Phytochemical analysis of Berberis lyceum methanolic extract and its antiviral activity through the restoration of MAPK signaling pathway modulated by HCV NS5A

Objective:To evaluate the antiviral activity and phytochemicals of selected plant extracts and their effect on the mitogen-activated protein kinase(MAPK)signaling pathway modulated by hepatitis C virus(HCV)nonstructural protein 5 A(NS5A).Methods:A total of ten plant extracts were initially screened for their toxicities against Hep G2 cells.The non-toxic plants were tested for their inhibitory effect on the expression of HCV NS5A at both m RNA and protein levels using real-time PCR and Western blotting assays,respectively.The differential expression of the genes associated with MAPK pathway in the presence of NS5A gene and plant extract was measured through real-time PCR.Subsequently,the identification of secondary metabolites was carried out by phytochemical and HPLC analysis.Results:The phytochemical profiling of Berberis lyceum revealed the presence of alkaloids,phenols,saponins,tannins,flavonoids,carbohydrates,terpenoids,steroids,and glycosides.Similarly,quercetin,myricetin,gallic acid,caffeic acid,and ferulic acid were identified through HPLC analysis.The methanolic extract of Berberis lyceum strongly inhibited HCV RNA replication with an IC50 of 11.44μg/m L.RT-PCR and Western blotting assays showed that the extract reduced the expression of HCV NS5A in a dosedependent manner.Berberis lyceum extract also attenuated NS5A-induced dysregulation of the MAPK signaling pathway.Conclusions:Our findings suggest that Berberis lyceum extract strongly inhibits HCV propagation by reducing HCV NS5A-induced perturbation of MAPK signaling.

Mutagenic and cytotoxic potential of Endosulfan and Lambda-cyhalothrin—In vitro study describing individual and combined effects of pesticides

Excessive use of pesticides poses increased risks to non target species including humans. In the developing countries, lack of proper awareness about the toxic potential of pesticides makes the farmer more vulnerable to pesticide linked toxicities, which could lead to diverse pathological conditions. The toxic potential of a pesticide could be determined by their ability to induce genetic mutations and cytotoxicity. Hence, determination of genetic mutation and cytotoxicity of each pesticide is unavoidable to legislate health and safety appraisal about pesticides. The objective of current investigation was to determine the genotoxic and cytotoxic potential of Endosulfan（EN） and Lambda-cyhalothrin（LC）; individually and in combination. 3-（4,5-Dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium Bromide（MTT） assay was utilized to determine cytotoxicity, while two mutant histidine dependent Salmonella strains（TA98, TA100） were used to determine the mutagenicity of EN and LC.Moreover, mutagenicity assay was conducted with and without S9 to evaluate the effects of metabolic activation on mutagenicity. Even though a dose dependent increase in the number of revertant colonies was detected with EN against both bacterial strains, a highly significant（p 〈 0.05） increase in the mutagenicity was detected in TA98 with S9. In comparison, data obtained from LC revealed less mutagenic potential than EN. Surprisingly,the non-mutagenic individual-concentrations of EN and LC showed dose dependent mutagenicity when combined. Combination of EN and LC synergistically induced mutagenicity both in TA98 and TA100. MTT assay spotlighted comparable dose dependent cytotoxicity effects of both pesticides. Interestingly, the combination of EN and LC produced increased reversion and cytotoxicity at lower doses as compared to each pesticide, concluding that pesticide exposure even at sub-lethal doses can produce cytotoxicity and genetic mutations, which could lead to carcinogenicity.

Sero-Epidemiological Survey of Crimean-Congo Hemorrhagic Fever among the Human Population of the Punjab Province in Pakistan

Dear Editor,Crimean-Congo hemorrhagic fever(CCHF),caused by the CCHF virus(CCHFV),is a tick-borne zoonotic infection characterized by myalgia,high-grade fever(>38℃),headache,nausea,bleeding from the body cavities,and in 10%–50%of cases,results in death(Swanepoel et al.1989).As CCHFV belongs to the Nairoviridae family,the virus can be transmitted to humans through the bite of infected ticks or by contact with the tissues or blood of infected animals(Bente et al.2013).