Effects of TAK-044, a nonselective endothelin receptor antagonist, on the spontaneous and indomethacin- or methylene blue-induced constriction of the ductus arteriosus in rats

We studied the effects of TAK-044, a nonselective endothelin (ET) receptor antagonist, on the indomethacin- or methylene blue-induced constriction of the ductus arteriosus (DA) in rats and compared them with the effects on spontaneous DA constriction. Injection of TAK-044 into 21-day-old fetuses in utero was performed through the uterine wall of laparotomized mother rats under light ether anesthesia. The fetuses were autopsied 3 hr after treatment with TAK-044 (10 mg/kg) in utero and simultaneous administration to the laparotomized mother rats of indomethacin (3 mg/kg, p.o.) or methylene blue (100 mg/kg, i.p.). In the second experiment, pregnant rats were decapitated on day 21 of gestation to obtain newborn rats by cesarean delivery. Newborn rats which were given TAK-044 (2, 10 mg/kg) immediately after or 1 hr before cesarean delivery were autopsied at various times after birth. In both experiments, pups were rapidly frozen in an acetone-dry ice mixture at autopsy to evaluate the DA constriction by the whole-body freezing and shaving method. TAK-044 injection into the fetus 3 hr before autopsy completely inhibited the DA constriction induced by maternal treatment with indomethacin or methylene blue. TAK-044 caused dose-dependent inhibition of the spontaneous closure of the DA after birth. The inhibitory effect was more pronounced in pups which were given TAK-044 in utero 1 hr before birth; however, the inhibitory effect was incomplete in newborn pups. These results, together with the previous finding that BQ-123, an ETA-specific receptor antagonist, inhibits the ductal constriction induced by oxygen in vitro [Coceani et al., 1992], indicate that the ETA receptor plays a significant role in the indomethacin- or methylene blue-induced DA constriction as well as in the spontaneous DA constriction after birth, and also indicate that the inhibition of ETA receptor by TAK-044 was more easily achieved in fetuses than in neonates.     

Exogenous expression of rat aquaporin-3 enhances permeability to water and cryoprotectants of immature oocytes in the zebrafish (Danio rerio)

Movement of water and cryoprotectants through the plasma membrane needs to be accelerated for successful cryopreservation of zebrafish oocytes/embryos, which are much larger than their mammalian counterparts. Aquaporin-3 is a water/solute channel that can transport not only water but also various cryoprotectants. In this study, we attempted to increase the permeability of immature zebrafish oocytes at stage III to water and cryoprotectants by exogenous expression of rat aquaporin-3. Immature zebrafish oocytes were injected with rat aquaporin-3 cRNA and cultured for 5-12 h. Permeability to water and cryoprotectants was then determined based on changes in the volumes of the oocytes in a hypertonic sucrose solution and various cryoprotectant solutions at 25 C. The permeability to water of the aquaporin-3 cRNA-injected oocytes was three times higher than that of intact and water-injected oocytes. The permeability of the aquaporin-3 cRNA-injected oocytes to ethylene glycol, glycerol, propylene glycol, and DMSO was also 2-4 times higher than that of intact oocytes. Thus, the permeability of immature zebrafish oocytes to water and cryoprotectants was enhanced by exogenous expression of aquaporin-3. Cryopreservation of teleost oocytes may be realized through a further increase in permeability.     

Molecular cloning and gene expression of two cytochrome P450s from permethrin-resistant Culex quinquefasciatus larvae

As part of a continuing study of factors influencing the development of pesticide resistance in insects, two new cytochrome P450s of the CYP6 family have been identified.     

Effect of acetic acid added to cooking water on the dissolution of proteins and activation of protease in rice

The effect of acetic acid on the dissolution of proteins in rice was studied to elucidate the mechanism for the textural change induced by the acid by chemical and SDS-PAGE analyses of the rice proteins in the soaking solution. More proteins were extracted with 0.2 M acetic acid (pH 2.7) than with water (pH 6.8). The effect of acetic acid on the protein dissolution increased with increasing temperature. Immunoblotting confirmed that, when rice was soaked in acetic acid, glutelin was dissolved into the soaking solution and degraded by aspartic proteinase. Aspartic proteinase degraded glutelin much more than it did albumin and globulin. It was found that the combined amount of albumin and globulin dissolved into the acetic acid solution was much larger than that of glutelin, despite the smaller amounts present of albumin and globulin than of glutelin. Metal ions were extracted more with acetic acid than with water. In addition, carboxypeptidase was activated under the acidic condition and resulted in an increase in the amount of free amino acids. The main effect of acetic acid on the dissolution of rice proteins was enhancement of the solubility of albumin, globulin, and glutelin, the effect of proteases being minor.     

Effect of Initial Stocking Density on Larviculture Performance of the Ocellate Puffer,Takifugu rubripes

Depending on the initial stocking density, ocellate puffer larvae exhibit the cannibalism that resulted in high mortality. We aimed to clarify the relationship between stocking density of larvae, mortality, and the effect of density on cannibalism. Ocellate puffer larvae were reared at five different stocking densities between 5 and 25 larvae/L with duplicate 1‐m³ tank for each treatment. Larvae were stocked at 0 d after hatch (d.a.h.), and the experimental cultures lasted for 55 d. In larvae stocked initially at more than 10 larvae/L, cannibalism was evident from 11 to 17 d.a.h., and the survival rate at 55 d.a.h. was less than 15%. The survival rate was 25.5% at 5 larvae/L, and cannibalism was observed from 31 to 32 d.a.h. Consequently, an initial density of 5 larvae/L was the most efficient in terms of survival and the prevention of cannibalism. On the other hand, when cannibalism is still observed at 5 larvae/L, it is necessary to lower the stocking density before the onset of cannibalism.     

Population genetic structure and variability of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> in the stocking area along the Pacific coast of northern Japan

ABSTRACT:   The genetic diversity of wild and hatchery-released Pacific herring Clupea pallasii collected from three brackish lakes and two bays in Honshu and Hokkaido, Japan was examined with five microsatellite loci. All loci showed high genetic variability with expected heterozygosities ranging from 0.815 to 0.945. Significant differences in genotypic and allelic distributions were detected among all locations except for between the two bays in Honshu Island. Pairwise population analysis based on the F _ST values showed close genetic relationships among the locations in Hokkaido Island, and the hierarchical analyses of molecular variance showed significant genetic difference between the two islands. Those results suggest the existence of subpopulations due to natal homing. In addition, stocked fish showed as much genetic diversity as the wild fish. The pairwise population analyses also showed close relationships between the hatchery fish and the wild fish in respective stocking areas, showing that no effects of stocking programs on genetic diversity of wild populations were detected.     

Distribution of mineral elements in the outer layer of rice and wheat grains,using electron microprobe X-Ray analysis

Electron microprobe X-ray analysis was used to determine the transversal microdistribution of P, K, Mg, Ca, Fe, and Mn, with special attention to the outer layers of rice and wheat grains. P, K, Mg, Fe, and Mn were concentrated in the aleurone layer in each case. In particular, P, Mg, and K were highly concentrated in the subcellular particles of the aleurone layer, and had very similar distribution patterns in the outer layers of the matured grain of rice and wheat. By contrast, Ca was abundant in the pericarp.     

Sporiolides A and B,New Cytotoxic Twelve-Membered Macrolides from a Marine-Derived Fungus Cladosporium Species

Two new cytotoxic twelve-membered macrolides, sporiolides A (1) and B (2), were isolated from the cultured broth of a fungus Cladosporium sp., which was separated from an Okinawan marine brown alga Actinotrichia fragilis, and the structures were elucidated by spectroscopic data. Sporiolides A (1) and B (2) exhibited cytotoxicity against murine lymphoma L1210 cells. Spoliolide A (1) showed antifungal activity against Cryptococcus neoformans and Neurospora crassa.