Virtual screening-based identification of novel fatty acid synthase inhibitor and evaluation of its antiproliferative activity in breast cancer cells.

Cancer cells activate de novo lipogenesis by overexpressing the lipogenic enzymes ACLY, ACC and FASN to support rapid cell division. FASN, previously known as oncogenic antigen-519 (OA-519) catalyzes seven sequential reactions to synthesize palmitic acid (C16) from substrates acetyl CoA, and malonyl CoA. The dependence of cancer cells on FASN-derived lipids and the differential expression of FASN in cancer cells compared to their normal counterparts make it an attractive metabolic drug target in cancer therapy. In the present study, an attempt has been made to identify potent FASN inhibitors from Asinex-Synergy compound database using structure-based virtual screening. The serial docking protocols of increasing precisions identified LEG-17649942, with glide score -10.34 kcal/mol as a promising compound which can directly interact with active site residues H293 and H331. LEG-17649942 possesses drug-like pharmacokinetic properties as predicted by Qikprop. LEG-17649942 exhibited cytotoxicity in breast cancer cell lines SK-BR-3, MCF-7 and MDA-MB-231 with maximum activity against MDA-MB-231 cells with IC50 of 50 µM. The study put forward LEG-17649942 as a novel drug-lead compound against triple negative breast cancer with an exquisite binding pattern to FASN-KS domain.

Comparative assessment of heat stress induced changes in carcass traits, plasma leptin profile and skeletal muscle myostatin and HSP70 gene expression patterns between indigenous Osmanabadi and Salem Black goat breeds.

The primary objective of the study was to compare the impact of heat stress on meat production characteristics of Osmanabadi and Salem Black breed goats based on changes in carcass characteristics, meat quality attributes, plasma leptin concentration, skeletal muscle myostatin and heat shock protein 70 (HSP70) gene expression patterns. The goats were randomly distributed into four groups: OSC (n = 6; Osmanabadi Control), OSHS (n = 6; Osmanabadi Heat Stress), SBC (n = 6; Salem Black Control) and SBHS (n = 6; Salem Black Heat Stress). The animals were slaughtered at the end of the study and their meat characteristics were assessed. This study established the impact of heat stress on a wide variety of carcass and meat quality characteristics in OS and SB goat breeds. The results from the study also provided some crucial evidence for a better resilience capacity of Salem Black breed as compared to Osmanabadi goats in maintaining the meat production during heat stress. The study also established plasma leptin and HSP70 genes to be the ideal biomarkers to reflect the impact of heat stress on meat characteristics in indigenous goats.

Comparative assessment of growth performance of three different indigenous goat breeds exposed to summer heat stress.

A study was conducted to assess comparatively the growth performance of three different indigenous goat breeds during exposure to summer heat stress. The primary objective of the study was to observe the heat stress impact on the growth performance based on the body weight changes, allometric measurements, growth hormone (GH) concentration and peripheral blood mononuclear cell (PBMC) Insulin-like growth factor-1 (IGF-1) mRNA expression pattern during the summer season in comparison with the local breed (Osmanabadi). Thirty-six ten-month- to one-year-old female goats of Osmanabadi, Malabari and Salem Black breeds were randomly divided into six groups, OC (n = 6; Osmanabadi control), OHS (n = 6; Osmanabadi heat stress), MC (n = 6; Malabari control), MHS (n = 6; Malabari heat stress), SBC (n = 6; Salem Black control) and SBHS (n = 6; Salem Black heat stress). Body weight was recorded at weekly intervals, whereas other growth and allometric measurements and blood collection were carried out at fortnightly intervals. Breed factor significantly (p < .05) influenced only few growth variables such as body weight, body mass index (BMI) and body condition score (BCS). However, heat stress treatment significantly (p < .05) reduced all growth parameters expect BMI. Further, the heat stress significantly (p < .01) increased plasma GH concentration in goats with significantly higher (p < .05) concentration recorded in OHS. Among the stress groups, the lower (p < .05) PBMC IGF-1 mRNA expression was recorded in OHS, while the higher (p < .05) expression was observed in SBHS indicating the extreme adaptive capability of Salem Black breed. Thus, the results indicated that the Salem Black breed performed much better compared to both Osmanabadi and Malabari breeds indicating the superior ability of this breed to adapt to heat stress challenges. The results also indicated that plasma GH and IGF-1 gene may act as ideal biomarkers for assessing the heat stress impact on growth performance in indigenous goats.

Modulation of gamma ray-induced genotoxic effect by thymol, a monoterpene phenol derivative of cymene.

The radioprotective effect of thymol (TOH), a monoterpene phenol, on radiation-induced DNA damage was analyzed in vitro. Chinese hamster lung fibroblast cells (V79) were treated with different concentrations of TOH (0-100 µg/mL) for 1 hour before exposure to 3 Gy gamma irradiation, and then cytokinesis-blocked micronucleus and single-cell gel electrophoresis (comet assay) assays were used to evaluate the radiation-induced cytogenetic damage and genotoxic effects. Furthermore, the modulating effect of TOH on radiation-induced cell death was assessed by apoptotic and necrotic cell detection by staining with ethidium bromide/acridine orange using fluorescence microscopy. To understand the mechanism of TOH-imparted cytoprotection, mitochondrial membrane potential (MMP) was detected by flow cytometry after staining the cells with Rhodamine 123. Pretreatment of V79 cells with various concentrations of TOH (0-100 µg/mL) for 1 hour reduced the radiation-induced micronuclei as well as percent tail DNA and mean Olive tail moment with a maximum protective effect observed at TOH (25 µg/mL). Apoptosis by microscopic, MMP measurements indicated that the V79 cells exposed to gamma radiation alone showed a maximal increase in the number of early and late apoptotic and necrotic cell death associated with a significant loss of the MMP. Pretreatment with TOH (25 µg/mL) showed a significant (P < .01) decrease in the level of apoptotic fraction as well as necrotic cells and suppressed the radiation-induced collapse of MMP when compared with the radiation alone group. These results suggest that TOH suppresses radiation-induced genotoxicity, apoptosis, and necrosis primarily by the free radical scavenging and modulation of oxidative stress.

Antioxidant and radiation antagonistic effect of Saraca indica.

The present study was undertaken to investigate the radiation mitigating effect of hydro-alcoholic extract of Saraca indica (SIE) against mice exposed to whole body gamma radiation. Free radical scavenging ability by SIE was demonstrated using hydroxyl, superoxide anion, ABTS*+ and DPPH radicals generated in vitro. A significant increase in the animal survival (dose reduction factor = 1.39), inhibition in the decline of hematological parameters as well as increased number of spleen colony-forming units was observed when SIE was administered prior to radiation. SIE pretreatment increased the levels of glutathione, glutathione S- trans-ferase, catalase and lowered lipid peroxidation. Our findings for the first time demonstrate the protective potential of SIE against radiation induced syndromes with an optimal dose of 400 mg/kg b.wt. The radiation mitigating effect may be attributed to the mechanisms such as free radical scavenging, elevation in antioxidant status, and reduction in lipid peroxidation.

Thymol, a naturally occurring monocyclic dietary phenolic compound protects Chinese hamster lung fibroblasts from radiation-induced cytotoxicity.

The effect of thymol (TOH), a dietary compound was investigated for its ability to protect against radiation-induced cytotoxicity in Chinese hamster lung fibroblast (V79) cells growing in vitro. Treatment of V79 cells with 25 microg/ml of TOH prior to 10 Gy gamma radiation resulted increase in the cell viability than that of radiation alone as evaluated by MTT assay. Similarly, there was a significant increase in the surviving fraction observed with 25 microg/ml of TOH administered 1h prior to graded doses of gamma radiation. Further, 25 microg/ml TOH treatment before irradiation significantly decreased the percentage of radiation-induced apoptotic cells (sub-G(1) population) analyzed by flow cytometry as well as DNA ladder assay. TOH was found to inhibit various free radicals generated in vitro, viz., DPPH, O(2), ABTS(+) and OH in a concentration dependent manner. TOH also inhibited the radiation-induced decrease in intracellular glutathione, superoxide dismutase and catalase enzyme levels in V79 cells accompanied by the reduction in lipid peroxides. Our study demonstrated antagonistic potential of TOH against radiation-induced oxidative stress, lipid peroxidation resulting in increased cell viability.