Immunolocalization of steroidogenic enzymes (P450scc, P450c17, P450arom) in gonad of Japanese eel

Antibodies against P450scc, P450c17 and P450arom were generated using recombinant proteins. In eel testis, P450scc and P450c17 were immunolocalized as clusters in Leydig cells. In vitellogenic eel ovary, P450scc and P450c17 immunoreactive cells were localized as clusters in the outer layer of the ovarian follicle. In contrast, P450arom seemed to be immunolocalized in the innermost follicle layer.     

Nutritional parameters in the blood of dams during late gestation and immediately after calving,in the umbilical vein at calving,and in the blood of calves immediately following birth in Holstein heifers pregnant with either Holstein or beef breed fetuses

Dairy cattle management lacks consideration of fetal breed, the effect of which on fetal growth and nutrition are unclear. We investigated blood parameters in 12 late-pregnant Holstein heifers with similar (Holstein, n = 5) or different (Japanese Black [n = 4] or F1 cross [n = 3]; Holstein × Japanese Black) fetus breeds and in their umbilical cords and calves. Samples were obtained from dams 1 week before calving (−1 week) and immediately after calving, from the umbilical vein at calving, and from calves immediately after birth. Dams with beef fetuses had higher serum glucose levels (−1 week; p < .05) than those with Holstein fetuses. Plasma total amino acid, total essential amino acid, total nonessential amino acid, and other amino acid concentrations were lower in the umbilical veins of dams with calves of the beef breeds than in those of the Holstein breeds (p < .05). Furthermore, serum glucose and plasma amino acid levels were lower in the beef calves than in the Holstein calves (p < .05). Overall, nutrient supply from dams to beef fetuses was lower than that to Holstein fetuses. Our findings may facilitate feeding management of dairy cattle pregnant with beef breeds for appropriate fetal growth and nutrition.     

Interspecies nuclear transfer embryos reconstructed from cat somatic cells and bovine ooplasm

This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage.     

Epidemiology of hepatitis E virus in indoor-captive cynomolgus monkey colony

A serological survey of hepatitis E virus (HEV) antibody was conducted using 202 adult captive cynomolgus monkeys, who did not show any clinical signs of acute hepatitis. Out of these, 44 monkeys were sero-positive for anti-HEV IgG and all monkeys were negative for anti-HEV IgM. All positive monkeys came from either Vietnam or China, but none from the Philippines, Indonesia, or our facility. Selected 12 monkeys out of positive monkeys from Vietnam, including 9 positive and 3 negative, revealed mostly within the reference ranges for alanine aminotransferase (ALT) and asparatate aminotransferase (AST) by serum biochemistries. Their titers of anti-HEV IgG did not correlate with the concentrations of ALT and AST. Moreover, HEV-RNA could not be detected from any fecal specimens of the 12 monkeys. Thus, monkeys with anti-HEV IgG sero-positive did not seem to be source of the HEV-pollution, because 1) sero-positive monkeys did not excrete HEV-RNA from their feces, and 2) monkeys from the Philippines and Indonesia have remained to be sero-negative for anti-HEV IgG, even if the monkeys were kept in same animal room of our facility. From these results, it could be inferred that primary infection of HEV occurred in the exported countries, but not in our colony. The contamination of HEV in indoor-captive monkeys could be prevented by precise quarantine tests, including ELISA for detecting anti-HEV and RT-PCR for HEV RNA.     

Variation in seed production among years and among individuals in 11 broadleaf tree species in northern Japan

Seed production is the most problematic part of tree regeneration and is the least amenable to control by silvicultural management. Understanding variability in seed production among years and among trees will allow better planning of seed collection for seedling production and natural regeneration. We estimated the extent of variability in seed production among years and within years among individual trees. Specifically, we measured individual annual seed production in 11 woody species in Hokkaido, northern Japan. We analyzed the coefficient of variation (CV) of seed production among years and among individual trees. We used population- and individual-based CVs (CV_pt and ) to estimate the variability in seed production among individual trees. Alnus hirsuta, in which these CVs among trees were maximal, will require further analyses of the spatial patterns of seed production among individual trees. Additionally, we used population- and individual-based CVs (CV_py and ) to estimate the variability in seed production among years. The was statistically higher than in 2 of the 11 species: Betula maximowicziana and Sorbus alnifolia. Activities for the regeneration of these species should regard the annual variation in seed production as more important than individual variation. For the nine species in which was similar to even if seed production by specific trees was not sufficient for regeneration in a particular year, other trees or stands often had high seed production in that year. We discuss the relative importance of annual and individual variability in determining efficient methods for artificial and natural regeneration of these woody plant species.     

Low-frequency spin dynamics in a canted antiferromagnet

Resistively detected nuclear spin relaxation measurements in closely separated two-dimensional electron systems reveal strong low-frequency electron-spin fluctuations in the quantum Hall regime. As the temperature is decreased, the spin fluctuations, manifested by a sharp enhancement of the nuclear spin-lattice relaxation rate 1/T1, continue to grow down to the lowest temperature of 66 millikelvin. The observed divergent behavior of 1/T1 signals a gapless spin excitation mode and is a hallmark of canted antiferromagnetic order. Our data demonstrate the realization of a two-dimensional system with planar broken symmetry, in which fluctuations do not freeze out when approaching the zero temperature limit.     

Decreases in serum apolipoprotein B-100 and A-I concentrations in cows with milk fever and downer cows

Milk fever occurring during the peripartum period has been suggested to be caused by fatty liver developed during the nonlactating stage because diseased cows have increased serum concentrations of non-esterified fatty acids (NEFA) and show hepatic lipidosis. In cows with fatty liver and related diseases such as ketosis, serum concentrations of apolipoprotein (apo) B-100 and apoA-I are decreased. The purpose of the present study was to examine whether apoB-100 and apoA-I concentrations are similarly decreased in cows with milk fever. Apolipoprotein concentrations were also measured in cows with downer syndrome, which has been suggested to be related, at least in part, to milk fever. Compared with healthy cows during early lactation, apoB-100 and apoA-I concentrations were decreased in cows with milk fever and also in downer cows. In cows with milk fever, the decreases in apoB-100 and apoA-I concentrations were associated with increased NEFA and decreased cholesterol and phospholipid concentrations. However, in downer cows, serum lipid concentration changes were not as distinct as in cows with milk fever. These results, coupled with previous findings on the decreases in apoB-100 and apoA-I concentrations of cows with fatty liver-related diseases, suggest that fatty liver is involved in the development of milk fever and partly in that of downer cow syndrome.     

DNA sequence analysis of ribosomal ITS region 1 of Phytophthora vignae f. sp. adzukicola, the pathogen that causes stem rot of adzuki bean

Sequences of the internal transcribed spacer (ITS) region 1 were used to examine the phylogenetic relationships among races of 19 isolates of Phytophthora vignae f. sp. adzukicola and between this forma specialis and three isolates of the closely related P. vignae f. sp. vignae. The ITS 1 sequences were highly conserved (> 98.7% similarity) among representatives of both formae speciales groups. The results of this study indicate that P. vignae is a monophyletic group. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession nos. AB120062–AB120080 and AB120122     

Effects of added protein (bovine serum albumin) on the rate and enantiotopic selectivity of oxidative O-demethylation of methoxychlor in rat liver microsomes

The addition of bovine serum albumin (BSA) to rat liver microsomal suspensions during the O-demethylation of methoxychlor (1,1-bis(4-methoxyphenyl)-2,2,2-trichloroethane) produced a good substrate concentration-reaction rate relationship, and allowed us to determine the rate parameters for this reaction. The apparent K_m became larger when the albumin content was increased, whereas V_max was not changed. The enantiotopic selectivity of the reaction dramatically changed when the substrate concentration was increased in the absence of BSA in phenobarbital-treated rat liver microsomes.     

Airborne basidiospores as an inoculum source of <Emphasis Type="Italic">Typhula variabilis</Emphasis> and the effect of hilling on the incidence of Typhula winter rot of carrots

Typhula winter rot on overwintering carrots caused by Typhula variabilis is a newly confirmed disease, and no practical control measure is yet available. To develop a control method, here we researched the infection period of T. variabilis and the time that winter rot appeared on carrots. Using spore traps, we found that basidiospore rain occurred from September to November before snowfall in Memuro, Hokkaido. In addition, carrot leaves collected in autumn had already been infected by T. variabilis. These epidemiological investigations revealed that the pathogen releases basidiospores to infect carrot leaves before snow cover, resulting in root decay under snow. An effective control method was then developed to avoid direct contact of basidiospores of T. variabilis with plant tops by covering the plants with soil in autumn. Thus, the percentage of rotted roots was reduced to about half.