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.     

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.     

0–7–21 Radiation Therapy for the Treatment of Canine Oral Melanoma

Eighteen dogs with malignant melanoma of the oral cavity were treated with high-dose per fraction (0–7–21) radiation therapy. Eight hundred cGy was administered on days 0, 7, and 21 for a total dose of 2,400 cGy in 3 weeks. Of 17 dogs evaluated, 9 (53%) had a complete remission and 5 (30%) achieved a partial remission with an overall response rate of 83%. Local failure occurred in 2 of the 9 dogs where a complete response was initially observed. One dog died of intercurrent disease, and one died of metastatic disease without evidence of local recurrence. Five dogs are alive and free of disease 9 to nineteen months from the initiation of therapy. The 0–7–21 protocol was well-tolerated, and acute radiation reactions were low-grade and limited to the skin. The results of this study demonstrate that oral melanomas in dogs are responsive to radiation. 0–7–21 radiation therapy offers a viable alternative to radical excision, especially when tumor volume or location would require cosmetically or functionally debilitating surgery.     

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.     

Effect of Split Nitrogen Application on Growth and Yield of Wheat (T. aestivum L.) Genotypes with Different N-Assimilation Potential

A field experiment was conducted with different nitrogen regimes to assess the growth and yield performance of wheat genotypes which differ in nitrate assimilation potential. Genotypic differences in biomass accumulation were observed at different growth stages. The nitrogen treatment had little effect on biomass accumulation at early stages of growth, while at later stages of growth there was enhanced biomass accumulation when N was applied in more than two splits. On an average, genotypes with high nitrate reductase activity (the 'HNR' genotypes) accumulated 14.2 % more biomass than the genotypes with low nitrate reductase activity ('LNR' genotypes) when an extra dose (40 kg N ha⁻¹) of nitrogen was given at the time of anthesis. The application of nitrogen in more than two splits increased grain yield of both 'HNR' and 'LNR' genotypes mainly by increasing grain weight per ear. The application of an extra dose of nitrogen (40 kg N ha⁻¹) at the time of anthesis increased grain yield of 'HNR' genotypes by 38.5 % as compared to 'LNR' genotypes.