Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries

Bovine granulosa cells (GC) vary in their morphological aspect during different stages of folliculogenesis. In this study, 10 morphologically normal bovine ovaries were collected to study the structural aspects of different stages of GC using intermediate filament protein antibodies including cytokeratin AE1/AE3 (AE1/AE3), vimentin, nectin‐4 and desmin. Hormonal immunolocalization was assessed using the immunomarkers anti‐Müllerian hormone (AMH) and inhibin alpha. In addition, tumour markers and proliferation markers using c‐erbB‐2 oncoprotein and proliferating cell nuclear antigen, respectively, were investigated. The immunolabelling of AE1/AE3 in GC was strongest in the early follicle stage and gradually decreased when reaching the Graafian follicle stage. Its immunolabelling increased again as the stage progressed from stage I to stage III. The immunolabelling of inhibin alpha was inversely proportional to that of AE1/AE3 in the developing ovarian follicles as their immunolabelling is opposite to each other during folliculogenesis. AMH was immunopositive in almost all GC stages in different intensities and percentages, except for some negative staining in the atretic IV follicles. The atretic IV follicle is a unique type of atretic follicle that shows Call‐Exner body formation, which was mainly found in older cows in this study. The distinct patterns of immunoreactivity for various types of immunomarkers in the different GC stages will play an important role in diagnostic assistance of various follicle conditions, including cystic ovaries and GC tumours.     

An enzyme-linked immunosorbent assay for detection of antibodies to exogenous avian leukosis virus

A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to avian leukosis virus (ALV) of subgroups A and B in infected chickens was developed with the use of Rous-associated virus (RAV)-1 (subgroup A) and RAV-2 (subgroup B) antigens purified by sucrose-gradient centrifugation. The antigen was used for ELISA after treatment with Triton X-100. In the ELISA, the subgroup viral antigen reacted strongly with homologous antiserum but also reacted with heterologous antiserum. Tests with serum absorbed with purified homologous and heterologous virus and tests for antigen-blocking by group-specific antibodies to ALV revealed that the reaction was caused mainly by subgroup-specific antibodies. The ELISA was 8 to 32 times more sensitive than the virus-neutralization (VN) test and detected antibodies to ALV earlier than the VN test in chickens infected experimentally with RAV-1 and RAV-2. In field application of the ELISA, 44.2% of 484 chicken sera were positive for RAV-1 and/or RAV-2 antigen, and 80.4% of flocks were positive. These findings indicate that ELISA is superior to the VN test in sensitivity, simplicity, rapidity, and applicability for large-scale field surveys for ALV infection.     

Monosaccharide composition of sweetpotato fiber and cell wall polysaccharides from sweetpotato, cassava, and potato analyzed by the high-performance anion exchange chromatography with pulsed amperometric detection method

The cell wall materials (CWMs) from sweetpotato (Ipomoea batatas cv. Kokei 14), cassava (Manihot esculenta), and potato (Solanum tuberosum cv. Danshaku) and commercial sweetpotato fiber as well as their polysaccharide fractions were analyzed for sugar composition by the high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) method. The separation of arabinose and rhamnose, and xylose and mannose, by this method has been improved using a CarboPac PA 10 column. Pretreatment of the CWMs and cellulose fractions with 12 M H(2)SO(4) was required for complete hydrolysis to occur. Commercial sweetpotato fiber was found to be mainly composed of glucose (88.4%), but small amounts of other sugars were also detected. Among the root crops, sweetpotato CWM had the highest amount of pectin and galacturonic acid. Fucose was detected only in cassava CWM and its hemicellulose fraction, while galactose was present in the highest amount in potato CWM. Among the polysaccharide fractions, it was only in the hemicellulose fraction where significant differences in the sugar composition, especially in the galactose content, were observed among the root crops.     

Quantitative structure-activity studies of benzoylphenylurea larvicides: I. Effect of substituents at aniline moiety against Chilo suppressalis walker

The larvicidal activity of a series of N-2,6-difluoro- and N-2,6-dichlorobenzoyl-N′-(4-substituted phenyl)ureas against nondiapause larvae of the rice stem borer, Chilo suppressalis Walker, was measured by topical application and oral administration methods under conditions with and without piperonyl butoxide as an inhibitor of oxidative metabolism. The effects of substituents at the anilide moiety on the larvicidal activity were analyzed quantitatively using physicochemical substituent parameters and regression analysis. The results indicate that the oxidative metabolism in the larval body which is favored by electron-donating substituents is significant in determining the activity. The activity, when the metabolic factor is eliminated, is enhanced by electron-with-drawing and hydrophobic substituents and lowered by bulky groups. The inhibitory activity against new cuticle formation of the same series of compounds was also measured using cultured integument of the rice stem borer diapause larva. The comparison of the quantitative analyses between larvicidal and integument-level activities shows that inhibition of cuticular development is the most important factor governing larvicidal activity.     

Release of Bound Lipids in Cereal Starches Upon Hydrolysis by Glucoamylase

The raw starch granules from corn, rice, and wheat were hydrolyzed by practically pure glucoamylase (Rhizopus niveus). The bound lipids remaining in the residual starches were investigated, of which the major components of the lipids, free fatty acids (FFA) in corn starch, FFA and phospholipids (PL) in rice starch, and PL in wheat starch were determined. In each case, the bound FFA and PL were decreased to some extent during the initial stage of hydrolysis. During the later stages, the FFA continued to gradually decrease, while the level of PL stabilized. It was interesting that some of the bound lipids were released from the granules upon glucoamylase hydrolysis, differing from the model amylose-lipid complexes. Furthermore, the structures of the residual starches were investigated. The blue value and λ_max of the starches were increased by partial hydrolysis of the starch granules using practically pure glucoamylase. Two gel-permeation chromatography analyses revealed that the relative amount of amylose fraction was increased by glucoamylase hydrolysis, and also that the increments were reduced by the defatting of bound lipids. The results suggest that the increase in amylose fraction is attributable to the existence of bound lipids in the granules.     

Effect of height-to-diameter ratio on stem stiffness of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

The lumber from sugi plantations in Japan displays large intra- and intertree variation in mechanical properties, even within a stand. These variations seem to be induced by the effects of the characteristics of cultivars as well as the effects of growth traits on mechanical properties. Therefore, the effects of growth traits on mechanical properties per cultivar need to be precisely examined. In this study, we focused on the effects of growth traits, especially height-to-diameter ratio (H/D ratio), on stem stiffness per cultivar. Sixteen cultivars were classified into three groups according to the relationships between stem stiffness and growth traits. In cultivars that showed a close correlation between growth traits and stem stiffness, it was assumed that stem stiffness could be controlled to a certain extent by silvicultural practices using the H/D ratio as an indicator. In cultivars that showed a weak correlation between growth traits and stem stiffness, selecting cultivars for the production of logs with higher mechanical properties seemed to be effective; in this study, Kumotoshi, Tanoaka, and Edanaga were found to be suitable. Tree age and site index may be important factors for producing wood with higher mechanical properties.     

Effect of vegetation types on oviposition preference of the giant purple emperor, Sasakia charonda

To determine the effect of vegetation types on the oviposition by the butterfly, Sasakia charonda, the host plants were planted in a meadow (site A), a small broadleaf forest (site B), a large broadleaf forest (site C), and at the edge of a Cryptomeria japonica forest adjacent to the large broadleaf forest (site D). The number of S. charonda eggs per planted host tree was higher at sites C and D than at site B, and no eggs were found at site A. The densities of both male and female adults were significantly higher at site C than at the other sites, and no adults were found at site A. A mark-and-recapture method estimated that the dispersal distances of wild adults were within 100m, although these adults already had heavily torn wings when they were initially captured. In one case, a female adult was caught 730m from the release point, a distance greater than that between sites A and C. When the wild adults were initially captured and when recaptured, they sipped the sap of Quercus acutissima. The capture and recapture points of most of the recaptured wild adults were in the Q. acutissima stand in the large broadleaf forest, where site C was located. Our observations at site A and the results of the mark-and-recapture study suggested that the flight range of mature adults was limited to large broadleaf forests containing Q. acutissima stands and long-distance dispersal seldom occurred or that the flyways of female adults searching for host trees were restricted to the vicinity of broadleaf forest areas, resulting in no visits to isolated host trees. Finally we discussed the selection of suitable host tree-planting sites for the conservation of this butterfly.     

Geographical variation in age–height relationships for dominant trees in Japanese cedar (Cryptomeria japonica D. Don) forests in Japan

In this study, we analyzed Japanese National Forest Inventory data to investigate the geographical variation in the relationship between tree height and age for dominant trees, and the effects of climatic conditions on these relationships. Our analysis focused on Cryptomeria japonica forests in 13 regions of Japan. The age–height relationships were classified into two regional groups that were distinguished by their climatic conditions. Several categories of climatic variables (warmth, solar radiation, precipitation, and snow depth) were significantly correlated with the parameters of a model for the age–height relationships. Our results also suggest the existence of a latitudinal cline for the maximum tree height of C. japonica in Japan. In regions with cold temperatures, deep snow, low solar radiation, and low summer precipitation, C. japonica shows a late-maturity pattern for height increase, with slow initial growth and a large maximum size. In regions with the opposite climatic conditions, it shows an early-maturity pattern with fast initial growth and small maximum size.