BACKGROUND:Several pathogenic conditions leading to morbidity, including cancer, aging, diabetes, reperfusion injury, cardiovascular disease, and neurological disorders, are known to be exacerbated by oxidative stress. Antioxidant therapy is effective in the treatment of such disorders and appears to be a potential therapeutic technique to reduce oxidative stress. The aim of our study is to investigate the antioxidant effects of L-ascorbic acid and nitric oxide (NO) modulators on rats suffering from oxidative stress induced by acute restraint stress (RSx1).METHODOLOGY:In this in vivo study, Wistar rats were subjected to one hour of restraint stress on day 21 to induce oxidative stress. Superoxide dismutase (SOD), total antioxidant capacity (TAC), catalase, glutathione (GSH), and malondialdehyde (MDA) were used to assess the antioxidant effects. IBM Corp. Released 2013. IBM SPSS Statistics for Windows, Version 22.0. Armonk, NY: IBM Corp. was used for data analysis.RESULTS:Compared to vehicle groups, acute restraint stress (RSx1) dramatically increased MDA levels while decreasing GSH, SOD, total antioxidant capacity, and catalase. L-NAME, 7-NI, AG (50 mg/kg each), and L-ascorbic acid (200 mg/kg) reversed the changes in SOD, MDA, GSH, total antioxidant capacity, and catalase levels. The NO precursor L-arginine (1000 mg/kg) and NO synthase inhibitors followed the same trend.CONCLUSION:Our study findings highlight the complex role of antioxidants and NO modulators in the pathogenesis of diseases, as evidenced by the reversal of oxidative stress indicators. Antioxidant therapy, with its potential to mitigate oxidative stress, emerges as a viable treatment option for a range of pathological conditions associated with oxidative stress.

Background: High sensitivity C-reactive protein (hs-CRP), a sensitive marker of inflammation and tissue damage is an acute phase reactant. It is raised in hypertension and predicts cardiovascular outcome. Moreover elevated levels of inflammatory markers such as hs-CRP and altered lipid profile are commonly seen in hypertension which may develop cardiovascular events, hence, hs-CRP, lipid profile and nitric oxide (NO) have been incorporated in the study. The aims of this study were to find out a relationship of serum hs-CRP and dyslipidemia in hypertensives and to find out an association of serum hs-CRP with the risk for Cardiovascular disease (CVD). Methods: A case-control study was done among the patients visiting the outpatient department (OPD) of BP Koirala Institute of Health Sciences, Dharan, Nepal in which forty seven newly diagnosed hypertensives as cases and fifty age and sex matched healthy normotensives as controls, were enrolled in the study with the prior informed consent. hs-CRP, nitric oxide (NO) and lipid profile were estimated in both the cases and controls. Results: Hypertensives have significantly raised levels of hs-CRP, non-high density lipoprotein cholesterol (non-HDL-C) and NO compared to that in controls (P< 0.05). hs-CRP has a significant positive correlation with systolic as well as diastolic blood pressure, BMI, and with triglyceride (TG) (P< 0.05). However, the correlation of hs-CRP with NO is negative but statistically significant (0.000). Conclusion: The level of hs-CRP, which is thought to be a marker of inflammation, is significantly raised in hypertensives. Moreover, the majority of hypertensives are dyslipidemic suggesting hypertensives to be at an increased risk for the development of CVD. Key words: Cardiovascular diseases, dyslipidemia, hs-CRP, hypertension, inflammation.

Ship voyage to Antarctica is a stressful journey for expedition members. The response of human gut microbiota to ship voyage and a feasible approach to maintain gut health, is still unexplored. The present findings describe a 24-day long longitudinal study involving 19 members from 38th Indian Antarctic Expedition, to investigate the impact of ship voyage and effect of probiotic intervention on gut microbiota. Fecal samples collected on day 0 as baseline and at the end of ship voyage (day 24), were analyzed using whole genome shotgun sequencing. Probiotic intervention reduced the sea sickness by 10% compared to 44% in placebo group. The gut microbiome in placebo group members on day 0 and day 24, indicated significant alteration compared to a marginal change in the microbial composition in probiotic group. Functional analysis revealed significant alterations in carbohydrate and amino acid metabolism. Carbohydrate-active enzymes analysis represented functional genes involved in glycoside hydrolases, glycosyltransferases and carbohydrate binding modules, for maintaining gut microbiome homeostasis. Suggesting thereby the possible mechanism of probiotic in stabilizing and restoring gut microflora during stressful ship journey. The present study is first of its kind, providing a feasible approach for protecting gut health during Antarctic expedition involving ship voyage.

The purpose of this study was to examine the free radical scavenging and antioxidant activities of ellagic acid (EA) and ellagic acid peracetate (EAPA) by measuring their reactions with the radicals, 2,2-diphenyl-1-picrylhydrazyl and galvinoxyl using EPR spectroscopy. We have also evaluated the influence of EA and EAPA on the ROS production in L-6 myoblasts and rat liver microsomal lipid peroxidation catalyzed by NADPH. The results obtained clearly indicated that EA has tremendous ability to scavenge free radicals, even at concentration of 1 µM. Interestingly even in the absence of esterase, EAPA, the acetylated product of EA, was also found to be a good scavenger but at a relatively slower rate. Kinetic studies revealed that both EA and EAPA have ability to scavenge free radicals at the concentrations of 1 µM over extended periods of time. In cellular systems, EA and EAPA were found to have similar potentials for the inhibition of ROS production in L-6 myoblasts and NADPH-dependent catalyzed microsomal lipid peroxidation.

•Study demonstrated reversal of oxidative stress markers by L-Arginine and L- Ascorbic acid.•L-Arginine and L- Ascorbic acid can be used therapeutically in disease associated with high oxidative stress.•Can be used in pathophysiological conditions associated with high oxidative stress.

BACKGROUND:The Indian peafowl (Pavo cristanus) is native to South Asia and is the national bird of India. Here we present a draft genome sequence of the male blue peacock using Illumina and Oxford Nanopore technology (ONT). RESULTS:ONT sequencing gave ∼2.3-fold sequencing coverage, whereas Illumina generated 150-base pair paired-end sequence data at 284.6-fold coverage from 5 libraries. Subsequently, we generated a 0.915-gigabase pair de novo assembly of the peacock genome with a scaffold N50 of 0.23 megabase pairs (Mb). We predict that the peacock genome contains 23,153 protein-coding genes and 75.3 Mb (7.33%) of repetitive sequences. CONCLUSIONS:We report a high-quality assembly of the peacock genome using a hybrid approach of sequences generated by both Illumina and ONT. The long-read chemistry generated by ONT was useful for addressing challenges related to de novo assembly, particularly at regions containing repetitive sequences spanning longer than the read length, and which could not be resolved with only short-read-based assembly. Contig assembly of Illumina short reads gave an N50 of 1,639 bases, whereas with ONT, the N50 increased by >9-fold to 14,749 bases. The initial contig assembly based on Illumina sequencing reads alone gave 685,241 contigs. Further scaffolding on assembled contigs using both Illumina and ONT sequencing reads resulted in a final assembly of 15,025 super-scaffolds, with an N50 of ∼0.23 Mb. Ninety-five percent of proteins predicted by homology matched with those in a public repository, verifying the completeness of our assembly. Like other phylogenetic studies of avian conserved genes, we found P. cristatus to be most closely related to Gallus gallus, followed by Meleagris gallopavo and Anas platyrhynchos. Compared with the recently published peacock genome assembly, the current, superior, hybrid assembly has greater sequencing depth, fewer non-ATGC sequences, and fewer scaffolds.

Adenosine is a signaling molecule which is produced in high concentrations during airway inflammation. Airway inflammation is a characteristic feature of COPD. Therefore, the current study was designed to evaluate the changes in adenosine metabolism in COPD and correlate these changes with severity of the disease. The study was conducted on 50 healthy controls (25 healthy non-smokers and 25 healthy smokers) and 46 COPD patients (21 moderate, 15 severe and 10 very severe). The patients were sub-divided into moderate, severe and very severe categories as per the GOLD spirometric classification. Blood was collected from each subject and serum, lymphocytes and erythrocytes were separated. The adenosine levels and activities of 5'-nucleotidase, adenosine deaminase and its isoenzymes were assessed in serum, lymphocytes and erythrocytes. The data were analyzed statistically. A p value < 0.05 was considered as significant. In healthy smokers and COPD patients the adenosine levels increased. In COPD patients 5'-nucleotidase activity increased significantly in serum, lymphocytes and erythrocytes. The activities of ADA and isoenzymes decreased significantly in serum of healthy smokers and COPD patients, in lymphocytes and erythrocytes of very severe COPD patients and of ADA and ADA2 in lymphocytes and erythrocytes of moderate and severe COPD patients. The FEV1 (% of predicted) showed a significant negative correlation with adenosine levels and 5'-nucleotidase activity in serum, lymphocytes and erythrocytes and significant positive correlation with ADA and isoenzymes activity in serum and lymphocytes of COPD patients. We conclude that the adenosine metabolism changes in COPD. The adenosine levels and 5'-nucleotidase activity increase, and ADA activity decreases with severity of the disease.

Tumour Necrosis Factor (TNF-α) is one of the most commonly studied cytokine of TNF superfamily. Patho-physiologically generation of high levels of TNF-α is a development of inflammatory responses for many of the chronic diseases. Simultaneously in pulmonary diseases increased level of TNF-α in blood serum relates with the cytotoxicity. The Study was conducted on 100 patients of Rajan Babu Institute for Pulmonary Medicine & Tuberculosis (RJPIMT) Hospital. Patient diagnosis was confirmed via Spirometry. The result reveals that the serum TNF-α level increases with the COPD severity. The mean values of TNF were significantly increased with the decreased FEV/FVC. The present study showed that serum TNF-α level correlates with severity of airway obstruction in Spirometry among the COPD patients. It is worth mentioning that serum TNF-α level may plays an important role to diagnose chronic obstructive pulmonary disease as a useful marker to monitor the disease severity in addition to spirometric parameters like FVC, FEV1 and FEV1/FVC ratio.

Hypertension in pregnancy is one of the potential causes of maternal and fetal morbidity and mortality. It complicates 7–10% of pregnancies. As of today, prediction of pregnancy hypertension is not possible.

Abstract Content: The increased oxidative stress in chronic obstructive pulmonary disease (COPD) patients is the result of increased inhaled oxidants, generated by various cells of the airways. Objective: The investigation included measurements of malondiadehyde (MDA), uric acid, ascorbic acid, and matrix metalloproteinase-12 (MMP-12) in COPD patient. We also performed genetic analysis for protein–protein interaction (PPI) network. Materials and methods: The study was conducted on healthy subjects with normal lung function (NS, 14 subjects) and 28 patients (Global Initiative for Chronic Obstructive Lung Disease (Gold) 1 and Gold 2) with COPD. Results: There was significant (p < .001) increase in MMP-12, MDA and uric acid levels as compared to healthy controls. A significant (p < .001) decline in ascorbic acid level was observed in COPD patients. The PPI was found to be 0.833 which indicated that proteins present in COPD are linked. Discussion and conclusion: This study suggests oxidative stress plays an important role in COPD and the PPI provide indication that proteins present in COPD are linked.

Introduction: Pregnancy hypertension is reported to affect 2-12% women in India. Currently available diagnostic criteria is applicable after 20 th weeks of gestation, this delayed diagnosis is the possible cause of the associated adverse obstetrics outcomes, which can be minimized by early detection. In this study the role of MMP-9 and PAPP-A was assessed in the first trimester (11 +0 -13 + 6 ) for early prediction of pregnancy hypertension. Methodology: This study was a part of ongoing project of first trimester preeclampsia screening. In that project 2000 women were screened. Samples were taken in first trimester and women were followed till delivery. Out of 2000 women, 199 developed Gestational Hypertension, 1454 remained normotensive till term and remaining 347 were excluded. In the present study we have included 199 women who developed hypertension and 199 controls, who were normotensive till term (selected from cohort of 1454 women who were normotensive). Results: The first trimester serum levels of PAPP-A (p<0.001) and MMP-9 (0.039) were significantly low in women who developed hypertension as compared to women who were normotensive till term. A significant positive correlation (p=0.03) was observed between MMP-9 and PAPP-A. The Combined sensitivity, specificity and Positive predictive value (PPV) of PAPP-A and MMP-9 was found to be 74.5%, 70.2% and 75% respectively. Conclusion: Inadequate expression of PAPP-A and MMP-9 may result in incomplete or poor trophoblast invasion and subsequent complication of pregnancy hypertension. In our population we have found that first trimester assessment of PAPP-A and MMP-9 may potentially predict pregnancy hypertension.

Background: Chronic Obstructive Pulmonary Disease (COPD) is characterized by airway obstruction and destruction of lung tissue, the disease for which at present there is no cure, although the treatment can slow worsening. Nearly 90% of COPD is caused by long term cigarette smoking; however, only 25% of chronic tobacco smokers develop COPD, the exact cause of this predisposition of some smokers to acquire COPD is still poorly understood at the molecular level. Lot of studies are available showing the correlation of MMP1 and asthma but not much work has been done on its correlation with COPD especially in North India. The outcome of our study was also expected to discover MMP1 as a new target for treating COPD and its progression in smokers.Aim and Objectives: The association of COPD and smoking with MMP1 gene product and its SNP in the North Indian population was intended to be studied.Material and Methods: The proposed study was a case control study. In this study adults aged 30 years and above fulfilling the inclusion criteria (Please see Annexure I) were considered and a total of 180 subjects were taken after determining the sample size. Pulmonary function test (PFT) was performed by Spirometry. Three main groups each consisting of 60 subjects were formed on the basis of smoking history and Spirometry: Ist group: Smokers without co-morbidity with normal PFT, IInd group: Smokers with Spirometry proved COPD and without any other co-morbidity, IIIrd group: Healthy Non-smoker controls. Quantification of metalloproteinase MMP1 in all the groups were performed in the serum with ELISA kits and Single Nucleotide Polymorphism (SNP 1G-1607 2G, ID rs1799750) studies in the gene encoding MMP1 linked to COPD susceptibility in smokers was performed by DNA Sequencing Analysis. The correlation between the SNPs, gene product, smoking and COPD was studied. Results: MMP1 concentration was seen increased in serum of COPD and smokers when compared to healthy controls. And there is high negative correlation between *FEV1/FVC % (post-bronchodilator) and MMP1 in COPD compared to Smokers and Healthy Controls. However, from the results of present analysis, we could show negative correlation between MMP1 and *FEV1/FVC % in all the 3 groups but the correlation proves to be much more negative in Group II i.e. Smokers with COPD. Statistically Significant Positive Correlation of Pack Years with MMP1 conc., and Statistically Significant Negative Correlation between Pack Years and *FEV1/FVC % was seen.Conclusion:In conclusion, our present analyses did show significant association between MMP1 and COPD risk in North Indian population. MMP1 levels of COPD patients were significantly increased in smokers and may contribute to or be a marker of the pathophysiology of COPD. Asian Journal of Medical Sciences Vol.8(1) 2017 5-14

The potential role of polyphenolic acetate (PA) in causing diverse biological and pharmacological actions has been well studied in our laboratory. Our investigations, for the first time, established the role of calreticulin transacetylase (CRTAase) in catalyzing the acetylation of nitric oxide synthase (NOS) by Pas leading to robust activation of NOS. 7, 8- Diacetoxy-4-methylcoumarin (DAMC) and other acetoxycoumarins augmented the expression of thioredoxin (TRX) and vascular endothelial growth factor (VEGF) in human peripheral blood mononuclear cells (PBMCs). These findings substantiated our earlier observations that DAMC was a superb inducer of angiogenesis. The enhanced expression of thioredoxin reductase (TRXR) and diminished expression of thioredoxin interacting protein (TRXIP) leading to increased expression and activity of TRX in PBMCs due to the action of DAMC was revealed by real time RT-PCR analysis. The possible activation of TRX due to acetylation was confirmed by the fact that TRX activity of PBMCs was enhanced by various acetoxycoumarins in tune with their affinities to CRTAase as substrates. DAMC caused enhanced production of NO by way of acetylation of NOS as mentioned above and thereby acted as an inducer of VEGF. Real time RT-PCR and VEGF ELISA results also revealed the overexpression of TRX. DAMC and other PAs were found to reduce the oxidative stress in cells as proved by significant reduction of intracellular ROS levels. Thus, the crucial role of TRX in DAMC-induced angiogenesis with the involvement of VEGF was established.

Objective: Role of Polyphenolic acetates (PAs) was studied to explore their ability to impart acetylation of histone protein by the novel mechanism of acetylation in Ehrlich Ascites tumour (EAT) mice model. Effect of HDAC inhibitor Valproic acid (VA) alone and in combination with Polyphenolic acetates and also Polyphenolic acetates along with Calreticulin (CAL) induced hyperacetylation causing modulation of apoptosis was also investigated in view of possibility to develop target oriented combination therapy in cancer.Materials and Methods: EAT mice model was established. EAT mice were treated with different PAs, VA and purified, recombinant calreticulin and their combinations by Intra peritoneal route (I.P) which induced transacetylation of histones resulting in apoptosis. After 26 hrs of the above treatment, 2ml of peritoneal fluid was aspirated from mice of all the above groups. The peritoneal fluid was studied for the amount of acetylated histone proteins by Western blotting using commercially available specific Anti- Acetyl Histone (Ac-Lys) Antibodies and extent of apoptosis in peritoneal cells by Flow-Cytometry and fluorescence microscopy.Results: The number of apoptotic cells were represented by the Median (25th, 75th percentile) clearly illustrates significant increase in the no. of apoptotic cells in both PAs viz. 7,8-Diacetoxy-4-Methyl Coumarin (DAMC) and 6-Acetoxy Quinolone (6-AQ) and their combinations with CAL and VA as compared to DMSO control group and the maximum no. of apoptotic cells were observed in Group DAMC+CAL+VA. Increased extent of histone protein acetylation was observed by Western blotting using specific Anti- Acetyl Histone (Ac-Lys) Antibodies.Conclusion: PAs alone and also synergistically with CAL and VA are potential drug candidates for Cancer therapy.Asian Journal of Medical Sciences Vol.7(2) 2015 13-20

Extensive research carried out in our group on polyphenolic acetates (PAs) substantiated the potential role of PAs in causing diverse biological and pharmacological actions. Our earlier investigations firmly established the calreticulin transacetylase (CRTAase) catalyzed activation of nitric oxide synthase (NOS) by PAs. In this report, we have studied the effect of 7,8-diacetoxy-4-methylcoumarin (DAMC, a model PA) and other acetoxy coumarins on the thioredoxin and VEGF expression in human peripheral blood mononuclear cells (PBMCs), with a view to substantiate our earlier observation that DAMC was a superb inducer of angiogenesis. Real time RT-PCR analysis revealed the enhanced expression of thioredoxin reductase (TRXR) and diminished expression of thioredoxin interacting protein (TRXIP) leading to the increased expression and activity of thioredoxin (TRX) in PBMCs due to the the action of DAMC. The fact that TRX activity of PBMCs was enhanced by various acetoxy coumarins in tune with their affinity to CRTAase as substrate, suggested the possible activation of TRX due to acetylation. The overexpression of thioredoxin was found to correlate with that of VEGF as proved by real time RT-PCR and VEGF -ELISA results, apart from the DAMC-caused enhanced production of NO acting as an inducer of VEGF. Moreover, the intracellular ROS levels were also found to be reduced drastically, by DAMC thus reducing the oxidative stress in cells. These observations strongly evidenced the crucial role of TRX in DAMC-induced tissue angiogenesis with the involvement of VEGF.

Background: Infertility is one of the medial, social, and psychological burdens in this part of world. Thyroid dysfunction can lead to menstrual disturbance, anovulatory cycles, and decreased fecundity. Proper management of thyroid dysfunction can result in restoration of normal fertility. Therefore it is very important to screen thyroid abnormalities among women with infertility. This study aimed to determine association of thyroid dysfunction among infertile women. This study comprises total of 735 primary infertile women with age ranging from 20 to 35 years. Blood samples were collected and subjected for estimation of thyroid hormones. Out of 635 cases 447 (74.4%) were thyroid. A number of 56 (7.6%) have primary hypothyroidism, 31 (4.2%) have primary hyperthyroidism, 87 (11.8%) have subclinical hypothyroidism, and 8 (1.1%) have subclinical hyperthyroidism.

Background: Dyslipidemia has been noted to play an integral role in the pathogenesis and progression of micro and macrovascular complications in diabetes mellitus patients. The complications exemplified by renal vascular and cardiovascular disease cause the most morbidity and mortality in this group of patients. This study is aimed at understanding the pattern of dyslipidemia among type 2 diabetic patients. Patients and Methods: A total of 108 consenting adult type 2 diabetic patients seen in medical unit of Nepalgunj medical college and Teaching Hospital, Banke, Nepal from January 2012 to November 2012 were evaluated in this cross sectional study. Their fasting lipid profile, fasting blood glucose, weight, height and blood pressure were evaluated. Results: The prevalence of dyslipidemia (at least one abnormal lipid profile) was 90.7%. The 24.1% had single dyslipidemia while 66.6% had combined dyslipidemia. Reduced HDL constituted the highest single abnormality (62%) followed by hypertriglyceridemia (56.5%), hypercholesterolemia (53.7%) and high LDL in (44.4%). The duration of DM was not significantly associated with dyslipidemia (p > 0.05). Conclusion: Dyslipidemia is highly prevalent among type 2 diabetic patients in Nepal with the majority of the patients having combined dyslipidemia. We recommend that aggressive treatment of lipidemia and hyperglycemia can be instituted to reduce the risk of macro and microvascular complications.

This is the first study characterizing spectrum of beta-thalassemia mutations in Nepalese population. Mutations were analyzed in 22 patients using 10 sets of allele-specific primers. Five of the mutations, namely F.S 41/42 (--TCTT), IVS1 nt5 (G-->C), IVS1 nt1 (G-->T), 619 bp deletion and F.S 8/9 (+G), were found to constitute 87.82% of total alleles studied. F.S 41/42 (--TCTT) was the commonest mutation. -88 (C-->T), Codon 16 (--C) and Codon 15 (G-->A), had a combined frequency of 12.18%. Distribution of mutations causing beta-thalassemia in different ethnic Nepalese groups was analyzed. The mutational profile in Nepal share several similarities with that from the two neighboring countries, India and China. Detection of more than one mutation in three cases of thalassemia trait raises the likelihood of existence of multiple mutations in cis in Nepalese thalassemic carriers. Such possibility has to be carefully considered while developing prenatal screening program for Nepalese population.