A model of sterile vesicoureteric reflux in the sheep

Eighteen Coopworth ewe lambs were divided into three groups based on the initial cystourethrogram and cystometry findings at 5-7 weeks of age: group 1, 6 lambs with spontaneous low-pressure bilateral vesicoureteric reflux (VUR) on bladder filling were used to study the natural history of reflux; group 2, 5 lambs with no VUR detected were used to establish an experimental model of bilateral VUR using an unroofing surgical procedure; group 3, 7 lambs with spontaneous VUR detected during micturition had the same surgical procedure to increase the degree of VUR. All three animal groups were followed for 4-10 months. Spontaneous VUR was demonstrated in 13 of 18 lambs (25/36 ureters). The presence and severity of spontaneously occurring reflux in group 1 lambs diminished with increasing age. VUR was created successfully in group 2 and increased in degree in group 3 animals. The only significant histological finding in all three animal groups with grades II and III VUR was distal renal tubular dilatation. The sheep is a useful and readily available animal for studying VUR. During 4-10 months of follow-up, sterile reflux without bladder outflow obstruction resulted in distal renal tubular dilatation, but no renal parenchymal damage.     

Entry of Borrelia burgdorferi into macrophages is end-on and leads to degradation in lysosomes

Blastocyst-derived pluripotent mouse embryonic stem cells can differentiate in vitro to form so-called embryoid bodies (EBs), which recapitulate several aspects of murine embryogenesis. We used this in vitro model to study oxygen supply and consumption as well as the response to reduced oxygenation during the earliest stages of development. EBs were found to grow equally well when cultured at 20% (normoxia) or 1% (hypoxia) oxygen during the first 5 days of differentiation. Microelectrode measurements of pericellular oxygen tension within 13- to 14-day-old EBs (diameter 510-890 micron) done at 20% oxygen revealed efficient oxygenation of the EBs' core region. Confocal laser scanning microscopy analysis of EBs incubated with fluorescent dyes that specifically stain living cells confirmed that the cells within an EB were viable. To determine the EBs' capability to sense low oxygen tension and to specifically respond to low ambient oxygen by modulating gene expression we quantified aldolase A and vascular endothelial growth factor (VEGF) mRNAs, since expression of these genes is upregulated by hypoxia in a variety of cells. Compared with the normoxic controls, we found increased aldolase A and VEGF mRNA levels after exposing 8- to 9-day-old EBs to 1% oxygen. We propose that EBs represent a powerful tool to study oxygen-regulated gene expression during the early steps of embryogenesis, where the preimplantation conceptus resides in a fluid environment with low oxygen tension until implantation and vascularization allow efficient oxygenation.     

Carcinoma of the stomach following the Chernobyl nuclear accident

Medical consequences of many nuclear accidents on humans are well studied, but the results pertaining to gastric cancer patients who were exposed to radiation as a result of the Chernobyl nuclear accident have not been analysed. In this study, the outcome of the surgical treatment of 68 gastric cancer patients who were exposed to radiation as a result of the Chernobyl nuclear accident was compared with that of 117 consecutive gastric cancer patients from uncontaminated areas of the Ukraine. Patients in the study group was significantly younger than that of the control group. Comparative analysis showed the same frequency of regional metastases (65.7% versus 71.1%, P > 0.05), but a smaller number of distant metastases (23.8% versus 38.1%, P < 0.05) in the study group. 41.2% of patients in the study group underwent total gastrectomy compared to 19.6% of patients in the control group (P = 0.002). Postoperative complications developed in 13.2% of patients in the study group, while postoperative mortality in the study group was 7.3% compared to 1.7% in the control group. A significant decrease in CD16 cells was noted in patients from the study group following the operative procedure. Young age, invasive tumours with smaller number of distant metastases, frequent necessity for total gastrectomy and combined operations with adjacent organs, a higher level of postoperative morbidity and mortality and low levels of natural killer cells (CD16+) with a tendency to decrease after surgery are characteristic of patients with carcinoma of the stomach affected by the Chernobyl accident.     

Mortality studies of machining fluid exposure in the automobile industry. V: A case-control study of pancreatic cancer

MANOVA and repeated measures ANOVA approaches have provided evidence of a number of limitations in several event-related potential (ERP) studies due to violations of their statistical assumptions and the typically moderate size of the available sample. Alternative, computer-intensive methods based on permutation principles have recently been developed. Up to now this methodology has focused mostly on magnitude differences between scalp distributions as measured by t statistics. In this paper the scope of permutation techniques in ERP analysis was widened. A new statistic (D statistic) is introduced to compare the shapes of scalp distributions of ERPs. Additionally a general non-parametric combinatory technique is introduced to evaluate, by means of multivariate permutation tests, several time points and/or recording sites in ERP data. The methodology described here was used to test if two ERP components elicited during word-pair matching tasks to semantic or phonological incongruences had different scalp distributions.     

Secretion of cardiac plasminogen activator during hypoxia-induced right ventricular hypertrophy

In the circulation, fibrinolytic activity is determined to a large degree by the relative levels of tissue plasminogen activator (tPA) and its major inhibitor (PAI-1). Vascular beds in different organs secrete tPA and PAI-1 into the circulation, and the total secretory rate of each protein is balanced by its half-life in the bloodstream. We are testing the hypothesis that in the heart, ventricular hypertrophy will alter the rates of formation of tPA and/or PAI-1 and the rates of their release into the cardiac vasculature. In this study, we have examined the effects of continuous hypoxia on PA activity in extracts of rat heart ventricles, on the activity secreted into the cardiac vasculature of perfused hearts, and on the levels of mRNAs for tPA and PAI-1. Rats were subjected to hypobaric hypoxia at 0.5 atm for 1-21 days. The treatment caused polycythemia within 1-3 days, and right ventricular hypertrophy by 3 days. PA activity in extracts of both right and left ventricles was significantly elevated after 3 days of hypoxia, continued to increase for 4 additional days, and remained elevated for 3 weeks. The actions of inhibitors of urokinase and tPA indicated that the PA activity in heart extracts was exclusively tPA. Fibrin zymography confirmed that result. The mRNAs for tPA and for PAI-1 were elevated after 1 day of hypoxia and then returned to near control levels on days 2 and 3. After 7 days, hearts from hypoxic rats secreted more tPA activity into perfusates than did hearts from controls. The difference in secretory rates was proportional to the differences in the levels of tPA in the corresponding heart extracts.     

Pharmacology and intracellular signaling mechanisms of the native human orphan receptor BRS-3 in lung cancer cells

Neither the native ligand nor the cell biology of the bombesin (Bn)-related orphan receptor subtype 3 (BRS-3) is known. In this study, we used RT-PCR to identify two human lung cancer lines that contain sufficient numbers of native hBRS-3 to allow study: NCI-N417 and NCI-H720. In both cell lines, [DPhe6,betaAla11,Phe13, Nle14]Bn(6-14) stimulates [3H]inositol phosphate. In NCI-N417 cells, binding of 125I-[DTyr6,betaAla11,Phe13,Nle14]Bn(6-14) was saturable and high-affinity. [DPhe6,betaAla11,Phe13,Nle14]Bn(6-14) stimulated phospholipase D activity and a concentration-dependent release of [3H]inositol phosphate (EC50 = 25 nM) and intracellular calcium (EC50 = 14 nM); the increases in intracellular calcium were primarily from intracellular stores. hBRS-3 activation was not coupled to changes in adenylate cyclase activity, [3H]-thymidine incorporation or cell proliferation. No naturally occurring Bn-related peptides bound or activated the hBRS-3 with high affinity. Four different bombesin receptor antagonists inhibited increases in [3H]inositol phosphate. Using cytosensor microphysiometry, we found that [DPhe6,betaAla11,Phe13, Nle14]Bn(6-14) caused concentration-dependent acidification. The results show that native hBRS-3 receptors couple to phospholipases C and D but not to adenylate cyclase and that they stimulate mobilization of intracellular calcium and increase metabolism but not growth. The discovery of human cell lines with native, functional BRS-3 receptors, of new leads for a more hBRS-3-specific antagonist and of the validity of microphysiometry as an assay has yielded important tools that can be used for the identification of a native ligand for hBRS-3 and for the characterization of BRS-3-mediated biological responses.     

Evaluation of clothing systems to determine heat strain

This article describes the basic evaluation process and test methodology employed when temperature extremes for clothing systems must be considered as part of the U.S. Army's Health Hazard Assessment for material in the development and acquisition process. The goals of the evaluation are to select clothing systems that minimize the hazards of heat strain and to predict the heat strain for persons wearing such clothing. Clothing evaluations begin with biophysical assessments that determine the thermal characteristics (vapor permeability and insulation) for textiles via guarded hot plate tests and for clothing systems via thermal manikin tests. The results from biophysical tests can be used to select the textile and/or clothing with the best thermal characteristics. The data from manikin evaluations also can be used in prediction modeling. Human physiological testing is best done in a controlled laboratory environment, although for realism and user acceptability field trials may also be conducted. Proven test and measurement methods must be employed, and tests must control for confounding variables; subjects serve as their own controls, and test environment and procedures are consistent between trials. The process and test methodology described can be applied to the evaluation of civilian clothing systems as well as to the military systems for which they were developed.