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.     

Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens

Lycopersicon esculentum cultivar Micro-Tom is a miniature tomato with many advantages for studies of the molecular biology and physiology of plants. To evaluate the suitability of Micro-Tom as a host plant for the study of pathogenesis, Micro-Tom plants were inoculated with 16 well-known fungal, bacterial, and viral pathogens of tomato. Athelia rolfsii, Botryotinia fuckeliana, Oidium sp., Phytophthora infestans, and Sclerotinia sclerotiorum caused typical symptoms and sporulated abundantly on Micro-Tom. Micro-Tom was resistant to Alternaria alternata, Corynespora cassiicola, and Fusarium oxysporum. When Micro-Tom was inoculated with 17 isolates of Ralstonia solanacearum, many isolates induced wilt symptoms. Agrobacterium tumefaciens also was pathogenic, causing crown galls on stem tissue after needle prick inoculation. In Micro-Tom sprayed with Pseudomonas syringae pv. tomato, P. s. pv. tabaci, or P. s. pv. glycinea, bacterial populations did not increase, and yellow lesions appeared only on leaves sprayed with P. s. pv. tomato. Tomato mosaic virus, Tomato aspermy virus, and Cucumber mosaic virus systemically infected Micro-Tom, which developed symptoms characteristic of other cultivars of tomato after infection with the respective virus. These results indicated that Micro-Tom was generally susceptible to most of the important tomato pathogens and developed typical symptoms, whereas certain pathogens were restricted by either hypersensitive resistance or nonhost resistance on Micro-Tom. Therefore, an assortment of Micro-Tom–pathogen systems should provide excellent models for studying the mechanism of susceptible and resistant interactions between plants and pathogens.     

Sward characteristics,nutritive value and choice by cattle of conterminous monocultures of centipedegrass (Eremochloa ophiuroides) and bahiagrass (Paspalum notatum)

In south‐western Japan, centipedegrass (Eremochloa ophiuroides; CG) offers a novel option for a warm‐season perennial for grazing use in areas where bahiagrass (Paspalum notatum; BG) can be grown. However, the potential of CG as a forage has not been fully explored because of the short history as a forage crop. We conducted four experiments to evaluate CG (cv. TifBlair) in comparison with BG (cv. Pensacola) in terms of sward characteristics, nutritive value and choice by animals. In each experiment, four Japanese Black cows (Bos taurus) were individually allowed to graze conterminous monocultures of CG and BG (5 × 10 m each) for 30 min. Irrespective of regrowth durations and fertilizer rates, CG was consistently shorter, leafier and denser, contained lower acid detergent fiber and cellulose, and was preferred or equally selected by cows, as compared with BG. Furthermore, CG maintained sufficient levels of crude protein (80–89 g/kg DM) to ensure voluntary intake of ruminant animals under extended regrowth^? and without fertilizer, whereas BG failed to do so (65 g/kg DM). CG provided higher digestible dry matter than BG when crude protein concentration exceeded 86 g/kg DM. The results indicate advantages of CG as a forage.     

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.     

Validation of the DNDC-Rice model by using CH4 and N2O flux data from rice cultivated in pots under alternate wetting and drying irrigation management

The DNDC (DeNitrification-DeComposition)-Rice model, one of the most advanced process-based models for the estimation of greenhouse gas emissions from paddy fields, has been discussed mostly in terms of the reproducibility of observed methane (CH₄) emissions from Japanese rice paddies, but the model has not yet been validated for tropical rice paddies under alternate wetting and drying (AWD) irrigation management, a water-saving technique. We validated the model by using CH₄ and nitrous oxide (N₂O) flux data from rice in pots cultivated under AWD irrigation management in a screen-house at the International Rice Research Institute (Los Baños, the Philippines). After minor modification and adjustment of the model to the experimental irrigation conditions, we calculated grain yield and straw production. The observed mean daily CH₄ fluxes from the continuous flooding (CF) and AWD pots were 4.49 and 1.22?kg?C?ha^?1?day^?1, respectively, and the observed mean daily N₂O fluxes from the pots were 0.105 and 34.1?g?N?ha^?1?day^?1, respectively. The root-mean-square errors, indicators of simulation error, of daily CH₄ fluxes from CF and AWD pots were calculated as 1.76 and 1.86?kg?C?ha^?1?day^?1, respectively, and those of daily N₂O fluxes were 2.23 and 124?g?N?ha^?1?day^?1, respectively. The simulated gross CH₄ emissions for CF and AWD from the puddling stage (2 days before transplanting) to harvest (97 days after transplanting) were 417 and 126?kg?C?ha^?1, respectively; these values were 9.8% lower and 0.76% higher, respectively, than the observed values. The simulated gross N₂O emissions during the same period were 0.0279 and 1.45?kg?N?ha^?1 for CF and AWD, respectively; these values were respectively 87% and 29% lower than the observed values. The observed total global warming potential (GWP) of AWD resulting from the CH₄ and N₂O emissions was approximately one-third of that in the CF treatment. The simulated GWPs of both CF and AWD were close to the observed values despite the discrepancy in N₂O emissions, because N₂O emissions contributed much less than CH₄ emissions to the total GWP. These results suggest that the DNDC-Rice model can be used to estimate CH₄ emission and total GWP from tropical paddy fields under both CF and AWD conditions.     

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.     

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。