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.     

Characterisation of the 33kDa piroplasm surface antigen of Theileria orientalis/sergenti/buffeli isolates from West Java, Indonesia.

The immunodominant 33/35kDa antigen of a Theileria isolate from West Java, Indonesia, was characterised and immuno-affinity purified by use of a monoclonal antibody, KUL-a4, and was shown to be representative of the T. orientalis/sergenti/buffeli group. The aminoterminal sequence of the purified 35kDa peptide (20 residues) was determined by automated Edman degradation and found to correspond to the predicted amino acid sequence of a prospective p33 gene previously sequenced from the same isolate. The cleavage site of a putative signal peptide was identified and conforms the (-3, -1) rule for signal peptidases. The existence of dimeric and trimeric forms of the p33/35 antigen is hypothesised from Western blot profiles. KUL-a4 appeared specific for the T. orientalis/sergenti/buffeli group. It did not recognise in indirect fluorescence antibody test (IFAT), intraerythrocytic bodies of Anaplasma marginale or piroplasms and schizonts of T. mutans, T. parva and T. annulata, whereas cattle antisera raised to these species showed cross-reactivity in IFAT. It however, appeared weakly cross-reactive in Western blot and ELISA, with the 34kDa piroplasm antigen of one T. annulata (Gharb) isolate. The present study indicates that the isolated antigen belongs to the p33/34 antigen family described within the T. sergenti/orientalis/buffeli group, and documents the group-specificity of one of its epitopes.     

Cutaneous vascular neoplasia in 15 cats: clinical, morphologic, and immunohistochemical studies.

Seven cases of cutaneous hemangioma and nine of cutaneous hemangiosarcoma were diagnosed from biopsy specimens of 15 Domestic Shorthairs of 5,091 cats that were examined by necropsy or biopsy during the 5-year period from 1 January 1986 through 31 December 1990. All but three cats were male. Tumor cells of both hemangiomas and hemangiosarcomas were immunoreactive for factor VIII-related antigen and for vimentin by the avidin biotin peroxidase complex method. In cats with a median age of 10 years, hemangiomas occurred in skin with pigmented hair in six of seven cases without apparent site predilection. These solitary tumors did not recur after excision although one cat (No. 3) subsequently developed cutaneous hemangiosarcoma at another site. Seven of nine hemangiosarcomas occurred in dermis and subcutis of the head, usually on the pinna. All five hemangiosarcomas of the head, for which cutaneous pigmentation could be determined, occurred in unpigmented skin. Cats with hemangiosarcoma had a median age of 12.5 years at the time of diagnosis. Metastasis has not been documented, but hemangiosarcoma has recurred, from 1 month to 2 years after excision, in 6/7 cats that were studied.     

Histochemical studies on the dependence of secretory function of the major vestibular gland (Bartholin's gland) on ovarian steroid hormones in the cat.

The dependency of the secretory function of the feline major vestibular gland (Bartholin's gland) on the ovarian steroid hormones was studied histochemically. After estrogen (estradiol-17 beta or diethylstilbestrol), progesterone or a combination of these was administered to cats which were previously ovariohysterectomized, the major vestibular glands were removed, embedded in paraffin, sectioned, and stained by alcian blue, periodic acid-Schiff or peroxidase-labeled lectin (peanut agglutinin and wheat germ agglutinin). The vividly positive reactions to the stainings applied were observed in the secretory epithelial cells of the major vestibular glands in the estrogen treated animals. The dependency of the secretory function of the major vestibular glands on the estrogen was demonstrated in this study.     

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.     

Uroendoscopy. Evaluation of the lower urinary tract.

Recent advances in uroendoscopy have allowed diagnostic evaluation of the lower urinary tract in most of our canine and feline patients. By providing a magnified view of the luminal surfaces of the lower urinary tract, uroendoscopy provides useful diagnostic information that is not readily available even by more invasive techniques.