A Triazophos-degrading strain, Klebsiella sp. E6, was identified by soil enrichment technology from the soil sampled from the vicinity of a factory manufacturing Triazophos (TAP). The nutrient requirement of the strain is simple. It can use TAP as the sole sources of carbon, nitrogen and phosphorus. Comparison of the degradation rates revealed that the strain degraded TAP most effectively when TAP was used as a sole nitrogen source. No inhibition effect occurred when TAP concentration was high as 1000 mg/L in the case of TAP was used as the sole nitrogen source. Analysis of the intermediates of TAP metabolism indicated that TAP is firstly hydrolyzed into 1-phenyl-3-hydroxy-1,2,4-triazole and O,O-diethyl phosphorothioic acid. 1-phenyl-3-hydroxy-1,2,4-triazole was further mineralized into inorganic compounds. A degradation pathway of TAP was proposed. The experiment results demonstrated that the strain has potential in biodegradation of TAP pollutions.
Biodegradation, Environmental ; Environmental Pollutants ; metabolism ; Klebsiella ; isolation & purification ; metabolism ; Organothiophosphates ; metabolism ; Pesticides ; metabolism ; Soil Microbiology ; Triazoles ; metabolism

In order to establish a fast and accurate method for novel DMIs fungicide screening, lanosterol 14alpha-demethylase of Magnaporthe grisea expressed in E. coli was used as target enzyme and the DMI fungicides diniconazole, tebuconazole, triadimenol and triadimefon were used as representative fungicides, the effects of enzyme activity, enzyme purity and concentration on the binding spectra were investigated. The results showed that active enzyme, elimination of interference of other P450s and proper enzyme concentration were necessary for obtaining accurate binding spectra. The Kd values of diniconazole, tebuconazole, triadimenol and triadimefon were 0.143 micromol/L, 0.24 micromol/L, 0.257 micromol/L and 0.307 micromol/L respectively, which significantly correlated to their 120h-EC50 values on the growth of Magnaporthe grisea. The results indicated that the binding spectra of fungicide and lanosterol 14alpha-demethylase can serve as a reliable and fast method for novel fungicide screening.
Cytochrome P-450 Enzyme System ; metabolism ; Fungicides, Industrial ; pharmacology ; Spectrophotometry ; methods ; Sterol 14-Demethylase ; Triazoles ; pharmacology

OBJECTIVETo investigate the safe use of 10% difenoconazole in planting Gentiana scabra.

METHODThe degradation dynamics of 10% ifenoconazole in the stems and leaves of G. scabra collecting in different time were determined by GC with ECD detection, and the half life of difenoconazole in the plant was calculated, and then the safe use method of 10% difenoconazole was formulated.

RESULTUnder the local climatic conditions, the half life of 10% difenoconazole was 6.84-6.90 days.

CONCLUSIONIn the good agricultural practice (GAP) of G. scabra, the maximal concentration of 10% difenoconazole is 400 g x ha(-1), the safety interval of using 10% difenoconazole is 40 days.

Agriculture ; methods ; Dioxolanes ; pharmacokinetics ; Gentiana ; metabolism ; Half-Life ; Plant Leaves ; metabolism ; Plant Stems ; metabolism ; Time Factors ; Triazoles ; pharmacokinetics

The ability of triadimefon (TDM), a triazolic fungicide, to alter the biochemical constituents and thereby minimizing the days required for sprouting in white yam (Dioscorea rotundata Poir.) tubers during storage under (30+/-2) degrees C in the dark, was studied. TDM at 20 mg/L was given to tubers by dipping the tubers in treatment solution containing 20 mg/L TDM on 10, 25 and 40 d after storage (DAS). Starch, sugars, protein, amino acid contents as well as protease and alpha-amylase activities were estimated on 15, 30 and 45 DAS from two physiological regions viz., apical and basal regions of the tubers. In normal conditions (control) sprouting occurred on 70 to 80 DAS. The starch content decreased, while protein, amino acid, sugar contents and protease and alpha-amylase activities were increased due to TDM treatment and led to early sprouting.
Dioscorea ; drug effects ; growth & development ; metabolism ; Food Preservation ; Plant Tubers ; drug effects ; growth & development ; metabolism ; Temperature ; Time Factors ; Triazoles

OBJECTIVETo observe the metabolism-based interaction of diphenytriazol and flavone compounds.

METHODSFlavone compounds kaempferol, isoharmnten and Elsholtzia blanda benth extract were chosen as the substrate of glucuronidation in the phase II metabolism. The metabolism was investigated in different rat liver microsome incubates pretreated with beta-naphthoflavone (BNF), diphenytriazol and tea oil (control). The concentrations of residual substrate were determined by HPLC. Quercetin and kaempferol were coincubated with diphenytriazol in control microsome to evaluate the inhibition for phase I metabolism. The concentration of diphenytriazol was determined by HPLC.

RESULTThe phase II metabolic activity of kaempferol, isoharmnten and Elsholtzia blanda benth extract in diphenytriazol-treated microsome was more potent than that in BNF-treated microsome (P<0.01). The phase I metabolism of diphenytriazol was markedly inhibited by quercetin and kaempferol, with the inhibition constants (Ki) (12.41 +/-0.26)microg . ml(-1) and (7.97 +/-0.08)microg . ml(-1), respectively.

CONCLUSIONDiphenytriazol demonstrates metabolism-based interaction with flavone compounds in vitro.

Abortifacient Agents ; metabolism ; pharmacology ; Animals ; Drug Interactions ; Female ; Flavones ; metabolism ; pharmacology ; Kaempferols ; metabolism ; pharmacology ; Plant Extracts ; pharmacology ; Quercetin ; metabolism ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Triazoles ; metabolism ; pharmacology

AIMTo establish a RP-HPLC method for determination of diphenytriazol (DL111-IT) in rat hepatic microsomes.

METHODSDL111-IT in rat hepatic microsomal incubates was extracted with chloroform, using diazepam as internal standard. The determination was performed on a Lichrospher ODS-C18 reversed column (25 cm x 0.46 cm ID) with mobile phase of methanol-pH 7.5 phosphate buffer (70:30) at a flow-rate of 1.0 mL.min-1. A UVVIS detector was operated at 235 nm.

RESULTSThe assaywas linear from 1.01-101.0 micrograms.mL-1 for DL111-IT. The limit of detection was 0.15 microgram.mL-1 (signal-to-noise ratio 3) and the limit of quantification was 1.01 micrograms.mL-1(RSD < 10%, n = 4). The method afforded average recoveries of (100.3 +/- 1.9)% (n = 5), and intra-day and inter-day RSD were less than 5.0%(n = 5). The method allowed study of the in vitro phase I metabolism of DL111-IT in rat liver microsomal incubates. The microsomes induced by beta-naphthoflavone showed high enzymatic activity for DL111-IT phase I metabolism.

CONCLUSIONThe method is simple, accurate and can be used to study the metabolism of DL111-IT in rat hepatic microsomes.

Abortifacient Agents, Nonsteroidal ; analysis ; metabolism ; Animals ; Cell Separation ; Chromatography, High Pressure Liquid ; methods ; Female ; Microsomes, Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triazoles ; analysis ; metabolism

AIMTo observe the metabolic interaction between diphenytriazol and steroid hormone drugs, and provide some useful information for clinical medication.

METHODSThe steroid hormone drugs which may be co-administrated with diphenytriazol were selected, such as mifepriston, estradiol, medroxyprogesterone acetate, progesterone, norethisterone and so on. Diphenytriazol was incubated with each drug in rat liver microsome. The residual concentration of diphenytriazol or steroid hormone drugs in the microsomal incubates was determined by reversed-phase high-performance liquid chromatography, separately. The inhibition constants (K(i)) for each of them were calculated.

RESULTSThe inhibition constant K(is) of diphenytriazol for the metabolism of mifepristone, estradiol, medroxyprogesterone acetate, progesterone and norethisterone were (201.3 +/- 1.0), (94 +/- 4), (128.7 +/- 2.2), (64 +/- 5) and (80 +/- 4) micromol x L(-1), respectively. The inhibition constants K(i) of steroid hormone drugs for the metabolism of diphenytriazol was (66.9 +/- 2.2) micromol x L(-1) for estradiol, (60.0 +/- 2.3) micromol x L(-1) for medroxyprogesterone acetate, (163 +/- 10) micromol x L(-1) for progesterone and (88 +/- 5) micromol x L(-1) for norethisterone, respectively.

CONCLUSIONDiphenytriazol shows metabolism interaction with steroid hormone drugs such as estradiol, medroxyprogesterone acetate, progesterone and norethisterone.

Abortifacient Agents, Nonsteroidal ; metabolism ; pharmacology ; Abortifacient Agents, Steroidal ; metabolism ; Animals ; Contraceptives, Oral, Synthetic ; metabolism ; Drug Interactions ; Estradiol ; metabolism ; Female ; In Vitro Techniques ; Medroxyprogesterone ; metabolism ; Microsomes, Liver ; metabolism ; Mifepristone ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triazoles ; metabolism ; pharmacology

BACKGROUNDThe prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document (M38-A) in 2002 by the Clinical and Laboratory Standards Institute (CLSI), dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay-glucose consumption method (GCM) for dermatophytes.

METHODSIn this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole (ITC), voriconazole (VOC), econazole nitrate (ECN) and terbinafine (TBF) by glucose consumption method (GCM), in comparison to the Clinical and Laboratory Standards Institute (CLSI) M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum (T. rubrum) (n = 14) and Trichophyton mentagrophytes (T. mentagrophytes) (n = 14), were tested. In the GCM, the minimum inhibitory concentrations (MICs) were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually.

RESULTSComparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM (0.125-16 microg/ml) but not M38-A method (0.5-1 microg/ml). The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively.

CONCLUSIONMeasurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Antifungal Agents ; pharmacology ; Econazole ; pharmacology ; Glucose ; metabolism ; Itraconazole ; pharmacology ; Microbial Sensitivity Tests ; Naphthalenes ; pharmacology ; Pyrimidines ; pharmacology ; Triazoles ; pharmacology ; Trichophyton ; drug effects ; metabolism ; Voriconazole

OBJECTIVETo establish a human breast cancer MCF-7 cell model stably overexpressing the aromatase gene (MCF-7-aromatase) and aromatase inhibitor letrozole-resistant MCF-7 cell model (MCF-7-LR).

METHODSWe utilized the lentivirus-mediated gene transfer approach to establish MCF-7-aromatase cell and MCF-7 cell model stably overexpressing green fluorescent protein (GFP) (MCF-7-GFP). The expression of aromatase in the MCF-7-aromatase and MCF-7-GFP cells was determined by reverse transcription polymerase chain reaction (RT-PCR), real time quantitative PCR (RT-qPCR), Western blot and immunoprecipitation (IP) assay. The proliferative ability in vitro of MCF-7-aromatase and MCF-7-GFP cells treated with testostorone and β-estradiol (E2) was determined by WST-1 cell proliferation assay. The proliferative ability of MCF-7-aromatase cells treated with letrozole was determined by WST-1 assay. The half maximal inhibitory concentration (IC50 value) for letrozole was calculated from the nonlinear regression line of the plot of cell viability (percentage of control) versus letrozole concentration using Graphpad Prism software. MCF-7-aromatase cells were continuously cultured in the presence of testosterone and letrozole, thus letrozole-resistant MCF-7-LR cells were obtained. WST-1 assay was performed to determine their chemoresistance to letrozole.

RESULTSRT-PCR and RT-qPCR results revealed that the mRNA expression of aromatase was significantly increased in the MCF-7-aromatase cells compared with that in the MCF-7-GFP cells. Both Western blot and IP assays showed that the expression of aromatase protein was drastically increased in the MCF-7-aromatase cells, compared with that in the MCF-7-GFP cells. WST-1 assay showed that the cell proliferation rate of MCF-7-aromatase cells treated with 1 and 10 nmol/L testosterone was 1.43- and 1.53-fold higher than that of the control cells, respectively. The proliferation rate of MCF-7-aromatase cells treated with 1 and 10 nmol/L E2 was 1.41- and 1.55-fold higher than that of the control cells, respectively. In contrast, the proliferation rate of MCF-7-GFP cells treated with 10 nmol/L testosterone was 1.12-fold higher than that of the control cells, and the proliferation rate of MCF-7-GFP cells treated with 1 and 10 nmol/L E2 was 1.41- and 1.51-fold higher than that of the control cells, respectively. Letrozole treatment significantly inhibited the testosterone-induced proliferation ability of MCF-7-aromatase cells in a dose-dependent manner and the IC50 value was 5.3 nmol/L. In contrast, letrozole treatment showed no inhibitory effect on the proliferative ability of MCF-7-LR cells and the IC50 value was >1000 nmol/L.

CONCLUSIONSMCF-7-aromatase and MCF-7-LR cells exhibit different response to letrozole treatment, which provides an important basis for further investigating the mechanism of letrozole resistance.

Antineoplastic Agents ; pharmacology ; Aromatase ; metabolism ; Aromatase Inhibitors ; pharmacology ; Breast Neoplasms ; Cell Proliferation ; Drug Resistance, Neoplasm ; Humans ; MCF-7 Cells ; Models, Biological ; Nitriles ; pharmacology ; Triazoles ; pharmacology

OBJECTIVE: To investigate the effects and mechanism of bromodomain and extra-terminal (BET) inhibitor JQ1 on the double-expressor lymphoma (DEL) cell lines. METHODS: Protein expressions of cMyc and BCL-2 in 3 lymphoma cell lines were checked by Western blot so as to identify DEL cell lines. CCK-8 assay was used to detect the effects of JQ1 on anti-proliferation in the DEL cell lines. Western blot and RT-PCR were used to measure the protein and mRNA expressions of cMyc, BCL-2 and BCL-6 in DEL cell lines which treated by JQ1. Flow cytometry was used to detect the effect of JQ1 on cell apoptosis. RESULTS: Based on the expressions of cMyc and BCL-2, the SU-DHL6 and OCILY3 cell lines were confirmed as DEL cell lines. CCK-8 assay showed that the proliferation of DEL cell lines was inhibited by JQ1, which was similar to non-DEL cell lines and mainly regulated the expression of cMyc and BCL-6 but not BCL-2. JQ1 had no effects on apoptosis in the DEL cell lines. CONCLUSION BET inhibitor JQ1 has anti-tumor effect in the DEL cell lines, thus providing evidence for the therapeutic potential of BET inhibitor JQ1.
Apoptosis ; Azepines/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Proto-Oncogene Proteins c-myc/metabolism* ; Sincalide/pharmacology* ; Triazoles/pharmacology* ; Xenograft Model Antitumor Assays