Participation of H1-receptors in histamine-induced contraction and relaxation of horse coronary artery in vitro.

The mechanisms of histamine-induced contraction and relaxation were investigated in rings isolated from a middle part of the left descending coronary arteries of horses. Intact and endothelium-denuded preparations were compared. Rings of horse coronary arteries contracted in response to histamine in a concentration dependent manner, but some of them relaxed with lower concentrations and contracted with higher concentrations. Removal of the endothelium abolished the relaxation and potentiated the contraction. The pD2 values were 4.70 +/- 0.08 in the rings with intact endothelium and 4.95 +/- 0.08 in endothelium-denuded rings. Histamine-induced contractions in intact and denuded preparations were not affected by an H2-antagonist, cimetidine, but were inhibited by an H1-antagonist, diphenhydramine in non-competitive manner in the rings with endothelium and in competitive manner in denuded rings. After precontraction with PGF2 alpha or norepinephrine, histamine relaxed preparations with intact endothelium (pD2 value, 7.80 +/- 0.11), although histamine-induced relaxations were not observed in denuded preparations. The relaxation was competitively inhibited by diphenhydramine. Relaxing response was significantly attenuated by methylene blue, quinacrine, L-nitro-arginine, gossypol and AA861 but not by indomethacin. These results suggest that the histamine-induced contraction and relaxation in horse coronary arteries are mediated mainly by H1-receptors in the smooth muscle and endothelium, respectively, and H1-receptor activation of endothelial cells may liberate vasodilator substances.     

MCPB-ethyl疏花对富士苹果授粉受精及胚珠发育的影响

通过在花期用MCPB-ethyl处理,对富士苹果花粉的发芽、花粉管的伸长以及胚珠的发育等进行了形态方面的观察和探讨,以阐明MCPB-ethyl的疏花机制。结果表明,MCPB-ethyl对花粉的发芽及花粉管的伸长没有影响,整个受精过程与对照相同,没有发现异常。但受精后胚乳核只进行了数次分裂便停止生长,此后珠皮、珠心细胞迅速解体。根据以上结果,认为MCPB-ethyl的疏花效果不是通过影响花粉的发芽或花粉管的伸长阻碍受精所致,而是使胚和胚珠的发育停止,形成离层导致了落花。     

Seasonal changes in semen characteristics, composition of seminal plasma and frequency of acrosome reaction induced by calcium and calcium ionophore A23187 in Large White boars

This study attempted to explain the mechanisms regulating boar fertility by examining seasonal changes in semen characteristics, the composition of seminal plasma and responsiveness of sperm acrosomes to Ca(2+) and the Ca(2+) ionophore A23187 (Ca(2+)/A23187). Sperm-rich and sperm-poor fractions were separately collected from 3 mature fertile Large White boars once a month over a one-year period. During the period of study, ambient temperature and relative humidity were recorded for within the stall in which the boars were kept and the semen characteristics, composition of the seminal plasma of sperm-rich fractions, and occurrence of the acrosome reaction in response to Ca(2+) (3 mM)/A23187 (0.3 microM) were examined. The highest mean maximum and minimum ambient temperatures were recorded in August-September, whereas the lowest mean maximum and minimum ambient temperatures were recorded in December and January, respectively. There was a moderate peak in relative humidity from July to October. The lowest percentages of motile spermatozoa and of spermatozoa with intact acrosomes and highest percentage of spermatozoa with abnormal morphology and strongest agglutination were seen in August-September. The total protein and albumin concentrations were lowest in August-September. Testosterone levels increased gradually as day length decreased after the summer solstice (June) and peaked in October-November. The percentage of acrosome reactions in response to Ca(2+)/A23187 was highest with the quickest response in August-September, as shown by the shortest time required for 50% of relative acrosome reactions. The farrowing rates were lowest in these same 2 months. These results suggest that seasonal infertility in Large White boars may be due, at least in part, to a combination of low motility, abnormal morphology including acrosomal abnormality, and early occurrence of the acrosome reaction in response to stimulus, possibly resulting from a decrease in acrosomal stabilizing proteins in the seminal plasma during summer. These changes may be modulated by heat/humidity stress and/or photoperiod-regulated testosterone.     

Gene silencing of cyclooxygenase-2 mRNA by RNA interference in bovine cumulus-granulosa cells

Inhibition of specific gene expression using RNA interference (RNAi) is a valuable tool for functional analysis of a target gene. However, there is little information available concerning RNAi for analysis of gene function in relation to the reproductive physiology of follicular cells in ruminants. Thus, the aim of this study was to evaluate the interfering effect of small interference RNA (siRNA) on expression of cyclooxygenase-2 (Cox-2) mRNA and prostagrandin F(2alpha) (PGF(2alpha)) production in bovine cumulus-granulosa (CG) cells. Bovine CG cells were collected from aspirated follicles and cultured. After reaching confluency, two experiments were conducted. In experiment 1, to investigate the effective concentration of siRNA, 0, 100, 250 and 500 pM of Cox-2 siRNA was introduced into the CG cells, respectively. After 24 h, the amount of Cox-2 mRNA expression was measured by RT-PCR and real-time PCR. In experiment 2, to investigate the time required for effective interference of siRNA and Cox-2 activity, 250 pM siRNA was introduced for 0, 3, 6, 12 and 24 h. After culture, the amount of Cox-2 mRNA expression was measured and the culture medium was collected to determine the PGF(2alpha) concentration by enzyme immunoassay. The Cox-2 mRNA expression was not affected by introduction of 100 pM siRNA into CG cells for 24 h, but 250 and 500 pM Cox-2 siRNA significantly reduced the Cox-2 mRNA expression. Moreover, the significant suppressive effect of 250 pM siRNA was observed 6 h after introduction, and the reduction of mRNA expression by RNAi became more obvious over 12 h. On the other hand, the PGF(2alpha) concentration in the culture medium was not significantly different 12 h after siRNA introduction; however, the PGF(2alpha) concentration 24 h after siRNA introduction was significantly decreased compared with the control at the same time point. These results suggest that gene silencing of Cox-2 with siRNA is capable of analyzing the function and expression of specific genes in bovine CG cells.     

Investigations on the radioactive contamination of crop plants as a result of H-bomb detonation (Part 2) Root and foliage uptake of “Bikini ash”

The Foliar Uptake by Squash Plant

The radioactive ash for experimental use, hereafter referred to as “Bikini ash”, was prepared by igniting the heavily contaminated substances on board No. 5 Fukuryu Maru at about 650°C, followed by sifting through a 100 mesh sieve. On ignition some parts of the fission products, particularly iodine, ruthenium and tellurium would have possibly been lost to the air.     

Cutting-grafts as a means to propagate greenhouse roses

A propagation method for rose nursery plants was studied by using a 2-node scion tongue-grafted on to an unrooted stock cutting. Uniform and disease-free nursery plants could be produced efficiently in a short period all the year round.Under greenhouse production trials, such cutting-grafts of ‘Sonia’ on R. multiflora ‘K-1’, R. indica ‘Major’, R. ‘Manetti’ and R. wichuraiana produced flowers of the same number and quality as when budded on to R. multiflora seedling root stock.     

Trophinin is expressed in the porcine endometrium during the estrous cycle

We investigated endometrial expression of trophinin mRNA and protein, homophilic cell adhesion molecules, during the estrous cycle of gilts. An immunopositive reaction for trophinin was observed in the luminal and glandular epithelia of the endometrium at all stages of the estrous cycle, but not in endometrial stromal cells or the myometrium. A partial coding sequence of porcine trophinin was similar to sequences in humans and mice, with homologies of 75% and 70%, respectively. As in humans and mice, the trophinin gene is expressed in the endometrium. Trophinin, however, is expressed in the endometrium of the pig throughout the estrous cycle, higher expression levels were observed at some points of the luteal phase, as in humans. These findings suggest that regulation of trophinin gene expression in the pig is different from that in mice, but similar to that in humans. Furthermore, the present results suggest that the pig might be a suitable model for studying the physiological importance of trophinin in early pregnancy in humans.     

Localization of inhibin alpha-, betaA- and betaB-subunits and aromatase in ovarian follicles with granulosa theca cell tumor (GTCT) in 6 mares

To clarify the morphological and immunohistochemical characteristics in mares with granulosa theca cell tumor (GTCT), the localization of inhibin subunits (alpha, betaA, betaB) and aromatase in the granulosa cell layers and theca layers in the ovarian follicles were determined by immunohistochemical staining. The follicles were obtained from the ovaries of 6 mares with GTCT and 4 normal mares as controls. Immunohistochemically, inhibin alpha-subunit was localized in the granulosa cells of all follicles showing different sizes in all GTCT cases and betaA- subunit was localized in two GTCT cases in all sized follicles. But inhibin betaB- subunit and aromatase were not localized in GTCT cases. On the other hand, inhibin alpha-, betaA-, and betaB-subunits and aromatase were localized in the large and medium sized follicles, but inhibin betaA- and betaB-subunits and aromatase were not stained in the small sized follicles in normal cases. These findings suggest that some mares with GTCT can secrete dimeric inhibin (inhibin A), but all GTCT cases cannot secrete inhibin B. By the results of aromatase staining it is clear that testosterone is not converted into estradiol due to the lack of aromatase in the GTCT follicles.     

Relation between phenotype of tumor cells and clinicopathology in bovine leukosis

Thirty-three cases of enzootic bovine leukosis (EBL) and 14 cases of sporadic bovine leukosis (SBL) were examined by immunohistochemistry using 6 monoclonal antibodies against leukocyte differentiation molecules of bovine leukocytes. There were 17 cases of B-1a cell type, 10 cases of B-1b cell type and 6 cases of B-2 cell type in EBL, and 5 cases originating from B cells (B-2 cell type) and 9 cases originating from immature T cells in SBL. The average age for the EBL cases of B-1a cell type was 8.6 years, B-1b cell type was 6.5 years, and of B-2 cell type was 4.5 years. In cases of SBL, immature T cell type patients were younger than B-2 cell type ones. The lymphoma originating from B cells differed from that originating from T cells in morphology. In T cell tumors, the nucleus of tumor cells was round, the edge of the cytoplasm obvious, and tumor cells were sporadically present and proliferated. When compared with T cells, the region among B cells was obscure. But, there was no relation between phenotype and the histologic classification of tumor cells. In EBL, beyond the lymph node, tumors of B-1a and B-1b types had developed in the heart and abomasum, and those of the B-2 type tended to occur in liver. In SBL, B-2 type and T type cells formed tumors in the liver, kidney, thymus, and one case of T-cell type tumor formed on the skin. We would like to propose a new classification of bovine leukosis as EBL, calf type B-cell lymphoma, juvenile T-cell lymphoma and skin type T-cell lymphoma.     

Flow cytometric analysis of chicken NK activity and its use on the effect of restraint stress

Immune system is organized by the influence of both neural and endocrine systems. NK activity plays an important role in the innate immunity. In this study, we observed the effects of restraint stress on chicken peripheral blood NK activity. Viability of FITC-labeled RP9 was measured with PI after treatment with the effector cells. Chicken peripheral blood CD8alpha+ cells expressed strong cytotoxic activity, in contrast to thrombocytes, while peripheral blood CD3+ CD8alpha+ cells and CD4+ cells had little cytotoxic activity. Con A supernatant enhanced the cytotoxic activity of CD8alpha+ cells. Therefore, it is considered that these cytotoxic activities measured by flow cytometry (FCM) analysis are NK activity. When chickens were exposed to restraint stress, the levels of serum corticosterone increased transiently over a short period of time while the NK activity decreased. The decreased NK activity, however, did not recover to the intact levels for a long time, even once the serum corticosterone levels had recovered. These data indicate that chicken NK activity is able to be measured by flow cytometric analysis and that restraint stress causes severe damage to the chicken NK activity.