Energy decomposition and waterswapping analysis to investigate the SNP associated DPD mediated 5-FU resistance.

5-fluorouracil is an essential component of systemic chemotherapy for colon, breast, head, and neck cancer patients. However, tumoral overexpression of the dihydropyrimidine dehydrogenase has rendered 5-FU clinically ineffective by inactivating it to 5'-6'-dihydro fluorouracil. The responses to 5-FU in terms of efficacy and toxicity greatly differ depending upon the population group, because of variability in the DPD activity levels. In the current study, key active site amino acids involved in the 5-FU inactivation were investigated by modelling the 3D structure of human DPD in a complex with 5-FU. The identified amino acids were analyzed for their possible missense mutations available in dbSNP database. Out of 12 missense SNPs, four were validated either by sequencing in the 1000 Genomes project or frequency/genotype data. The recorded validated missense SNPs were further considered to analyze the effect of their respective alterations on 5-FU binding. Overall findings suggested that population bearing the Glu611Val DPD mutation (rs762523739) is highly vulnerable to 5-FU resistance. From the docking, electrostatic complementarity, dynamics, and energy decomposition analyses it was found that the above mutation showed superior scores than the wild DPD -5FU complex. Therefore, prescribing prodrug NUC-3373 or DPD inhibitors (Gimeracil/3-Cyano-2,6-Dihydroxypyridines) as adjuvant therapy may overcome the 5-FU resistance.

An insight into the possible mechanisms of antispermatogenic action of Dalbergia sissoo in male mice.

This study investigated the possible mechanisms of antispermatogenic action of Dalbergia sissoo in Parkes male mice. Mice were orally administered aqueous leaf extract of Dalbergia sissoo (50 and 100 mg kg-1 body weight day-1 for 35 days) and various testicular indices such 3ß- and 17ß-hydroxysteroid dehydrogenase (HSD) activities, Western blot analyses of StAR, cytochrome P450scc and caspase-3, germ cell apoptosis by TUNEL, and lipid peroxidation and antioxidant enzymes activities were assessed. A significant increase in lipid peroxidation level and a marked decrease in activities of superoxide dismutase, catalase, 3ß- and 17ß-HSD were noted in the testis of Dalbergia-treated mice compared to controls. The treatment also had adverse effect on expression levels of StAR and cytochrome P450scc in the testis. There was an increase in the number of TUNEL-positive germ cells and in expression level of caspase-3 in testes of Dalbergia-treated mice, especially in those treated with 100 mg dose compared to controls. By 56 days of withdrawal therapy, the alterations induced in the above parameters recovered to control levels. Our results thus suggest that Dalbergia treatment interferes with steroidogenesis and produces oxidative stress in the testis, which may induce germ cell apoptosis leading to suppression of spermatogenesis.

Effect of magnesium supplementation on type 2 diabetes associated cardiovascular risk factors: a systematic review and meta-analysis.

BACKGROUND: Cardiovascular disorders remain the leading cause of death in type 2 diabetic patients. In the present study, a systematic review and a meta-analysis of randomised controlled trials (RCTs) were conducted aiming to evaluate the effect of magnesium supplementation on type 2 diabetes (T2D) associated cardiovascular risk factors in both diabetic and nondiabetic individuals. METHODS: PubMed, Scopus, Cochrane, Web of Science and Google Scholar databases were searched from inception to 30 June 2016 aiming to identify RCTs evaluating the effect of magnesium supplementation on T2D associated cardiovascular risk factors. The data were analysed using a random effect model with inverse variance methodology. Sensitivity analysis, risk of bias analysis, subgroup analysis, meta-regression and publication bias analysis were also conducted for the included studies using standard methods. RESULTS: Following magnesium supplementation, a significant improvement was observed in fasting plasma glucose (FPG) [weighted mean difference (WMD) = -4.641 mg dL-1 , 95% confidence interval (CI) = -7.602, -1.680, P = 0.002], high-density lipoprotein (HDL) (WMD = 3.197 mg dL-1 , 95% CI = 1.455, 4.938, P < 0.001), low-density lipoprotein (LDL) (WMD = -10.668 mg dL-1 , 95% CI = -19.108, -2.228, P = 0.013), plasma triglycerides (TG) (WMD = -15.323 mg dL-1 , 95% CI = -28.821, -1.826, P = 0.026) and systolic blood pressure (SBP) (WMD = -3.056 mmHg, 95% CI = -5.509, -0.603, P = 0.015). During subgroup analysis, a more beneficial effect of magnesium supplementation was observed in diabetic subjects with hypomagnesaemia. CONCLUSIONS: Magnesium supplementation can produce a favourable effect on FPG, HDL, LDL, TG and SBP. Therefore, magnesium supplementation may decrease the risk T2D associated cardiovascular diseases, although future large RCTs are needed for making robust guidelines for clinical practice.

Characterisation and diagnosis of frangipani mosaic virus from India.

Frangipani mosaic virus (FrMV) is known to infect frangipani tree (Plumeria rubra f. acutifolia) in India but the virus has not been characterized at genomic level and diagnosis is not available. In the present study, an isolate of FrMV (FrMV-Ind-1) showing greenish mosaic and vein-banding symptoms in P. rubra f. acutifolia in New Delhi was characterized based on host reactions, serology and genome sequence. The virus isolate induced local symptoms on several new experimental host species: Capsicum annuum (chilli), Nicotiana benthamiana, Solanum lycopersicum and S. melongena. N. benthamiana could be used as an efficient propagation host as it developed systemic mottle mosaic symptoms all round the year. The genome of FrMV-Ind-1 was 6643 (JN555602) nucleotides long with genome organization similar to tobamoviruses. The Indian isolate of FrMV shared a very close genome sequence identity (98.3 %) with the lone isolate of FrMV-P from Australia. FrMV-Ind-1 together with FrMV-P formed a new phylogenetic group i.e. Apocynaceae-infecting tobamovirus. The polyclonal antiserum generated through the purified virus preparation was successfully utilized to detect the virus in field samples of frangipani by ELISA. Of the eight different tobamoviruses tested, FrMV-Ind-1 shared distant serological relationships with only cucumber green mottle mosaic virus, tobacco mosaic virus, bell pepper mottle virus and kyuri green mottle mosaic virus. RT-PCR based on coat protein gene primer successfully detected the virus in frangipani plants. This study is the first comprehensive description of FrMV occurring in India.

BDP-30, a systemic resistance inducer from Boerhaavia diffusa L., suppresses TMV infection, and displays homology with ribosome-inactivating proteins.

Root extract of Boerhaavia diffusa L. induced systemic resistance in tobacco against Tobacco mosaic virus. A 30 kDa protein was isolated as the active component, called BDP-30 on the basis of the molecular weight and source plant. BDP-30, a glycoprotein, was found to be temperature and protease resistant. It was basic, possessing a pI greater than 9.0. In-gel proteolytic digestion of BDP-30 generated two peptides that possessed the amino acid sequence KLYDIPPLR and KVTLPYSGNYER by LC/MS/MS. Both peptides shared absolute sequence identity with trichosanthin, a ribosome-inactivating protein from Trichosanthes kirilowii, and a 78 percent and 100 percent homology respectively with an RIP from Bryonia dioica, bryodin. Further, effort was made to look at the fate of TMV in induced resistant Nicotiana tabacum cv. Xanthi, a systemic host of the virus, at specified days after inoculation in control and treated plants. TMV coat protein (CP) was detected by immunoblot 7 days post inoculation up to 21 days in the control set, but not in treated resistant plants. TMV RNA was detected by RT-PCR using TMV-CP specific primers. Resistant tobacco did not show presence of TMV RNA up to 21 days of inoculation. This suggests that BDP-30 may be suppressing TMV replication.

Antiviral activity of the Indian medicinal plant extract Swertia chirata against herpes simplex viruses: a study by in-vitro and molecular approach.

PURPOSE: The antiviral activity of Indian Medicinal plant extract Swertia chirata was tested against Herpes simplex virus (HSV) type-1, using multiple approaches both at cellular and molecular level. METHODS: Cytotoxicity, plaque reduction, virus infectivity, antigen expression and polymerase chain reaction (PCR) assays were conducted to test the antiviral activity of the plant extract. RESULTS: Swertia plant crude extract (1 gm/mL) at 1:64 dilution inhibited HSV-1, plaque formation at more than 70% level. HSV antigen expression and time kinetics experiments conducted by indirect immunofluorescence (IFA) test, revealed a characteristic pattern of small foci of single fluorescent cells in Swertia extract treated HSV-1 infected cells at 4 hours post infection dose, suggested drug inhibited viral dissemination. Infected cell cultures treated with Swertia extract at various time intervals, tested by PCR, failed to show amplification at 12, 24-72 hours. HSV-1 infected cells treated with Acyclovir (antiviral drug) did not show any amplification by PCR. CONCLUSIONS: In this preliminary study, the Indian medicinal plant extract, Swertia chirata showed antiviral properties against Herpes simplex virus type-1.