Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

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.     

Diversity in the carotenoid profiles and the expression of genes related to carotenoid accumulation among citrus genotypes

Carotenoids are not only important to the plants themselves but also are beneficial to human health. Since citrus fruit is a good source of carotenoids for the human diet, it is important to study carotenoid profiles and the accumulation mechanism in citrus fruit. Thus, in the present paper, we describe the diversity in the carotenoid profiles of fruit among citrus genotypes. In regard to carotenoids, such as β-cryptoxanthin, violaxanthin, lycopene, and β-citraurin, the relationship between the carotenoid profile and the expression of carotenoid-biosynthetic genes is discussed. Finally, recent results of quantitative trait locus (QTL) analyses of carotenoid contents and expression levels of carotenoid-biosynthetic genes in citrus fruit are shown.     

Differences in lectin-binding properties between the common mucosal epithelium and follicle-associated epithelium in the rabbit small intestine

Differences in sugar distribution between the villous epithelium and follicle-associated epithelium (FAE) were compared using lectins in the rabbit small intestine. In every portion, villous columnar epithelial cells primarily exhibited a positive reaction to the GalNAc, GlcNAc, galactose, and oligosaccharide. In the ileal Peyer's patch (PP), whereas microvillous epithelial cells exhibited positive reactions, M cells tended to be negative. The villous epithelial reaction to the fucose group was negative, but M cells and microvillous epithelial cells showed a positive to the fucose. No epithelium had a positive reaction to the mannose and glucose. The variety of lectin-binding properties of villous epithelial cells and M cells may reflect specificity for the recognizing luminal substances such as antigenic molecules and bacterial elements.     

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.     

Relationship between the ultrasonographic findings of suspected superficial digital flexor tendon injury and the prevalence of subsequent severe superficial digital flexor tendon injuries in Thoroughbred horses: a retrospective study

The onset of severe injury to the superficial digital flexor tendon (SDFT) is extremely difficult to predict from slight changes in ultrasonographic findings in cases with no apparent clinical signs. This study investigated the relationship between an increased cross-sectional area (CSA) or edema in the subcutaneous tissue around the tendon and the subsequent onset of severe SDFT injury in Thoroughbred racehorses. Horses were classified into three groups based on ultrasound diagnosis (USD) findings: Group A included cases with enlarged tendons; Group B included cases with tendons of normal size but with prominent edema in the peritendinous tissue; and Group C (control group) included cases with no abnormal USD findings. The incidence of subsequent severe tendon injury was significantly higher in the horses in Groups A (25.7%, 28/101) and B (28.3%, 65/212) than in those in Group C (4.9%, 2/41). There were no significant differences in the median period and the median number of races from the first examination to the subsequent tendon injury between Groups A (140 days, 1 race) and B (120 days, 1 race). The results of this study revealed that horses with increased CSA and peritendinous edema are likely to suffer a subsequent severe tendon injury. Also, these two USD findings, i.e., increased CSA and peritendinous edema, indicate the risk of onset of severe SDFT injury.     

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。     

Effect of nitrogen application during ripening period on photosynthesis and dry matter production and its impact on yield and yield components of semidwarf iniica rice varieties under water culture conditions

Abstract

A water culture pot experiment was conducted to analyze the effects of N application during the ripening period (RP) on photosynthesis, dry matter production, and its impact on grain ripening and yield in two semidwarf indica type varieties viz. Gui Zhao 2 (GZ) and BR3 (BR) compared with a japonica type variety Koganemasari (KO) under four N rates viz. 0 (N₀), 10 (N₁₀), 20 (N₂₀), and 40 (N₄₀) mg L^?1. Results showed that N application enabled to maintain a higher leaf area and delay leaf senescence in both indica and japonica type varieties but the decrease in the rate, of leaf area was higher in the former than in the latter and the rate was reduced with increasing N rates. Flag leaf photosynthesis and SPAD values of N treated plants were higher throughout the RP, showing the presence of a significant correlation either for each variety or all the varieties together. Higher photosynthetic rate was supported by higher leaf chlorophyll (SPAD value basis) content, stomatai conductance, and N concentration in leaf blades. Top dry matter content increased with increasing N rates mainly due to mean leaf area rather than NAR except for BR during RP but it was higher in KO than in GZ and BR. Reduction of shoot weight due to translocation of dry matter to panicles during RP was suppressed by N rates both in GZ and BR while shoot weight increased in the N-treated plants in KO. The dependency of KO on current photosynthates for panicle weight was found to be almost hundred percent while the contribution of stored carbohydrates in shoot before heading to panicle weight in GZ and BR was in the range of 4-27 and 33-54%, respectively and the rest was contributed by current photosynthates. The percentage increased with increasing N rates. Percentage of ripened grains (PRG) increased with increasing N rates in GZ and BR due to the increase in dry matter production and in the photosynthetic rates of apex leaves, despite the larger spikelet number and larger hull size. However, KO showed almost no variation although it had the highest PRG among the varieties. Brown rice yield followed the same pattern as that of PRG in GZ and BR and the highest yield was produced by BR followed by GZ and KO. These results suggest that N application during RP was more effective in increasing yield in the semidwarf indica type varieties than in the japonica type variety.