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921.
Hirasawa K Uchimura K Kashiwa M Grant WD Ito S Kobayashi T Horikoshi K 《Antonie van Leeuwenhoek》2006,89(2):211-219
An endoglucanase was purified to homogeneity from an alkaline culture broth of a strain isolated from␣seawater and identified
here as Bacillus agaradhaerens JAM-KU023. The molecular mass was around 38-kDa and the N-terminal 19 amino acids of the purified enzyme exhibited 100% sequence
identity to Cel5A of B. agaradhaerens DSM8721T. The enzyme activity increased around 4-fold by the addition of 0.2–2.0 M NaCl in 0.1 M glycine–NaOH buffer (pH 9.0). KCl,
Na2SO4, NaBr, NaNO3, CH3COONa, LiCl, NH4NO3, and NH4Cl also activated the enzyme up to 2- to 4-fold. The optimal pH and temperature values were pH 7–9.4 and 60 °C with 0.2 M
NaCl, but pH 6.5–7 and 50 °C without NaCl; enzyme activity increased approximately 6-fold at 60 °C with 0.2 M NaCl compared
to that at 50 °C without NaCl in 0.1 M glycine–NaOH buffer (pH 9.0). The thermostability and pH stability of the enzyme were
not affected by NaCl. The enzyme was very stable to several chemical compounds, surfactants and metal ions (except for Fe2+ and Hg2+ ions), regardless whether NaCl was present or not.
* The nucleotide sequence of 16S rRNA of this strain has been submitted to DDBJ, EMBL, and GenBank databases under accession
no. AB211544. 相似文献
922.
Chen YL Hirabayashi H Akhtar S Pelzer M Kobayashi M 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2006,830(2):330-341
An ammonium-adduct based liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous determination of three isomeric metabolites of tacrolimus (FK506), 13-O-demethylated (M1), 31-O-demethylated (M2) and 15-O-demethylated (M3) tacrolimus in human whole blood and plasma. These metabolites and the internal standards were extracted from biological matrix by methylbutyl ether (MTBE). Separation was achieved on a Genesis C(18) column with a gradient mobile phase elution. Ammonium-adduct ions formed by a Turbo Ionspray in positive ion mode were used to detect each analyte and internal standard. The MS/MS detection was by monitoring the fragmentation of 807.5-->772.4 (m/z) for M1, 807.5-->754.5 (m/z) for both M2 and M3, 795.5-->760.5 (m/z) for IS1 (FR298701) and 961.5-->908.5 (m/z) for IS2 (FR290198) on a triple quadrupole mass spectrometer (Sciex API 3000). The retention times were approximately 4.1 min for M1, 6.8 min for M2, 6.0 min for M3, and 3.9 min for IS1 and 6.4 min for IS2, respectively. The validated dynamic range was 0.2-20 ng/ml for all three metabolites based on a sample volume of 0.25-ml. The linearity of calibration curves for M1, M2, and M3 in both matrices had a correlation coefficient of >/=0.9984. In whole blood, validation data showed intra-batch (n=6) CVs of =5.9% and REs between -4.9 and 3.6% and inter-batch (N=18) CVs of =4.9% and REs between -3.5 and 1.5% for all three metabolites. In human plasma, validation data showed intra-batch (n=6) CVs of =7.3% and REs between -5.1 and 7.6% and inter-batch (N=18) CVs of =6.6% and REs between -0.3 and 4.7% for all three metabolites. Extraction recoveries were 72% for M1, 87% for M2, 69% for M3, 79% for IS1, and 74% for IS2 from blood; and 94% for M1, 96% for M2, 98% for M3, 92% for IS1, and 93% for IS2 from plasma. All three metabolites in human blood and plasma were stable for three freeze-thaw cycles, or 24-h ambient storage, or 12 months storage at approximately -80 degrees C. Extracted samples were stable for at least 50h at room temperature (RT). This method has been successfully used to analyze whole blood and plasma samples from human pharmacokinetic studies. Several key factors affecting the performance of the assay methods have also been addressed briefly. 相似文献
923.
Negative regulation of the alpha interferon-induced antiviral response by the Ras/Raf/MEK pathway
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Interferon (IFN) is one of the molecules released by virus-infected cells, resulting in the establishment of an antiviral state within infected and neighboring cells. IFN-induced antiviral response may be subject to modulation by the cellular signaling environment of host cells which impact the effectiveness of viral replication. Here, we show that cells with an activated Ras/Raf/MEK signaling cascade allow propagation of viruses in the presence of IFN. Ras-transformed (RasV12) and vector control NIH 3T3 cells were infected with vesicular stomatitis virus (VSV) or an IFN-sensitive vaccinia virus (delE3L) in the presence of alpha interferon. While IFN protected vector control cells from infection by both viruses, RasV12 cells were susceptible to viral infection regardless of the presence of IFN. IFN sensitivity was restored in RasV12 cells upon RNA interference (RNAi) knockdown of Ras. We further investigated which elements downstream of Ras are responsible for counteracting IFN-induced antiviral responses. A Ras effector domain mutant that can only stimulate the Raf kinase family of effectors was able to suppress the IFN response and allow VSV replication. IFN-induced antiviral mechanisms were also restored in RasV12 cells by treatment with a MEK inhibitor (U0126 or PD98059). Moreover, by using RNAi to MEK1 and MEK2, we determined that MEK2, rather than MEK1, is responsible for suppression of the IFN response. In conclusion, our results suggest that activation of the Ras/Raf/MEK pathway downregulates IFN-induced antiviral response. 相似文献
924.
Raloxifene increases proliferation and up-regulates telomerase activity in human umbilical vein endothelial cells 总被引:3,自引:0,他引:3
925.
The aim of the present study was to investigate the responses of Merkel cells that are numerous in the palatine rugae, due to the continuous mechanical stimulation exerted by the palatal plate. Forty golden hamsters were used in this experiment. The palatal plate was made of adhesive resin and it was set on the palate of the animal. To exert a continuous pressure, a 0.8 mm elevation on the internal surface of the palatal plate was created at the middle portion of the fourth palatine ruga. Thereafter, the number of Merkel cells in the mucosa was calculated by immunohistochemical observation. Morphological changes of Merkel cells were examined by electron microscopy. There was significant difference among the control and any of the treated groups on the number of CK20 positive Merkel cells (p < 0.05) and that numbers were decreased at the sites where continuous mechanical stimulation was exerted. Degeneration of the cytoplasm mitochondria and nerve endings, and a decrease in both the number of neurosecretory granules and cytoplasmic processes were observed. Furthermore, the presence of nuclear chromatin aggregation and fragmentation was recognized. The continuous mechanical stimulation by the palatal plate affected the responses of Merkel cells and nerve endings, thus inducing a decrease in the number of Merkel cells. A portion of these changes was also associated with the expression of apoptosis. 相似文献
926.
We examined the elongation rate, water status and solute accumulation in the seminal roots of wheat seedlings (Triticum aestivum L.) that were growing in vermiculite with a water potential (Ψw) ranging from −0 03 to −1 10 MPa. The elongation rate of the primary seminal root was similar to that of the first pair of
seminal roots but that of the second pair of seminal roots was lower at all values of Ψw tested. The elongation rate was highest in vermiculite with a Ψw of −0.03 MPa but did not decrease significantly until the Ψw was reduced to −0.15 MPa. Further reductions in Ψw reduced the elongation rate markedly. The Ψw of mature tissues was always similar to that of vermiculite. The osmotic potential (Ψo) decreased to the same extent as the decrease in Ψw. Thus, the turgor pressure (Ψp) remained unchanged even in vermiculite with a low Ψw. In elongating tissues, Ψw and Ψo were far lower than they were in mature tissues and, thus, reductions in turgor were not significant. Even when the Ψw of vermiculite changed, there were no consistent changes in terms of a difference in Ψw between elongating plus mature tissues and vermiculite. There were also no consistent changes in levels of osmotica, calculated
using the van’t Hoff’s law, in the elongating tissues but the levels in mature tissues increased in vermiculite with a low
Ψw. Our results suggest that (1) reductions in root elongation in vermiculite with a low Ψw were caused by reductions in the extensibility and/or increases in the yield threshold of cell walls and by reductions in
the hydraulic conductivity of the tissues; and (2) a seminal root regulates its growth to keep turgor pressure unchanged. 相似文献
927.
In proteomic analysis, one of the major limitations is the detection of low-abundance proteins. To detect low-abundance RNA-binding
proteins in mature dry seeds of rice, fractionation by single stranded DNA (ssDNA) affinity column chromatography was carried
out before analysis by two-dimensional gel electrophoresis (2-DE). Proteomic analysis of the ssDNA-binding fraction revealed
the existence of three types of RNA-binding proteins, including a K homology (KH) domain containing protein, a putative RNA-binding
protein and a glycine-rich RNA-binding protein, in mature seeds. In addition, decreases in the putative RNA-binding protein
and glycine-rich RNA-binding protein after absorbing water in seeds appear to be associated with seed germination. 相似文献
928.
Koyama K Hirasawa Y Zaima K Hoe TC Chan KL Morita H 《Bioorganic & medicinal chemistry》2008,16(13):6483-6488
Five new alkaloids, alstilobanines A (1)-E (5) were isolated from Alstonia angustiloba (Apocynaceae) and their structures were determined by MS and 2D NMR spectral analysis. Alstilobanines A-E showed a moderate vasorelaxant activity against phenylephrine-induced contraction of isolated rat aorta. 相似文献
929.
The processing and presentation of lipid antigens by antigen presenting cells (APC) is important for defense against infection, tumor immunosurveillance, and autoimmunity. CD1, a family of cell surface glycoproteins, is responsible for the binding and presentation of lipid antigens to receptors expressed on the surface of T lymphocytes. Among the several (glyco)lipids identified to cause T-cell stimulation in complex with CD1, alpha-galactosyl ceramide (alpha-GalCer) is one of the most well studied. A combination of structure-activity relationship (SAR), crystallographic studies, and discovery of new 'natural' antigens has led to greater understanding of the structural requirements for optimal natural killer T-cell activation. 相似文献
930.
Nishizawa M Shiota M Moore MC Gustavson SM Neal DW Cherrington AD 《American journal of physiology. Regulatory, integrative and comparative physiology》2008,294(4):R1197-R1204
We examined whether intraportal delivery of neuropeptide Y (NPY) affects glucose metabolism in 42-h-fasted conscious dogs using arteriovenous difference methodology. The experimental period was divided into three subperiods (P1, P2, and P3). During all subperiods, the dogs received infusions of somatostatin, intraportal insulin (threefold basal), intraportal glucagon (basal), and peripheral intravenous glucose to increase the hepatic glucose load twofold basal. Following P1, in the NPY group (n = 7), NPY was infused intraportally at 0.2 and 5.1 pmol.kg(-1).min(-1) during P2 and P3, respectively. The control group (n = 7) received intraportal saline infusion without NPY. There were no significant changes in hepatic blood flow in NPY vs. control. The lower infusion rate of NPY (P2) did not enhance net hepatic glucose uptake. During P3, the increment in net hepatic glucose uptake (compared with P1) was 4 +/- 1 and 10 +/- 2 micromol.kg(-1).min(-1) in control and NPY, respectively (P < 0.05). The increment in net hepatic fractional glucose extraction during P3 was 0.015 +/- 0.005 and 0.039 +/- 0.008 in control and NPY, respectively (P < 0.05). Net hepatic carbon retention was enhanced in NPY vs. control (22 +/- 2 vs. 14 +/- 2 micromol.kg(-1).min(-1), P < 0.05). There were no significant differences between groups in the total glucose infusion rate. Thus, intraportal NPY stimulates net hepatic glucose uptake without significantly altering whole body glucose disposal in dogs. 相似文献