Identification of the major soluble cuticular glycoprotein of lymphatic filarial nematode parasites (gp29) as a secretory homolog of glutathione peroxidase

We have cloned and identified the major cuticular glycoprotein (gp29) of lymphatic filarial nematode parasites as a homolog of the antioxidant enzyme glutathione peroxidase. The derived amino acid sequence predicted a protein of 25.8 kDa, with an amino-terminal hydrophobic signal peptide and two sites for N-linked glycosylation, consistent with the documented properties of gp29. Transcription of a full-length cDNA in an SP65 vector and subsequent translation of the RNA in reticulocyte lysates in vitro generated a protein of 27 kDa, which was glycosylated upon the addition of pancreatic microsomal membranes. A postulated role for this secreted enzyme could be inhibition of the oxidative burst of leukocytes and neutralization of secondary products of lipid peroxidation, thus providing one explanation for the resistance of these parasites to immune effector mechanisms and their persistence in the mammalian host

Biochemical analysis of Caenorhabditis elegans surface mutants

A collection of Caenorhabditis elegans mutants that show ectopic surface lectin binding (Srf mutants) was analyzed to determine the biochemical basis for this phenotype. This analysis involved selective removal or labeling of surface components, specific labeling of surface glycans, and fractionation of total protein with subsequent detection of wheat germ agglutinin (WGA) binding proteins. Wild-type and mutant nematodes showed no differences in their profiles of extractable surface glycoproteins or total WGA-binding proteins, suggesting that the ectopic lectin binding does not result from the novel expression of surface glycans. Instead, these results support a model in which ectopic lectin binding results from an unmasking of glycosylated components present in the insoluble cuticle matrix of wild-type animals. To explain the multiple internal defects found in some surface mutants, we propose that these mutants have a basic defect in protein processing. This defect would interfere with the expression of the postulated masking protein(s), as well as other proteins required for normal development

Energy and protein nutritional requirements for Nellore bulls

The objective of this study was to determine the nutritional requirements of energy and protein and estimate the efficiencies of metabolizable energy utilization for fat and protein deposition, as well as for maintenance (k m) and growth (k g). An experiment of comparative slaughter was carried out with thirty-seven 14-month-old (±1 month) Nellore bulls with 259±24.9 kg. Animals were divided as follows: five to reference, four to maintenance level and twenty-eight bulls feeding ad libitum. Bulls were also grouped in 4 different feedlot periods (42, 84, 126 and 168 days) for slaughter. The diet was composed of corn silage and concentrate, at a 55:45 ratio. After the slaughter, the left half carcasses were totally dissected for determination of body composition. The energy requirements for maintenance were obtained by exponentially relating the heat production and the metabolizable energy intake, while the energy requirements for gain (NEg) were obtained according to empty body weight (EBW) and EBW gain (EBG). The net protein requirements for gain (NPg) were estimated according to EBG and retained energy (RE). The net (NEm) and metabolizable (MEm) energy requirements for maintenance were 76.5 and 113.84 kcal/EBW0.75/day, respectively. The k m was 0.67. The equations for NEg and NPg were: NEg (Mcal/day) = 0.0555 × EBW0.75 × EBG1.095 and NPg (g/day) = 263.37 × EBG - 23.21 × RE. The k g was 0.33. The efficiencies to deposition of energy as protein and fat were 0.18 and 0.71, respectively. The model obtained for the percentage of retained energy as protein (%REp) was %REp = 2.4221 × (RE/EBG)-1.6472. The net and metabolizable energy requirements for maintenance of Nellore bulls were 76.5 and 113.84 kcal/EBW0.75/day. The energy and protein requirements for gain could be obtained by the respective equations: NEg (Mcal/day) = 0.0555 × EBW0.75 × EBG1.095 and NPg (g/day) = 263.37 × EBG - 23.21 × RE

Operon Conservation and the Evolution of trans-Splicing in the Phylum Nematoda

The nematode Caenorhabditis elegans is unique among model animals in that many of its genes are cotranscribed as polycistronic pre-mRNAs from operons. The mechanism by which these operonic transcripts are resolved into mature mRNAs includes trans-splicing to a family of SL2-like spliced leader exons. SL2-like spliced leaders are distinct from SL1, the major spliced leader in C. elegans and other nematode species. We surveyed five additional nematode species, representing three of the five major clades of the phylum Nematoda, for the presence of operons and the use of trans-spliced leaders in resolution of polycistronic pre-mRNAs. Conserved operons were found in Pristionchus pacificus, Nippostrongylus brasiliensis, Strongyloides ratti, Brugia malayi, and Ascaris suum. In nematodes closely related to the rhabditine C. elegans, a related family of SL2-like spliced leaders is used for operonic transcript resolution. However, in the tylenchine S. ratti operonic transcripts are resolved using a family of spliced leaders related to SL1. Non-operonic genes in S. ratti may also receive these SL1 variants. In the spirurine nematodes B. malayi and A. suum operonic transcripts are resolved using SL1. Mapping these phenotypes onto the robust molecular phylogeny for the Nematoda suggests that operons evolved before SL2-like spliced leaders, which are an evolutionary invention of the rhabditine lineage