Norsalsolinol uptake into secretory vesicles via vesicular monoamine transporter and its secretion by membrane depolarization or purinoceptor stimulation in PC12 cells.

The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter.     

Effect of heterogeneous digesta chemical composition on the accuracy of measurements of fiber flow in dairy cows.

The objective of the current study was to evaluate the effect of unrepresentative sampling of digesta particulate matter entering the omasal canal on the accuracy of fiber flow measurements. The experimental design comprised one period, one diet, and three cows as experimental units. Within each cow, the physical and chemical composition of digesta particulate matter was assessed at seven sites within the digestive tract. Three Finnish Ayshire dairy cows, equipped with ruminal and simple-T duodenal cannulas, in extended lactation were offered grass silage twice daily on an ad libitum basis. Digesta samples were collected from the rumen (dorsal and ventral sac), reticulum, omasal canal, omasum, duodenum, and rectum to determine particle size distribution in digesta, chemical composition of various particle size fractions, and distribution of two flow markers (Cr-labeled straw and indigestible NDF [INDF]) among particle size fractions. Digesta samples were wet-sieved using sieves of 2.50, 1.25, 0.630, 0.315, 0.160, and 0.080 mm. Particulate matter was analyzed for OM, NDF, and Cr concentrations, and INDF concentration was determined based on 12-d ruminal incubation. The particle size of digesta entering the omasal canal was larger compared with the omasum or the duodenum, suggesting that omasal canal samples were not representative of particle size distribution truly escaping the rumen. The concentration of potentially digestible NDF (PDNDF) decreased with decreasing particle size. The PDNDF concentration of particulate matter retained on all sieves was greatest in the rumen and gradually decreased along the digestive tract. From the reticulorumen to the omasum, the decrease was associated with decreased particle size, reflecting selective passage of particulate matter. In contrast, from the omasum to the duodenum and rectum, the PDNDF concentration decreased within each particle size fraction without effect on particle size, indicating a nonselective passage of particulate matter between these sites. Variation between particle size fractions was slightly greater for Cr concentration than for INDF concentration, indicating that unrepresentative sampling of particulate matter had a greater effect on Cr concentration compared with that of INDF. Owing to unrepresentative sampling, NDF entering the omasal canal was overestimated by 5% using INDF and underestimated by 7% using Cr as a particle phase marker. Of total NDF digestibility, proportionally 0.90, 0.07, and 0.03 occurred in the reticulorumen, omasum, and intestines, respectively. The current results indicate that, despite unrepresentative sampling of digesta particulate matter entering the omasal canal, the errors in determined NDF flow were small. The omasum may have a greater role in postruminal NDF digestion than the intestines.     

The Effect of Exogenous Hormone Treatment on Spermiation in Rhynchocypris oxycephalus (Sauvage and Dabry)

For the evaluation of hormonal control of spermiation in fish, a method to quanify the spermiation response of mature Rhynchocypris oxycephalus (Sauvage and Dabry) to hormonal therapy is described. Spermatocrit was determined after 7 min centrifugation at 18,000 ± g and sperm density was estimated by a standard hemocytomer method. Sperm density can be predicted from spermatocrit since their relationship is linear as described by the regression equation, Y = 3.68X - 27.18 ( R ² = 0.82, N = 50). where Y is spermatocrit and × is sperm density. Milt production by mature R. oxycephalus was highest at 24 h after injection of 1,000 IU human chorionic gonadotropin (HCG) and 50 μg luteinizing hormone-releasing hormone analogue (LHRHa) per kg body weight. Increased milt production coincided with low spermatocrit and sperm density levels. These results demonstrate that spermiation in mature R. oxycephalus can be reliably evaluated by a spermatocrit method and that HCG and LHRHa are effective in stimulating of spermiation in this species.     

Morphological study on the stomach of the lesser mouse deer (Tragulus javanicus) with special reference to the internal surface.

The stomach of the lesser mouse deer (Tragulus javanicus) was observed macroscopically. It consisted of only three compartments, rumen, reticulum and abomasum without omasum. The rumen was S-shaped with large ventral and caudoventral blind sacs and the reticulum was larger than the abomasum. Internally, the rumen was covered with numerous ruminal papillae even on the pillars and the ruminoreticular fold. These papillae were leaf- or tongue-like shaped and varied in size and density. The reticulum had honey-combed crests and the secondary crests were found rarely. The lips of the reticular groove were prominent and more developed in the aboral part than in the oral one. A sac-like transition zone, which had more prominent mucosal folds than had the floor of the reticular groove, was observed between the caudal end of the reticular groove and the abomasum. Mucosal folds of the abomasum were spiral, low but rather thick. These findings were discussed in view of comparison with other ruminants and of possible functional implications.     

ULTRASONOGRAPHY OF ALIMENTARY LYMPHOSARCOMA IN THE CAT

Clinical, radiological and ultrasonographic findings from 22 cats with gastrointestinal lymphosarcoma were reviewed. The most common clinical findings were anorexia, weight loss, vomiting, lethargy, depression, fever, anemia and a palpable abdominal mass. Abdominal radiographs of 12 cats revealed a mass associated with the gastrointestinal tract and/or mesentery, and decreased serosal detail, suggestive of peritoneal effusion. The most common ultrasonographic finding was transmural gastric or intestinal thickening associated with loss of normal wall layering, reduced wall echogenicity and localized hypomotility. As is noted in people, other patterns such as transmural-segmental, transmural-nodular, transmural-bulky and mucosal infiltration were represented. In 3 cats, mesenteric lymph-adenopathy was responsible for most of the mass-effect. Fine-needle aspiration biopsy, automated microcore biopsy, endoscopic biopsy or necropsy confirmed the diagnosis of lymphosarcoma in all cats. Chemotherapy was instituted in 19 cats. Only 6 cats had a complete response to treatment. The results of this study indicate that ultrasonography is a valuable tool for the diagnosis of feline alimentary lymphosarcoma and that biopsies can be obtained under ultrasonographic guidance.     

Evaluation of an enzyme-linked immunosorbent assay for diagnosis of trichinellosis in swine.

Experimental and field trials were conducted to evaluate an ELISA for its ability to detect Trichinella-infected domestic swine and to compare ELISA results with muscle-digestion test results. The ELISA used was a commercial double-antibody kit, containing an excretory-secretory antigen, and was evaluated principally for epidemiologic use. Experimentally induced infection in swine (4 groups of 3 pigs each; inoculated with 0, 50, 500 or 5,000 larvae) was detected as early as postinoculation week 4, with seroconversion of all inoculated swine by postinoculation week 8. The rate of seroconversion appeared to be affected by initial larval dose, time after inoculation, and immunocompetence of the individual host. Determination of antibody kinetics generally revealed rapidly increasing antibody titer, followed by its steady decrease in most pigs. Once seropositive, however, all pigs remained seropositive for the duration of the 10-week study. Presence of muscle larvae was confirmed in all infected pigs at termination of the study. We recognize that the experimental conditions may not be truly representative of those under which natural infection develops in pigs; however, the ELISA detected an infected pig with muscle larval density of 0.87 larvae/g of tissue. Results of a field trial (n = 310) indicated no muscle digestion test-positive pigs (35 g of diaphragm muscle digested/pig), but 3 samples tested positive by ELISA for specificity of 99.0%.     

Skeletal muscle characteristics in young trained and untrained standardbred trotters.

Muscle biopsies were taken from the middle gluteal muscle of 28 Standardbred trotters, 3-4 years of age. The 13 horses in Group T were trained consistently from 18 months of age, whereas the 15 horses in Group UT were not exposed to any systematic training before 3 years of age. Group T horses had a lower percentage of Type IIB fibres (31%) than did Group UT horses (39%). Citrate synthase (CS) activity, representing oxidative capacity, was higher in Group T (72 mmol kg-1 min-1) than in Group UT (47 mmol kg-1 min-1). Biopsies were taken from 4 horses in each group when they were foals and then annually until 3-4 years of age. Results from this study indicate that regular training of Standardbreds from 18 months of age resulted in increased CS activity and a decrease in the percentage of Type IIB fibres. This study shows that training, not growth, is the main factor that induces a high oxidative capacity and a high Type IIA/IIB fibre ratio in muscle of Standardbred trotters.