A new assay for diaphorase activity in reagent formulations, based on the reduction of thiazolyl blue.

1979 ◽  
Vol 25 (12) ◽  
pp. 2040-2042 ◽  
Author(s):  
R S Boethling ◽  
T L Weaver
Keyword(s):  
Nonlinear Kinetics ◽  
Tetrazolium Bromide ◽  
Diaphorase Activity ◽  
Blue Tetrazolium ◽  
Maximum Absorbance ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Assay Procedure ◽  
Thiazolyl Blue Tetrazolium

Abstract This new assay procedure for diaphorase eliminates problems of high blank rates and nonlinear kinetics associated with other methods. The dye thiazolyl blue tetrazolium bromide is reduced in the presence of NADH and diaphorase to yield a colored formazan, which as maximum absorbance at 560 nm.

Colorimetry of diaphorase in commercial preparations and clinical chemical reagents by use of tetrazolium salts.

1981 ◽  
Vol 27 (10) ◽  
pp. 1686-1689 ◽  
Author(s):  
F J Gella ◽  
M T Olivella ◽  
F Pegueroles ◽  
J Gener
Keyword(s):  
Enzyme Concentration ◽  
Tetrazolium Salts ◽  
Chemical Reagents ◽  
Diaphorase Activity ◽  
Linear Kinetics ◽  
Assay Procedure ◽  
Clinical Chemical ◽  
Iodonitrotetrazolium Chloride

Abstract We describe a procedure for assay of diaphorase activity in commercial purified preparations and in clinical chemical reagents by use of iodonitrotetrazolium chloride or other tetrazolium salts. The method is based on measurement of the formazan produced by enzymic reduction of tetrazolium salts in the presence of NADH. The assay procedure has been optimized for linear kinetics, simplicity of operation, nondetectable blank rates, and extended activity/enzyme concentration proportionality. The proposed method has several advantages over the older assay by use of dichlorophenolindophenol.

In vitro cytotoxicity investigations of chloroform extract of Anisomeles malabarica on vero and lung cancer [A-549] cell lines

2021 ◽  
Vol 16 (12) ◽  
pp. 95-99
Author(s):  
Duguluri Sajusha ◽  
Sivagnanam Selvakumar
Keyword(s):  
Lung Cancer ◽  
Cell Lines ◽  
Human Lung ◽  
Chloroform Extract ◽  
In Vitro Cytotoxicity ◽  
Human Lung Cancer ◽  
Lung Cancer Cell ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Thiazolyl Blue Tetrazolium

The main objective of this study is to investigate the cytotoxic potential of chloroform extract of medicinal plant Anisomeles malabarica. Successive solvent extraction of Anisomeles malabarica in chloroform was done. The extracts were tested against normal cell lines (Vero) human lung cancer cell lines (A-549) using the thiazolyl blue tetrazolium bromide test (MTT) assay. The results of the present investigation revealed that the chloroform extract of Anisomeles malabarica shown anti-lung cancer activity. The evaluation of the toxicity of medicinal plants and their herbal preparations is essential to determine the applicability of the samples as pharmacological drugs. Further studies are necessary for more extensive pharmacological and toxicological evaluations.

scholarly journals Synthesis, Crystal Structure and Bioactivity of Phenazine-1-carboxylic Acylhydrazone Derivatives

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5320
Author(s):  
Shouting Wu ◽  
Xi Liang ◽  
Fang Luo ◽  
Hua Liu ◽  
Lingyi Shen ◽  
...  
Keyword(s):  
Carboxylic Acid ◽  
Condensation Reaction ◽  
Acid Hydrazide ◽  
13C Nmr Spectroscopy ◽  
X Ray Diffraction ◽  
1H And 13C Nmr ◽  
X Ray ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Thiazolyl Blue Tetrazolium

A phenazine-1-carboxylic acid intermediate was synthesized from the reaction of aniline and 2-bromo-3-nitro-benzoic acid. It was then esterified and reacted with hydrazine hydrate to afford phenazine-1-carboxylic hydrazine. Finally, 10 new hydrazone compounds 3a–3j were obtained by the condensation reaction of phenazine-1-carboxylic acid hydrazide and the respective aldehyde-containing compound. The structures were characterized by 1H and 13C NMR spectroscopy, MS and single crystal X-ray diffraction. The antitumor activity of the target compounds in vitro (HeLa and A549) was determined by thiazolyl blue tetrazolium bromide. The results showed that compound (E)-N′-(2-hydroxy-4-(2-(piperidine-1-yl) ethoxy) benzyl) phenazine-1-carbonyl hydrazide 3d exhibited good cytotoxic activity.

scholarly journals Structural Properties, Cytotoxicity, and Anti-Inflammatory Activity of Silver(I) Complexes with tris(p-tolyl)Phosphine and 5-Chloro-2-Mercaptobenzothiazole

2010 ◽  
Vol 2010 ◽  
pp. 1-12 ◽  
Author(s):  
L. Kyros ◽  
N. Kourkoumelis ◽  
M. Kubicki ◽  
L. Male ◽  
M. B. Hursthouse ◽  
...  
Keyword(s):  
Inflammatory Activity ◽  
Flow Cytometry Assay ◽  
X Ray Crystallography ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Diphenyltetrazolium Bromide ◽  
Anti Inflammatory ◽  
Mtt Assays ◽  
Thiazolyl Blue Tetrazolium

The synthesis and characterization of the silver(I) chloride complex of formula{[AgCI(CMBZT)(TPTP)2]⋅(MeOH)}(1) (CMBZT = 5-chloro-2-mercaptobenzothiazole, TPTP = tris(p-tolyl)phosphine) is described. Also the structure of the hydrate derivative{[AgCI(TPTP)3]⋅(0.5⋅H2O)}(2) of the corresponding known anhydrous silver complex (Zartilas et al., 2009), and the polymorph3of the known[AgI(TPTP)3]complex (Zartilas et al., 2009) were determined and compared with the known ones. In addition, the structure of the known one silver(I) cluster{[AgI(TPTP)]4}(4) (Meijboom et al., 2009) was re-determined at 120(2) K and possible Ag-Ag interactions were analyzed. The compounds1–4were characterized by X-ray crystallography at r.t (1) and 120 K (2–4). All these complexes and{[(Et3NH)+]2⋅[Ag6(μ3-Hmna)4(μ3-mna)2]2−⋅(DMSO)2⋅(H2O)}(5) (Hmna = 2-mercaptonicotinic acid) were evaluated for cytotoxic and anti-inflammatory activity. Thein vitrotesting of cytotoxic activity of1–5against leiomyosarcoma cancer cells (LMS), were evaluated with Trypan Blue and Thiazolyl Blue Tetrazolium Bromide or 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assays. The flow cytometry assay for complex1and showed that at 15 μMof1, 62.38% of LMS cells undergo apoptosis, while 7% of LMS cells undergo cell necrosis. The antitumor activity of3is comparable with that of its reported polymorph (Zartilas et al., 2009). The anti-inflammatory, activity of complexes1–3and5was also studied. The activity towards cell viability was2>3>5>1>4, while the order of the inhibitory activity in cell growth proliferation follows the order,2>3>1>4>5. The anti-inflammatory activity on the other hand is1>2>5>⋯>3.

scholarly journals Screening of some Cameroonian Medicinal Plants against Bacterial and Yeasts involved in Gastrointestinal Disorders

2017 ◽  
Vol 9 (3) ◽  
pp. 56
Author(s):  
Laure Brigitte Kouitcheu Mabeku ◽  
Tchouangueu Thibau Flaurant ◽  
Kouam Jacques
Keyword(s):  
Colorimetric Assay ◽  
Folk Medicine ◽  
Antimicrobial Activities ◽  
Gastrointestinal Disorders ◽  
Plant Sample ◽  
Phytochemical Screening ◽  
Mimosa Pudica ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Thiazolyl Blue Tetrazolium ◽  
Ethyl Acetate Extracts

The present study was designated to evaluated the antimicrobial activities of methanol and ethyl acetate extracts of Carcia arereh, Nelsonia canescens, Ficus thonningu, Bryophillum pinnatum, Cyclosorus striatus, Euphorbia cordifolia, Euphorbia hirta, Erygium foetidum and Mimosa pudica. The selected plants species are used as traditional folk medicine in Cameroon for the treatment of various diseases. Thiazolyl blue tetrazolium bromide colorimetric assay was used to evaluate the antimicrobial activity against 16 microbial species. Preliminary phytochemical screening of plant sample was also carried out. Alkaloids and anthraquinones were the most frequent secondary metabolite in the tested plant extracts. The MIC values obtained ranging from 32 to >1024

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

2021 ◽  
pp. 1-9
Author(s):  
Jie Wang ◽  
Jialing You ◽  
Ding Gong ◽  
Ying Xu ◽  
Bo Yang ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Collagen Synthesis ◽  
Akt Signaling ◽  
Control Group ◽  
Type I ◽  
Akt Signaling Pathway ◽  
Western Blots ◽  
Tetrazolium Bromide ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
The Impact

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.

scholarly journals microRNA-204-5p participates in atherosclerosis via targeting MMP-9

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 231-239 ◽  
Author(s):  
Na Wang ◽  
Yuliang Yuan ◽  
Shipeng Sun ◽  
Guijian Liu
Keyword(s):  
Transwell Assay ◽  
Blood Samples ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Promising Therapeutic Target ◽  
Matrix Metallopeptidase ◽  
Matrix Metallopeptidase 9 ◽  
And Migration ◽  
Thiazolyl Blue Tetrazolium ◽  
Normal Controls ◽  
Proliferation And Migration

AbstractThe aim of the present study was to investigate the role and mechanism of microRNA-204-5p (miR-204-5p) in atherosclerosis (AS)-related abnormal human vascular smooth muscle cells (hVSMCs) function. Firstly, we analyzed the expression of miR-204-5p and found that the miR-204-5p expression level was clearly downregulated in atherosclerotic plaque tissues and blood samples compared to the normal controls. Then, matrix metallopeptidase-9 (MMP-9) was predicted to be the potential target of miR-204-5p by TargetScan and this prediction was confirmed by luciferase assays. Besides, we observed that miR-204-5p could negatively regulate the expression of MMP-9 in hVSMCs. Subsequently, Thiazolyl Blue Tetrazolium Bromide (MTT) assay, transwell assay and flow cytometry were performed to detect the proliferation, migration and apoptosis of hVSMCs. Down-expression of miR-204-5p led to the promotion of proliferation and migration accompanied with the suppression of apoptosis in hVSMCs, and these effects were reversed by MMP-9-siRNA. In addition, overexpressed miR-204-5p could inhibit hVSMC proliferation and migration and promote the apoptosis of hVSMCs. However, the effects were also abrogated by overexpressed MMP-9. Together, our findings showed that miR-204-5p plays an important role in the growth and migration of hVSMCs by targeting MMP-9, which might be a novel biomarker and promising therapeutic target for AS.

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