Distribution of Cytochrome P450-side Chain Cleavage in the Theca Interna Layers of Bovine Small Antral and Cystic Follicles

Cystic follicle is anovulatory follicular structure that is caused by an endocrine imbalance. The activity of cytochrome P450‐side chain cleavage (P450scc) is essential for the initiation of steroidogenesis in the follicle. The present study was designed to compare the frequency of cells containing P450scc between healthy and atretic small antral follicles, and among several types (I, II and III, classified based on the presence of granulosa layer) of cystic follicles. Paraffin sections of healthy (2–5 mm in diameter), atretic (2–5 mm) and cystic follicles (>25 mm) were immunohistochemically stained with rabbit polyclonal antibody to bovine P450scc. The P450scc‐positive cells were counted in four different regions of the follicles from the apical to the basal side. In small antral follicles and cystic follicles, P450scc‐positive cells were localized in the theca interna layers but not granulosa layers. The P450scc‐positive cell populations decreased in the late atretic follicles compared with the early and advanced atretic follicles at all the regions of follicle. Type III cystic follicles showed significantly lower frequencies of P450scc‐positive cells than those in the types I and II cystic follicles. These results suggest that in both small and cystic follicles in cows, total loss of granulosa cells may be associated with the reduction of frequency of P450scc‐positive cells in theca interna layer.     

Contribution of insulin to the ascorbate recycling system in the chicken liver

The effect of insulin on the ascorbate recycling system in the chicken liver was examined. First, insulin was injected subcutaneously into the chicken, and liver glutathione‐dependent dehydroascorbate reductase (GSH‐DHAR) activity was determined. Insulin increased liver GSH‐DHAR activity, but did not affect plasma and liver ascorbate concentration. Dehydroascorbate increased plasma and liver ascorbate levels, and liver GSH‐DHAR activity. However, distinct changes in plasma insulin level were not observed by dehydroascorbate injection. In addition, reduction of external dehydroascorbate in cultured chicken hepatocytes could not be observed in an insulin‐deprived culture, although the cells reduced external dehydroascorbate in a serum‐free culture with insulin. We concluded that insulin affects the ascorbate recycling system as an essential factor in the chicken liver.     

Changes in the expression of toll-like receptor mRNAs during follicular growth and in response to lipopolysaccharide in the ovarian follicles of laying hens

The aim of this study was to determine the changes in the mRNA expression of Toll-like receptors (TLRs) in hen ovarian follicles during follicular growth and in response to lipopolysaccharide (LPS). White follicles and the fifth largest to largest follicles (WF and F(5)-F(1), respectively) were collected from laying hens. To examine the effects of LPS, the laying hens were treated intravenously with LPS (1 mg/kg BW) 0, 3, 6, 12 and 24 h before examination. Expressions of TLRs and IL-1beta in the theca and granulosa layers were examined by semi-quantitative RT-PCR. Immunocytochemistry was performed to identify immunoreactive TLR-4. The theca layer expressed TLR-2, TLR-4, TLR-5 and TLR-7, whereas the granulosa layer expressed only TLR-4 and TLR-5. The expression of TLR-4 and TLR-5 in the theca layer increased significantly during follicular growth. In the granulosa layer, the expression of TLR-5 increased, but that of TLR-4 was unchanged. Expression of TLR-4 increased significantly during the period of 6 to 12 h of LPS treatment in the theca layer and during the period of 12 to 24 h in the granulosa layer of F(3). Immunoreaction products for TLR-4 were observed in theca interna and granulosa layers of WF and F(5)-F(1), with the greater amount observed in the theca interna. LPS treatment significantly increased expression of IL-1beta in the theca layer after 3 h and in the granulosa layer during the period of 12 to 24 h. These results suggest that TLRs are expressed in ovarian follicles and that TLR-4 and TLR-5 expression increase with the growth of follicles. Enhanced expression of TLR-4 and IL-1beta by LPS in the theca and granulosa layers suggests possible roles of TLR in recognition of microorganisms.     

松油烯与马来酸酐的环加成反应的条件研究

研究了松油烯与马来酸酐的D iels-A lder环加成反应和松油烯的异构转化率。使用相对于α-松油烯过量的马来酸酐,在65~70℃、100~105℃和135~140℃时α-松油烯的异构转化率分别为15%、38%和69%。采用异核远距离关联(HMBC)等核磁共振测试手法解析了环加成产物的结构,归属了各原子的化学位移。     

Turnover time of microbial biomass carbon in Japanese upland soils with different textures

We investigated the turnover time of microbial biomass-C in Japanese upland soils with various textures and examined the soil physicochemical properties influencing their turnover time. Samples from five different soil types (sand-dune regosol, light-colored Andosol, humic Andosol, brown forest soil, and dark red soil) were taken from upland concrete-frame plots in the experimental field of Chiba University. Each soil amended with [U -¹³C] glucose was incubated for 80 d at 25°C. Microbial biomass-C and -¹³C in soil were periodically determined by the fumigation-extraction method. The longest turnover time of microbial biomass-C was observed in the dark red soil (215 d) followed by the humic Andosol (134 d), brown forest soil (97 d), and light-colored Andosol (83 d) and the shortest in the sand-dune regosol (45 d). The turnover time of microbial biomass-C was significantly correlated with the value of soil clay (R: 0.917*), CEC (R: 0.921*), and macroaggregate (R: 0.907*) contents, but not with the total-C content. The amount of microbial biomass-C showed a close correlation with the turnover time of microbial biomass-C, suggesting that the turnover time of microbial biomass-C is an important factor influencing the accumulation of microbial biomass-C in soil.     

Analysis of QTL for Seed Dormancy and Their Response to Dry Heat Treatment in Rice(Oryza sativa L.)

Quantitative trait loci(QTL)controlling seed dormancy in rice were identified using recombinant inbred lines(RILs)population derived from the cross between a japonica variety Kinmaze and an indica variety DV85. Seeds of two parental cultivars and each RIL were harvested in 35d after heading.. The germination percentage of these seeds at 30℃for 7 days were measured as the degree of seed dormancy.. QTL analysis was performed with Windows QTL Cartographer 1.13a program by composite interval mapping.. A total of four QTL for seed dormancy were detected on chromosome 2(two regions),5 and 11,respectively.Phenotypic variation explained by each QTL ranged from 8.37 to 17.40%.. Responses of such loci to a dormancy-breaking treatment with dry heat were further detected. The results showed that two alleles of qDOR-2-1 and qDOR-5 from DV85 as well as the allele of qDOR11 from Kinmaze increased the seed dormancy,which seemed to be easily broken by dry heat treatment. Such loci of seed dormancy may be applied to rice genetic improvement.The allele of qDOR-2-2 from DV85 increased the seed dormancy,which could not be broken by dry heat treatment.     

Effective Synthesis and Antifouling Activity of Dolastatin 16 Derivatives

Some derivatives of dolastatin 16, a depsipeptide natural product first obtained from the sea hare Dolabella auricularia, were synthesized through second-generation synthesis of two unusual amino acids, dolaphenvaline and dolamethylleuine. The second-generation synthesis enabled derivatizations such as functionalization of the aromatic ring in dolaphenvaline. The derivatives of fragments and whole structures were evaluated for antifouling activity against the cypris larvae of Amphibalanus amphitrite. Small fragments inhibited the settlement of the cypris larvae at potent to moderate concentrations (EC₅₀ = 0.60-4.62 μg/mL), although dolastatin 16 with a substituent on the aromatic ring (24) was much less potent than dolastatin 16.     

Synthesis,insecticidal activity and pyrethroidal mode of action of new tin ether derivatives

A novel range of trimethylstanniomethyl ethers of well known pyrethroid alcohols were synthesised, and their insecticidal activities and modes of action as insecticides were investigated. Among them, ethers from three types of alcohol (3-phenoxybenzyl, 4-fluoro-3-phenoxybenzyl and 6-phenoxy-2-pyridylmethyl) showed remarkable insecticidal activities against rice stem borers, houseflies and German cockroaches. According to electrophysiological studies on the abdominal nerve cords of German cockroaches, trimethylstanniomethyl 6-phenoxy-2-pyridylmethyl ether induced a rapid decline in spontaneous firing similar to that from tetramethrin. However, insecticidal trimethyltin chloride caused an entirely different response. These observations suggest that the present tin ether derivatives resemble pyrethroids, rather than the insecticidal tin compounds known so far.     

Susceptibility of Djungarian hamsters (Phodopus sungorus) to Neospora caninum infection

Djungarian hamsters were examined for the susceptibility to Neospora caninum infection. After 29 Djungarian hamsters were intraperitoneally inoculated with 5 x 10(6) N. caninum tachyzoites of JPA1 strain, some animals showed symptoms such as ataxia, and many tissue cysts were detected in the brain and a cyst in the muscular tunics of stomach. Especially, more than 100 cysts per head were observed after 5 weeks post inoculation. It is suggested that the Djungarian hamster is a model useful to examine neosporosis.