Imran Altaf

Assessment of the cytotoxic and anti-viral potential of aqueous extracts from different parts of Acacia nilotica (Linn) Delile against Peste des petits ruminants virus

Peste des petits ruminants virus (PPRV); a negative sense single stranded RNA enveloped virus that causes Peste des petits ruminants (PPR), is dramatically affecting the health status of ruminants all around the world resulting in extensive economical losses in livestock sector. Acacia nilotica (Linn) Delile; a tannin-rich medicinal plant distributed throughout subcontinent, is traditionally used as food for ruminants and possesses anti-viral potential against different RNA viruses. In the current study, aqueous extracts from the bark, leaves and pods of A. nilotica (Linn) Delile indica were evaluated for their cytotoxicity and anti-viral activities against PPRV by adopting MTT colorimetric assay and anti-viral assay using Vero cell line. Aqueous extract from the leaves presented significantly better (P<0.05) anti-PPRV activities in comparison to pods extract. On the contrary, bark extract did not show any anti-viral activity. The data presented in the study could pave a way toward the discovery of novel anti-viral chemicals in the plants against PPRV and other viral diseases.

In vitro toxic action potential of anti tuberculosis drugs and their combinations

Tuberculosis (TB) is one of the leading infectious causes of death due to single infectious agent after HIV/AIDS. Rifampicin (RIF), Isoniazid (INH), Ethambutol (EMB), Pyrazinamide (PZA) and/or their combinations are extensively prescribed to treat TB. Despite several therapeutic implications, these drugs also produce several toxic effects at cellular level. MTT assay and Ames test were adopted in this study for the determination of cytotoxic and mutagenic potential of these anti-TB drugs. Among all tested drugs, cytotoxic potential of RIF was strongest with highly significant decline (p<0.001) in cell numbers at the concentration of 250μg/ml with LC50 at 325μg/ml, while significant decline (p<0.01) in cell count was observed in INH treated group at the concentration 500μg/ml with LC50 at 1000μg/ml. Moreover, combination RIPE demonstrated significant reduction (p<0.01) in cell number at the concentration of 25-500-500-500μg/ml with LC50 at 60-1200-1200-1200μg/ml. It is apparent from the data that almost all drugs represented identical mutagenic pattern i.e., more significant results were achieved in TA100 with metabolic activation (+S9). RIF proved to be highly mutagenic of all tested drugs with significant mutagenicity (p<0.01) at 0.0525μg/plate against TA98 strain with S9. The combination RIPE exhibited highly significant mutagenic activity (p<0.01) at concentration 0.125-3-3-3μg/plate without S9, while addition of S9 resulted in similar activity at lower doses, i.e., 0.0525-1-1-1μg/plate. It was concluded from the data that all anti-TB drugs possess significant cytotoxic and mutagenic potential, especially in combination, making TB patient more vulnerable to cytotoxic and mutagenic effects of anti-TB drugs, which could produce further health complications in TB patients.

Comparative study to evaluate the anti-viral efficacy of Glycyrrhiza glabra extract and ribavirin against the Newcastle disease virus

Background: The Newcastle disease represents as one of the most infectious viral disease, which afflicts almost every species of the birds. The causative agent of the disease is a single-stranded RNA virus with rapid replication capability. Objective: This study was performed to evaluate the comparative anti-viral efficacy and toxicity of Glycyrrhiza glabra aqueous extract and ribavirin against the Newcastle disease virus. Materials and Methods: The embryonated eggs were divided into six groups (A, B, C, D, E and F). Groups A, B, C, and D were further subdivided into three subgroups. The virus was identified by hemagglutination inhibition test. Spot hemagglutination test and viability of embryos were also evaluated. Three different concentrations i-e., 30 mg/100 ml, 60 mg/100 ml, and 120 mg/100 ml of the Glycyrrhiza aqueous extract and 10 μg/ml, 20 μg/ml, and 40 μg/ml ribavirin in deionized water were evaluated for their toxicity and anti-viral activity in the embryonated eggs. Results: 60 mg/100 ml concentration of Glycyrrhiza extract did not produce any toxicity in the embryonated eggs and showed anti-viral activity against the virus. Similarly, 20 μg/ml ribavirin was non-toxic in the embryonated eggs and contained anti-viral activity. Conclusion: It may conclude from the presented study that 60 mg/100 ml Glycyrrhiza extract inhibits replication of Newcastle disease virus and is non-toxic in the embryonated eggs. So, Glycyrrhiza glabra extract may be further evaluated in future to determine the potentially active compounds for their anti-viral activity against Newcastle disease virus. Furthermore, the mechanism of action of these active phytochemicals as an antiviral agent would be helpful to elucidate the pathogenesis of the disease.

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.

Evaluation of antifungal and antioxidant potential of two medicinal plants: Aconitum heterophyllum and Polygonum bistorta

ABSTRACT Objective To focus on the evaluation of antimicrobial and antioxidant activity of two endangered medicinal plants Aconitum heterophyllum (A. heterophyllum) and Polygonum bistorta (P. bistorta). Materials Plant extracts were obtained by using microwave assisted extraction method. The in vitro antifungal activity of A. heterophyllum and P. bistorta extracts were determined by measuring diameters of inhibitory zones of these extracts against Aspergillus niger and Alternalia solani. Results Methanolic extract of A. heterophyllum showed significant (P≤0.05) antifungal activity against both the tested organisms. It was also observed that ethanolic extracts of P. bistorta also had good antifungal activity against the tested fungal strains as compared to the methanolic extracts. It showed significant antifungal activity (P≤0.05) against both the tested strains. Antioxidant activity of methanolic and ethanolic extracts of A. heterophyllum and P. bistorta were also measured using a radical scavenging method. Ascorbic acid was used as a standard. Conclusions It was observed that A. heterophyllum and P. bistorta have significant antioxidant activity. Higher antioxidant activity was recorded in methanolic extract of A. heterophyllum as compared to its ethanolic extract. However, in case of P. bistorta ethanolic extract of the plant exhibited higher antioxidant potential than methanolic extracts. Hence both of these plants have significant antimicrobial as well as antioxidant potential.

In vitro evaluation of mutagenicity and genotoxicity of sitagliptin alone and in combination with artificial sweeteners

Purpose: To determine the in vitro genotoxicity and mutagenicity of sitagliptin alone and in combination with three commonly used artificial sweeteners (saccharin, aspartame and acesulfame-k). Methods: The in vitro genotoxicity and mutagenicity of Sitagliptin alone and in combination with three popular artificial sweeteners (saccharin, aspartame and acesulfame-k) were evaluated by Comet and Ames assays, respectively. Results: Sitagliptin demonstrated mutagenic potential only to TA 98 with S9 mix at a concentration of 3040 μg/plate. The mutagenicity of sitagliptin was enhanced when tested in combination with the artificial sweeteners. Furthermore, sitagliptin also caused pronounced DNA fragmentation at higher doses compared with negative control. Conclusion: At higher doses, sitagliptin showed both mutagenicity and genotoxicity. Thus, long-term use of artificial sweeteners with sitagliptin may lead to increase in both mutagenicity and genotoxicity. Keywords: Sitagliptin, Artificial sweeteners, Comet assay, DNA damage, Ames assay, Genotoxicity, Mutagenicity