Improved Formazan Dissolution for Bacterial MTT Assay

Reduction of the water-soluble tetrazolium salt 3-(4,5-dimethylthiazol)-2,5 diphenyl-tetrazolium bromide (MTT) to purple, water-insoluble formazan is commonly used for assessment of cell viability and proliferation. Spectrophotometric detection of formazan requires its solubilization.

INFLUENCE OF CHITOSAN SOLUTION ON THE VIABILITY OF ARBUSCULAR MYCORRHIZAL SPORES

The study was aimed to fortify the Arbuscular Mycorrhizal Fungal (AMF) spores, widely used in Bio-fertilizers with chitosan and check the inuence of chitosan on the viability of AMF spores. Chitosan was prepared from shrimp shells using chemical method involving demineralization, deproteinization and deacetylation.AMF spores were kept in three different concentrations of chitosan (0.1%, 0.5 %and 1%) which was prepared in 0.1% acetic acid and 0.1% ascorbic acid. Spore viability was checked by the MTT 3-(4, 5 th th th dimethylthiazol-yl-2, 5-diphenyl-2H-tetrazolium bromide) after a day, 10 day, 20 day and 30 day. Highest number of viable spores was observed in chitosan dissolved in 0.1% ascorbic acid as compared to chitosan in acetic acid.

PDGF-BB induces conversion, proliferation, migration, and collagen synthesis of oral mucosal fibroblasts through PDGFR-β/PI3K/ AKT signaling pathway

OBJECTIVE: To explore the pathogenesis of oral submucosal fibrosis (OSF) by analyzing the impact of Platelet Derived Growth Factor (PDGF)-BB on oral mucosal fibroblasts (FB) and PDGFR-β/Phosphoinositide 3-kinase (PI3K)/serine/threonine protein kinase (AKT) signaling pathway. METHODS: The isolated and purified oral mucosal fibroblasts were divided into four groups: the control group (CON, 10% FBS DMEM), the PDGF-BB group (40 ng/ml PDGF-BB), the PDGF-BB+IMA group (40 ng/ml PDGF-BB and 60 μmol/L IMA), and the PDGF-BB+LY294002 group (40 ng/ml PDGF-BB and 48 μmol/L LY294002). Primary human FB cells were isolated and cultured for detecting the effects of PDGF-BB on α-smooth muscle actin (α-SMA) by indirect immunofluorescence. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H -tetrazolium bromide, Thiazolyl Blue Tetrazolium Bromide (MTT) method and scratch test were used to detect the proliferation and migration of FB. Western Blots were used to detect the synthesis of type I collagen (Col I) and the expression of PDGFR-β/PI3K/AKT signaling pathway-related proteins. The effects of PDGFR-β inhibitor and PI3K inhibitor were observed. RESULTS: Compared with group CON, group IMA, and group LY294002, α-SMA was upregulated in group PDGF-BB (p< 0.05), with higher OD490 nm value (p< 0.05), narrower average scratch width, and higher relative cell migration rate (p< 0.05). The expression levels of Col I, p-PDGFR-β, p-PI3K, and p-AKT were higher in group PDGF-BB (p< 0.05). CONCLUSIONS: PDGF-BB induces FB to transform into myofibroblasts (MFB) through the PDGFR-β/PI3K/AKT signaling pathway, and promotes the proliferation, migration, and collagen synthesis.

The use of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide–based colorimetric assay in the viability analysis of the filamentous cyanobacterium Arthrospira platensis

ABSTRACT The applicability of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to an industrially valuable filamentous cyanobacterium Arthrospira platensis was examined. When it was applied to A. platensis NIES-39, as few as 10 viable trichomes were quantitatively detected. However, depending on the experimental conditions, it also generated artifactual viability signals. The results should help clarify the scope and limits of the MTT assay in viability analysis.

Antiproliferative Activity of Ethanol Extract of Boswellia dalzielii (Hutch) Stem Bark in Breast Cancer Cell Line

Aim and Objectives: The present study aimed to examine the antiproliferative activity of ethanol extract derived from the Boswellia dalzielii stem bark in breast carcinoma. Methodology: Ethanolic extract of stem bark of Boswellia dalzielii were prepared. Antiproliferative activity was assessed in estrogen receptor (ER)-negative breast carcinoma (MDA-MB-231) cells by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. Results: Obtained results in 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay indicated that ethanol extract of Boswellia dalzielii stem bark showed significant antiproliferative activity in MDA-MB-231 cells in a dose-dependent manner. IC50 of Boswellia dalzielii stem bark ethanol extract in MDA-MB231 was 98. 12 μg/mL. Conclusion: The results obtained suggest that Boswellia dalzielii stem bark extract possesses significant antiproliferative potential which could be mediated by the chemical constituents present in the plant. However, further research needs to be carried out to determine the effect of the extract on the cell cycle and also to determine the type of cell death produced by the extract.

Design, Synthesis, and Preliminary Antitumor Activity Evaluation of Novel Alkaloid Derivatives

A novel alkaloid scaffold was designed through scaffold-hopping strategy based on the active pyrazines alkaloid isolated previously. A total of 25 derivatives were synthesized based on this scaffold and evaluated for their antitumor activities. Among all these tested compounds, 9f exhibited most excellent antitumor activities toward H460 cells, TMD-8 cells, and MV4-11 cells in vitro by 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide assay with IC50 values of 29.8, 14.9, and 18.8 μM, respectively.

Inhibitory Activity of HEp-2 Cells by Honey from Indonesia

Indonesian local honey contains active compounds that have the potential as an antioxidant and anticancer, primarily as a laryngeal anticancer through the inhibition of HEp-2 cells. This study aims to determine the anticancer activity of several types of honey in Indonesia through the inhibition of HEp-2 cells. Samples used in the form of Trigona, Longan, Rambutan, and Kaliandra honey obtained from honey farmers in Sulawesi and Java, Indonesia. Honey samples extracted by using methanol, then liquids partition was carried out consecutively using n-hexane and ethyl acetate. The cytotoxicity test for HEp-2 cells was carried out using the MTT method (3- (4,5-dimethyl thiazol-2-il) -2,5-diphenyl tetrazolium bromide). The results showed that all honey samples were active against preventing HEp-2 cells with the highest inhibition activity from longan honey with ethyl acetate extract at 65.18% at 100 ppm. Longan honey has decreased HEp-2 cell inhibitory activity after fractionation. Indonesian local honey, namely trigona honey, kaliandra honey, rambutan honey, and longan honey, can be used as a supplementary supplement for patients with laryngeal cancer.