Protein requirements of Texel crossbred lambs

A comparative slaughter trial was conducted to determine the protein requirements for maintenance and growth of feedlot Texel crossbred lambs. Thirty 11/16 Texel × 5/16 Ile de France crossbred noncastrated male lambs weaned at 42 days of age (16.2 ± 2.1 kg of shrunk body weight; SBW) were used. Five lambs were chosen randomly and slaughtered after 10 days of experimental management and diet adaptation (baseline group). Fifteen lambs were fed ad libitum and slaughtered at 25, 30 or 35 kg of SBW. The remaining 10 lambs were then assigned randomly to two levels of dry matter intake, either 70 or 55% of the ad libitum intake, and were slaughtered concomitantly with lambs slaughtered at 35 kg of SBW but given free choice access to feed. Total body N content and retention were determined. Additionally, six Texel × Ile de France crossbred lambs (30.4 ± 2.6 kg of SBW) housed in individual metabolic cages were used in a replicated 3 × 3 Latin square digestibility trial to evaluate diet digestibility and microbial protein synthesis at the different levels of feed intake. The endogenous N loss was 243 ± 29 mg/kg^0.75 of SBW, corresponding to a net protein requirement for maintenance of 1.52 ± 0.18 g/kg^0.75 of SBW. The metabolizable protein (MP) requirement for maintenance was 2.31 g/kg^0.75 of SBW, and the efficiency of MP use for maintenance was 0.66. Fleece-free body protein content decreased from 163 to 155 g/kg of empty body weight (EBW) as EBW increased from 13 to 28 kg. However, when protein in fleece was considered the whole-body protein content remained nearly constant. Net protein requirements for body weight gain and wool growth of lambs at 15 and 35 kg of SBW, and an average daily gain of 250 g, were 28.7 and 27.3 g/day and 3.8 and 5.8 g/day, respectively. Estimated efficiencies of MP use for body weight gain (k_pg) and wool growth (k_pw) were, respectively, 0.71 and 0.46. Growth pattern of the wool has a high influence on protein requirements of lambs. Texel crossbred growing lambs used in our study showed protein requirements for growth lower than those reported by the most nutritional systems.     

Comparative evaluation of PCR in Ziehl-Neelsen stained smears and PCR in tissues for diagnosis of Mycobacterium avium subsp. paratuberculosis

Thirty six tissues from sheep, previously diagnosed with paratuberculosis, were tested by PCR in positive Ziehl-Neelsen staining smears of tissues, and PCR in tissues targeting IS900 specific for Mycobacterium avium subsp. paratuberculosis. DNA amplification was achieved in 33.3% Ziehl-Neelsen smears, and in 61.1% tissue samples. Combination of both techniques found 66.7% samples as positive. Combination of techniques would, therefore, increase the sensitivity of diagnosis.     

Clomipramine Treatment and Repeated Restraint Stress Alter Parameters of Oxidative Stress in Brain Regions of Male Rats

This study aimed to compare the effects of repeated restraint stress alone and the combination with clomipramine treatment on parameters of oxidative stress in cerebral cortex, striatum and hippocampus of male rats. Animals were divided into control and repeated restraint stress, and subdivided into treated or not with clomipramine. After 40 days of stress and 27 days of clomipramine treatment with 30 mg/kg, the repeated restraint stress alone reduced levels of Na⁺, K⁺-ATPase in all tissues studied. The combination of repeated restraint stress and clomipramine increased the lipid peroxidation, free radicals and CAT activity as well as decreased levels of NP-SH in the tissues studied. However, Na⁺, K⁺-ATPase level decreased in striatum and cerebral cortex and the SOD activity increased in hippocampus and striatum. Results indicated that clomipramine may have deleterious effects on the central nervous system especially when associated with repeated restraint stress and chronically administered in non therapeutic levels.     

Seasonal changes in fish community structure of intermittent streams in the middle reaches of the Guadiana basin, Portugal

Seven sites on three tributaries with intermittent flow regimes in the middle reaches of the Guadiana, i.e. Xévora, Caia and Degebe, were sampled approximately every 3 months from April 1995 to April 1997. The density and biomass of fish were greater in downstream rivers and sites. Along the Xévora River, a series of point abundance samples showed evidence of fish zonation along the river. Canonical correspondence analysis (CCA) suggested a simple relationship between the different species and their preferred habitat. The small-sized Rutilus alburnoides and Leuciscus pyrenaicus used most or all of the habitats with some cover and flowing water. Greater concentrations of larger fish were found in downstream reaches which generally had greater depths as stream width increased: large Barbus microcephalus , B. steindachneri and B. comiza preferred such sites on the Degebe River. Droughts in seasonal Mediterranean streams such as the Guadiana River are a predictable natural disturbance. In the summer, flow ceases and some stretches of river consist of isolated pools. This results in intense aggregations of fish and possible competition for food and/or space. Although some species are well adapted to natural droughts, habitat degradation and possibly the introduction of exotic species contribute to marked variability in species composition.     

An improved synthesis of an umbelliferyl 5-thioxylopyranoside, precursor of the antithrombotic drug Iliparcil.

4-Ethyl-2-oxo-2H-1-benzopyran-7-yl 2,3,4-tri-O-acetyl-5-thio-beta-D-xylopyranoside, a synthetic intermediate of the orally active antithrombotic compound Iliparcil, has been prepared in 44-47% isolated yield. Different conditions were used for the glycosylation of 4-ethyl-2H-7-hydroxy-1-benzopyran-2-one 6 applying 2,3,4-tri-O-acetyl-5-thio-D-xylopyranosyl bromide (2), the analogous beta-chloride 3 or the alpha-trichloroacetimidate 5 as donors. With halides 2 and 3, the reaction was carried out in the presence of ZnO-ZnCl2 or ZnO alone. Both promoters are cheap, safe and therefore compatible with large-scale industrial processes.     

Hierarchical genetic and spatial structure among varieties and populations of Hymenaea stigonocarpa (Fabaceae) in Brazilian savannah

Tree Genetics & Genomes - Tree breeders must almost always address multiple traits. That entails choosing selection methods and weighing up known or assumed economic-worth functions for traits....     

Olfactory responses of Anaphes nitens (Hymenoptera,Mymaridae) to host and habitat cues

The eucalyptus weevil, Gonipterus spp. Schoenherr, 1833 (Coleoptera, Curculionidae) is considered a major pest of eucalyptus plantations. In regions where control is achieved, success is usually brought by the action of the solitary egg parasitoid Anaphes nitens (Girault, 1928) (Hymenoptera, Mymaridae). Research was conducted to ascertain which cues might mediate female wasp host location and selection. In Petri dish arenas, females were attracted to Gonipterus platensis Marelli, 1927 egg capsules, to G. platensis mated female faeces and to leaves of Eucalyptus globulus Labillardière, 1799. Gas chromatography–mass spectrometry electroantennographic detection analysis was conducted using extracts obtained from leaves of E. globulus, from G. platensis egg capsules, as well as from adults of both sexes and their faeces, in order to detect and identify compounds perceived by the wasp's olfactory system. The parasitoids were shown to detect a wide range of compounds emitted by each one of these sources, and for 31 compounds, antennal response was confirmed by dose-response tests. Further behavioural trials were conducted in Petri dishes in order to decode the effect, on parasitoid behaviour, of selected compounds emitted by E. globulus and of the pheromones, emitted by the weevils on parasitoid behaviour. Attraction was observed to two compounds emitted by E. globulus, namely 1,8-cineole and γ-terpinene, and to the main component of the male sex/aggregation pheromone, cis-verbenol. To our knowledge, this is the first report of attraction of a parasitoid from the family Mymaridae to a component of its host's sexual/aggregation pheromone. Similarly, to other egg parasitoid species, A. nitens females are likely to use the host plant volatiles as long-range host location cues and to adopt the ‘infochemical detour’ strategy in order to get in the vicinity of their hosts.     

Fluorometric determination of isocitrate dehydrogenase (EC 1.1.1.42; 1; NADP+ dependent) in ruminant milk

The enzyme isocitrate dehydrogenase (EC 1.1.1.42; 1; NADP⁺ dependent) located in the mammary cell cytosol mediates the synthesis of the majority of reducing equivalents for the energetically demanding milk fat and cholesterol synthesis in mammary cell cytosol. The present article presents a novel fluorometric method for quantification of the activity of this enzyme (IDH) in ruminant milk without pretreatment of the sample. Further, 493 goat milk samples – harvested before, during and after a nutritional restriction – were analysed for IDH activity i) with addition of extra substrate (isocitrate), and ii) with the intrinsic isocitrate solely. The IDH activity ranged from 0.22 to 15.4 units [nano moles product/(ml * min)] (un-supplemented) and from 0.22 to 45.6 units (isocitrate supplemented). The IDH activity increased considerably in milk during the nutritional restriction period concomitant with the increase in the metabolite isocitrate concentration and somatic cell count and returned to the initial level shortly after restriction period. The present ‘high through-put’ analytical method may be beneficial in future studies to phenotype modifications in mammary energy metabolism and milk fat synthesis, for which IDH activity may be a biomarker.     

Molecular evolution of the period gene in Drosophila athabasca

We measured nucleotide variability within and between the three semispecies of the Drosophila athabasca complex, at the period (per) gene by using a polymerase chain reaction-based four-cutter restriction- enzyme analysis. The levels of polymorphism varied considerably between the three semispecies. Our results for per, combined with previous data for X-linked allozymes, suggest that the X chromosome in the western- northern semispecies is less variable than expected under an equilibrium-neutral model. Both the pattern of divergence between the semispecies and a cladistic clustering of per haplotypes support the previously hypothesized grouping of eastern A and eastern B as the two most recently diverged semispecies. A 21-bp in-frame segment in the region of per which shares sequence similarity with the neuronal development gene single minded is deleted in all eastern A and eastern B flies examined but is present in all of the western-northern flies and all other published per sequences. Despite these hints that there may be significant differences at the per gene between the semispecies, especially the western-northern group versus the two eastern groups, there is no compelling evidence that per is involved in the mating song differences between the semispecies.      

Human LC3 and GABARAP subfamily members achieve functional specificity via specific structural modulations

ABSTRACT

Autophagy is a conserved adaptive cellular pathway essential to maintain a variety of physiological functions. Core components of this machinery are the six human Atg8 orthologs that initiate formation of appropriate protein complexes. While these proteins are routinely used as indicators of autophagic flux, it is presently not possible to discern their individual biological functions due to our inability to predict specific binding partners. In our attempts towards determining downstream effector functions, we developed a computational pipeline to define structural determinants of human Atg8 family members that dictate functional diversity. We found a clear evolutionary separation between human LC3 and GABARAP subfamilies and also defined a novel sequence motif responsible for their specificity. By analyzing known protein structures, we observed that functional modules or microclusters reveal a pattern of intramolecular network, including distinct hydrogen bonding of key residues (F52/Y49; a subset of HP2) that may directly modulate their interaction preferences. Multiple molecular dynamics simulations were performed to characterize how these proteins interact with a common protein binding partner, PLEKHM1. Our analysis showed remarkable differences in binding modes via intrinsic protein dynamics, with PLEKHM1-bound GABARAP complexes showing less fluctuations and higher number of contacts. We further mapped 373 genomic variations and demonstrated that distinct cancer-related mutations are likely to lead to significant structural changes. Our findings present a quantitative framework to establish factors underlying exquisite specificity of human Atg8 proteins, and thus facilitate the design of precise modulators.

Abbreviations: Atg: autophagy-related; ECs: evolutionary constraints; GABARAP: GABA type A receptor-associated protein; HsAtg8: human Atg8; HP: hydrophobic pocket; KBTBD6: kelch repeat and BTB domain containing 6; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MD: molecular dynamics; HIV-1 Nef: human immunodeficiency virus type 1 negative regulatory factor; PLEKHM1: pleckstrin homology and RUN domain containing M1; RMSD: root mean square deviation; SQSTM1/p62: sequestosome 1; WDFY3/ALFY: WD repeat and FYVE domain containing 3