Sabhiya Majid

ROLE OF HEMATOLOGICAL PARAMETERS IN DIAGNOSIS AND PROGNOSIS OF GASTRI C CARCINOMA IN KASHMIR , INDIA

Gastric cancer is the second leading cause of death in the world . More than two - thirds of the patients are dia gnosed at an advanced stage. Metastatic gastric cancer has poor prognosis with a median 5 years - survival rate of 7 %. Hematological parameters including leukocyte count, platelet count and their ratios have been used as prognostic indicators in several tum or types. The aim of the study was to examine an association of the difference in haematological parameters between gastric cancer patients and normal controls of Kashmir valley. We enrolled 210 subjects of which 110 were newly diagnosed gastric cancer ca ses and 100 were healthy controls. Participants were re cruited from hospitals, clinics and radiology department of Sh ri Maharaja Hari Singh ( SMHS) hospital Srinagar, India from May 2011 to Apr 2012 . After informed consent, all patient s were interviewed and examined and demographic and clinical information was collected. Blood samples were drawn for examination of hematological measures and for measurement of carcino embryonic antigen (CEA) . W e found the hematological parameters like: Hb (10 ± 2, P = 0.004 ) , MCV (84.25 ± 5, p = 0.01), Granulocyte % (70.04 ± 10.63, p = 0.001), Lymphocyte % (26.12 ± 10.7, p < 0.0001) , RDW (48 ± 10, p = 0.004 ) in gastric cancer patients and these were found to be highly decreased as compared to normal healthy controls where hema tological paramete rs was in normal range . Our study was statistically significant (p < 0.005). The study suggests that the hematological parameters like HB, MCV, Granuloc yte %, Lymphocyte % and RDW are de creased in gastric cancer patients and acts as a n ea rly marker in the prognosis and diagnosis of gastric cancer.

Polymorphism of the XRCC3 gene and risk of gastric cancer in a Kashmiri population: a case-control study.

DNA repair plays a critical role in protecting the genome of the cell from the insults of cancer-causing agents. Genetic polymorphisms in DNA repair genes may influence individual variation in DNA repair capacity, which may be associated with the risk of developing cancer. Inherited polymorphisms of DNA repair genes may contribute to variations in DNA repair capacity and genetic susceptibility to different cancers. The X-ray repair complementing defective repair in Chinese hamster cells 3 (XRCC3) gene is a member of the RAD51 gene family. It encodes an important protein that functions in the homologous recombination repair of a DNA double-strand break. For gastric cancer, the importance of mutations in mismatch repair genes has been well documented, but less is known about other DNA repair pathways in gastric carcinogenesis. In this study, we have focused on the XRCC3 gene, involved in homologous recombinational repair. The Kashmir valley has an increased incidence of gastric cancer and its etiology has not been understood fully as yet. As the Kashmiri population is ethnically and demographically different from that in other parts of the world, the aim of this study was to determine whether a single nucleotide polymorphism of the XRCC3 gene (Thr241Met) of exon 7 can influence the risk of gastric cancer in the population. As many as 80 histopathologically confirmed gastric cancer cases and 70 healthy controls, age, sex, and ethnicity matched for known genotypes of XRCC3 exon 7 were studied. We genotyped for this variant using PCR-restriction fragment length polymorphisms. The XRCC3 genotype and allele frequencies were not significantly different between cases and controls (P=0.92 for the genotype; P=0.72 for the allele). The XRCC3 241Met allele frequency (6.6%) was significantly lower in healthy Kashmiri controls than reported previously in healthy US White controls (38.9%). Compared with the XRCC3 241Thr/Thr genotype, the variant XRCC3 241Thr/Met and Met/Met genotypes were not associated with an increased risk of gastric cancer (adjusted odds ratio=1.19; 95% confidence interval=0.44–3.18). These findings suggest that polymorphisms of XRCC3 Thr241Met may not play a role in the etiology of gastric cancer. Further studies with a larger number of participants and simultaneous measurement of different polymorphisms in DNA repair genes in the same pathway are needed.

Type 2 diabetes and metabolic syndrome – adipokine levels and effect of drugs

Abstract Type 2 diabetes mellitus (T2DM) is a consequence of complex interactions among multiple genetic variants and environmental risk factors. This complex disorder is also characterized by changes in various adipokines. In this study, our objective was to estimate the levels of adiponectin, leptin, and resistin (ALR) in T2DM patients, besides studying the effect of various drugs on their levels. Study participants included 400 diabetic and 300 normal patients from the Department of Endocrinology and Department of Biochemistry, Govt Medical College Srinagar. Subjects were categorized under various groups, i.e., Group 1 (metformin treated) and Group 2 (glimepiride treated), and cases were also categorized as obese with T2DM (Group A), obese without T2DM (Group B), and T2DM only (Group C). The serum ALR levels were estimated by ELISA (Alere), and biochemical parameters were also evaluated before and after treatment. Adiponectin levels were found to be significantly lower in T2DM cases as compared to controls (12 ± 5.5 versus 22.5 ± 7.9 μg/ml), while leptin and resistin levels were found to be significantly higher than controls (14.3 ± 7.4 versus 7.36 ± 3.73 ng/ml) (13.4 ± 1.56 versus 7.236 ± 2.129 pg/ml). Taking the effect of drugs into consideration, the effect on adiponectin and resistin levels was found to be highly significant in Group 2 before and after treatment (11 ± 5 versus 19.2 ± 4.5 μg/ml) (13.6 ± 2.5 versus 7.3 ± 2.9 pg/ml), while more effect was observed in leptin among Group 1 (metformin)-treated cases (27 ± 15 ng/ml versus 15 ± 15 ng/ml). Further the adiponectin levels were found to be significantly lower in Group B, while leptin and resistin levels were found to be significantly higher among obese cases when compared to T2DM cases only. Glimepiride also shows more effect on FBG, HbA1c% levels, while metformin shows more effect on Lipid profile levels. From the study, it can be concluded that ALR levels are affected by use of antidiabetic drugs among which glimepiride shows more effect on adiponectin and resistin levels, while leptin gets affected more by metformin. It can also be proposed that ALR levels are not affected by diabetes only, suggesting that their alterations in T2DM may be due to obesity as we observed more ALR changes in obese cases when compared to T2DM cases, and so there might be an important link between adiposity and insulin resistance.

Increased prevalence of subclinical hypothyroidism in females in mountainous valley of Kashmir.

Background: Iodine-rich diet is necessary for proper thyroid gland function. Subclinical hypothyroidism (SCH) is associated with serious complications. Substantial numbers of patients have risk of SCH getting converted into primary hypothyroidism. Objectives: The objectives of the present study are to survey dietary iodine intake pattern in ethnic population of Kashmir and to study the prevalence of SCH. Materials and Methods: A retrospective, cross-sectional referral hospital study was conducted. Sample size comprised of 2550 patients who were referred to Department of Biochemistry, Government Medical college, Srinagar diagnostic laboratory from OPD and IPD of associated SMHS hospital. Assessment of thyroid function over a period of one year from March 2010 to March 2011 in the serum has been performed by electro-chemiluminescence immunoassay method on ECLIA 2010 fully automatic analyzer. Interview cum questionnaire methods were used to record the patient history and dietary iodine intake pattern. Iodine status of these patients was assessed by measuring urinary iodine excretion. Results: Total patients were 2550 comprising of 44.6% males and 56.4% females. Subjects with elevated and normal thyroid stimulating hormone (TSH) levels in the serum were 30.51 and 69.4% respectively. About 550 patients (21.56%) had subclinical hypothyroidism which includes both males and females. Prevalence of SCH was more in females (81.8%) than in males (18.2%). Most of the patients presenting with SCH were in the age group of 20–65 years. Conclusion: The percentage of SCH amongst the study sample patients was 21.56%, which is much higher as compared to other parts of the world. The highest percentage of SCH was found in females (81.8%) as compared to males (18.2%). On the basis of the present study, we suggest that routine screening of selected populations, especially women between 20 and 65 years of age, may be advocated. Further community level awareness programs need to be organized wherein people in mountainous valley of Kashmir are motivated to take salt in iodized form and diet rich in iodine to ensure proper thyroid gland functioning.

Association of Vitamin D Receptor Gene Polymorphism in Patients with Type 2 Diabetes in the Kashmir Valley

OBJECTIVES Approx 1 billion people across various ethnic and age groups have vitamin D deficiency. The high prevalence of such a deficiency is an imperative public health issue because hypovitaminosis D is an autonomous risk factor for mortality in the general population. Beyond bone integrity and calcium homeostasis, it is involved in numerous physiologic and pathologic processes. The role of vitamin D in the pathogenesis and prevention of type 2 diabetes mellitus has sparked universal interest. METHODS This hospital-based case-control study was designed to study the association between 25-hydroxy vitamin D (25[OH]D) levels and the vitamin D receptor (VDR) gene polymorphism with diabetes and to evaluate their roles as risk factors for diabetes. 100 cases and controls were taken. 25(OH)D levels were analyzed by the chemilumenescence method using a Siemens ADVIA Centaur analyzer. Genomic DNA was extracted and Taq-1 and Bsm-1 genotyping in the VDR gene was done by using the polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP). RESULTS 25(OH)D levels of patients with diabetes were significantly lower than those of controls (19.26±0.95 ng/mL vs. 25.49±1.02 ng/mL; p=0.001). 25(OH)D levels were found to be inversely associated with glycated hemoglobin percentages in cases (r2=0.74). The results suggested that the single nucleotide polymorphisms Taq-1 t(T) allele and b (G allele) in Bsm-1 might be a susceptibility allele for diabetes in the Kashmiri population. CONCLUSIONS VDR gene polymorphisms appear to be an important genetic determinant in the progression of diabetes. Considering the important predisposition risk factor, we observed that Taq-1 and Bsm-1 were strongly associated with diabetes in northern Indians. But requires further study as a probable genetic risk marker for diabetes.

Diminished expression of MGMT & RASSF1A genes in gastric cancer in ethnic population of Kashmir

BACKGROUND Cancer initiation and progression are accompanied by profound changes in DNA. DNA methylation that was the first epigenetic alterations identified in cancer. DNA hypermethylation at promoter sites is closely associated with down regulation of protein and as major participant in the development and progression of series of human tumors. Therefore we hypothesized that promoter hypermethylation of RASSF1A & MGMT gene could influence susceptibility to gastric cancer (GC) as well, and we conducted this study to test the hypothesis in Kashmiri population. METHODS A hospital based case-control study; including 200 GC cases and 200 matched controls from patients who went surgical resection. Promoter hypermethylation was determined by Methylation Specific Polymerase chain reaction. The expression of MGMT & RASSF1A protein was examined by Western blotting technique. RESULTS Frequency of promoter region hypermethylation of MGMT gene were 46.5% in cases and 5.5% in controls (P<0.05) while as in case of RASSF1A frequency was 44% in cases and 4.5% in controls (P<0.05). Further, frequency of hypermethylation of both genes was found predominant in males, aged and advanced pathological stage subjects. Loss of MGMT expression was found in 46.5% cases (P<0.05) while as loss of RASSF1A expression was found in 40.5% cases (P<0.05). In both genes a positive correlation was observed between promoter CpG island hypermethylation and down regulation of respective proteins. CONCLUSIONS These findings indicate that promoter hypermethylation at CpG island may be responsible for reduction of expression at protein level which may be an initial event in carcinogenesis and the progression of GC.

Prevalence and prognostic relevance of BrafV600E mutation in colorectal carcinomas from Kashmir (North India) valley

Molecular studies have implicated mutant B-type Raf kinase (BRAFMut) of MAP-kinase signalling pathway in the pathogenesis of several cancers including colorectal cancer. Recently, the prognostic and therapeutic relevance of the most frequent BRAFV600E mutation also has been highlighted in colorectal carcinomas (CRC). Thus, the aim of this study was to investigate the prevalence of BRAFV600E mutation and to determine the correlation between this mutation and indicators of poor prognosis and outcome in patients with CRCs from Kashmir, North India. Here, we developed a highly sensitive technique, mutation allele-specific multiplex PCR (MASMP), for detection of BRAFV600E/BRAFc.1799T>A mutation, the results of which were confirmed by sequencing the product and compared to direct DNA sequencing. In total, BRAFV600E mutation status was analyzed in 57 colorectal tumour samples and an equal number of adjacent normal tissues. A high frequency of BRAFV600E mutation 21% (12/57) was identified in tumour tissues by MASMP compared to only 5.2% by direct DNA sequencing. Statistical analysis indicated that compared to BRAF-negative colorectal tumours, patients with BRAFV600E colorectal tumours were more likely to be >50 years old (61%) (P < 0.03). These tumours were more likely to be of clinical tumour stages III and IV (63%) (P < 0.04) with lymph node metastasis (52%) (P < 0.02) and characterised by a high-grade histology (63%) (P < 0.04). Colorectal patients harbouring BRAFV600E mutation experience more relapse/recurrence (52%) (P < 0.02). We, therefore, conclude that BRAFV600E mutation can be used as an indicator of poor prognosis to predict the outcome for CRC patients from Kashmir. MASMP proved to be a simple, sensitive and reliable technique for screening patients for BRAFV600E mutation. Testing for this mutation may be useful for selecting initial therapy mode and for follow-up monitoring in CRC patients.

Adipocytokines: unravelling the missing link in diabetes and metabolic syndrome

Adipose tissue composed of adipocytes, fibroblasts, immune cells, and various other cell types, once known as passive reservoir is now considered an endocrine organ secreting bioactive molecules including hormones now termed adipokines with various known and unknown endocrine functions in addition to regulating fat mass and nutrient homeostasis. Due to the dramatic rise in obesity and its metabolic sequelae during the past decades, adipose tissue gained tremendous scientific interest. Adipose tissue secretes variety of products known as ‘adipokines’, including leptin, adiponectin, resistin and visfatin, as well as cytokines and chemokines such as tumor necrosis factorAlpha (TNF-α), interleukin-6 and monocyte chemoattractant protein-1. These adipokines helps in the regulation of hemostasis, blood pressure, lipid and glucose metabolism, inflammation, and atherosclerosis. The release of these cytokines causes a chronic sub inflammatory state that could play a central role in the development of insulin resistance and type 2 diabetes, and the increased risk of cardiovascular disease associated with obesity, together referred as metabolic syndrome [4]. This chronic low-grade inflammation causes increase in macrophage infiltration, leads to increased adipocyte secretion of pro-inflammatory molecules such as resistin, tumor necrosis factorAlpha, and interleukin-6 (IL-6) [4]. These adipocytokines can act as key regulators of response to insulin in peripheral tissues [5]. Their role in causing insulin resistance via various mechanisms involving the flux of in prooxidant and antioxidant state has been seen [6,7]. Adipokines have a great potential for clinical use as potential therapeutics for obesity, obesity related metabolic, cardiovascular and other diseases [8]. Several extracellular factors cause obesity, related adipocyte metabolism and macrophage infiltration. Interestingly, recent research provides increasing evidence of the importance of regulating adipocyte function, adipose tissue metabolism and inflammation. In this mini review, we will briefly highlight roles of various adipokines (adiponectin, leptin, resistin, TNF-α, and IL-6) in regulating insulin sensitivity and resistance. Main Body

Biochemical Profile and Genetic Polymorphism of MTHFRC677T in Risk of Type 2 Diabetes Mellituss

Diabetes mellitus (DM) is a common endocrine metabolic disorder and a leading cause of death worldwide Diabetes type-2 is a multicausal disease which develops slowly and in a stepwise order. Our study showed there was no significant difference in serum high density lipoprotein (HDL) and Tri glyceride (TG) of patients and controls (0.90±0.59 vs 1.15±0.39 p>0.05) and (1.19±0.70 vs 1.01±0.52 p>0.060) respectively. Low density lipoprotein (LDL) and total cholesterol (TC) are significantly higher in patients than control group (4.09±1.14 vs 3.01±1.02 p<0.0002) and (4.21±1.28 vs 3.78±1.29 p<0.05). However, HDL/TC ratio is significantly higher in patients than controls (0.21±0.91 vs 0.30±0.99 p<0.05). Serum levels of all liver enzymes (ALT, AST, and ALP) analyzed are significantly higher in patients than controls (12.69±10.80 vs 4.95±2.66, p<0.0002), (15.99±10.70 vs 6.95±3.84, p<0.0002) and 68.29±27.78 vs 21.27±7.77, p<0.0001) respectively. On genetic level the role of MTHFR C677T polymorphisms our results showed 63% of the cases showed homozygous mutant condition. The allelic association of polymorphism of controls with cases was found to be significant (P=0.007). Homozygous mutant condition of MTHFRC677T gene was found to be certainly higher in Diabetes Mellitus 2 Cases of above 60 years of age (80%), than ages below 60 years and in controls (16.6%) and was significant as p=0.005, compared to below 60 years of age (33.3%) and in controls (0%) and association was insignificant as p=0.4667. Our data suggest that there is an important role of LDL, TC, HDL/TC, ALT, AST, and ALP in type-2 Diabetes, also gene polymorphisms of MTHFR C677T gene may act synergistically to increase the risk of type 2 diabetes.