The Effects of Adjuvants on Autoimmune Responses Against Testicular Antigens in Mice

Experimental autoimmune orchitis (EAO) is a model of immunologic male infertility and pathologically characterized by lymphocytic inflammation, which causes breakdown of the testicular immune privilege with spermatogenic disturbance. Generally, murine EAO is induced by immunization with testicular homogenate (TH) from the testes of donor mice + complete Freund''s adjuvant (CFA) + Bordetella pertussigens (BP), and it has been considered that treatment with these two adjuvants is required to enhance the immune response against testicular antigens. However, there remains a possibility that CFA and BP may affect autoimmune responses against the testicular antigens without TH. In the present study, we examined this possibility using real-time RT-PCR, Western blotting and immunohistochemical staining. The results demonstrated that immunization with TH in combination with CFA and BP evoked more severe EAO than that with only TH. Real-time RT-PCR analyses revealed that Fas mRNA expression in TH+CFA+BP-induced EAO was significantly higher than that in TH-induced EAO. Interestingly, IL-6 mRNA expression dramatically increased in TH+CFA+BP-induced EAO; however, no apparent change in IL-6 mRNA expression occurred in TH-induced EAO. It was also noted that treatment with CFA and BP alone augmented autoimmune reactions against some testicular autoantigens. These results indicates that these adjuvants are helpful in evoking severe EAO, and treatment with the adjuvants alone can evoke autoimmune reactions against some testicular autoantigens despite the use of no TH.     

Isolation and characterization of a novel simazine-degrading beta-proteobacterium and detection of genes encoding s-triazine-degrading enzymes

A moderately persistent herbicide, simazine, has been used globally and detected as a contaminant in soil and water. The authors have isolated a simazine-degrading bacterium from a simazine-degrading bacterial consortium that was enriched using charcoal as a microhabitat. The isolate, strain CDB21, was gram-negative, rod-shaped (0.5-0.6 microm x 1.0-1.2 microm) and motile by means of a single polar flagellum. Based on 16S rRNA sequence analysis, strain CDB21 was identified as a novel beta-proteobacterium exhibiting 100% sequence identity with the uncultured bacterium HOClCi25 (GenBank accession number AY328574). PCR using primers that were specific for the genes of the atrazine-degrading enzymes (atzABCDEF) of Pseudomonas sp. strain ADP showed that strain CDB21 also possessed the entire set of genes of these enzymes. Nucleotide sequences of the atzCDEF genes of strain CDB21 were 100% identical to those of Pseudomonas sp. strain ADP. Sequence identity of the atzA genes between these bacteria was 99.7%. The 398-nucleotide upstream fragment of the atzB gene of strain CDB21 was 100% identical to ORF30 of Pseudomonas sp. strain ADP, and the 1526-nucleotide downstream fragment showed 99.8% sequence similarity to the atzB gene of the pseudomonad.     

Biodegradation of methylthio-s-triazines by Rhodococcus sp. strain FJ1117YT, and production of the corresponding methylsulfinyl, methylsulfonyl and hydroxy analogues

A novel bacterial strain FJ1117YT was isolated from an enrichment culture with the herbicide simetryn. The isolate was capable of degrading the herbicide supplied as the sole sulfur source in an aquatic batch culture. The strain FJ1117YT was identified as that belonging to Rhodococcus sp. on the basis of comparative morphology, physiological characteristics and comparison of the 16S rRNA gene sequence. The biodegradation pathway of simetryn was established by isolating the methylsulfinyl analogue as the first metabolite and by identification of the methylsulfonyl intermediate and the hydroxy analogue by liquid chromatography-mass spectrometry (LC-MS) and/or nuclear magnetic resonance (NMR) analysis. The results indicate that the methylthio group was progressively oxidised and hydrolysed by the strain FJ1117YT. The same strain is also able to metabolise other methylthio-s-triazines such as ametryn, desmetryn, dimethametryn and prometryn through similar pathways.     

Breeding of new rice cultivar ‘Tohoku 194’ with ‘Sasanishiki’-type good eating quality of cooked rice

Cooked rice of ‘Sasanishiki’ is soft and not as sticky as those of Japanese leading cultivars ‘Koshihikari’ and ‘Hitomebore’. As a method for efficient selection of a breeding line having a good eating quality like that of ‘Sasanishiki’, the use of physical properties of cooked rice and cooking quality was examined. There were differences of physical properties of the surface layer, starch-iodine blue value per solid substance weight in cooking water and volume expansion of cooked rice between ‘Sasanishiki’ and ‘Hitomebore’, these properties being considered to be usable for the selection of breeding lines. Using these traits as selection targets, one line, named ‘Tohoku 194’, which has eating quality highly similar to that of ‘Sasanishiki’ and cold tolerance derived from ‘Hitomebore’, was selected from progeny of a cross between ‘Sasanishiki’ and ‘Hitomebore’. An application for registration as a new variety has been submitted for ‘Tohoku 194’ under the Japanese Plant Variety Protection Act, and is expected to become a recommended cultivar in Miyagi Prefecture. ‘Tohoku 194’ may fulfill various demands of consumers and companies in the food industry.     

Genetic characterization of rainfed upland New Rice for Africa (NERICA) varieties

A total of 18 rainfed upland New Rice for Africa (NERICA) varieties were categorized as the heavy panicle and low tillering types and early heading, in compared with 32 different varieties. These chromosome components were clarified using 243 SSR markers which showed polymorphism among NERICA varieties and their parents, CG 14 (O. glaberrima Steud.) and one of the recurrent parents, WAB-56-104 (O. sativa L.). NERICA varieties were classified into three groups, which corresponded with these parents’ continuation including two exceptions, NERICAs 14 and 17, by a cluster analysis using polymorphism data of SSR markers and 14 differential markers among them were selected to classify NERICA varieties. However, three groups: NERICAs, 3 and 4, NERICAs, 8, 9 and 11 and NERICAs, 15 and 16 were not distinguishable. Association analysis was carried out for characterization of NERICA varieties by using SSR markers genotype and phenotype of agronomic traits. A total of 131 quantitative trait loci between SSR markers and 11 agronomic traits were detected. The characteristics of early maturity and heavy panicle of upland NERICA varieties were succeeded from Asian rice varieties and the characteristics of high dry matter production and late heading were introduced from CG 14 and the other varieties.     

Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. The nucleotide sequences of VsNTPase1 and VsNTPase2 in this article have been submitted to DDBJ as accession numbers AB196769 and AB196770, respectively.     

Immunohistological study on bovine, swine and ovine skeletal muscle fibers for the localization of fatty acid translocase FAT/CD36

The present immunohistological study was conducted to investigate the localization of fatty acid translocase CD36 (FAT/CD36) in the skeletal muscle (Biceps femoris) fibers of bovine, swine and ovine. The results showed that CD36 was mainly localized in type I muscle fiber of these animals. In contrast, FAT/CD36 localization in type II fiber was insignificant in the types of muscle in the present experiments, suggesting that type II fiber of bovine, swine and ovine might lack fatty acid translocase FAT/CD36.     

Histopathological Characteristics of an Ossifying Fibroma Formed in the Maxilla of a Racehorse

A 1-year-old male thoroughbred racehorse experienced swelling of the left upper lip. The swelling was attributable to enlargement around the incisive bone of the interdental space posterior to the third incisor in the left maxilla. Even after two operations to reduce the bulk of the mass, it continued to increase in size. Dyspnea caused by stenosis of the nasal cavity forced us to perform euthanasia, and a pathological examination was conducted. Macroscopic examination of a section of the mass revealed the formation of multiple areas of solid fibrous tissue, and trabeculae within the incisive bone which had displaced the cortical bone. On histology, the mass was composed of trabecular bone-like structures due to the proliferation and aggregation of fibroblasts. Therefore, we diagnosed it as an ossifying fibroma. Equine ossifying fibroma is characterized by development in the mandible, but was formed in the maxilla in this case. Equine ossifying fibroma has not been reported previously in Japan. This is the first case of equine ossifying fibroma identified in Japan.