Comparison of retinal pigment epithelium cell preparations from the bovine eye

Summary Retinal pigment epithelium (RPE) cells were collected from, bovine eyes using a new method. The cells were harvested by vortexing the RPE and underlying choroid in 0.05 M citrate phosphate buffer, pH 5. RPE cells recovered by this method were compared to a standard method by microscopic examination of cell integrity, estimation of total protein, and assay of 11-cis and all-trans retinyl ester hydrolase (REH) activities. Results suggest that this method collects RPE cells of good integrity and with a significantly higher protein yield than the conventional method. Additionally, a much higher retinyl ester hydrolase activity was noted. Therefore we propose that this procedure offers a new and convenient method in the collection of RPE proteins for certain purposes such as enzyme purification.     

Development of venous occlusions in mice transgenic for the plasminogen activator inhibitor-1 gene

The fibrinolytic potential of the vasculature is modulated primarily by the availability and activity of plasminogen activators, which convert the zymogen plasminogen into the active fibrin-degrading enzyme plasmin. The activities of these key regulatory enzymes are directly neutralized by their primary endogenous inhibitor, plasminogen activator inhibitor-1 (PAI-1). Although some individuals with a tendency to develop thrombotic disorders exhibit elevated levels of PAI-1 in their plasma, the cause-and-effect relationship between increased PAI-1 and thrombosis is still unclear. Specifically, it is not known whether chronic depression of fibrinolytic activity results in the development of thrombosis. To address this question we developed transgenic mice in which the contribution of PAI-1 to thrombus formation could be evaluated. The results presented in this report indicate that elevated levels of PAI-1 contribute to the development of venous but not arterial occlusions.     

Effect of equal channel angular pressing on aging treatment of Al-7075 alloy

The effect of aging treatment on microstructure and mechanical properties of equal channel angular pressed Al-7075 alloy was examined.Commercial Al-7075 alloy in the solid solution heat-treated condition was processed by equal channel angular pressing through route BCat both the room temperature and 120 1C. Only three passes of equal channel angular pressing was possible due to the low ductility of the alloy at both temperatures. Followed by equal channel angular pressing, the specimens have been aged at 120 1C for different aging times. Mechanical properties were measured by Vickers microhardness and tensile tests and microstructural observations were undertaken using transmission electron microscopy, X-ray diffractometer as well as optical microscopy. Microstructural investigations showed that ultrafine-grained materials with grain size in the range of 200–350 nm and 300–500 nm could be obtained after three passes of equal channel angular pressing at room temperature and 120 1C, respectively. Equal channel angular pressing of solid solution heat-treated Al-7075 alloy accelerates precipitation rate and subsequently leads to a significant decrease in aging time to attain maximum mechanical properties. Furthermore, it is possible to achieve maximum mechanical properties during equal channel angular pressing at 120 1C as a result of dynamic aging and formation of small η′ phase.     

Adrenaline and the electrogenic sodium pump in Rana catesbeiana sympathetic ganglion cells.

The effect of adrenaline on the Na+-pump in bullfrog (Rana catesbeiana) sympathetic ganglion cells was studied by use of electrophysiological methods. The rate of removal of excess Na+ injected into a ganglion cell was increased by adrenaline. The K+-activated hyperpolarization of cell membrane, which might be produced by an electrogenic Na+-pump, was also increased by adrenaline. These results suggested that adrenaline was able to accelerate the Na+-pump, possibly the electrogenic Na+-pump.     

Selective agenesis of the dorsal pancreas in mice lacking homeobox gene Hlxb9.

The initial stages of pancreatic development occur early during mammalian embryogenesis, but the genes governing this process remain largely unknown. The homeodomain protein Pdx1 is expressed in the developing pancreatic anlagen from the approximately 10-somite stage, and mutations in the gene Pdx1 prevent the development of the pancreas. The initial stages of pancreatic development, however, still occur in Pdx1-deficient mice. Hlxb9 (encoding Hb9; ref. 6) is a homeobox gene that in humans has been linked to dominant inherited sacral agenesis and we show here that Hb9 is expressed at early stages of mouse pancreatic development and later in differentiated beta-cells. Hlxb9 has an essential function in the initial stages of pancreatic development. In absence of Hlxb9 expression, the dorsal region of the gut epithelium fails to initiate a pancreatic differentiation program. In contrast, the ventral pancreatic endoderm develops but exhibits a later and more subtle perturbation in beta-cell differentiation and in islet cell organization. Thus, dorsally Hlxb9 is required for specifying the gut epithelium to a pancreatic fate and ventrally for ensuring proper endocrine cell differentiation.     

Radiation hybrid map of the mouse genome.

Radiation hybrid (RH) maps are a useful tool for genome analysis, providing a direct method for localizing genes and anchoring physical maps and genomic sequence along chromosomes. The construction of a comprehensive RH map for the human genome has resulted in gene maps reflecting the location of more than 30,000 human genes. Here we report the first comprehensive RH map of the mouse genome. The map contains 2,486 loci screened against an RH panel of 93 cell lines. Most loci (93%) are simple sequence length polymorphisms (SSLPs) taken from the mouse genetic map, thereby providing direct integration between these two key maps. We performed RH mapping by a new and efficient approach in which we replaced traditional gel- or hybridization-based assays by a homogeneous 5'-nuclease assays involving a single common probe for all genetic markers. The map provides essentially complete connectivity and coverage across the genome, and good resolution for ordering loci, with 1 centiRay (cR) corresponding to an average of approximately 100 kb. The RH map, together with an accompanying World-Wide Web server, makes it possible for any investigator to rapidly localize sequences in the mouse genome. Together with the previously constructed genetic map and a YAC-based physical map reported in a companion paper, the fundamental maps required for mouse genomics are now available.     

Regional assessment of wadable streams in Idaho, USA

There has been a resurgence in applying bioassessment techniques for evaluating and monitoring the biological integrity of stream ecosystms. In all cases biological metrics have been refined to account for regional variation in aquatic habitats and fauna. This study evaluated environmental and macroinvertebrate properties for wadable streams in 3 major ecoregions of Idaho: Northern Basin and Range, Snake River Plain, and Northern Rocky Mountain. These 3 ecoregions constitute > 80% of the land area in Idaho. Reference streams were delineated from test streams in each ecoregion using standard habitat assessment protocols. (Plafkin et al. 1989). Multiple discriminant analysis effectively determined habitat (quantified measures) and macroinvertebrate differences between reference and test streams within ecoregions, although the results suggested that quantifiable habitat measures (e.g., water chemistry and nutrients) and biotic measures based on taxonomic groups (e.g., % Elmidae) improved the discriminatory power of evaluation procedures. Our results support the contention of a multi-metric approach for assessing differences among streams within an ecoregion. Lastly, individual metrics differed in their importance for evaluating stream condition among ecoregions, further emphasizing the importance of regionally stratifying metric selection or scoring procedures.     

Factors affecting selection of winter food and roosting resources by porcupines in Utah

Ecological and phytochemical factors potentially affecting winter dietary discrimination by porcupines ( Erethizon dorsatum ) in the mountain brush zone of Utah were studied. Porcupines utilized gambel oak ( Quercus gambelii ) as their primary winter food and roosting resource. Big-tooth maple ( Acer grandidentatum ) was the most common tree species in the study area but was rarely utilized by porcupines. Conifer species were used as a food and roosting resource significantly less often than they occurred in the study area, despite thermal advantages provided by their relatively dense canopies. Oak feed trees were successfully separated from conifer feed trees by discriminant analysis 100% of the time. Oak trees were correctly classified as feed and nonfeed trees 71% of the time. Gambel oak contained higher amounts of crude protein, fiber, and tannins, but was lower in either extract fractions and fatty acid content than conifers. A layer of adipose tissue used as an energy reserve by porcupines may have relaxed energy intake demands sufficiently to permit them to concentrate on a diet of oak tissue, which is high in protein, rather than a high-fat conifer diet. A diet relatively high in protein may have facilitated digestion of food material high in fiber. Temperature did not affect selection of tree species for roosting. Rock and snow caves were utilized infrequently and the study population ranged widely. Three of 15 study animals were eaten by predators.