Thymocyte and peripheral blood T lymphocyte subpopulation changes in piglets following in utero infection with porcine reproductive and respiratory syndrome virus

Piglets infected in utero with porcine reproductive and respiratory syndrome virus (PRRSV) are born severely immunocompromised. In this article we more closely examine the effects of in utero PRRSV infection on circulating and thymic T cell populations. Numbers of CD4+, CD8+, and dual-positive lymphocytes were quantitated in circulation and in the thymus during the 2 weeks following birth. At birth we found that the number of circulating lymphocytes was suppressed by 60%. Lymphocyte numbers were also suppressed by 42% at day 7, but by day 14 the number of lymphocytes had rebounded and was actually 47% greater than controls. At birth and day 7, a drop in the number of CD4+ cells could partially explain the suppression we observed, while the rebound in total lymphocyte numbers seen at day 14 was due to a nearly fourfold increase in the number of circulating CD8+ cells. As a result, the normal CD4+:CD8+ ratio of between 1.4 and 2.2 for neonatal pigs was reduced to 0.1-0.5. The thymuses of infected piglets were found to be 50% smaller than those of control pigs and were characterized by cortical involution and severe cortical depletion of thymocytes. Analysis of the population of thymocytes revealed that double-positive thymocytes were suppressed to a greater degree than either single positive subpopulation. In addition, we show that the number of thymocytes undergoing apoptosis was increased twofold in piglets infected with PRRSV. Taken together, these results help explain the dramatic immunosuppression observed in neonatal animals infected in utero with PRRSV.     

Initiating arbovirus surveillance in Arkansas, 2001

Migratory birds could introduce West Nile (WN) virus to Arkansas. The purpose of this study was to establish a cooperative arbovirus surveillance program to monitor mosquitoes and birds in Arkansas for arboviruses. Our objectives were to: 1) perform routine, multicounty collections of mosquitoes and test them for eastern equine encephalitis, St. Louis encephalitis, and WN viruses; and 2) conduct passive surveillance by testing dead wild birds for WN virus. Arbovirus surveillance was organized by the Arkansas Department of Health, University of Arkansas, and Vector Disease Control Incorporated. None of the 14,560 mosquitoes (425 pools) tested were virus positive. Two hundred forty-two dead birds from 62 counties were tested for WN virus. Four blue jays in three counties were positive. These infections are the first reported incidences of WN virus in Arkansas. Sera from five horses with suspected encephalitis all tested negative for WN, eastern equine encephalitis, and western equine encephalitis viruses.     

Teaching medical students about medical errors and patient safety: evaluation of a required curriculum.

PURPOSE: To assess the effectiveness of a brief curriculum about patient safety and medical errors with third-year medical students. METHOD: From 2000-03, third-year medical students at New York Medical College, Valhalla, New York, were required to participate in a new curriculum on patient safety and medical errors during their family medicine clerkships. Five hundred seventy-two students participated in a four-hour curriculum that included interactive discussion, readings, a videotape session with a standardized patient, and a small-group debriefing facilitated by a family physician. Before and after participating in the curriculum, students were asked to complete questionnaires on self-awareness about patient communication and safety. Curriculum evaluations and follow-up surveys were also distributed. Responses to each statement on the before and after questionnaires were compared using the Wilcoxon signed-rank test for matched data. RESULTS: Five hundred eleven (89%) students reported that the opportunity to present an error to a patient increased their confidence about discussing this issue with patients, and 537 (94%) students reported that they strongly agreed or agreed that the standardized patient and feedback exercise was a useful learning experience. A total of 535 before and after questionnaires were used in the analysis. A comparison of before and after questionnaire data revealed statistically significant increases in the self-reported awareness of students' strengths and weaknesses in communicating medical errors to patients (p 相似文献   

Phosphorylation of a C-SRC-related protein in macrophages activated in vitro with lymphokine

Treatment of proteose peptone elicited peritoneal macrophages from C3H/HeN mice or the macrophage cell line B6MP102 with a T-cell lymphokine preparation induces cytotoxicity for SV3T3 tumor cells. The Triton X-100 (TX-100) insoluble fractions from activated macrophages possessed kinase activity for an endogenous 53 kDa phosphoprotein (pp53) which was markedly greater than extracts from untreated macrophages. Addition of the tyrosine phosphatase inhibitor, Na₃,VO₄ to the cytotoxicity assay also enhanced tumor cell lysis and Na₃VO₄ treated macrophages showed increased phosphorylation of pp53. Moreover, addition of Na₃VO₄ to the cytoskeleton kinase assay enhanced the phosphorylation of pp53 in a dose dependent manner. Pp53 was immunoprecipitated from the in vitro phosphorylated TX-100 insoluble fraction with monoclonal antibody to pp60^v-src. Anti-pp60^v-src also precipitated a 53 and a 60 kDa phosphoprotein from whole cell extracts and from TX-100 cytoskeleton extracts of macrophages phosphorylated as viable intact cells. Addition of a known tyrosine kinase inhibitor, quercetin, to the macrophage cytoskeleton kinase assay inhibited phosphorylation of pp53, and the in vitro phosphorylated pp53 was resistant to 1 N NaOH hydrolysis, indicating phosphorylation of tyrosine residues. Immune complex kinase assays of anti-pp60^c-src precipitated TX-100 insoluble macrophage fractions revealed strong phosphorylation for α-casein which was inhibited by quercetin. These data suggest that macrophage pp53 is a c-src-related gene product that is inducible by stimuli that activate macrophages to cytotoxicity.     

Identification of stimuli that mediate experience-dependent modification of homosexual courtship inDrosophila melanogaster

A naive, sexually matureD. melanogaster male tested with a young, sexually immature male will perform vigorous courtship, but the mature male will perform much less courtship if he is subsequently tested with a second young male. This phenomenon is called experience-dependent courtship modification (EDCM). We have shown that exposure to either or both of the two courtship-stimulating pheromones that immature males synthesize is sufficient to induce EDCM.     

Diffusive-convective mass transfer rates for solutes present on both sides of a dialyzer membrane

The transport (J) of waste products across dialyzer membranes is known to be proportional to the blood inlet concentration (Cbi) according to J = KCbi, where K is the clearance. For solutes present on both sides of the membrane, like sodium chloride, it has been shown that under certain conditions the transport rate will depend linearly also upon the dialysis fluid inlet concentration Cdi according to J = KbCbi -KdCdi. Kb and Kd are generalized clearances, which depend upon flow rates and membrane permeability but are independent of the concentrations. We have extended the results of Ross et al. in three ways. First, they only considered ultrafiltration (UF) that is equally distributed along the dialyzer. This is an unrealistic assumption, especially in hemodiafiltration and hemofiltration treatments with large UF rates (Quf) leading to large pressure drops along the dialyzer. Our approach allows for an arbitrary UF distribution. Second, it was possible to incorporate the more realistic model of Villaroel et al. for the local combination of diffusion and convection. Finally, we allow an arbitrary distribution of blood among the different fibers. All of these results are valid in both cocurrent and countercurrent configurations. With a sieving coefficient of 1, a good approximation for small solutes, we were also able to show that Kd = Kb - Quf, irrespective of the UF distribution along the dialyzer. This is an important result that, for example, provides a theoretical foundation for allowing a nonzero Quf in conductivity based clearance measurements.     

Interstrain variability of larval photokinesis inDrosophila melanogaster

Larvae from seven laboratory strains and eight isofemale lines ofDrosophila melanogaster differ significantly with regard to their responses to light in a photokinesis assay in which the larvae are tested en masse. Larvae from the CA-2 laboratorystock fail to disperse on assay plates, although observations of individual CA-2 larvae suggest that the larvae are repelled by light. Larvae from all of the other laboratory stocks and all of the isofemale lines (except LI2 and NC5) avoid light in the photokinesis assay. Larvae from some stocks are much more strongly repelled by light than larvae from other stocks. LI2 larvae are unresponsive to light in most replicates of the photokinesis assay, while NC5 larvae are consistently unresponsive to light. Observations of F₁ heterozygotes suggest that the allele(s) that affects the vision of LI2 and NC5 larvae has net effects on the animals' behavior that are partially dominant and recessive, respectively.     

Amino acid supplementation improves cell-specific functions of the rat hepatocytes exposed to human plasma

Maintaining hepatocyte function during plasma exposure is critical for the successful development of hepatocyte-based bioartificial liver assist systems. Past attempts to culture hepatocytes in plasma yielded discouraging results. Using a stable culture model based on sandwiching hepatocytes between two layers of collagen gel, we investigated the effect of hormone and amino acid supplementation during exposure of rat hepatocytes to heparin-treated human plasma for 1 week. Morphology and hepatocyte-specific functions were evaluated for hepatocytes cultured in Dulbecco's Modified Eagle medium (DMEM), nonsupplemented plasma, plasma supplemented with hormones, or with hormones plus amino acids. Amino acids were supplemented at four-fold concentration of Basal Medium Eagle with 4 mM glutamine, whereas hormones included 7.5 microg/mL of hydrocortisone and 50 microU/mL of insulin. Cuboidal structure and bile canaliculi formation were observed throughout the 1-week exposure period for control hepatocytes in DMEM and for hepatocytes cultured in hormone supplemented plasma. Albumin and urea synthesis rates of hepatocytes in hormone plus amino acid supplemented plasma during the last day of plasma exposure were 60.4 +/- 13.7 and 75.6 +/- 6.5 (microg/day per 1 x 10(6) cells, mean +/- SD), respectively, comparable to cultures in standard culture medium. On the other hand, hepatocytes exposed to nonsupplemented plasma suffered significant morphological and functional damage. The results of this study indicate that hormone plus amino acid supplementation help to restore function in hepatocytes exposed to plasma.