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Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring14C-oleic acid release from14C-triolein.14C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that Vmax=0.016 nmol/h/mg protein and that Km=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg2+. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.  相似文献   
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Details of the management, feeding, level of activity and routine health care of horses in Scotland and the five northernmost counties in England were recorded through a stratified random sample of horse owners who had responded to a previous survey. Sixty-eight per cent of the horses were kept where their owners resided, and 32 per cent were kept away from the owner's home. More than 99 per cent were turned out to grazing for at least part of the year and 81 per cent were stabled for at least part of the time, most commonly bedded on straw (50 per cent) or shavings (34 per cent). Hay was fed to 87 per cent, sugar beet pulp to 64 per cent and commercially prepared concentrate mixes to 60 per cent of the horses. Hacking was the most popular activity (52 per cent of horses) followed by riding/pony club events (28 per cent) and showing (21 per cent). The majority of the horses were involved in more than one activity. There were an estimated 0.88 veterinary visits per horse per year and 29 per cent of the horses were reported to suffer from at least one permanent or recurrent health disorder. The median annual numbers of administrations of vaccines (influenza and tetanus) and anthelmintics were one and seven respectively per horse, and each horse was shod a median seven times. There were significant differences in the management of horses kept in different types of premises and in areas of different human population density.  相似文献   
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Two mature breeding stallions were evaluated because of specific ejaculatory dysfunction, and each was found to have aortic-iliac thrombosis occluding 60 to 70% of the aortic lumen. In each case, the stallion had strong libido, normal mounting, and vigorous initial thrusting. With continued exertion, thrusting became weak and dismount was awkward. Treatment aimed at maximizing sexual arousal before mounting and reducing hind limb pain and exertion during breeding allowed these stallions to continue breeding. A program of gradually increasing daily exercise also was associated with improved breeding performance during subsequent breeding seasons. In cases of specific ejaculatory dysfunction, aortic-iliac disease should be included among the differential diagnoses.  相似文献   
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Equine herpesvirus abortion in Australia 1977 to 1982   总被引:1,自引:0,他引:1  
Until 1977 no case of abortion caused by equine herpesvirus 1 (EHV1) had been recorded in Australia although the virus, called equine rhinopneumonitis virus, had been known to have been present at least since 1962. Outbreaks of EHV1 abortion occurred in New South Wales in 1977 and in 1981. Sporadic cases of EHV1 abortion had been confirmed in some parts of Australia each year since 1975. It was concluded that an abortigenic subtype of EHV1 had been introduced to Australia in 1977 and that the previously endemic respiratory subtype occasionally caused abortion. Virus isolation in a variety of cell cultures and histopathological examination of tissue were shown to be satisfactory methods of diagnosis of EHV1 abortion. Lung proved to be the specimen of choice. Slight serological differences between "abortigenic" and "respiratory" subtypes of EHV1 were found in cross neutralisation tests. A serological survey of 219 Sydney horses of various ages revealed that most yearlings had already acquired neutralising antibody to both subtypes.  相似文献   
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Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (VL) and heavy chain (VH) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.  相似文献   
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