The Role of Serum Erythropoietin (EPO) and Vascular Endothelial Growth Factor (VEGF) in Pulse Wave Velocity (PWV) Among Hypertensive Patients: A Cross-Sectional Study.

Background and objective While hypertension (HTN) is a major health-related threat globally, it is often an under-reported clinical condition as most of the stage I hypertensive patients do not present with any symptoms. The relationship between endogenous oxygen-sensing protein [erythropoietin (EPO) and vascular endothelial growth factor (VEGF)] levels and vascular stress in hypertensive patients is not fully understood as the mechanistic pathway by which these oxygen-sensing proteins alter the vascular physiology and cause hypertension is still a matter of debate. In light of this, we explored the role of these two proteins in the development of vascular stress including increased pulse wave velocity (PWV). We aimed to examine the correlation between oxygen-sensing proteins and vascular stress markers including PWV in hypertensive patients. Materials and methods We conducted a cross-sectional study involving age-matched participants classified into three groups (group 1: normotensive persons, n=36; group 2: stage I hypertensive patients, n=36; and group 3, stage II hypertensive patients, n=36). Adiposity-related parameters such as waist circumference (WC), hip circumference (HC), BMI, and waist-hip ratio (WHR) were measured. BP was recorded manually in resting posture by using a sphygmomanometer. PWV, which predicts the progression of BP and the development of HTN, was recorded using a periscope, which works based on the oscillometric method. Vascular stress-induced oxidative stress parameters [serum malondialdehyde (MDA) and serum nitric oxide (NO)] were also estimated by using a UV spectrophotometer. Quantitative estimations of oxygen-sensing proteins (serum EPO and serum VEGF) were done by using the ELISA kit method. The results were expressed as mean ± standard deviation (SD). The correlation between the variables was done using Spearman's correlation. A p-value <0.05 was considered statistically significant. Results Adiposity indices and vascular stiffness parameters were found to be significantly (p <0.05) increased in group 2 and group 3 compared to group 1. The levels of serum MDA were found to be significantly (p<0.05) increased in group 2 and group 3 than group 1, whereas the levels of serum NO were significantly (p<0.05) decreased in group 3 and group 2 than group 1. A significant (p<0.05) positive correlation was observed between the PWV and EPO (r=0.492) while a significant (p<0.05) negative correlation was observed between PWV and VEGF (r=-0.406) among the study population. Conclusion The results are indicative of the influence of vascular stress in stage I and II hypertensive patients. Furthermore, the relationship between oxygen-sensing proteins and vascular stress in hypertensive patients has also been established.

Effect of Mucuna pruriens on brain NMDA receptor and tau protein gene expression in cerebral ischemic rats.

Present study aimed to assess effect of pre-treatment with Mucuna pruriens seed extract and its bioactive molecule(s) on NMDAR and Tau protein gene expression in cerebral ischemic rodent model. Methanol extract of M. pruriens seeds was characterized by HPLC, and ß-sitosterol was isolated by flash chromatography. In vivo studies to observe the effect of pre-treatment (28 days) with methanol extract of M. pruriens seed and ß-sitosterol on the unilateral cerebral ischemic rat model. Cerebral ischemia induced by left common carotid artery occlusion (LCCAO) for 75 min (on day 29) followed by reperfusion for 12 h. Rats (n = 48) divided into four groups. GroupI (control,Untreated + LCCAO)-No pre-treatment + cerebral ischemia; GroupII(ß-sitosterol + Sham)-pre-treatment with ß-sitosterol, 10 mg/kg/day + sham-operated; GroupIII(ß-sitosterol + LCCAO)-pre-treatment with ß-sitosterol, 10 mg/kg/day + cerebral ischemia; GroupIV(methanol extract + LCCAO)-pre-treatment with methanol extract of M. pruriens seeds, 50 mg/kg/day + cerebral ischemia. Neurological deficit score was assessed just before sacrifice. Experimental animals were sacrificed after 12 h reperfusion. Brain histopathology was performed. Gene expression of NMDAR and Tau protein of left cerebral hemisphere (occluded side) was performed by RT-PCR. Results revealed that the neurological deficit score was lower in groups III and IV compared to group I. NMDAR and tau protein mRNA expression in left cerebral hemisphere were upregulated in Group I, downregulated in groups III and IV. Histopathology of left cerebral hemisphere (occluded side) in Group I showed features of ischemic brain damage. Groups III and IV, left cerebral hemisphere showed less ischemic damage compared GroupI. Right cerebral hemisphere showed no areas of ischemia-induced brain changes. Pre-treatment with ß-sitosterol and methanol extract of M. pruriens seeds may reduce ischemic brain injury following unilateral common carotid artery occlusion in rats.

Ubiquitin Mediated Degradation of EGFR by 17 ß-estradiol in Triple Negative MDA-MB-231 (TNBC) Breast Cancer Cells Line.

BACKGROUND: Triple Negative Breast Cancer (TNBC) commonly displays Epidermal growth factor receptor (EGFR). Effective EGFR degradation results in the suppression of tumor in various models. Studies have addressed the relevance of this strategy in the treatment of TNBC. In the present study, we examined the effect of 17 ß- estradiol on EGFR expression in MDA-MB-231 (TNBC) cell line and assessed whether 17 ß-estradiol degrades EGFR by ubiquitination pathway. OBJECTIVES: The objective of this study is to treat MDA-MB-231 cell lines with Cycloheximide with or without 17ß-estrdiol to observe whether 17ß-estradiol leads to EGFR degradation and to treat with MG-132 to assess whether degradation occurs through ubiquitination pathway. METHODS: MDA-MB-231 cells were treated with 17ß-estradiol (E2) and EGFR expression was studied by western blotting at different intervals by using Cycloheximide chase. To assess ubiquitination pathway of degradation of EGFR in MDA-MB-231 cell line, MG-132 was used. RESULTS: EGFR expression was reduced with ß-estradiol treatment in MDA-MB-231 cell line with Cycloheximide chase. Upon Treatment with MG-132 and E2, EGFR expression did not reduce, suggesting that Estrogen degrades EGFR by ubiquitination pathway. CONCLUSION: Estrogen degrades EGFR in MDA-MB-231 cells and this degradation occurs by ubiquitination.

Screening and Development of Transglutaminase-2 Inhibitors and their Derivative as Anti-lung Cancer Agent by in silico and in vitro Approaches.

AIM: This study aimed at screening and development of TG2 inhibitors as anti lung cancer agent. BACKGROUND: Transglutaminase 2 (TG2) is multifunctional and ubiquitously expressed protein from the transglutaminase family. It takes part in various cellular processes and plays an important role in the pathogenesis of autoimmune, neurodegerative diseases, and also cancer. OBJECTIVE: The proposed study focused on screening potent inhibitors of TG2 by in-silico method and synthesize their derivative as well as analyse its activity by utilizing an in-vitro approach. MATERIALS AND METHODS: Molecular docking studies have been carried out on the different classes of TG2 inhibitors against the target protein. Nearly thirty TG2 inhibitors were selected from literature and docking was performed against transglutaminase 2. The computational ADME property screening was also carried out to check their pharmacokinetic properties. The compounds which exhibited positive ADME properties with good interaction while possessing the least binding energy were further validated for their anti-lung cancer inhibition property against A549 cell lines using cytotoxicity studies. RESULTS: The results of the present study indicate that the docked complex formed by cystamine showed better binding affinity towards target protein, so this derivative of cystamine was formed using 2,5 dihydrobenzoic acid. Invitro results revealed that both molecules proved to be good cytotoxic agents against A549 lung cancer (875.10, 553.22 µg/ml), respectively. Further, their activity needs to be validated on TG2 expressing lung cancer. CONCLUSION: Cystamine and its derivative can act as a potential therapeutic target for lung cancer but its activity should be further validated on TG2 expressing lung cancer.

Maternal Myocardial Performance in Second Trimester of Pregnancy With Iron Deficiency Anaemia.

INTRODUCTION: Anaemia affects various organs in body including the heart. In anaemia, oxygen carrying capacity of blood decreases. Iron depletion and the amount of stored iron are reduced in iron deficiency anaemia which limits red cell production. However, the studies which show the effect of anaemia on myocardial function during pregnancy are few in India. AIM: To study the effect of iron deficiency anaemia on myocardial function by ECG during second trimester of pregnancy and to compare ECG changes with normal pregnant women in second trimester. MATERIALS AND METHODS: The study was conducted at antenatal OPD between Oct 2014 to Jul 2015. Hundred pregnant women were selected and divided into 2 groups. A total of 50 normal pregnant women (control group) in 2(nd) trimester (10-14 weeks of gestation) were compared with equal number of pregnant women with anaemia (study group) in 2(nd) trimester, aged between 20-30 years. Electrocardiogram was recorded using Philips twelve channel ECG machine model TC20 in both control and study groups to evaluate myocardial performance. Haematological parameters were analysed by SYSMEX auto analyser. Analysis of Variance (One way ANOVA) was used for comparison between study and control groups and the data was analysed by t-tests. RESULTS: In our study a significant decrease in QRS duration and increase in QTc were observed in study group (p<0.05). T-wave abnormalities like flat and negative T-waves in lead II, III, avF, V2 - V4 were more frequent (p<0.05). 90% of subjects in study group had tachycardia and ECG abnormalities. There was a negative correlation between Hb level, serum ferritin and tachycardia, ECG abnormalities. CONCLUSION: Pregnancy with Iron deficiency anaemia brings about various changes in ECG, suggesting that anaemia and volume overload in pregnancy is a risk factor that may lead to cardiac hypertrophy.

Different anthropometric adiposity measures and their association with cardiovascular disease risk factors in middle aged women.

A cross sectional study of 136 women age group40-55 years was conducted to study which anthropometric measure had the strongest association with cardiovascular disease risk factors in middle aged women. In accordance with their BMI measurement subjects were divided into three groups namely: Normal weight (Group1) BMI 18.5-24.9 kg/m2, Overweight (Group2) - 25 kg/m2 - 29.9 kg/m2, Obese (Group3) - BMI > 30 kg/m2. Each group was further divided into premenopausal and postmenopausal women BMI, WHR, WHtR, SBP, DBP were recorded. TG, TC, HDL and FBS values were estimated. Results showed that both pre and post menopausal middle aged obese women with higher BMI, WHR and WHtR have more chances of having cardiovascular diseases. BMI, WHR and WHtR are theeasy and practical methods to diagnose obesity and together can be used as simple measures to predict cardiovascular risk factors in middle aged women.

Role of alpha-tocopherol in cardiopulmonary fitness in endurance athletes, cyclists.

Exercise increases oxygen consumption and causes a disturbance of intracellular pro-oxidant-antioxidant homeostasis. Athletes are exposed to acute and chronic stress that may lead to increased generation of oxidative species. Hence oxidative stress increases in athletes. Administration of antioxidant like alpha-tocopherol as supplementation may reduce the cell damage caused due to oxidative stress. In the present study, our aim was to study the effects of alpha-tocopherol supplementation on the cardiopulmonary fitness in endurance athletes (cyclists) and non-athletes. Our study included 40 cyclists who were trained under District Youth Service & Sports Office. 40 controls were randomly selected from student group of B.L.D.E.A's Medical College. Alpha-tocopherol (Vitamin E) 200 mg/day for 21 days wasgiven to study group and placebo was given to placebo group. Various physiological parameters like heart rate, blood pressure, respiratory rate were recorded, for assessing cardiopulmonary fitness: Physical Fitness Index (PFI) and VO2 max ml/min/kg were recorded before and after supplementation of vitamin E in athletes, and were compared with placebo group before and after supplementation of placebo and also with non-athletes. The results obtained from present study indicate that antioxidant like alpha-tocopherol supplementation did not contribute significantly to improve the cardiopulmonary fitness of endurance athletes.