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61.
Peshwa MV Page LA Qian L Yang D van Schooten WC 《Biotechnology and bioengineering》1996,50(5):529-540
CD8(+) cytotoxic T-lymphocytes (CTLs) have been proven, in multiple animal models, to be the most powerful antiviral and antitumor components of the immune system. We have developed a protocol to activate and expand tumor and virus peptide-specific CD8(+) T-lymphocytes from the peripheral blood of healthy, human trophic leukemia virus-1 (HTLV-1) seronegative human leucocyte antigen (HLA)-A*0201 individuals. A combination of density-based separation and culture conditions was employed to isolate dendritic cells (DCs), which are the most potent antigen-presenting cells (APCs), and T-lymphocytes. The DCs were pulsed with HLA-A*0201 binding peptides and cultured with autologous T-lymphocytes to generate peptide-specific CTLs. The CTLs were generated against a nine-amino-acid peptide from the Tax protein of HTLV-1. The CTLs were expanded according to a restimulation schedule employing peptide-pulsed autologous monocytes and low-dose interleukin-2 (IL-2) to numbers in excess of 100 x 10(6) cells following 5 weeks of culture. Expanded cells contained primarily CD3(+) T-cells, of which CD8(+) T-lymphocytes constituted greater than two-thirds of the cell population. Obtained CTLs exhibited potent antigen-specific lysis of peptide-pulsed target cells in a dose-dependent fashion in in vitro (51)Cr release cytotoxicity assay. This antigen-specific killing was shown to be HLA class I restricted and mediated by CD8(+) T-lymphocytes. Since the T-lymphocytes were obtained from HTLV-1 seronegative donors, the generation of peptide-specific CTLs represents reliable and reproducible elicitation of a primary immune response in vitro against naive antigens and subsequent expansion of generated CTLs for adoptive immunotherapy. (c) 1996 John Wiley & Sons, Inc. 相似文献
62.
Predictive models for phosphorus retention in wetlands 总被引:1,自引:0,他引:1
C. J. Richardson S. Qian C. B. Craft R. G. Qualls 《Wetlands Ecology and Management》1996,4(3):159-175
The potential of wetlands to efficiently remove (i.e., act as a nutrient sink) or to transform nutrients like phosphorus under high nutrient loading has resulted in their consideration as a cost-effective means of treating wastewater on the landscape. Few predictive models exist which can accurately assess P retention capacity. An analysis of the north American data base (NADB) allowed us to develop a mass loading model that can be used to predict P storage and effluent concentrations from wetlands. Phosphorus storage in wetlands is proportional to P loadings but the output total phosphorus (TP) concentrations increase exponentially after a P loading threshold is reached. The threshold P assimilative capacity based on the NADB and a test site in the Everglades is approximately 1 g m–2 yr–1. We hypothesize that once loadings exceed 1 g m–2 yr–1 and short-term mechanisms are saturated, that the mechanisms controlling the uptake and storage of P in wetlands are exceeded and effluent concentrations of TP rise exponentially. We propose a One Gram Rule for freshwater wetlands and contend that this loading is near the assimilative capacity of wetlands. Our analysis further suggests that P loadings must be reduced to 1 g m–2 yr–1 or lower within the wetland if maintaining long-term low P output concentrations from the wetlands is the central goal. A carbon based phosphorus retention model developed for peatlands and tested in the Everglades of Florida provided further evidence of the proposed One Gram Rule for wetlands. This model is based on data from the Everglades areas impacted by agricultural runoff during the past 30 years. Preliminary estimates indicate that these wetlands store P primarily as humic organic-P, insoluble P, and Ca bound P at 0.44 g m–2 yr–1 on average. Areas loaded with 4.0 g m–2 yr–1 (at water concentrations>150 g·L–1 TP) stored 0.8 to 0.6 g m–2 yr–1 P, areas loaded with 3.3 g m–2 yr–1 P retained 0.6 to 0.4 g m–2 yr–1 P, and areas receiving 0.6 g m–2 yr–1 P retained 0.3 to 0.2 g m–2 yr–1. The TP water concentrations in the wetland did not drop below 50 g·L–1 until loadings were below 1 g m2 yr–1 P. 相似文献
63.
应用SDS-聚丙烯酰胺凝胶电泳及免疫转印技术对流行性出血热患才血清中免疫合物组分进行了分析。流行性出血热循环免疫复合物经SDS-PAGE分离,考马斯亮兰染色,显色主要有7条带,分子量分别为23kD,50kD,52kD,65kD,72kD,80kD及100kD。采用该病毒特异性抗血清、单克隆抗体以及人免疫球蛋白、补体成分抗血清识别,在其特环免疫复合物中可检出特异性病毒抗在及相应的免疫球状蛋白和补体成 相似文献
64.
本文报道人疱疹病豢-6型(HHV-6)pSTY28DNA片段的序列测定。应用分子克隆、缺损突变体(Dcletionmutant)制备和序列测定等技术,完成了3.9kbHHV-6pSTY28DNA片段的全序列测定。经DNASIS核酸蛋白软件分析,该片段含有两个开读框架(ORF)核糖核苷酸还原酶(RIR)ORF有2414个核苷酸,可编码805个氨基酸;P41蛋白由1100个核苷酸组成。与其他疱疹病毒作氨基酸同源性比较,HHV-6RiR与人巨细胞病毒(HCMV)有高度同源性,最适记分(Optimizedscore)达459。实验结果支持Esftathiou提出的论点,HHV-6属于β-疱疹病毒。 相似文献
65.
Retinoic acid-mediated activation of HNF-3 alpha during EC stem cell differentiation. 总被引:2,自引:0,他引:2 下载免费PDF全文
A Jacob S Budhiraja X Qian D Clevidence R H Costa R R Reichel 《Nucleic acids research》1994,22(11):2126-2133
66.
Purification and characterization of two phosphoglucomutases from Lactococcus lactis subsp. lactis and their regulation in maltose- and glucose-utilizing cells. 下载免费PDF全文
Two distinct forms of phosphoglucomutase were found in Lactococcus lactis subsp. lactis, strains 19435 and 65.1, growing on maltose: beta-phosphoglucomutase (beta-PGM), which catalyzes the reversible conversion of beta-glucose 1-phosphate to glucose 6-phosphate in the maltose catabolism, and alpha-phosphoglucomutase (alpha-PGM). beta-PGM was purified to more than 90% homogeneity in crude cell extract from maltose-grown lactococci, and polyclonal antisera to the enzyme were prepared. The molecular mass of beta-PGM was estimated by gel filtration to be 28 kDa; its isoelectric point was 4.8. The corresponding values for alpha-PGM were 65 kDa and 4.4, respectively. The expression of both PGM enzymes was investigated under different growth conditions. The specific activity and amount of beta-PGM per milliliter of cell extract increased with time in lactococci grown on maltose, but the enzyme was absent in lactococci grown on glucose, indicating enzyme synthesis to be induced by maltose in the growth medium. When glucose was added to maltose-grown lactococci, both the specific activity and amount of beta-PGM per milliliter of cell extract decreased rapidly. This suggests that synthesis of beta-PGM is repressed by glucose in the medium. Although the specific activity of alpha-PGM did not change during growth on maltose or glucose, lactococcal strain 19435 showed a much higher specific activity of both alpha- and beta-PGM than strain 65.1 when grown on maltose. 相似文献
67.
John A. Lowe III Weimin Qian Pamela J. Scott Stafford McLean Dianne K. Bryce Rosemary T. Crawford Jon Bordner 《Bioorganic & medicinal chemistry letters》1994,4(24):2877-2882
A series of 5,7-diphenyl-3-ureidohexahydroazepin-2-one cholecystokinin-B (CCK-B) receptor antagonists was synthesized using Beckmann ring expansion of a suitable 2,4-diphenylcyclohexanone as a key step. SAR studies revealed the importance of the 5-aryl group for high and selective CCK-B receptor affinity, as illustrated in compound (−)-10i (CCK-B IC50 = 6.8 nM). 相似文献
68.
采用Rotofor等电聚焦和分子筛技术纯化大肠杆菌表达的重组链激酶(r—SK),其纯度为97%。比活性1×106IU/mg,回收率41%。分析所纯化的r—SK N端氨基酸顺序证实其1—15个氨基酸顺序与SK基因的核苷酸顺序所示3联密码子一致。以纯化r—SK为抗原,通过杂交瘤技术构建了分泌抗r—SK单克隆抗体(McAb)杂交瘤细胞(4D11),该McAb属IgG1亚型.特异地作用于r-SK和C组口溶血性链球菌分泌的SK。用底物显色法测定该McAb可抑制SK对纤溶酶原的激活。Western blot法证实该McAb能识别分子量为16 250Da的r—SK的CNBr裂解片段。推测SK蛋白中第71残基至第237残基间的氨基酸顺序参与SK与纤溶酶原的结合。 相似文献
69.
R. Hao D. R. Cerutis H. S. Blaxall J. F. Rodriguez-Sierra R. F. Pfeiffer M. Ebadi 《Neurochemical research》1994,19(6):761-767
Metallothionein (MT) isoforms I and II were first identified and characterized in our laboratories in several regions of brain, in hippocampal neurons in primary culture, and in retinoblastoma and neuroblastoma cell lines. In this study, by having employed the MT-I cDNA as a probe, we sought to gain additional insight about the function of MT by discerning the regional distribution of its mRNA. Northern blot analyses of brain mRNA revealed that the administration of zinc enhanced dramatically MT-I mRNA (570 bp). The in situ hybridization study revealed that MT-I mRNA was located in several areas of brain, with the highest concentrations found in the cerebellum, hippocampus, and ventricles. The results of these studies are interpreted to suggest that zinc enhances the synthesis of MT mRNA and MT in turn may participate in zinc associated functions in neurons.Abbreviations MT-I
Metallothionein I isoform
- mRNA
Messenger ribonucleic acid
-
35S dCTP
35S Deoxycytidine triphosphate
-
32P dCTP
32P Deoxycytidine triphosphate
- icv
Intracerebroventricularly
- IP
Intraperitoneally
- PBS
Paraformaldehyde phosphate buffered saline solution
- Tris
2 amino-2-hydroxymethylpropane-1,3 diol
- EDTA
Ethylenediaminetetraacetic acid
- cDNA
Complimentary deoxyribonucleic acid
- bp
Base pair 相似文献
70.
Gary L. Johnson Anne M. Gardner Carol Lange-Carter Nan-Xin Qian Marijane Russell Sim Winitz 《Journal of cellular biochemistry》1994,54(4):415-422
Serpentine receptors coupled to the heterotrimeric G protein, Gi2, are capable of stimulating DNA synthesis in a variety of cell types. A common feature of the Gi2-coupled stimulation of DNA synthesis is the activation of the mitogen-activated protein kinases (MAPKs). The regulation of MAPK activation by the Gi2-coupled thrombin and acetylcholine muscarinic M2 receptors occurs by a sequential activation of a network of protein kinases. The MAPK kinase (MEK) which phosphorylates and activates MAPK is also activated by phosphorylation. MEK is phosphorylated and activated by either Raf or MEK kinase (MEKK). Thus, Raf and MEKK converge at MEK to regulate MAPK. Gi2-coupled receptors are capable of activating MEK and MAPK by Raf-dependent and Raf-independent mechanisms. Pertussis toxin catalyzed ADP-ribosylation of αi2 inhibits both the Raf-dependent and-independent pathways activated by Gi2-coupled receptors. The Raf-dependent pathway involves Ras activation, while the Raf-independent activation of MEK and MAPK does not involve Ras. The Raf-independent activation of MEK and MAPK most likely involves the activation of MEKK. The vertebrate MEKK is homologous to the Ste11 and Byr2 protein kinases in the yeast Saccharomyces cerevisiae and Schizosaccharomyces pombe, respectively. The yeast Ste11 and Byr2 protein kinases are involved in signal transduction cascades initiated by pheromone receptors having a 7 membrane spanning serpentine structure coupled to G proteins. MEKK appears to be conserved in the regulation of G protein-coupled signal pathways in yeast and vertebrates. Raf represents a divergence in vertebrates from the yeast pheromone-responsive protein kinase system. Defining MEKK and Raf as a divergence in the MAPK regulatory network provides a mechanism for differential regulation of this system by Gi2-coupled receptors as well as other receptor systems, including the tyrosine kinases. 相似文献