Role of curcumin on chloroquine phosphate-induced reproductive toxicity.

The present study was conducted to screen the efficacy of curcumin against chloroquine phosphate (CQ)-induced reproductive toxicity in adult male Swiss albino mice. Animals were given oral doses of 100, 200, 300 mg/kg body weight (b.w.), and high dose of CQ (300 mg/kg b.w.) + curcumin (80 mg/kg b.w.) for 45 days. Animals of the withdrawal group were given high dose of CQ (300 mg/kg b.w.) for 45 days and, at day 46, were kept for another 45 days. Effects were observed on some key enzymes, such as alkaline phosphatase, which was found to be decreased, whereas acid phosphatase was increased and succinate dehydrogenase and adenosine triphosphatase were decreased. Oxidative parameters, such as superoxide dismutase declined, whereas thiobarbituric acid-reactive substances were found to be elevated. Protein level was also decreased. Gravimetric indices were also recorded. Results obtained indicated adverse effects of CQ in a dose-dependent manner. The presence of curcumin with CQ alleviated its toxic effects. Hence, it can be concluded that curcumin has beneficial influences and appears able to ameliorate CQ toxicity.

Controlling non small cell lung cancer progression by blocking focal adhesion kinase-c-Src active site with Rosmarinus officinalis L. phytocomponents: An in silico and in vitro study.

Background: Non small cell lung cancer (NSCLC) is a global, fatal oncological malady to which conventional and targeted therapies proved less effective with consequent side effects; hence, phytocomponents from herbal sources may provide potent alternative and should be tested for cancer intervention. Activation and overexpression of proto-oncogene tyrosine kinase Src (c-Src) and focal adhesion kinase (FAK) lead to cell proliferation and invasion. Hence, in the present investigation, in silico analysis was carried out to identify molecular intervention of phytocomponents in blocking the active site and thus inhibiting c-Src and FAK activation, which in turn could control progression of NSCLC. Materials and Methods: In silico analysis was carried out using Molegro Virtual Docker, Molegro Molecular Viewer, and ClusPro server for ligand-protein and protein-protein interaction study. Phytochemical analysis and assay for antioxidant activity of hydroalcoholic extract of Rosmarinus officinalis L. were carried out using standard phytochemical tests, high-performance thin-layer chromatography, and 2, 2-diphenyl-1-picrylhydrazyl assay. Effectiveness of extract in arresting cell proliferation was confirmed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay on A549 cell line. Results: In silico analysis indicated effective binding of rosmarinic acid to the active site of target proteins FAK and c-Src, blocking their activity. MTT assay revealed potent antiproliferative activity of hydroalcoholic extract which acted in dose-dependent manner. Phytochemical analysis confirmed that the extract was rich in phytocomponents and had antioxidant activity of 94.9%, which could therefore effectively eliminate free radicals and inhibit cell progression. Conclusion: In silico and in vitro studies confirmed that phytocomponents present in hydroalcoholic extract of R. officinalis L. could effectively block the active site of target proteins and thus controlled cell proliferation on NSCLC cells, suggesting herb as an effective alternative medicine for the treatment of NSCLC.

Phyllanthus fraternus manifests potent anti-proliferative activity on cultured Daudi cells.

OBJECTIVE: The aim of this study is to observe the apoptosis of Phyllanthus fraternus Webster against Daudi cells and to study its primary mechanism. MATERIALS AND METHODS: Antiproliferative activity of cultured Daudi cells was evaluated using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in a dose- and time-dependent manner after treatment with the hydroalcoholic extract of P. fraternus . Trypan blue viability assay was also performed. Apoptosis induction in the cells posttreatment was determined by DNA fragmentation assay, Agarose gel electrophoresis, and Acridine orange/Ethidium bromide dual staining. Protein isolation and analysis was carried out using the standard polyacrylamide gel electrophoresis protocols. RESULTS: The extracts inhibited the growth and proliferation of Daudi cells through induced cell death, which was dose-dependent and time-dependent. The IC50 value was found to be 220 µg/ml after 72 h of treatment. The induction of DNA fragmentation and increase in a number of apoptotic cells posttreatment suggest the possibility of apoptosis induction. A significant decrease in protein level was also observed. CONCLUSION: The results raise the possibility that the hydroalcoholic extract of P. fraternus could be a potent chemotherapeutic agent for the treatment of various cancers. Further evaluation of its potency as a chemotherapeutic agent is imperative.

Ficoll-400 density gradient method as an effective sperm preparation technique for assisted reproductive techniques.

BACKGROUND: Infertility being a burning issue, the male itself contributes about 40% as a cause, as evident by statistical data. However, Assisted Reproductive Technology (ART) has emerged as a powerful tool in the management of infertility. Sperm preparation techniques govern the selection procedure to separate functional spermatozoa which can then be used in IUI, IVF, and ART and for cryopreservation. AIM: The present study was aimed at evaluation of sperm preparation techniques for reliability, performance and to determine the most effective, feasible and economical technique. MATERIALS AND METHODS: The subjects under study includes males with normal proven fertility (n=40) and the males with unexplained infertility (n=40). Four sperm separation techniques, viz., Swim-up, Swim-down, Sucrose and Ficoll-400 density gradient techniques were evaluated for their efficacy in separation of good quality fraction of spermatozoa. Sperm viability, morphology and maturation status of spermatozoa were taken as evaluation parameters following the standard methods (WHO 2010). STATISTICAL ANALYSIS: Data was analyzed using student's t-test and the four selected techniques were compared with the normal semen samples for scoring the efficiency of the techniques. RESULTS AND DISCUSSION: Out of the several techniques used, Ficoll-400 was found to be more efficient method for separation of spermatozoa. The percentage of change in each parameter was calculated and taken as the index for recovery of potent sperm from the original sperm. Ficoll-400 density gradient yielded higher percentage of live, mature, morphologically normal spermatozoa in an isolated fraction as compared to other three techniques. CONCLUSION: It was observed that a combination of Ficoll-400 gradient separation with Swim-up technique could give quality spermatozoa which in-turn would directly have an impact on the success of IVF and other ART techniques.

Molecular interaction of selected phytochemicals under the charged environment of Plasmodium falciparum chloroquine resistance transporter (PfCRT) model.

Phytochemicals of Catharanthus roseus Linn. and Tylophora indica have been known for their inhibition of malarial parasite, Plasmodium falciparum in cell culture. Resistance to chloroquine (CQ), a widely used antimalarial drug, is due to the CQ resistance transporter (CRT) system. The present study deals with computational modeling of Plasmodium falciparum chloroquine resistance transporter (PfCRT) protein and development of charged environment to mimic a condition of resistance. The model of PfCRT was developed using Protein homology/analogy engine (PHYRE ver 0.2) and was validated based on the results obtained using PSI-PRED. Subsequently, molecular interactions of selected phytochemicals extracted from C. roseus Linn. and T. indica were studied using multiple-iterated genetic algorithm-based docking protocol in order to investigate the translocation of these legends across the PfCRT protein. Further, molecular dynamics studies exhibiting interaction energy estimates of these compounds within the active site of the protein showed that compounds are more selective toward PfCRT. Clusters of conformations with the free energy of binding were estimated which clearly demonstrated the potential channel and by this means the translocation across the PfCRT is anticipated.

Ameliorative Effects of Curcumin on Artesunate-Induced Subchronic Toxicity in Testis of Swiss Albino Male Mice.

India is one of the endemic areas where control of malaria has become a formidable task. Artesunate is the current antimalarial drug used to treat malaria, especially chloroquine resistant. The objective of the present study was to investigate the dose-dependent effect of oral administration of artesunate on the oxidative parameters in testes of adult male Swiss albino mice and ameliorative efficacy of curcumin, a widely used antioxidant. An oral dose of 150 mg/kg body weight (bwt; low dose) and 300 mg/kg bwt (high dose) of artesunate was administered for a period of 45 days to male mice, and ameliorative efficacy of curcumin was also assessed. The results revealed that artesunate caused significant alteration in oxidative parameters in dose-dependent manner. Administration of artesunate brought about significant decrease in activities of superoxide dismutase, glutathione, glutathione peroxidase, and glutathione reductase, whereas lipid peroxidation and glutathione-S-transferase activity were found to be significantly increased. The results obtained show that oxidative insult is incurred upon the intracellular antioxidant system of testis tissue by artesunate treatment. Further, administration of curcumin at the dose level of 80 mg/kg bwt along with both doses of artesunate attenuated adverse effects in male mice.

A Computational Approach towards the Understanding of Plasmodium falciparum Multidrug Resistance Protein 1.

The emergence of drug resistance in Plasmodium falciparum tremendously affected the chemotherapy worldwide while the intense distribution of chloroquine-resistant strains in most of the endemic areas added more complications in the treatment of malaria. The situation has even worsened by the lack of molecular mechanism to understand the resistance conferred by Plasmodia species. Recent studies have suggested the association of antimalarial resistance with P. falciparum multidrug resistance protein 1 (PfMDR1), an ATP-binding cassette (ABC) transporter and a homologue of human P-glycoprotein 1 (P-gp1). The present study deals about the development of PfMDR1 computational model and the model of substrate transport across PfMDR1 with insights derived from conformations relative to inward- and outward-facing topologies that switch on/off the transportation system. Comparison of ATP docked positions and its structural motif binding properties were found to be similar among other ATPases, and thereby contributes to NBD domains dimerization, a unique structural agreement noticed in Mus musculus Pgp and Escherichia coli MDR transporter homolog (MsbA). The interaction of leading antimalarials and phytochemicals within the active pocket of both wild-type and mutant-type PfMDR1 demonstrated the mode of binding and provided insights of less binding affinity thereby contributing to parasite's resistance mechanism.