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11.
Chung-Ming Chi Leah N. Frautschy Frank B. Cerra Wei-Shou Hu 《Biotechnology Techniques》1993,7(2):99-104
Summary The measurement of the specific oxygen uptake rate (OUR) of slow growing organisms using a small sample size is often hampered by the consumption of oxygen by the electrode used. Using a optochemical sensor we measured the OUR of carrot embryos with approximately 1000 cell clusters and of hybridoma with approximately 106 cells. An OUR as low as 0.02 mol/h can be accurately measured. 相似文献
12.
Kazi Mirajul Hoque Owen M. Woodward Damian B. van Rossum Nicholas C. Zachos Linxi Chen George P.H. Leung William B. Guggino Sandra E. Guggino Chung-Ming Tse 《The Journal of general physiology》2010,135(1):43-58
Intestinal Cl− secretion is stimulated by cyclic AMP (cAMP) and intracellular calcium ([Ca2+]i). Recent studies show that protein kinase A (PKA) and the exchange protein directly activated by cAMP (Epac) are downstream targets of cAMP. Therefore, we tested whether both PKA and Epac are involved in forskolin (FSK)/cAMP-stimulated Cl− secretion. Human intestinal T84 cells and mouse small intestine were used for short circuit current (Isc) measurement in response to agonist-stimulated Cl− secretion. FSK-stimulated Cl− secretion was completely inhibited by the additive effects of the PKA inhibitor, H89 (1 µM), and the [Ca2+]i chelator, 1,2-bis-(o-aminophenoxy)-ethane-N,N,N’,N’-tetraacetic acid, tetraacetoxymethyl ester (BAPTA-AM; 25 µM). Both FSK and the Epac activator 8-pCPT-2’-O-Me-cAMP (50 µM) elevated [Ca2+]i, activated Ras-related protein 2, and induced Cl− secretion in intact or basolateral membrane–permeabilized T84 cells and mouse ileal sheets. The effects of 8-pCPT-2’-O-Me-cAMP were completely abolished by BAPTA-AM, but not by H89. In contrast, T84 cells with silenced Epac1 had a reduced Isc response to FSK, and this response was completely inhibited by H89, but not by the phospholipase C inhibitor or BAPTA-AM. The stimulatory effect of 8-pCPT-2’-O-Me-cAMP on Cl− secretion was not abolished by cystic fibrosis transmembrane conductance (CFTR) inhibitor 172 or glibenclamide, suggesting that CFTR channels are not involved. This was confirmed by lack of effect of 8-pCPT-2’-O-Me-cAMP on whole cell patch clamp recordings of CFTR currents in Chinese hamster ovary cells transiently expressing the human CFTR channel. Furthermore, biophysical characterization of the Epac1-dependent Cl− conductance of T84 cells mounted in Ussing chambers suggested that this conductance was hyperpolarization activated, inwardly rectifying, and displayed a Cl−>Br−>I− permeability sequence. These results led us to conclude that the Epac-Rap-PLC-[Ca2+]i signaling pathway is involved in cAMP-stimulated Cl− secretion, which is carried by a novel, previously undescribed Cl− channel. U73122相似文献
13.
Chuin-Shee Shang Chi-Tai Fang Chung-Ming Liu Tzai-Hung Wen Kun-Hsien Tsai Chwan-Chuen King 《PLoS neglected tropical diseases》2010,4(8)
Background
Travelers who acquire dengue infection are often routes for virus transmission to other regions. Nevertheless, the interplay between infected travelers, climate, vectors, and indigenous dengue incidence remains unclear. The role of foreign-origin cases on local dengue epidemics thus has been largely neglected by research. This study investigated the effect of both imported dengue and local meteorological factors on the occurrence of indigenous dengue in Taiwan.Methods and Principal Findings
Using logistic and Poisson regression models, we analyzed bi-weekly, laboratory-confirmed dengue cases at their onset dates of illness from 1998 to 2007 to identify correlations between indigenous dengue and imported dengue cases (in the context of local meteorological factors) across different time lags. Our results revealed that the occurrence of indigenous dengue was significantly correlated with temporally-lagged cases of imported dengue (2–14 weeks), higher temperatures (6–14 weeks), and lower relative humidity (6–20 weeks). In addition, imported and indigenous dengue cases had a significant quantitative relationship in the onset of local epidemics. However, this relationship became less significant once indigenous epidemics progressed past the initial stage.Conclusions
These findings imply that imported dengue cases are able to initiate indigenous epidemics when appropriate weather conditions are present. Early detection and case management of imported cases through rapid diagnosis may avert large-scale epidemics of dengue/dengue hemorrhagic fever. The deployment of an early-warning surveillance system, with the capacity to integrate meteorological data, will be an invaluable tool for successful prevention and control of dengue, particularly in non-endemic countries. 相似文献14.
Imai M Shiota T Kataoka K Tarby CM Moree WJ Tsutsumi T Sudo M Ramirez-Weinhouse MM Comer D Sun CM Yamagami S Tanaka H Morita T Hada T Greene J Barnum D Saunders J Myers PL Kato Y Endo N 《Bioorganic & medicinal chemistry letters》2004,14(21):5407-5411
N,N'-Disubstituted homopiperazine derivatives have been discovered as CC-chemokine receptor 2b (CCR2b) inhibitors with submicromolar activity in the CCR2b binding assay. A 4-substituted benzyl group on one homopiperazine nitrogen was an important moiety for binding affinity to the CCR2b receptor. The SAR for CCR2b binding affinity correlated inversely with the sigma factor of the functional group on this benzyl moiety. Introduction of hydroxy groups to appropriate positions in the 3,3-diphenylpropyl group on the other homopiperazine nitrogen increased CCR2b binding activity. The synthesis of an informer library to search for alternative substructures is also described. 相似文献
15.
Half-lives of plasma membrane Na+/H+ exchangers NHE1-3: plasma membrane NHE2 has a rapid rate of degradation 总被引:2,自引:0,他引:2
Cavet Megan E.; Akhter Shafinaz; Murtazina Rakhilya; Sanchez de Medina Fermin; Tse Chung-Ming; Donowitz Mark 《American journal of physiology. Cell physiology》2001,281(6):C2039
The Na+/H+ exchangers NHE2 and NHE3 areinvolved in epithelial Na+ and HCO absorption. To increase insights into the functions of NHE2 vs. NHE3,we compared their cellular processing with each other and with thehousekeeping isoform NHE1. Using biotinylated exchanger, we determinedthat the half-life of plasma membrane NHE2 was short (3 h) comparedwith that of NHE1 (24 h) and NHE3 (14 h) in both PS120 fibroblasts andCaco-2 cells. NHE2 transport and plasma membrane levels were reduced by3 h of Brefeldin A treatment, whereas NHE1 was unaffected. NHE2was degraded by the lysosomes but not proteosomes, as demonstrated byincreasing levels of endocytosed NHE2 protein after inhibition of thelysosomes, but not with proteosome inhibition. Unlike that of NHE3,basal NHE2 transport activity was not affected by phosphatidylinositol 3-kinase inhibition and did not appear to be localized in the juxtanuclear recycling endosome. Therefore, for NHE2, protein degradation and/or protein synthesis probably play important roles inits basal and regulated states. These results suggest fundamental differences in the cellular processing and trafficking of NHE2 andNHE3. These differences may underlie the specialized roles that theseexchangers play in epithelial cells. 相似文献
16.
Tzu-Wen Huang Te-Li Chen Ying-Tsong Chen Tsai-Ling Lauderdale Tsai-Lien Liao Yi-Tzu Lee Chien-Pei Chen Yen-Ming Liu Ann-Chi Lin Ya-Hui Chang Keh-Ming Wu Ralph Kirby Jui-Fen Lai Mei-Chen Tan Leung-Kei Siu Chung-Ming Chang Chang-Phone Fung Shih-Feng Tsai 《PloS one》2013,8(4)
The genetic features of the antimicrobial resistance of a multidrug resistant Klebsiella pneumoniae strain harboring bla
NDM-1 were investigated to increase our understanding of the evolution of NDM-1. The strain, KPX, came from a Taiwanese patient with a hospitalization history in New Delhi. Complete DNA sequencing was performed; and the genes responsible for antimicrobial resistance were systematically examined and isolated by library screening. KPX harbored two resistance plasmids, pKPX-1 and pKPX-2, which are 250-kb and 141-kb in size, respectively, with bla
NDM-1 present on pKPX-1. The plasmid pKPX-1 contained genes associated with the IncR and IncF groups, while pKPX-2 belonged to the IncF family. Each plasmid carried multiple antimicrobial resistance genetic determinants. The gene responsible for resistance to carbapenems was found on pKPX-1 and that for resistance to aztreonam was found on pKPX-2. To our surprise, we discovered that bla
NDM-1 exists on pKPX-1 as multiple copies in the form of tandem repeats. Amplification of bla
NDM-1 was found to occur by duplication of an 8.6-kb unit, with the copy number of the repeat varying from colony to colony. This repeat sequence is identical to that of the pNDM-MAR except for two base substitutions. The copy number of bla
NDM-1 of colonies under different conditions was assessed by Southern blotting and quantitative PCR. The bla
NDM-1 sequence was maintained in the presence of the antimicrobial selection; however, removal of antimicrobial selection led to the emergence of susceptible bacterial populations with a reduced copy number or even the complete loss of the bla
NDM-1 sequence. The dynamic nature of the NDM-1 sequence provides a strong argument for judicious use of the broad-spectrum antimicrobials in order to reduce the development and spread of antimicrobial resistance among pathogens. 相似文献
17.
Govindarajan R Bakken AH Hudkins KL Lai Y Casado FJ Pastor-Anglada M Tse CM Hayashi J Unadkat JD 《American journal of physiology. Regulatory, integrative and comparative physiology》2007,293(5):R1809-R1822
To better understand the role of human equilibrative (hENTs) and concentrative (hCNTs) nucleoside transporters in physiology and pharmacology, we investigated the regional, cellular, and spatial distribution of two hCNTs (hCNT1 and hCNT2) and two hENTs (hENT1 and hENT2) in four human tissues. Using in situ hybridization and immunohistochemical techniques, we found that the duodenum expressed hCNT1 and hCNT2 mRNAs in enterocytes and hENT1 and hENT2 mRNAs in crypt cells. In these cells, the hCNT and hENT proteins were predominantly localized in the apical and lateral membrane, respectively. Hepatocytes expressed higher levels of mRNAs of hENT1, hCNT1, and hENT2 than of hCNT2 and expressed all these proteins at hepatocyte cell borders and in the cytoplasm. While the kidney expressed hCNT1 and hCNT2 mRNAs in the proximal tubules, hENT1 and hENT2 mRNAs were present in the distal tubules, glomeruli, endothelial cells, and vascular smooth muscle cells. Proximal tubules adjacent to corticomedullary junctions expressed hENT1, hCNT1, and hCNT2 mRNA. Immunolocalization studies revealed predominant localization of hCNTs in the brush-border membrane of the proximal tubular epithelial cells and hENTs in the basolateral membrane of the distal tubular epithelial cells. Chorionic villi sections of human term placenta expressed mRNAs and proteins for hENT1 and hENT2 but only mRNA for hCNT2. Immunolocalization studies showed presence of hENT1 in the brush-border membrane of the syncytiotrophoblasts. These data are critical for a better understanding of the role of nucleoside transporters in the physiological and pharmacological effects of nucleosides and nucleoside drugs, respectively. 相似文献
18.
Yu CM Peng HP Chen IC Lee YC Chen JB Tsai KC Chen CT Chang JY Yang EW Hsu PC Jian JW Hsu HJ Chang HJ Hsu WL Huang KF Ma AC Yang AS 《PloS one》2012,7(3):e33340
Protein-protein interactions are critical determinants in biological systems. Engineered proteins binding to specific areas on protein surfaces could lead to therapeutics or diagnostics for treating diseases in humans. But designing epitope-specific protein-protein interactions with computational atomistic interaction free energy remains a difficult challenge. Here we show that, with the antibody-VEGF (vascular endothelial growth factor) interaction as a model system, the experimentally observed amino acid preferences in the antibody-antigen interface can be rationalized with 3-dimensional distributions of interacting atoms derived from the database of protein structures. Machine learning models established on the rationalization can be generalized to design amino acid preferences in antibody-antigen interfaces, for which the experimental validations are tractable with current high throughput synthetic antibody display technologies. Leave-one-out cross validation on the benchmark system yielded the accuracy, precision, recall (sensitivity) and specificity of the overall binary predictions to be 0.69, 0.45, 0.63, and 0.71 respectively, and the overall Matthews correlation coefficient of the 20 amino acid types in the 24 interface CDR positions was 0.312. The structure-based computational antibody design methodology was further tested with other antibodies binding to VEGF. The results indicate that the methodology could provide alternatives to the current antibody technologies based on animal immune systems in engineering therapeutic and diagnostic antibodies against predetermined antigen epitopes. 相似文献
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20.
Extensive landslides triggered by the Chi-Chi earthquake of 21 September 1999 introduced debris into the middle section of Tachia River, while debris flow was subsequently induced by Typhoon Toraji on 30 July 2001. We compared population size, species composition, and diversity of the fish community during a six-year period before and after these disturbances. The dominant taxa were Acrossocheilus paradoxus, Crossostoma lacustre, Hemimyzon formosanum, Rhinogobius brunneus, Varicorhinus barbatulus and Zacco pachycephalus. H. formosanum and R. brunneus increased then decreased suddenly in abundance following the earthquake and the typhoon. Our results suggest that both resistance and resilience were important in maintaining long-term fish community structure. Fish community resistance at station 1 (downstream of the disturbance) was lower than at station 2 and station 3 (at and upstream of the disturbance). Fish communities recovered quickly after a few months, possibly reflecting a correlation between assemblage composition and seasonal variation. Our study illustrates the ecological variability that can be induced by hydrologic and evolutionary processes in a stream. Relative positions of habitats provide a spatial framework for evaluating stress effects of abiotic and biotic factors in regulating population size and community succession patterns. 相似文献