Temporal-spatial expression of ANG-(1-7) and angiotensin-converting enzyme 2 in the kidney of normal and hypertensive pregnant rats.

We recently demonstrated that renin-angiotensin system (RAS) overactivity during late gestation in rats is associated with increased kidney and urine levels of ANG-(1-7) and enhanced kidney immunostaining of ANG-(1-7) and angiotensin-converting enzyme 2 (ACE2). To understand the temporal-spatial changes in normal and hypertensive pregnancies, the renal distribution of ANG-(1-7) and ACE2 in association with kidney angiotensin peptides and ACE2 activity was examined in virgin, normal pregnant (NP; gestational days 5, 15, and 19) and reduced uterine perfusion pressure (RUPP at day 19) pregnant Sprague-Dawley rats. ANG-(1-7) and ACE2 immunocytochemical staining increased 1.8- and 1.9-fold and 1.7- and 1.8-fold, respectively, at days 15 and 19 of NP, compared with virgin rats. ANG-(1-7) and ANG II concentrations were increased in the kidney at 19 days of gestation. ACE2 activity measured using a fluorescent substrate was increased 1.9- and 1.9-fold in the cortex and 1.9- and 1.8-fold in the medulla at days 15 and 19 of NP. In the RUPP animals, ANG-(1-7) immunostaining and concentration were significantly decreased compared with 19-day NP rats. ACE2 activity was unchanged in the cortex and medulla of RUPP rats. In conclusion, during NP, the concurrent changes of ACE2 and ANG-(1-7) suggest that ACE2 plays an important role in regulating the renal levels of ANG-(1-7) at mid to late gestation. However, the decrease in renal ANG-(1-7) content in the absence of a concomitant decrease in ACE2 implicates the participation of other ANG-(1-7) forming or degrading enzymes during hypertensive pregnancy.

Redescription of Besnoitia bennetti (Protozoa: Apicomplexa) from the donkey (Equus asinus).

Besnoitia bennetti tissue cysts were found in four naturally-infected donkeys (Equus asinus) from the USA. Infectivity of its bradyzoites and tachyzoites to animals and cell culture was studied. The bradyzoites were not infectious to out-bred Swiss Webster mice, rabbits or gerbils. When fed tissue cysts, cats did not excrete oocysts. However, the parasite was infectious to interferon-gamma gene knock out mice. The parasite from tissues of two donkeys was grown successfully in bovine monocyte monolayers for the first time. Non-dividing, uninucleate tachyzoites were approximately 6 x 1.5 microm in size. Longitudinally-cut bradyzoites in tissue sections measured 8.7 x 1.9 microm. Ultrastructurally, tachyzoites and bradyzoites were similar to those in other Besnoitia species, and in particular to parasites described from cattle (Besnoitia besnoiti) and reindeer (Besnoitia tarandi), in that their bradyzoites lacked enigmatic bodies. Based on comparative analysis of three portions of nuclear ribosomal DNA (the small and large subunits and the first internal transcribed spacer) B. bennetti was found to be more closely related to the other congeners described from ungulates. The parasite was formally redescribed and specimens deposited in the US National Parasite Collections.

Effects of exercise on plasma high-density lipoprotein cholesteryl ester metabolism in male and female miniature swine.

We studied the effects of exercise on high-density lipoprotein (HDL) cholesteryl ester (CE) metabolism in 6 male and 6 female miniature pigs fed a commercial swine diet supplemented with cholesterol and fat. The diets were fed for a total period of 20 weeks. During the last 12 weeks of the feeding period, the pigs were exercised on a motorized treadmill 5 days per week for 45 min/d at a speed of 9.5 to 10.0 km/h at 0% grade. Homologous HDL preparations were radiolabeled with cholesteryl (1-14C)oleate and intravenously administered to the pigs, followed by blood sampling at the appropriate time points and measurement of radiolabeled HDL CE. This was performed while the animals were sedentary and after the exercise period. Plasma cholesterol increased after the exercise protocol from 7.21 +/- 1.90 to 8.50 +/- 2.81 mmol/L (mean +/- SD, n = 6) in the females and from 8.11 +/- 3.61 to 10.07 +/- 3.61 in the males. HDL CE transport rates in female pigs were significantly lower (23%) after the exercise protocol (118 +/- 14 v 91 +/- 14 micromol/h/L plasma). HDL CE transport rates in the males were also lower (11%) after exercise (90 +/- 20 v 80 +/- 18 micromol/h/L plasma), but this effect was not statistically significant. Further, the residence time or life span of HDL CE was significantly longer after the exercise protocol in both male and female pigs. Thus, the results of this study suggest that exercise reduces the transport rate of HDL CE and prolongs the life span of HDL CE in hypercholesterolemic pigs.

Asbestos and cigarette smoke cause increased DNA strand breaks and necrosis in bronchiolar epithelial cells in vivo.

Coexposures to asbestos and cigarette smoke cause increased risks of lung cancer in asbestos workers. Although these carcinogens cause DNA damage to epithelial cells in vitro via generation of reactive oxygen species (ROS), it is unclear whether they cause injury to bronchiolar epithelial cells (i.e., the target cells of lung cancers in vivo). We exposed rats to amosite asbestos, cigarette smoke, and the two agents in combination for 1, 2, and 14 d. Numbers of cells exhibiting DNA strand breaks in comparison to sham rats were then evaluated in lungs using the terminal deoxynucleotidyl transferase (TDT)-mediated dUTP-biotin nick end labeling (TUNEL) method and by transmission electron microscopy (TEM). Increases in TUNEL-positive, necrotic epithelial cells occurred after exposure to asbestos alone and in an additive fashion after smoke and asbestos in combination. These results indicate that DNA strand breakage and necrosis are prominent mechanisms of injury by asbestos fibers and cigarette smoke in vivo to epithelial cells of the respiratory tract, thus validating in vitro observations from a number of laboratories.

Disseminated Rhodococcus equi infection in two goats.

Rhodococcus equi infection was diagnosed in two goats from the same herd. At necropsy, numerous caseating granulomas were disseminated throughout the liver, lungs, abdominal lymph nodes, medulla of right humerus, and the right fifth rib of goat No. 1, and the liver of goat No. 2. Histopathologic examination confirmed the presence of multiple caseating granulomas in these organs. Numerous gram-positive and Giemsa-positive coccobacilli were identified within the cytoplasm of macrophages. Aerobic bacterial cultures of the liver and lung from both goats yielded a pure growth of R. equi. R. equi antigens were immunohistochemically identified in caseating granulomas from both goats. However, the 15- to 17-kd virulence antigens of R. equi were not detected, suggesting possible infection by an avirulent strain of this organism.

Simultaneous triple fluorescence detection of mRNA localization, nuclear DNA, and apoptosis in cultured cells using confocal scanning laser microscopy.

We describe a multifluorescence labeling technique for simultaneous detection of mRNA, nuclear DNA, and apoptosis in cultured cells. Digoxigenin-labeled cRNA probes were used to study proto-oncogene expression in rat pleural mesothelial cells undergoing apoptosis following exposure to crocidolite asbestos or hydrogen peroxide (H2O2). Hybridized cRNA probe was detected by immunolocalization with an anti-digoxigenin monoclonal primary and fluorophore-conjugated anti-mouse secondary antibody. Cells undergoing apoptosis were simultaneously identified by the TdT-mediated biotin-dUTP nick-end labeling (TUNEL) method and a streptavidin-conjugated far-red fluorophore, and nuclear DNA was stained with oxazole yellow dimer (YOYO-1). With confocal scanning laser microscopy, we demonstrated increased c-jun mRNA expression within the cytoplasm of both TUNEL-positive and non-apoptotic cells following exposure to either crocidolite asbestos or H2O2. Thus, this technique represents a useful in vivo approach for evaluating apoptosis-associated gene expression with confocal scanning laser microscopy.

Malignant cauda equina paraganglioma in a cat.

An 11-year-old spayed female domestic long-haired cat presented for surgical removal of a slowly growing and deeply invasive 2.5 x 3.5-cm mass cranial to the base of the tail. Light microscopic examination of surgical biopsy specimens revealed an encapsulated mass composed of packets of polygonal cells of various sizes separated by a delicate fibrovascular stroma. Gömöri's reticulum stain revealed a characteristic endocrine or "Zellballen" pattern. Tumor cells contained diffuse positive reactivity to synaptophysin and neuron-specific enolase, reactions consistent with a neuroendocrine neoplasm. S-100 protein-positive cells reminiscent of sustentacular (support) cells occurred singly or in small clusters within tumor packets. At postmortem examination 3 months later, a 9- x 5- x 4-cm multinodular raised tan mass involving the caudal pelvis, sacrum, and tail-head regions was found. The base of this neoplastic mass originated within the cauda equina region and involved approximately five caudal nerve roots. Numerous 1-3-mm metastatic nodules were identified disseminated throughout the pulmonary parenchyma. The tumor was diagnosed as a malignant paraganglioma of the cauda equina region with pulmonary metastasis.

Extreme clonal diversity and divergence in populations of a selfing hermaphroditic fish.

Recombination is unknown in natural populations of Rivulus marmoratus, a selfing hermaphrodite, and genetic variation is likely due to mutation alone. DNA fingerprinting with an array of microsatellite [e.g., (CT)9] and minisatellite (e.g., the 33.15 core sequence) probes reveals very high clonal diversity within samples of seven Floridian populations, of which five contain about as many clones as there are individuals. There are 42 clones among 58 individuals surveyed (mean, 1.4 individuals per clone), a level of genetic diversity unprecedented among clonal animals. Moreover, all of the probes recognize the same clones even though, at high hybridization stringencies, there is little overlap in the fingerprint patterns they generate. This suggests that most sympatric clones differ by multiple and independent mutational steps. In one population studied in detail, the average number of mutational steps separating two clones is estimated at 9 or 10 and may be substantially higher. The mutational discontinuities among sympatric clones make it unlikely that they evolved by accumulation of neutral mutations in populations that are otherwise genetically uniform. The data argue that the mixing of unrelated individuals from different local populations occurs to an extent previously unappreciated and/or that divergence of clones is mediated by natural selection. If confirmed, the latter would be a serious challenge to current ideas on the predominant role of recombination in promoting the evolution of biological novelty.

The effect of hypotonicity on the mean platelet volume of patients with thrombo-embolism.

When dipotassium EDTA anticoagulated platelet rich plasma was incubated against a range of hypotonic saline concentrations, platelet volume was seen to change in a particular and reproducible fashion. When platelets taken from patients suffering a thromboembolic episode were tested in the same manner, a different pattern of response was observed the platelets being far less tolerant to osmotic stress. Similar changes occurred in platelets from ante-natal subjects. Platelets from both groups regained their tolerance to hypotonicity following either the thromboembolic incident or parturition respectively.

Genetic variation in clonal vertebrates detected by simple-sequence DNA fingerprinting.

The measurement of clonal heterogeneity is central to understanding the evolutionary and population genetics of the roughly 50 species of vertebrates that lack effective genetic recombination. Simple-sequence DNA fingerprinting with oligonucleotide probes (CAC)5 and (GACA)4 is a sensitive and efficient means of detecting this heterogeneity in natural populations of two clonal fishes, Poecilia formosa, an apomictic unisexual, and Rivulus marmoratus, a selfing hermaphrodite. The fingerprints are clonally stable for at least three generations. The technique clearly differentiates allozymically identical laboratory lines of R. marmoratus that were previously distinguishable only by histocompatibility analysis. The technique also reveals apparent cases of shifts in clonal composition of a natural population of each species. Clonal variation in most natural populations is quite high. For example, a sample of 19 specimens of P. formosa from one station on the Rio Soto la Marina contained 16 clones (average clonal frequency = 0.07). This level of clonal diversity implies that mutation, subsequent to the founding of clonal lineages, is an important source of variation in these populations. It also suggests that chance (sampling error) has a previously unappreciated role in determining the clonal composition of populations even though some of the clones may be divergent in biologically significant features.

A cluster analysis of learning disabled children.

Cluster analysis was employed with the intent of describing more homogeneous subgroups of learning disabled (LD) children. The parents and teachers of 29 children placed in special education as learning disabled completed questionnaires concerning demographic information regarding the families; birth, medical, and developmental histories regarding the children were also included. Variables for the analysis were those found, by means of discriminant analysis in an earlier study, to be significantly related to LD as opposed to regular education children. The presence of developmental delays (speaking or walking later than expected), birth order, and sex of the children were influential in the definition of the clusters.

Hemispheric asymmetry in the processing of Stroop stimuli: a balanced design.

Based on results from earlier research, the Stroop test appears to be a means for gaining insight into the specialized functions of the cerebral hemispheres as related to sex differences. This study was designed to determine whether sex differences continue to be present in a balanced design. Sixty-two psychology students responded to color words printed in incongruent colors when presented by a slide projector. Four mean scores comprising 15 responses in each condition, grouped word, grouped color, random word, and random color were computed for each subject. One half of the subjects responded to the grouped condition first, while the other half responded to the random condition first. Two variables were considered, sex and the order of presentation of random and grouped modes. Analyses of covariance of color response times over the categories of sex and order of presentation with word scores as covariates were performed for all conditions. For all individuals, response latency was consistently shorter when reading the word than when reporting the color. As in earlier studies, the covariate (word scores) was significantly related to the color sores. Sex differences were not found; however a significant difference was found in the analysis of the random color responses. When means were adjusted for sex and the covariate, subjects having the experience of the grouped experiment first scored faster than those performing for the first time. It appears that in this balanced design, practice improves time on color responses for both sexes.

A systems approach to planning large-scale, integrated health care facilities.

This paper presents an overview of a systems approach being implemented within the Kaiser-Permanente Medical Care Program (KPMCP) for the planning of large-scale, integrated health care facilities. This approach was motivated by the increasing change and complexity of factors, both external and internal to KPMCP, that now require more systematic treatment and that before were handled by more intuitive approaches. Among others, these factors include dealing with increasing regulation from government agencies and accommodating the active participation of knowledgeable personnel to ensure the continuation of a progressive and innovative system. Described within, along with more general issues, are various computer models, based upon Kaiser concepts and data that have been developed to facilitate the effective allocation of resources.

An input-output model for resources planning in a medical care system.

As medical care delivery systems grow in complexity, the understanding of interaction between entities within the system becomes a key aspect in resource planning. The model presented in this paper is geared to aiding resource planning both at the community or regional level and at the medical facility level. At the regional level the problem is related to the elimination of duplication and the sharing of services. At the medical facility level the concern is the balancing of the supply and demand of services among the departments. The framework of the model is a directed graph with nodes representing the service entities and branches representing the interrelationships. At the regional level, entities are hospitals, clinics, and centralized supportive units, such as the data center or the central laboratory. At the medical center level, entities are generally departments within the facility. An iterative procedure is used to simulate the propagation effect of a change on all entities. The model at the medical center level, as implemented in a minicomputer system was applied to a real problem. The results strongly correlated with another, independent study.