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51.
To investigate whether human immunodeficiency virus (HIV)-1 and HIV-1 antigens modulate surface and cytoplasmic CD8 or CD3, as well as CD4, we used cell permeabilization reagents, surface/cytoplasmic fluorescent staining, multiparameter flow cytometric techniques and an in vitro culture system in which relatively few lymphocytes are actively infected with HIV. Human peripheral blood lymphocytes were: not stimulated, not stimulated but HIV-inoculated, phytohaemagglutinin (PHA)-stimulated, PHA/HIV-inoculated (PHA/HIV), or placed into media with soluble gp120, Rev or Nef. HIV inoculation and Nef had striking modulatory effects on CD8. The cytoplasmic CD8 median fluorescent intensity (MFI) of positive lymphocytes was lower for cells in unstimulated/HIV-infected cultures than unstimulated cultures (44 versus 62% of ex vivo value, P = 0.032) and lower for cells in PHA/HIV cultures than in PHA cultures (56 versus 100% of ex vivo, P = 0.041). The surface CD8 MFI values for Nef were significantly lower than the ex vivo value (75% of ex vivo, P = 0.006). At days 2-7 of culture, Rev was associated with slight reductions in surface CD4 MFI (58% of ex vivo versus 78% of ex vivo for unstimulated cultures, P = 0.047) and greater effects on cytoplasmic CD3 MFI (131 versus 179% of ex vivo for unstimulated cultures, P = 0.035), and surface CD8 MFI (70% of ex vivo, P = 0.006 versus ex vivo value). The globality of Rev's effects suggests these are related to a shared processing pathway, i.e. not due to direct interaction with CD3, CD4 and CD8; the effects of HIV inoculation and Nef on CD8 expression appear to be more CD8 specific. Because CD8 is essential for cytotoxic T-cell function, its down-modulation could inhibit this activity, including anti-HIV cytotoxicity. Given the critical roles of CD3 and CD8 in T-lymphocyte signal transduction and antigen responsiveness, the effects of HIV, Rev and Nef on these molecules have clinically significant implications concerning the pathogenesis and treatment of HIV. 相似文献
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53.
Traditional approaches to generating tissue-engineered arteries in vitro rely on expansion of cells in culture to seed appropriate scaffolds. In most envisioned applications, small autologous blood vessels would be harvested and used as a source for these cells. We propose that small autologous arteries, not the cells derived from them, may be an attractive starting point for engineered arteries. This approach capitalizes on the ability of intact arteries to grow and remodel in response to chronic changes in their mechanical environment. Carotid arteries from juvenile (approximately 30-kg) pigs were stretched longitudinally in an ex vivo perfusion system over 9 days. This resulted in a 40% increase in artery length at physiological longitudinal stress and a 20 +/- 3% increase when unstressed. Control arteries were perfused for 9 days ex vivo at their physiological loaded length. Control and elongated arteries displayed native appearance (macroscopic and histological), excellent viability (cellularity and mitochondrial activity), normal vasoactivity, and similar mechanical properties (ultimate stress and ultimate strain) as compared with freshly harvested arteries. Growth, as opposed to just redistribution of existing mass, contributed to elongation as evidenced by an increase in artery weight. Results on elongation of arteries from neonatal and adolescent pigs are also presented and discussed. 相似文献
54.
Cornel AJ McAbee RD Rasgon J Stanich MA Scott TW Coetzee M 《Journal of medical entomology》2003,40(1):36-51
Comparisons of five morphological characters, 12 enzyme electrophoresis profiles, and Wolbachia pipientis infection rates were used to characterize populations of members of the Culex pipiens L. complex in California and South Africa. In South Africa, male phallosome DV/D ratio, male maxillary palp index, branching of siphonal seta 1a, the enzyme locus Mdhp-1, and W. pipientis infection rates proved highly diagnostic for separating Culex quinquefasciatus from Cx. pipiens phenotypes. In Johannesburg, where sympatric members of the Cx. pipiens complex were analyzed as one population, a significant Wahlund Effect was observed in the enzyme loci such as Ao, 6-Pgdh, Mdh-2, and Pgm. In California, all populations of the Cx. pipiens complex were in Hardy Weinberg equilibrium at all polymorphic enzyme loci examined. Additionally, in California, all populations had similar W. pipientis infection rates and appeared morphologically identical (except for DV/D ratio, in extreme north and south). These findings indicate that in South Africa, Cx. pipiens and Cx. quinquefasciatus remain as genetically distinct populations and behave as separate species. Conversely, in California, there is considerable genetic introgression between Cx. pipiens and Cx. quinquefasciatus, and they behave as a single species. 相似文献
55.
Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus 总被引:7,自引:0,他引:7
Berry JD Jones S Drebot MA Andonov A Sabara M Yuan XY Weingartl H Fernando L Marszal P Gren J Nicolas B Andonova M Ranada F Gubbins MJ Ball TB Kitching P Li Y Kabani A Plummer F 《Journal of virological methods》2004,120(1):87-96
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development. 相似文献
56.
Schillerstrom JE Horton MS Schillerstrom TL Joshi KG Earthman BS Velez AM Royall DR 《Psychosomatics》2005,46(5):411-417
The purpose of this study was to determine the prevalence, course, and risk factors for executive impairment in patients hospitalized on a general medicine service. One hundred patients were administered the Executive Interview (EXIT25), the Executive Clock Drawing Task (CLOX), and the Mini-Mental State Examination at admission and discharge. Fifty-two percent of the patients at admission and 56% at discharge had scores indicating impairment on at least one measure of executive function. Median scores on every measure improved during hospitalization. Older patients and those with a cardiac or gastrointestinal disorder were more likely to have executive impairment. The prevalence of executive impairment on general medicine services is high. Although improvement in executive function occurs during hospitalization, many patients remained impaired. 相似文献
57.
58.
The purpose is to incorporate clinically relevant factors such as patient-specific and dosimetric information as well as data from clinical trials in the decision-making process for the selection of prostate intensity-modulated radiation therapy (IMRT) plans. The approach is to incorporate the decision theoretic concept of an influence diagram into the solution of the multiobjective optimization inverse planning problem. A set of candidate IMRT plans was obtained by varying the importance factors for the planning target volume (PTV) and the organ-at-risk (OAR) in combination with simulated annealing to explore a large part of the solution space. The Pareto set for the PTV and OAR was analysed to demonstrate how the selection of the weighting factors influenced which part of the solution space was explored. An influence diagram based on a Bayesian network with 18 nodes was designed to model the decision process for plan selection. The model possessed nodes for clinical laboratory results, tumour grading, staging information, patient-specific information, dosimetric information, complications and survival statistics from clinical studies. A utility node was utilized for the decision-making process. The influence diagram successfully ranked the plans based on the available information. Sensitivity analyses were used to judge the reasonableness of the diagram and the results. In conclusion, influence diagrams lend themselves well to modelling the decision processes for IMRT plan selection. They provide an excellent means to incorporate the probabilistic nature of data and beliefs into one model. They also provide a means for introducing evidence-based medicine, in the form of results of clinical trials, into the decision-making process. 相似文献
59.
Chen JJ Huang JC Shirtliff M Briscoe E Ali S Cesani F Paar D Cloyd MW 《Journal of leukocyte biology》2002,71(2):271-278
Recent studies have provided evidence that macrophages from Th1-prone mouse strains respond with an M1 profile, and macrophages from Th2-prone mouse strains respond with an M2 profile, characterized by the dominant production of NO or TGF-beta 1, respectively. We have shown that peritoneal macrophages from IL-12p40 gene knockout mice have a bias toward the M2 profile, spontaneously secreting large amounts of TGF-beta 1 and responding to rIFN-gamma with weak NO production. Moreover, IL-12p40KO macrophages are more permissive to Trypanosoma cruzi replication than their wild-type littermate cells. Prolonged incubation with rIL-12 fails to reverse the M2 polarization of IL-12p40KO macrophages. However, TGF-beta 1 is directly implicated in sustaining the M2 profile because its inhibition increases NO release from IL-12p40KO macrophages. IFN-gamma deficiency is apparently not the reason for TGF-beta 1 up-regulation, because rIFN-gamma KO macrophages produce normal amounts of this cytokine. These findings raise the possibility that IL-12 has a central role in driving macrophage polarization, regulating their intrinsic ability to respond against intracellular parasites. 相似文献
60.
Variation in response latency to identical sensory stimuli has been attributed to variation in neural activity mediating preparatory set. Here we report evidence for a relationship between saccadic reaction time (SRT) and set-related brain activity measured with event-related functional magnetic resonance imaging. We measured hemodynamic activation time-courses during a preparatory "gap" period, during which no visual stimulus was present and no saccades were made. The subjects merely anticipated appearance of the target. Saccade direction and latency were recorded during scanning, and trials were sorted according to SRT. Both the frontal (FEF) and supplementary eye fields showed pre-target preparatory activity, but only in the FEF was this activity correlated with SRT. Activation in the intraparietal sulcus did not show any preparatory activity. These data provide evidence that the human FEF plays a central role in saccade initiation; pre-target activity in this region predicts both the type of eye movement (whether the subject will look toward or away from the target) and when a future saccade will occur. 相似文献