K. Sri Manjari

Association of vitamin D receptor gene polymorphisms with BMD and their effect on 1, 25-dihydroxy vitamin D3 levels in pre- and postmenopausal South Indian women from Andhra Pradesh

BACKGROUND Osteoporosis is a multifactorial disorder with a strong genetic component and vitamin D receptor gene has been suggested as a candidate gene for osteoporosis. Therefore the present study was aimed to investigate the role of the VDR Fok 1 gene polymorphism and its influence on vitamin D3 levels and BMD in pre- and postmenopausal osteoporotic women of Indian ethnicity. METHODS 427 osteoporotic women and 460 age matched controls were included in the study. VDR FOK1 gene polymorphism was assessed by the PCR-RFLP method. Serum vitamin D3 was measured by the HPLC method. RESULTS The frequency of ff genotype and f allele was significantly high in pre- and postmenopausal osteoporotic women in comparison with controls (p<0.001).In each case individuals with ff genotype had significantly low BMD in comparison with Ff and FF genotypes. No significant association was found between the genotypes and vitamin D3 levels. CONCLUSION The VDR Fok 1 gene is associated with low bone mass in all our study subjects. The genotype ff of the VDR Fok 1 gene is associated with osteoporosis. Further ff genotype associated significantly with low bone mass. Therefore the f allele of VDR Fok1 gene is an important risk factor for osteoporosis.

A single-nucleotide polymorphism in tumor necrosis factor-α (− 308 G/A) as a biomarker in chronic pancreatitis

OBJECTIVE Chronic pancreatitis is a gradual, long-term inflammation of the pancreas that results in alteration of its normal structure and function. The study aims to investigate the role of -308 (G/A) polymorphism of TNF-α gene in chronic pancreatitis. MATERIAL AND METHODS A total of 200 subjects were included in this case-control study. A total of 100 in patients admitted in the Gastroenterology Unit of Gandhi Hospital and Osmania General Hospital, Hyderabad were included in the present study. An equal number of healthy control subjects were randomly selected for the study. The genotyping of TNF-α gene was carried out by tetra-primer ARMS PCR followed by gel electrophoresis. The TNF-α levels were assayed by enzyme-linked immunosorbent assay. RESULTS A significant variation with respect to the genotypic and allelic distribution in the disease group when compared to control subjects [OR=2.001 (1.33-3.005), p<0.0001**] was observed. Subjects homozygous for the A allele had higher TNF-α levels compared to G allele. CONCLUSION The present study revealed a significant association of the TNF-α gene promoter polymorphism with chronic pancreatitis. Thus, TNF-α genotype can be considered as one of the biological markers in the etiology of chronic pancreatitis.

Role of Plasma MMP 9 levels in the Pathogenesis of Chronic Pancreatitis

Pancreatic fibrosis is a key pathological feature in the etiology of chronic pancreatitis that leads to obliteration of exocrine and endocrine pancreatic tissues and its replacement by fibrous tissue resulting in clinical manifestations. Matrix metalloproteinase 9 is a member of the MMP family that is also known as gelatinase B, degrades type IV collagen of extracellular matrix and basal membrane. The present study is aimed at evaluating the clinical significance of plasma concentration of MMP-9 in chronic pancreatitis. The samples were obtained from 112 chronic pancreatitis patients and an equal number of age and sex matched healthy controls. MMP-9 levels were quantitatively measured by ELISA assay. Statistical analysis was applied to test the significance of results. The present study revealed a significant increase of plasma MMP 9 levels in chronic pancreatitis patients compared to control subjects. Elevated levels were also observed in all the patient groups compared to control subjects with regard to sex, age, addictions etc. MMP-9 degrades the type IV collagens in normal basement membrane, which in turn activates the pancreatic stellate cells which promote the development of pancreatitic fibrosis. Thus, elevated plasma levels of MMP-9 may act as a susceptibility factor for the development of chronic pancreatitis.

Interleukin 10 gene promoter polymorphisms in women with early-onset pre-eclampsia.

Pre‐eclampsia is one of the most serious disorders of human pregnancy and T helper type 1 (Th1)/Th2 imbalance plays a major role in its aetiology. The Th2 cytokine, interleukin (IL)‐10, plays a significant role in the maintenance of pregnancy. The present study is aimed at understanding the role of IL‐10 promoter polymorphisms (−1082 G/A; −592 A/C and −819 C/T) and their haplotypes in early‐onset pre‐eclampsia. A total of 120 patients and an equal number of women with normal pregnancy, from Government Maternity Hospital, Petlaburz, Hyderabad, India, were considered for the present study. A standard amplification refractory mutation system–polymerase chain reaction (ARMS–PCR) was carried out for genotyping followed by agarose gel electrophoresis. Appropriate statistical methods were applied to test for the significance of the results. It was found that the IL‐10 −819 C allele (P = 0·003) and −592 A (P = 0·005) allele frequencies increased significantly in patients compared to controls. No significant difference was found with regard to −1082 promoter polymorphism. Haplotype analysis of the IL‐10 single nucleotide polymorphisms (SNPs) revealed a significant association with ACC haplotype with a twofold increased risk in patients compared to controls. The frequencies of two common IL‐10 haplotypes (GCC and ATA) did not show any significant difference. Further, the diplotype analysis revealed five genotypes: −1082A with −819C (P = 0·0016); −1082G with −819C (P = 0·0018); −819C with −592C (P = 0·001); −1082A with −592C (P = 0·032); and −1082G with −592C (P = 0·005) associated with the disease. These findings support the concept of contribution of IL‐10 gene polymorphisms in the pathogenesis of early‐onset pre‐eclampsia.

Role of matrix metalloproteinase 3 gene promoter polymorphism in chronic pancreatitis

AimTo study the role of 5A/6A polymorphism of matrix metalloproteinase (MMP-3) and their levels in the pathogenesis of chronic pancreatitis (CP).MethodsOne hundred and twenty CP patients and an equal number of age and sex-matched healthy controls were included in the study. Genotypes were determined for 5A/6A allele of MMP-3 gene by allele specific PCR (AS-PCR). The serum MMP-3 levels were estimated using sandwich ELISA method.ResultsThe distribution of the genotypes of the 5A/6A polymorphism in both control and study patients was similar (p = 0.523). Within the disease group, patients with older age, early onset of the disease, and addictions such as smoking and alcohol consumption had higher levels as compared to those who did not have these features.ConclusionWe conclude that functional polymorphism of MMP-3 (5A/6A) is not associated with CP. However, the higher levels within the disease group indicate its possible role in the disease process.

Transferrin (rs3811647) gene polymorphism in iron deficiency anemia

Background Iron-deficiency anemia (IDA) is the most common type of anemia, caused by inadequate iron availability for hemoglobin production due to the lack of dietary iron or insufficient uptake of iron. Transferrin (TF) exerts a crucial function in the maintenance of systematic iron homeostasis. The expression of the TF gene is controlled by transcriptional mechanism, although little is known about its influence on IDA. Hence, the aim of the current investigation was to determine the functional polymorphism (rs3811647) of TF gene in iron deficiency anemia. Material and Methods A total of 207 school children of age from 12-16 years were selected from Government High School Seetaphalmandi, Secunderabad, Andhra Pradesh and screened for iron deficiency. Out of which 70 school children had iron deficiency anemia with hemoglobin levels less than 11.5 g/dl and 137 were normal. Demographic details and blood samples were obtained from all the subjects. Genotyping was carried out by tetra-primer ARMS PCR followed by agarose gel electrophoresis, and appropriate statistical analysis. Results The genotype distribution of TF (rs3811647) region were 4.3% (AA), 81.4% (AG) and 14.3% (GG) in iron deficient children compared to 8% (AA), 72.3% (AG), and 19.7% (GG) in normal children. No significant variation was observed with respect to the allelic distribution [OD = 1.035 (0.67 – 1.59), p = 0.917] in the IDA group when compared to normal group. Conclusions There was no significant association of the TF (rs3811647) gene polymorphism with iron deficiency anemia. Thus, our study highlights the importance of other genetic variants influencing the outcome of iron deficiency anemia. However, larger samples have to be analyzed to confirm the same.