,Andhra Pradesh, IN

Abstract

Objective: Many studies have showed that foot ulcer is the most frequent occurrence among diabetic patients in the south Indian origin. This study aimed to investigate on the hypothesis that in patients with diabetic foot ulcer in south Indian subjects there is a substitution within nucleotide sequence of coding region related to insulin gene and this may be signifying a link disequilibrium with the class III allele in the insulin gene locus, which can be considered due to its relevance with diabetes. We aim to examine this part of insulin gene to find out if there is any mutation within the insulin gene sequence that is relevant with diabetes mellitus. Materials and Methods: We used the PCR products of the amplified insulin gene from 40 patients diagnosed with foot ulcer seeking mutations, which might be in association with the class III allele and therefore indicates its relevance with the manifestation of diabetic foot ulcers. Results: We identified a β-chain mutant (i.e. insulin Chicago) using the restriction enzyme (MboII) in individuals afflicted with foot ulcer. A couple of patterns (α and β) were found indicating the presence of PstI polymorphism within the 3' un-translated part of the insulin gene. Besides, allele frequencies in subjects homozygous for α allele, homozygous for β allele and heterozygous were respectively 0.3 (12/40), 0.125 (5/40) and 0.575 (23/40). No significant variant regarding allelic frequency was characterized between the patients and normal controls, though a mutation in B-chain (insulin Chicago) was observed. Conclusion: Despite linkage disequilibrium between this polymorphism in the 3' un-translated portion of the insulin gene and the class III allele, it is unlikely to have advantage than the class III allele itself on prognosticating diabetes.

Keywords

Diabetes mellitus, Insulin Gene, Polymerase Chain Reaction.

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