An Insight into Survivin in Relevance to Hematological, Biochemical and Genetic Characteristics in Tobacco Chewers with Oral Squamous Cell Carcinoma.

BACKGROUND: Survivin is an inhibitor of apoptosis protein (IAP), encoded by the Baculoviral IAP Repeat Containing 5 (BIRC5) gene located on q arm (25.3) on chromosome 17. It is expressed in various human cancers and involved in tumor resistance to radiation and chemotherapy. The genetic analysis of the BIRC5 gene and its protein survivin levels in buccal tissue related to oral squamous cell carcinoma (OSCC) in South Indian tobacco chewers has not been studied. Hence, the study was designed to quantify survivin in buccal tissue and its association with pretreatment hematological parameters and to analyze the BIRC5 gene sequence. METHOD: In a single centric case control study, buccal tissue survivin levels were measured by ELISA. A total of 189 study subjects were categorized into Group 1 (n = 63) habitual tobacco chewers with OSCC, Group 2 (n = 63) habitual tobacco chewers without OSCC, and Group 3 (n = 63) healthy subjects as control. Retrospective hematological data were collected from Group 1 subjects and statistically analyzed. The BIRC5 gene was sequenced and data were analyzed using a bioinformatics tool. RESULTS: Survivin protein mean ± SD in Group 1 was (1670.9 ± 796.21 pg/mL), in Group 2 it was (1096.02 ± 346.17 pg/mL), and in Group 3 it was (397.5 ± 96.1 pg/mL) with significance (p < 0.001). Survivin levels showed significance with cut-off levels of absolute monocyte count (AMC), neutrophil/lymphocyte ratio (NLR), and lymphocyte/monocyte ratio (LMR) at (p = 0.001). The unique variants found only in OSCC patients were T → G in the promoter region, G → C in exon 3, C → A, A → G, G → T, T → G, A → C, G → A in exon 4, C → A, G → T, G → C in the exon 5 region. CONCLUSIONS: The tissue survivin level increased in OSCC patients compared to controls; pretreatment AMC, LMR, and NLR may serve as add-on markers along with survivin to measure the progression of OSCC. Unique mutations in the promoter and exons 3-5 were observed in sequence analysis and were associated with survivin concentrations.

Cellular Concentration of Survivin and Caspase 3 in Habitual Tobacco Chewers with and without Oral Squamous Cell Carcinoma in South Indian Rural Population-A Case Control Study.

Background: There is paucity of data on tissue levels of Survivin and Caspase 3 in south Indian tobacco chewers with oral Squamous cell carcinoma (OSCC). Oral cancer is a rapidly growing, highly prevalent head and neck malignancy; it involves a mucosal epithelium of a buccal cavity exposed to tobacco and other carcinogens. The basis of the survival of a tumor cell or transformed normal cell into a neoplastic cell is by the suppression of apoptosis regulation. Recently, researchers have focused on Survivin, an inhibitor of apoptosis family of proteins (IAP), involved in apoptosis regulation in cancer cells targeting the executioner Caspase 3. The current study aims to quantify the cellular levels of Survivin and Caspase 3 in tobacco chewers with OSCC and in habitual tobacco chewers without OSCC, in comparison to controls. Methods: A single centric case control study included 186 study subjects, categorized into: Group I (n = 63), habitual tobacco chewers with OSCC; Group 2 (n = 63), habitual tobacco chewers without OSCC; and Group 3 (n = 63), the controls. Resected tumor tissue from Group 1 and buccal cell samples from Groups 2 and 3 were collected into phosphate buffer saline (PBS) and assayed for Survivin and Caspase 3 levels by the ELISA sandwich method. Results: The mean ± SD of the Survivin protein in Group 1 was (1670.9 ± 796.21 pg/mL); in Group 2, it was (1096.02 ± 346.17 pg/mL); and in Group 3, it was (397.5 ± 96.1 pg/mL) with a significance of p < 0.001. Similarly, the level of Caspase 3 in Group 1 was (7.48 ± 2.67 ng/mL); in Group 2, it was (8.85 ± 2.41 ng/mL); and in Group 3, it was (2.27 ± 2.24 ng/mL) with a significance of p < 0.001. Conclusion: The progressive transformation of buccal cells to neoplastic cells is evident; in the case of OSCC, this indicates that the over-expression of Survivin compared to Caspase 3 confirms the suppression and dysregulation of apoptosis.

Maternal serum Apelin 13 and APLN gene promoter variant -1860T > C in preeclampsia.

OBJECTIVE: To evaluate the apelin (APLN) -1860 T > C (rs56204867) polymorphism and maternal serum apelin 13 levels in preeclampsia and its association with blood pressure. METHODS: This case-control study was conducted in department of Biochemistry, Sri Devaraj Urs Medical College, Karnataka, India. A total of 181 subjects were enrolled in the study from department of Department of Obstetrics and Gynecology. The recruited women were grouped as: Group-I (n = 91) cases with preeclampsia and Group-II (n = 90) normotensive healthy pregnant women as controls. Under aseptic conditions, the collected 5 mL blood was distributed for serum separation (3 mL) and genetic analysis (2 mL). Serum was stored at -80 °C after centrifugation at 3000 rpm for 10 min. The collected five mL urine sample was used for urinary protein analysis by dipstick method. The APLN gene -1860 T > C polymorphism and Apelin 13 levels were analyzed by molecular methods and ELISA technique respectively. Birth weight and demographic details were recorded. RESULTS: In the present study, no significant difference was observed for mean gestational age and maternal age. Systolic (158.7 ± 14.0 mmHg) and diastolic (104.9 ± 10.7 mmHg) blood pressure, and mean arterial pressure (MAP) (123.0 ± 11.1 mmHg) (p-value .001) were significantly increased in preeclamptic women compared with healthy pregnant women. Birth weight (2.4 ± 0.5 kg) (p-value .001) was significantly decreased in babies born to preeclamptic mothers. Birth weights were also expressed in centiles, according to Fenton Chart. Number of small for gestational age (SGA) babies were more in preeclampsia (n = 55) than healthy pregnant women (n = 28). Mean maternal serum apelin 13 (239.4 ± 126.3 pg/mL) (p-value .001) concentrations were significantly lower in preeclampsia compared with healthy controls. Maternal serum apelin 13 concentration in preeclampsia was negatively correlated with systolic blood pressure (r = -0.235), diastolic blood pressure (r= -0.172) and mean arterial pressure (r = -0. 206). However, maternal serum apelin 13 levels showed insignificant positive correlation with age, gestational age and birth weight. The genotype and allele frequencies of APLN gene were found significant between study groups as in preeclampsia (χ2 = 11.69; df = 2; p = .0028 and χ2 = 14.27; df = 1; p = .00013 respectively). CC genotype and C allele of APLN - 1860 T > C site was high in preeclampsia. CONCLUSION: Study concludes that preeclamptic women have low level of serum apelin 13 and -1860 T > C polymorphism at APLN gene promoter site with increased allelic frequency of CC genotype and C allele compared to normotensive pregnant women. And this evidence may link to cardiac complications in preeclamptic women after delivery in later stage.

Insulin resistance and decreased spexin in Indian Patients with Type 2 Diabetes Mellitus.

Spexin is novel biomarker, which plays a potential role in glucose and lipid metabolisms. However, there was paucity of serum spexin levels in obesity and diabetes mellitus subjects. Hence the current study was aimed to find the relationship between the serum spexin levels in type 2 Diabetes mellitus (type 2 DM) with extrapolation of cardiovascular disease (CVD) risk. A cross-sectional study included 330 participants, subdivided as control (n=110), type 2 DM (n=110) and type 2 DM with CVD groups (n=110). HbA1c, insulin, lipid profile, spexin & leptin including blood pressure and body mass index were analyzed from all the participants. The serum spexin levels (ng/ml) were significantly decreased in type 2 DM (mean ± sd: 0.65 ± 0.03) and type 2 DM with CVD (0.48 ± 0.02) groups compared to the control (0.79 ± 0.03) group (p<0.001). The decreased spexin levels were observed in type 2 DM, and further more decreased in type 2 DM with CVD patients compared to controls indicating that spexin levels could be served as an early prediction of obesity-induced T2DM with CVD risk.

Relationship Between Xanthine Oxidase, Ischemia Modified Albumin, Nitric Oxide with Antioxidants in Non Pregnants, Pre and Post-delivery of Normal Pregnants and Preeclampsia.

Preeclampsia is a multisystem disorder involves altered homeostasis of oxidants-antioxidants, inflammatory process and endothelial dysfunction. The present study aim was to determine the levels of oxidative stress parameters (malondialdehyde, protein carbonyl, ischemia modified albumin and xanthine oxidase), nutrient antioxidants (vitamin C and vitamin E), enzyme antioxidants (catalase, superoxide dismutase, glutathione peroxidase glutathione reductase), total antioxidant status (TAS) and its association with nitric oxide. The study population consists of three groups, non pregnants (Group 1, n = 57), normotensive pregnants (Group 2, n = 57) and Preeclampsia (Group 3, n = 57). Group 2 and 3 were followed after delivery within 48 h. In preeclampsia xanthine oxidase, malondialdehyde and uric acid levels were significantly increased (p < 0.001), while TAS decreased (p < 0.05) when compared to normotensive pregnant and non pregnant. Catalase, glutathione reductase levels were increased (p < 0.005) and vitamin E, super oxide dismutase levels were decreased (p < 0.001) in preeclampsia when compared to normal pregnants. Receiver operating characteristics curve analysis showed area under curve for xanthine oxidase (0.8), malondialdehyde (0.804), Uric acid (0.84), ischemia modified albumin (0.92) and catalase (0.88) which indicated as good markers in preeclampsia. Amongst, ischemia modified albumin is a better marker of intrauterine hypoxic reperfusion risk with sensitivity 87.7 % and specificity 91.2 %. The increased hydrogen peroxide from xanthine oxidase adds to oxidative stress and increased catalase activity in preeclampsia represents combating action. Increased oxidative stress, decreased TAS and its apparent reversible changes evinced within 48 h after delivery in preeclampsia illustrated that placental abnormality is the contributing factor in the pathogenesis.

Is Xanthine Oxidase, a Marker in Pre-eclampsia? A Case-Control Study.

INTRODUCTION: Pre-eclampsia is an obstetrics problem that affects multiple systemic functions and leads to the increased maternal and fetal morbidity and mortality. The objective of the study was to evaluate the plasma levels of Xanthine oxidase (XO) activity, uric acid and Nitric oxide (NO) levels in women with pre-eclampsia and normal pregnancy during antenatal and postpartum period. MATERIALS AND METHODS: A case control study was conducted in women with normal pregnancy (n=50) and pre-eclampsia (n=50) before and after delivery. XO activity, uric acid and NO levels were determined from samples at 30-39 weeks of gestation. The current study was conducted in association with Obstetrics and Gynecology Department of R.L. Jalappa Hospital and Research Center. The blood samples were analysed for assay of XO, uric acid and NO. The results were analysed by using SPSS software version 2013. P-value < 0.05 was considered as statistically significant. RESULTS: The plasma XO activity was elevated (p<0.001) in the pre-eclampsia compared to normotensive pregnant women before delivery and decreased after delivery (p<0.001) significantly. Uric acid level showed a significant increase in pre-eclampsia when compared to the control before delivery (p<0.001) however values were non-significant after delivery. CONCLUSION: Placenta plays a key role in the pathophysiology of pre-eclampsia. Placenta removal leads to decrease trend of xanthine oxidase activity, uric acid and elevation of Nitric oxide as reversible changes in pre-eclampsia patients within 48 hours after delivery.

Evidence of Paternal N5, N10 - Methylenetetrahydrofolate Reductase (MTHFR) C677T Gene Polymorphism in Couples with Recurrent Spontaneous Abortions (RSAs) in Kolar District- A South West of India.

INTRODUCTION: Recurrent spontaneous abortion (RSA) is a multifactorial clinical obstetrics complication commonly occurring in pregnancy. Many research studies have noted the mutations such as C677T in N5, N10 - Methylenetetrahydrofolate reductase (MTHFR)gene which is regarded as RSA risk factor. This study was carried out to determine the occurrence of frequency of C677T of the MTHFR gene mutations with RSA. AIM: The purpose of present study is to determine the frequency of MTHFR C677T polymorphisms in couples with recurrent pregnancy loss and the impact of paternal polymorphisms of MTHFR C677T in recurrent pregnancy loss in population of couples living in Kolar district of Karnataka with RSA. DESIGN: A total of 15 couples with a history of two or more unexplained RSA were enrolled as subjects in the study and a total of 15 couples with normal reproductive history, having two or more children and no history of miscarriages were enrolled as controls. MATERIALS AND METHODS: DNA extraction from samples case and control group couples and its quantification by Agarose gel electrophoresis, assessment of DNA purity, MTHFR C 677T gene mutation detection by PCR-RFLP method. STATISTICAL ANALYSIS: Carried out by web based online SPSS tool. RESULTS: The frequency of C677T genotype showed homozygous wild type CC (80%), heterozygous CT type (13.3%) and homozygous mutation TT type (6.67%) observed in males. Similarly from female's homozygous wild type CC (86.6%), heterozygous type (13.3%), and homozygous type mutations TT (0%) was recorded. In couple control groups, we observed homozygous wild type CC (86.6%), heterozygous CT type (13.3%) and homozygous type mutations TT type (0%). CONCLUSION: We noticed a high frequency of MTHFR specifically T allele associated with paternal side.Therefore, the present study indicated the impact of paternal gene polymorphism of MTHFR C677T on screening in couples with recurrent pregnancy loss.

The association of hypomagnesaemia, high normal uricaemia and dyslipidaemia in the patients with diabetic retinopathy.

CONTEXT: Diabetic retinopathy is fast becoming an important cause of a visual disability. The visual disability which results from diabetes is a significant public health problem; however, this morbidity is largely preventable and treatable. If it is managed with a timely intervention, the quality of life can be preserved. AIMS: The objective of this study was to investigate the association of serum uric acid, magnesium and the lipid profile in diabetic retinopathy with Normal subjects and Diabetes mellitus without retinopathy, among the south Indian population. SETTINGS AND DESIGN: The diabetic retinopathy patients were identified from the diabetic health camps which were held in rural areas, and they were compared with those with diabetes without complications and the normal subjects. MATERIAL AND METHODS: The diabetic retinopathy patients were compared with the healthy subjects and with diabetes without retinopathy. Furthermore, the Diabetic retinopathy patients were grouped as proliferative and non-proliferative, based on the fundoscopic findings. Magnesium, uric acid, FPG, fructosamine and the lipid profile were measured in the above groups and they were analyzed. STATISTICAL ANALYSIS: The statistical analysis was done by using the SPSS software, by applying the Student 't' test. RESULTS: The mean serum magnesium concentration was observed to be low in the diabetic retinopathy group (1.43mg/dl) as compared to those in the controls and the diabetic subjects. The serum Uric acid concentration was high normal (4.84mg/dl), which was associated with the dyslipidaemia in diabetic retinopathy. CONCLUSION: The poor glycaemic control in diabetes is associated with hypomagnesaemia, and increased uric acid concentration with dyslipidaemia, which can be an initial picture of the ongoing biochemical changes in the complication of diabetes, which can help in predicting the onset of diabetic retinopathy in diabetes.