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991.
Xiaojuan Sun Hui He Zhihui Xie Weiping Qian Haiyen E. Zhau Leland W. K. Chung Fray F. Marshall Ruoxiang Wang 《In vitro cellular & developmental biology. Animal》2010,46(6):538-546
Prostate stromal cells may play binary roles in the process of prostate cancer development. As the first to be encountered
by infiltrating prostate cancer cells, prostate stromal cells form the first defense line against prostate cancer progression
and metastasis. However, interaction between prostate cancer and stromal cells may facilitate the formation of a tumor microenvironment
favoring cancer cell growth and survival. To establish an experimental system for studying the interaction between cancer
and stromal cells, we isolated three matched pairs of normal and cancer-associated human prostate stromal clones. In this
report, we describe the morphologic and behavioral characteristics of these cells and their effect on LNCaP prostate cancer
cells in co-culture. Unlike LNCaP prostate cancer cells, the isolated prostate stromal clones are large fibroblast-like cells
with a slow proliferation rate. Growth and survival of these clones are not affected by androgens. The stromal cells display
high resistance to serum starvation, while cancer-associated stromal clones have differentiated survival ability. In co-culture
experiments, the stromal cells protected some LNCaP prostate cancer cells from death by serum starvation, and cancer-associated
stromal clones showed more protection. This work thus established a panel of valuable human prostate stromal cell lines, which
could be used in co-culture to study the interaction between prostate cancer and prostate stromal cells. 相似文献
992.
Yan Liu Guihua Gong Liping Xie Ning Yuan Chunbao Zhu Baoquan Zhu Youjia Hu 《Molecular biotechnology》2010,44(2):101-109
Cephalosporins are widely used as anti-infectious β-lactam antibiotics in clinic. For the purpose of increasing the yield
of cephalosporin C (CPC) fermentation, especially in an industrial strain, A. chrysogenum genes cefEF and cefG, which encode the ultimate and penultimate steps in CPC biosynthesis, cefT, which encodes a CPC efflux pump, and vgb, which encodes a bacterial hemoglobin gene were transformed in various combinations into an industrial strain of A. chrysogenum. Both PCR and Southern blotting indicated that the introduced genes were integrated into the chromosome of A. chrysogenum. Carbon monoxide difference spectrum absorbance assay was performed and the result showed that Vitreoscilla hemoglobin was successfully expressed in A. chrysogenum and had biological activity. HPLC analysis of fermentation broth of recombinant A. chrysogenum showed that most transformants had a higher CPC production level than the parental strain. Multiple transformants containing
an additional copy of cefG showed a significant increase in CPC production. However, cefT showed little effect on CPC production in this high producer. The highest improvement of CPC titer was observed in the mutant
with an extra copy of cefG + cefEF + vgb whose CPC production was increased by 116.3%. This was the first report that three or more genes were introduced simultaneously
into A. chrysogenum. Our results also demonstrated that the combination of these genes had a synergy effect in a CPC high producer. 相似文献
993.
Xiaodong Leng Bingguang Xiao Sheng Wang Yijie Gui Yu Wang Xiuping Lu Jiahua Xie Yongping Li Longjiang Fan 《Plant Molecular Biology Reporter》2010,28(1):152-161
Tobacco (Nicotiana tabacum) is an important cash crop and an ideal experimental system for studies on plant–pathogen interaction. The sequenced tobacco
genome provides an opportunity for examining resistance gene homologs (RGHs) in the tobacco genome. Thirty nucleotide-binding
site-type RGHs were annotated from genomic data, and another 281 putative RGHs were identified via PCR amplification from
wild and cultivated tobacco. The newly identified RGHs are similar to other known RGHs, and some were categorized into new
groups or branches that are different from known Nicotiana R genes or RGHs. Of the 281RGHs, 146 were identified from a single tobacco genome. We did not find any polymorphism at the
RGHs in cultivated accessions, implying that strong domestication selection and/or demographic effects might have caused a
sharp reduction in nucleotide diversity. Three positive selection sites were found in several RGH groups, while purifying
selection is pervasive in the RGH family. Our results provide a primary RGH pool and several positively selected sites for
the further functional validation of resistance genes in tobacco. 相似文献
994.
Trifluoperazine (TFP; Stelazine?) is an antagonist of calmodulin (CaM), an essential regulator of calcium‐dependent signal transduction. Reports differ regarding whether, or where, TFP binds to apo CaM. Three crystallographic structures (1CTR, 1A29, and 1LIN) show TFP bound to (Ca2+)4‐CaM in ratios of 1, 2, or 4 TFP per CaM. In all of these, CaM domains adopt the “open” conformation seen in CaM‐kinase complexes having increased calcium affinity. Most reports suggest TFP also increases calcium affinity of CaM. To compare TFP binding to apo CaM and (Ca2+)4‐CaM and explore differential effects on the N‐ and C‐domains of CaM, stoichiometric TFP titrations of CaM were monitored by 15N‐HSQC NMR. Two TFP bound to apo CaM, whereas four bound to (Ca2+)4‐CaM. In both cases, the preferred site was in the C‐domain. During the titrations, biphasic responses for some resonances suggested intersite interactions. TFP‐binding sites in apo CaM appeared distinct from those in (Ca2+)4‐CaM. In equilibrium calcium titrations at defined ratios of TFP:CaM, TFP reduced calcium affinity at most levels tested; this is similar to the effect of many IQ‐motifs on CaM. However, at the highest level tested, TFP raised the calcium affinity of the N‐domain of CaM. A model of conformational switching is proposed to explain how TFP can exert opposing allosteric effects on calcium affinity by binding to different sites in the “closed,” “semi‐open,” and “open” domains of CaM. In physiological processes, apo CaM, as well as (Ca2+)4‐CaM, needs to be considered a potential target of drug action. Proteins 2010. © 2010 Wiley‐Liss, Inc. 相似文献
995.
The MEK–ERK pathway plays a role in DNA damage response (DDR). This has been thoroughly studied by modulating MEK activation. However, much less has been done to directly examine the contributions of ERK1 and ERK2 kinases to DDR. Etoposide induces G2/M arrest in a variety of cell lines, including MCF7 cells. DNA damage-induced G2/M arrest depends on the activation of the protein kinase ataxia-telangiectasia mutated (ATM). ATM subsequently activates CHK2 by phosphorylating CHK2 threonine 68 (T68) and CHK2 inactivates CDC25C via phosphorylation of its serine 216 (S216), resulting in G2/M arrest. To determine the contribution of ERK1 and ERK2 to etoposide-induced G2/M arrest, we individually knocked-down ERK1 and ERK2 in MCF7 cells using specific small interfering RNA (siRNA). Knockdown of either kinases significantly reduced ATM activation in response to etoposide treatment, and thereby attenuated phosphorylation of the ATM substrates, including the S139 of H2AX (γH2AX), p53 S15, and CHK2 T68. Consistent with these observations, knockdown of either ERK1 or ERK2 reduced etoposide-induced CDC25C S216 phosphorylation and significantly compromised etoposide-induced G2/M arrest in MCF7 cells. Taken together, we demonstrated that both ERK1 and ERK2 kinases play a role in etoposide-induced G2/M arrest by facilitating activation of the ATM pathway. These observations suggest that a cellular threshold level of ERK kinase activity is required for the proper checkpoint activation in MCF7 cells. 相似文献
996.
Hai-Nan Su Bin-Bin Xie Xiu-Lan Chen Jin-Xia Wang Xi-Ying Zhang Bai-Cheng Zhou Yu-Zhong Zhang 《Journal of applied phycology》2010,22(1):65-70
Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple
and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding
hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed
by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude
phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0).
In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A
650/A
280 of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g−1 wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from
5.6 to 4.0. The absorbance ratio A
650/A
280 of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g−1 wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE. 相似文献
997.
Some strains of white rot fungi, non-lignolytic fungi and litter-decomposing basidiomycetes have been recognized as PAH degraders.
The purpose of our research was to enlarge the scope of PAH-degrading fungi and explore the huge endophytic microorganism
resource for bioremediation of PAHs. In this study, phenanthrene was used as a model PAHs compound. Nine strains of endophytic
fungi isolated from four kinds of plant from Eupharbiaceae were screened for degradation of phenanthrene. The endophytic fungus Ceratobasidum stevensii (strain B6) isolated from Bischofia polycarpam showed high degradation efficiency and was selected for further studies. Into the fungal culture, 100 mg l−1 phenanthrene was added, and after 10 days of incubation, about 89.51% of the phenanthrene was removed by strain B6. Extracellular
ligninolytic enzyme activities of strain B6 were tested. The results showed that manganese peroxidase [MnP] was the predominant
ligninolytic enzyme and that its production was greatly induced by the presence of phenanthrene. To confirm the involvement
of MnP in phenanthrene degradation, promotion and inhibition studies on MnP in different concentration level of Mn2+ and NaN3 were performed. Additionally, fungal mycelium-free and resuspended experiments were carried out. The results showed no apparent
correlation between MnP activity and phenanthrene degradation. The mycelium and fresh medium were the crucial factors affecting
the degradation of phenanthrene. To date, this is the first report on PAH degradation by Ceratobasidum stevensii. This study suggests that endophytic fungi might be a novel and important resource for microorganisms that have PAH-degrading
capabilities. 相似文献
998.
Parkinson's disease (PD) is a common neurodegenerative movement disorder that affects increasing number of elderly in the world population. The disease is caused by a selective degeneration of dopaminergic neurons in the substantia nigra pars compacta with the molecular mechanism underlying this neurodegeneration still not fully understood. However, various studies have shown that mitochondrial dysfunction and abnormal protein aggregation are two of the major contributors for PD. In fact this notion has been supported by recent studies on genes that are linked to familial PD (FPD). For instance, FPD linked gene products such as PINK1 and parkin have been shown to play critical roles in the quality control of mitochondria, whereas α-synuclein has been found to be the major protein aggregates accumulated in PD patients. These findings suggest that further understanding of how dysfunction of these pathways in PD will help develop new approaches for the treatment of this neurodegenerative disorder. 相似文献
999.
1000.
Girijavallabhan VM Chen L Dai C Feltz RJ Firmansjah L Li D Kim SH Kozlowski JA Lavey BJ Kosinski A Piwinski JJ Popovici-Muller J Rizvi R Rosner KE Shankar BB Shih NY Siddiqui MA Tong L Wong MK Yang DY Yang L Yu W Zhou G Guo Z Orth P Madison V Bian H Lundell D Niu X Shah H Sun J Umland S 《Bioorganic & medicinal chemistry letters》2010,20(24):7283-7287
Our research on hydantoin based TNF-α converting enzyme (TACE) inhibitors has led to an acetylene containing series that demonstrates sub-nanomolar potency (K(i)) as well as excellent activity in human whole blood. These studies led to the discovery of highly potent TACE inhibitors with good DMPK profiles. 相似文献