Inducible Arginase 1 Deficiency in Mice Leads to Hyperargininemia and Altered Amino Acid Metabolism

Arginase deficiency is a rare autosomal recessive disorder resulting from a loss of the liver arginase isoform, arginase 1 (ARG1), which is the final step in the urea cycle for detoxifying ammonia. ARG1 deficiency leads to hyperargininemia, characterized by progressive neurological impairment, persistent growth retardation and infrequent episodes of hyperammonemia. Using the Cre/loxP-directed conditional gene knockout system, we generated an inducible Arg1-deficient mouse model by crossing “floxed” Arg1 mice with CreER^T2 mice. The resulting mice (Arg-Cre) die about two weeks after tamoxifen administration regardless of the starting age of inducing the knockout. These treated mice were nearly devoid of Arg1 mRNA, protein and liver arginase activity, and exhibited symptoms of hyperammonemia. Plasma amino acid analysis revealed pronounced hyperargininemia and significant alterations in amino acid and guanidino compound metabolism, including increased citrulline and guanidinoacetic acid. Despite no alteration in ornithine levels, concentrations of other amino acids such as proline and the branched-chain amino acids were reduced. In summary, we have generated and characterized an inducible Arg1-deficient mouse model exhibiting several pathologic manifestations of hyperargininemia. This model should prove useful for exploring potential treatment options of ARG1 deficiency.     

Thirty-one flavors of Drosophila rab proteins

Rab proteins are small GTPases that play important roles in transport of vesicle cargo and recruitment, association of motor and other proteins with vesicles, and docking and fusion of vesicles at defined locations. In vertebrates, >75 Rab genes have been identified, some of which have been intensively studied for their roles in endosome and synaptic vesicle trafficking. Recent studies of the functions of certain Rab proteins have revealed specific roles in mediating developmental signal transduction. We have begun a systematic genetic study of the 33 Rab genes in Drosophila. Most of the fly proteins are clearly related to specific vertebrate proteins. We report here the creation of a set of transgenic fly lines that allow spatially and temporally regulated expression of Drosophila Rab proteins. We generated fluorescent protein-tagged wild-type, dominant-negative, and constitutively active forms of 31 Drosophila Rab proteins. We describe Drosophila Rab expression patterns during embryogenesis, the subcellular localization of some Rab proteins, and comparisons of the localization of wild-type, dominant-negative, and constitutively active forms of selected Rab proteins. The high evolutionary conservation and low redundancy of Drosophila Rab proteins make these transgenic lines a useful tool kit for investigating Rab functions in vivo.     

Extrusion of wheat or sorghum and/or addition of exogenous enzymes to pig diets influences the large intestinal microbiota but does not prevent development of swine dysentery following experimental challenge

A study was made of dietary influences on the large intestinal microbiota of pigs and on the incidence of swine dysentery (SD) after experimental infection with Brachyspira hyodysenteriae, the aetiological agent of SD. Animals were fed diets based either on wheat (expts 1 and 2) or sorghum (expt 2). Grains were ground and fed either raw or after high temperature and pressure extrusion and/or after addition of exogenous enzymes to the whole diet to reduce the starch and soluble non-starch polysaccharides available for fermentation in the large intestine. Limiting fermentation creates conditions that apparently reduce the incidence of SD after infection with B. hyodysenteriae. The diets were fed to weaned pigs for 4-6 weeks, then half the animals on each diet were killed and gut samples collected for microbiology. The treatments had little effect on bacterial numbers. In expt 1, dietary extrusion of wheat reduced lactobacilli in the large intestine. Addition of enzymes to extruded wheat-based diets in expt 2 reduced facultative anaerobes and increased non-sporing anaerobes. Addition of enzymes to a raw sorghum diet in expt 3 decreased numbers of facultative anaerobes, while extrusion of sorghum increased total anaerobes. Bacteroides spp. and Fusobacterium spp., which act in synergy with B. hyodysenteriae in SD, were isolated at a higher percentage in pigs fed the untreated wheat diet than in pigs fed the treated wheat diets. Following experimental infection the incidence of SD amongst pigs fed treated wheat diets was slightly lower than those fed the untreated diet, but with sorghum-based diets the opposite was found. Overall, the different dietary treatments used did not significantly reduce SD.     

A Comparison of the Phyllostomid Bat Assemblages in Undisturbed Neotropical Forest and in Forest Fragments of a Slash‐and‐Burn Farming Mosaic in Petén,Guatemala1

Using mist nets, we compared phyllostomid bat ensembles of continuous mature forest in Tikal National Park, Guatemala, and of forest fragments in the nearby farming landscape. Of 20 species captured, 13 were shared between treatments, 4 were unique to continuous forest, and 3 were unique to forest fragments. Dominance–diversity curves were similar for the two treatments except that Sturnira lilium comprised 43 percent of captures in the forest fragments, resulting in greater dominance there. Capture rates (and presumably relative abundance) differed significantly between continuous forest and forest fragments, both in terms of species and feeding guilds. Sturnia lilium and Dermanura sp. were captured significantly more often in forest fragments than in continuous forest, whereas Artibeus jamaicensis, A. lituratus, and Centurio senex were taken significantly more often in continuous forest. Large frugivores accounted for a higher proportion of total captures in continuous forest than in forest fragments, whereas small frugivores showed the opposite pattern. By their abundances, Carollia perspicillata and S. lilium are indicators of forest disturbance. The relative abundances of large frugivores, which feed on large fruits of mature forest trees, and small frugivores, which feed on small‐fruited plants occurring in early succession, are an indicator of forest disturbance. Other groups, such as large insect‐ and vertebrate‐eating bats, because of their low capture rates, are impractical as indicators for rapid assessment of forest disturbance based on mist netting, but may prove especially vulnerable to forest fragmentation.     

Effects of an application of granular carbaryl on nontarget forest floor arthropods

We evaluated the effects of a single application of granular carbaryl made against nymphal Ixodes scapularis Say on the diversity and abundance of forest arthropods taken in pitfall traps in oak and mixed oak-pine forest sites for 12 wk after treatment in central New Jersey. Significant short-term changes in arthropod assemblages were detected immediately posttreatment. Effects were not distributed equally across taxa. Seasonal changes in numbers and diversity of forest arthropods in the study areas may have affected the impact of the acaricide in the treatment area. Comparison with control areas indicated that reductions in abundance of some arthropod taxa in the treatment area were detectable 12 wk after treatment.     

Pyruvate dehydrogenase phosphatase 1 (<Emphasis Type="Italic">PDP1</Emphasis>) null mutation produces a lethal infantile phenotype

Pyruvate dehydrogenase phosphatase deficiency has previously only been confirmed at the molecular level in two brothers and two breeds of dog with exercise intolerance. A female patient, who died at 6 months, presented with lactic acidemia in the neonatal period with serum lactate levels ranging from 2.5 to 17 mM. Failure of dichloroacetate to activate the PDH complex in skin fibroblasts was evident, but not in early passages. A homozygous c.277G > T (p.E93X) nonsense mutation in the PDP1 gene was identified in genomic DNA and immunoblotting showed a complete absence of PDP1 protein in mitochondria. Native PDHC activity could be restored by the addition of either recombinant PDP1 or PDP2. This highlights the role of PDP2, the second phosphatase isoform, in PDP1-deficient patients for the first time. We conclude that the severity of the clinical course associated with PDP1 deficiency can be quite variable depending on the exact nature of the molecular defect.