Demonstration of protein tyrosine phosphatase activity in the second of two homologous domains of CD45

It has been reported that alteration of deletion of critical residues within one of the two homologous protein tyrosine phosphatase (PTPase)-like domains of CD45 completely abolishes all activity, suggesting that only the more N-terminal domain is catalytically active. However, we now demonstrate, by two independent techniques, that the second (C-terminal) domain is also a viable phosphatase. Limited proteolysis by endoproteinase Lys-C or trypsin increased the phosphatase activity toward reduced, carboxymethylated, and maleylated lysozyme approximately 8-fold. A 50-kDa fragment, isolated by ion exchange chromatography, was found to be responsible for this activity. N-terminal sequencing revealed that this fragment includes less than half of the first phosphatase domain and most, if not all, of the second. In a second experiment, 109 residues, including the presumed catalytic region, were removed from domain I by site-directed mutagenesis. Expression of this construct in a mammalian cell line resulted in increased PTPase activity over nontransfected control cells. Isolation of the recombinant CD45 by immunoprecipitation and immunoaffinity chromatography revealed that it had phosphatase activity. Both of these experimental approaches demonstrate that the second conserved PTPase domain of CD45 is a functioning PTPase, but that external regulation may be required to express its activity in the context of the native molecule.     

Patterns of health-compromising behaviors among Minnesota adolescents: sociodemographic variations

OBJECTIVES: This study compared prevalence rates of health-compromising behaviors among boys and girls from different ethnic backgrounds in early, middle, and late adolescence and compared co-occurrences of such behaviors across gender and ethnic groups. METHODS: The study population included 123 132 adolescents in grades 6, 9, and 12. Adolescents completed a classroom-administered statewide survey focusing on high-risk behaviors, including unhealthy weight loss, substance abuse, suicide risk, delinquency, and sexual activity. RESULTS: Prevalence rates of most health-compromising behaviors differed by gender, increased with age, and tended to be highest among American Indian youth and lowest among Asian Americans. Strong associations were found between substance abuse and delinquency across all ethnic groups. Substance abuse and delinquency were associated with suicide risk across most ethnic groups. Covariations with sexual activity and unhealthy weight loss behaviors showed more ethnic variation. CONCLUSIONS: Prevention interventions should take into account the tendency for health-compromising behaviors to co-occur and should be sensitive to demographic and socioeconomic differences in behavior patterns.     

Health-related quality of life in patients served by the Department of Veterans Affairs: results from the Veterans Health Study

In a curative resection for advanced sigmoid or rectal cancer, an extensive dissection of the regional lymph nodes is generally required. This often necessitates the removal of the autonomic nerves around the inferior mesenteric artery. The present study was done in an attempt to clarify the influence of a neurectomy around the inferior mesenteric ganglion and plexus on the motility of the colon. In eight dogs, we resected the ganglion and plexus around the inferior mesenteric artery, together with an implantation of strain gauge force transducers in various parts of the colon, and 7-10 days later, colonic motility was examined. The percentage of contractile states and contractile forces increased at both the distal colon in fasting dogs, as well as at the middle colon in the late postprandial period. At the distal colon, contractile forces were noted in the early and late postprandial periods. These contractile abnormalities at the middle and distal colon may thus explain the frequent bowel movements or diarrhea often observed after extensive surgery in patients with sigmoid or rectal cancer.     

Multiple developmental stage-specific enhancers regulate CD8 expression in developing thymocytes and in thymus-independent T cells

We and others have recently identified a CD8 locus enhancer (E8) that directs expression in mature CD8 single-positive thymocytes and peripheral CD8+ T cells and in extrathymically derived intestinal intraepithelial lymphocytes (IEL). In this study, we show that deletion of E8, by homologous recombination results in reduced CD8alphaalpha homodimer expression on IEL. Since CD8 expression on thymus-derived T cells was normal, other enhancers regulate CD8 expression in these cells. By exploiting a transgenic reporter expression assay, we identified three additional enhancers that directed expression in diverse thymocyte subsets and mature T cells but not in CD8alphaalpha+ IEL. The results suggest that CD8alpha expression is primarily regulated by E8, in IEL and by the novel enhancers in the thymus-dependent lineages.     

Evaluation of an ultramicroELISA system for the diagnosis of rubella

An indirect Ultramicro ELISA assay, previously standardized in our laboratory, for detecting antibodies IgG to the rubella virus was assessed in comparison to the hemagglutination inhibition technique. This assessment allowed to determine its efficacy in the National System for Epidemiological Surveillance of this entity. One hundred and ninety serum pairs of clinically suspected cases of rubella were studied and a high percent of coincidence (99.4%), specificity (99.4%), and sensitivity (100%) was found between both techniques. In addition, 73 serum samples of blood donors were processed using an indirect microELISA system (Berhing) which was compared to the Ultramicro ELISA technique for rubella and it showed a 100% of sensitivity, specificity, and coincidence.