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  • Coupling aerobic biodegradation of methanol vapors with heterologous protein expression of endochitinase Ech42 from Trichoderma atroviride in Pichia pastoris. 20709543

    Methanol is included among the most hazardous air pollutants, and an effort of vapors biofiltration by using microbial consortiums has been reported. The aim of this work was to couple the methanol vapors biodegradation with the production of recombinant endochitinase (ech42) from Trichoderma atroviride in Pichia pastoris transformed with the pPIC-ech42 plasmid. After carrying out batch experiments at 0.5% (w/v) of methanol concentration, the recombinant P. pastoris Mut(+) strain was selected because it showed methanol biodegradation rates similar to those of wild type GS115 strain (39 g/m(3)h), but 15% higher than the transformed Mut(S) strain. In addition, the recombinant Ech42 protein production was higher in Mut(+) than Mut(S). After various methanol vapor concentrations were evaluated, the maximum recombinant protein recovery was 317 mg/l and the volumetric methanol consumption rate was 88.7 g/m(3)h at 0.5% (w/v) of methanol concentration. This research underlines the promising application of linking methanol vapors biodegradation with the production of recombinant protein with high biotechnological interests.
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    ECM630
    Produktbezeichnung:
    Fibrin In Vitro Angiogenesis Assay
  • Independent exponential feeding of glycerol and methanol for fed-batch culture of recombinant Hansenula polymorpha DL-1. 14645999

    As a novel feeding strategy for optimizing human epidermal growth factor (hEGF) production with a recombinant Hansenula polymorpha DL-1 using the methanol oxidase (MOX) promoter in H. polymorpha DL-1, independent exponential feeding of two substrates was used. A simple kinetic model considering the cell growth on two substrates was established and used to calculate the respective feeding rates of glycerol and methanol. In the fedbatch culture with methanol-only feeding, the optimal set point of specific growth rate on methanol was found to be 0.10 h-1. When the fed-batch cultures were conducted by the independent feeding of glycerol and methanol, the actual specific growth rate on glycerol and methanol was slightly lower than the set point of specific growth rate. By the uncoupled feeding of glycerol and methanol the volumetric productivity of hEGF increased from 6.4 to 8.0 mg/(L.h), compared with methanol-only feeding.
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    03-111
    Produktbezeichnung:
    RIPAb+ AUF1 - RIP Validated Antibody and Primer Set
  • Water in Methanol

    Dokumententyp:
    Anwendung
    Produkbestellnummer:
    Mehrere
    Produktbezeichnung:
    Mehrere
  • Wasser in Methanol

    Dokumententyp:
    Anwendung
    Produkbestellnummer:
    Mehrere
    Produktbezeichnung:
    Mehrere
  • Chemoprevention and inhibition of P-glycoprotein in cancer cells by Chinese medicinal herbs. 18690658

    Many of the herbal extracts used in the Chinese clinical medical routine inhibit the growth of tumor cells. In the present work, extracts of 12 selected herbs were prepared with methanol, chloroform, ethyl acetate and water, and the effects of these on the multidrug resistance (MDR) and P-glycoprotein of mouse lymphoma cells transfected with the human mdr1 gene and on a human lung alveolar epithelial cell line were investigated. The extracts were tested for antiproliferative effects, and the reversal of MDR in mouse lymphoma cells. The possible chemopreventive effect of the chloroform extracts was studied on the expression of cytomegalovirus (CMV) immediate-early (IE) antigen in human lung cancer cells (A549). The antimicrobial effects of the extracts were tested on some representative micro-organisms. Certain of the chloroform extracts of the plant materials were the most effective compounds on the reversal of MDR. Two of the chloroform extracts enhanced the antiproliferative effect of doxorubicin on MDR mouse lymphoma cells. The selected extracts did not show any antibacterial effect with the agar diffusion method. Certain chloroform extracts decreased the intermediate IE antigen expression of CMV in A459 cells. Copyright (c) 2008 John Wiley Sons, Ltd.
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    MAB810
  • Amyloid-like aggregates of neuronal tau induced by formaldehyde promote apoptosis of neuronal cells. 17241479

    The microtubule associated protein tau is the principle component of neurofibrillar tangles, which are a characteristic marker in the pathology of Alzheimer's disease; similar lesions are also observed after chronic alcohol abuse. Formaldehyde is a common environmental contaminant and also a metabolite of methanol. Although many studies have been done on methanol and formaldehyde intoxication, none of these address the contribution of protein misfolding to the pathological mechanism, in particular the effect of formaldehyde on protein conformation and polymerization.We found that unlike the typical globular protein BSA, the natively-unfolded structure of human neuronal tau was induced to misfold and aggregate in the presence of ~0.01% formaldehyde, leading to formation of amyloid-like deposits that appeared as densely staining granules by electron microscopy and atomic force microscopy, and bound the amyloid-specific dyes thioflavin T and Congo Red. The amyloid-like aggregates of tau were found to induce apoptosis in the neurotypic cell line SH-SY5Y and in rat hippocampal cells, as observed by Hoechst 33258 staining, assay of caspase-3 activity, and flow cytometry using Annexin V and Propidium Iodide staining. Further experiments showed that Congo Red specifically attenuated the caspase-3 activity induced by amyloid-like deposits of tau.The results suggest that low concentrations of formaldehyde can induce human tau protein to form neurotoxic aggregates, which could play a role in the induction of tauopathies.
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    MAB3420
    Produktbezeichnung:
    Anti-Tau-1 Antibody, clone PC1C6
  • LC method for telithromycin in tablets: a stability-indicating assay. 18977423

    A liquid chromatographic (LC) method for the quantitative determination of telithromycin, the first member of the ketolides, which is a new class of macrolides, was developed. Analytical parameters were studied according to International Conference on Harmonization (ICH) guidelines. An Ace RP-18 octadecyl silane column (250 mm x 4.6 mm, 5 microm) maintained at 50 degrees C was used as the stationary phase, and methanol and 0.067 M potassium monobasic phosphate buffer pH 4.0 (55:45, v/v) were used as the mobile phase with UV detection at 265 nm. In forced degradation studies, the effects of acid, base, oxidation, UV light and temperature were investigated showing no interference in the drug peak. The method was linear (r=0.9999) at concentration ranging from 10.0 to 40.0 microg/mL, precise (intra-day relative standard deviation [RSD] and inter-day RSD values<2.0%), accurate (mean recovery=100.76%), specific and robust. Detection and quantitation limits were 0.0027 and 0.0082 microg/mL, respectively. The results showed the proposed method is suitable for its intended use. The validated method may be used to quantify telithromycin tablets and to determine the stability of the drug. The method is able to separate telithromycin from its degradation products and tablet excipients for its sensitivity and reproducibility. These results are in accordance with a previous microbiological assay study, which used the same tested conditions showing that the methods can be interchangeable.,
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    2752
    Produktbezeichnung:
    BrdU Cell Proliferation Kit
  • Flow cytometric assessment of estrogen receptor beta expression in bovine blood neutrophils. 17451738

    The optimisation of a flow cytometric protocol for the determination of the estrogen receptor beta (ERbeta) expression in bovine blood neutrophils is described. The following final incubation conditions were obtained: fixation with 0.25% formaldehyde and 70% methanol, both for 1 h; permeabilisation with 0.05% Triton X-100, overnight labelling at 4 degrees C with the primary antibody diluted at 10 microg/ml and subsequent labelling for 30 min on ice with the fluorescein isothiocyanate-conjugated secondary antibody at 8 microg/ml. Of the three anti-human or anti-rat ERbeta primary antibodies evaluated, only PA1-311 was found to cross-react with bovine cells. Immunoblot analysis supports the obtained results. The flow cytometric technique allows reproducible quantitative determination of the ERbeta protein in neutrophils and may be a valuable tool for future expression studies in these cells of the innate immune system.
    Dokumententyp:
    Referenz
    Produkbestellnummer:
    AB1410