Biochemical properties of the pathogenesis-related proteins from tobacco

This review describes the discovery and identification of the pathogenesis-related proteins (PRs) from tobacco. In crude leaf extracts the PRs are distinguished from the proteins in uninfected plants by their solubility at pH 3, resistance to a range of proteases, and mobility in polyacrylamide gels upon electrophoresis (PAGE) in non-denaturing conditions. PAGE has been used as a qualitative and semi-quantitative assay for PRs, and their migration in gels made from different acrylamide concentrations has been used to identify charge and size isomers and electrophoretically identical PRs in different tobacco cultivars. The subunit composition and molecular weight (mol. wt) of the four PRs identified first in Xanthi-nc were determined by SDS-PAGE; staining the gels has shown that these same four proteins in Samsun NN did not contain carbohydrate, lipid or nucleic acid, nor were they isozymic forms of twenty five enzymes known to increase in activity following infection with TMV. Evidence suggests that most of the PRs in Xanthi-nc and Samsun NN are extracellular.The purification of several PRs from Xanthi-nc, Samsun NN and other tobaccos is described, as well as their mol. wt, subunit and amino acid composition. PRs 1a, b and c consist of a single polypeptide and have similar mol. wt and amino acid compositions. Antisera prepared against purified Xanthi-nc b₁ protein have been used to determine serological relationships between PRs and form the basis of a very sensitive quantitative assay using ELISA. The regulation of synthesis of some PRs has been shown to involve translational control.     

The metabolism of R-20458 [(E)-6,7-epoxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene] in rat hepatocyte suspensions

The metabolism of R-20458 [(E)-6,7-epoxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene] by rat hepatocytes has been analyzed and compared with that of juvenile hormone I [methyl-(E,E)-cis-10,11-epoxy-7-ethyl-3,11-dimethyl-2,6-tridecadienoate] under identical conditions. The metabolism of R-20458 is characterized by the predominance of NADPH-dependent cytochrome P-450 and epoxide hydrolase reactions; whereas, JH I is metabolized mainly by carboxylesterase, epoxide hydrolase, and glutathione S-transferases. The metabolites of R-20458 have been shown to correspond to (E)-6,7-epoxy-1-(4-hydroxyethylphenoxy)-3,7-dimethyl-2-octene; (E)-6,7-epoxy-1-(4-acetylphenoxy)-3,7-dimethyl-2-octene; (E)-6,7-dihydroxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene; and, (E)-6,7-dihydroxy-1-(4-acetylphenoxy)-3,7-dimethyl-2-octene. The production of the α-hydroxyethyl, p-acetylphenoxy, and acetylphenoxy-6,7-diol metabolites is markedly inhibited by SKF 525-A. No dramatic effects are produced by diethylmaleate and 1,2-epoxy-3,3,3-trichloropropane.     

Endoscopic sex determination and gonadal manipulation in Gulf of Mexico sturgeon (Acipenser oxyrinchus desotoi).

Seventeen Gulf of Mexico sturgeons (Acipenser oxyrinchus desotoi) underwent endoscopic sex determination, gonadal biopsy, and various reproductive surgeries as part of a conservation development plan. The fish were anesthetized with tricaine methanesulfonate (MS-222) buffered with sodium bicarbonate and maintained on a recirculating water anesthesia circuit. A 6-mm Ternamian EndoTip Cannula, placed through the ventral midline, midway between pectoral and pelvic fins, permitted the introduction of a 5-mm telescope. Swim bladder aspiration and CO2 insufflation of the coelomic cavity provided excellent observation. Second and third cannulae were placed under direct visual control, lateral and cranial or caudal to the telescope cannula. Sex determination was successfully performed in all fish; however, five of 17 sturgeons (29%) required endoscopic gonadal biopsy to confirm sex. Bilateral ovariectomy or orchidectomy was successfully performed in three males and four females. Unilateral ovariectomy and bilateral ligation of the müllerian ducts using an extracorporeal suturing technique was accomplished in an additional three females. No apparent morbidity was associated with the anesthesia or endoscopic surgery in any fish. The ability to safely perform minimally invasive reproductive surgery in fish may have important management and conservation benefits.     

Evaluation of vegetable protein in canine diets: Assessment of performance and apparent ileal amino acid digestibility using a broiler model

Recent technological advances in the human food industry with respect to meat processing have decreased the availability of animal proteins to the pet food industry which typically formulates diets with an excess of animal protein. In the long term, this is not sustainable; thus, alternative protein sources need to be investigated. This study examined three canine diets, comparing a typical animal protein‐based diet (control) with two experimental diets where the animal protein was substituted in part with vegetable protein (formulated based either on total protein or amino acid content) using a broiler model. Each diet was fed to six cages each containing two birds from day 15, 18 cages in total (36 birds). Excreta were collected from days 19 to 21. On day 23, birds were euthanized and weighed, and their ileal digesta were collected and pooled for each cage. In addition, one leg per cage was collected for evaluation of muscle mass. Results showed no significant difference in animal performance (feed intake or live weight gain) or muscle to leg proportion across the diets. Birds fed the control diet and the diet balanced for amino acid content exhibited the greatest coefficients of apparent metabolizability for nitrogen (p < .001). Birds fed the diets that contained partial replacement of animal with vegetable protein generally had greater ileal digestibility of amino acids compared to birds fed the control (animal protein) diet. Analysis of excreta showed no dietary difference in terms of dry matter content; however, birds fed the diet balanced for total protein and the diet balanced for amino acid content had significantly greater excreta nitrogen than the control (p = .038). Overall, the study suggests vegetable proteins when formulated based on amino acid content are a viable alternative to animal proteins in canine diets.