Quantification of cell viability and proliferation form the fundamental for numerous in vitro assays in response to external factors. An MTT assay is a colorimetric assay based on assessing the cell metabolic activity. A549 Lung adenocarcinoma cell line was used to see the cytotoxic potential of a new drug for initial screening of apoptosis or necrosis. The biochemical mechanism behind the MTT assay involves NAD(P)H-dependent cellular oxidoreductase enzyme that converts the yellow tetrazolium MTT [3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] into insoluble (E,Z)-5-(4,5-dimethylthiazol-2-yl)-1,3-diphenylformazan (formazan). The formed formazan can be dissolved with dimethyl sulfoxide (DMSO) to give a purple color with characteristic absorption at 540 nm. Intensity of purple color is directly proportional to the cell number and thus indicating the cell viability.Video Clip of Methodology: 3 min 56 sec Full Screen Alternate

The objectives of this study were to determine phenolic content and antioxidant activities of chloroform, acetone, methanol and hot water extracts of Bauhinia vahlii leaves. The hot water extract afforded the highest yield (6.3%) while the lowest yield was obtained from the chloroform extract (2.1%). The methanol extract contains higher levels of total phenolics (48.7 ± 0.7 g GAE/100 g extract), tannins (21.7 ± 0.7 g GAE/100 g extract) and flavonoids (10.3 ± 0.2 RE/100 g extract). The extracts were subjected to assess their antioxidant potential using various in vitro systems such as DPPH(•), ABTS(•+), FRAP, OH(•), β-carotene linoleic acid bleaching system, phosphomolybdenum reduction and Fe(2+) chelation. It is concluded that the methanolic extract of B. vahlii leaves have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidants, which may serve as a potential source of natural antioxidants.

Bacterial resistance to antibiotics poses a serious threat to cure diseases associated with microbial infection. Among the resistant bacteria, extended-spectrum β-lactamase (ESBL)-producing bacteria are the most concerned one as they encode the enzyme β-lactamase that confers resistance to most β-lactam antibiotics. The present study was carried out to determine the antimicrobial potential and the principle mechanism of action of carvacrol against ESBL Escherichia coli isolated from ascitic fluid of a patient having a urinary tract infection. Carvacrol exhibited a minimum inhibitory concentration (MIC) of 450 μg/ml at which it reduced E. coli cell counts significantly in a time-dependent manner. Carvacrol completely diminished the growth of E. coli after 2 h of incubation at its MIC. Fluorescent imaging displayed the elevated reactive oxygen species level and bacterial membrane depolarization leading to E. coli cell death in presence of carvacrol at its MIC. Furthermore, carvacrol displayed a severe detrimental effect on bacterial membrane disruption and cellular material release. In addition, a significant effect of carvacrol at sub-inhibitory concentration was observed on motility of E. coli cells and invasion of human colon HCT-116 cells in an ex vivo model. Based on the results, we conclude a potential antimicrobial role of carvacrol against ESBL E. coli.

Carvacrol is present abundantly in the essential oils of many medicinal plants and well known for its numerous biological activities. Since partial solubility in water and physicochemical instability limits its industrial uses, the present study was performed to prepare a carvacrol nanoemulsion (CANE) using an ultrasonication technique and further evaluation of its anticancer potential against human lung adenocarcinoma A549 cells. The nanoemulsion formulation was optimized by varying carvacrol and polysorbate 80 ratios and characterized by dynamic light scattering (DLS), which revealed a negative surface charge with a mean droplet size between 105.5 ± 3.4 to 169.8 ± 4.9 nm. The CANE induced reactive oxygen species (ROS) production in A549 cells, leading to activation of key regulators of apoptosis such as p-JNK, Bax and Bcl2 as well as release of cytochrome C, and activation of the caspase cascade. Suppression of mitochondrial ROS using Mito-TEMPO reversed the apoptotic potential of CANE signifying involvement of mitochondrial ROS in cell death. Beside, CANE displayed a strong antitumor potential in vivo using an athymic nude mice model. The results strongly support that CANE induced apoptosis in A549 cells by induction of ROS and could be a promising candidate for lung cancer therapy.

Vitexin induces apoptosis by suppressing autophagy in multi-drug resistant colorectal cancer cells Vitexin induces apoptosis by suppressing autophagy in multi-drug resistant colorectal cancer cells