Development of a monoclonal antibody-based sandwich ELISA for detection of guinea pig interleukin-2     

Adachi S  Hashimoto T  Takeyoshi M  Kato H  Iwata H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(12):1281-1287

Interleukin 2 (IL-2) is a T cell proliferation factor released by Th0- and Th1-type helper T cells and is an essential cytokine for immune responses. In the present study, recombinant glutathione S-transferase (GST)-guinea pig IL-2 (GPIL-2) fusion protein was prepared by Escherichia coli (E. coli) and by using this protein as an immunogen, monoclonal antibodies (mAbs) against GPIL-2 were produced to establish a basis for a research on immune responses in guinea pigs. Three stable hybridoma cell lines were established, and specific binding of each mAb to recombinant GPIL-2 produced by E. coli and insect cells infected with recombinant baculovirus was shown by enzyme linked immunosorbent assay (ELISA) and/or immunoblot analysis. Isotype analyses of these mAbs revealed that all three mAbs were IgG1 and had kappa chain. Furthermore, assessment of their epitopes by competition binding assay indicated that the mAbs obtained in this study bound to three different epitopes. Thus, a sandwich ELISA based on the two mAbs specific to different GPIL-2 epitopes was developed for detection of GPIL-2, which had a sensitivity threshold of about 0.3 ng/ml of GPIL-2.  相似文献   

Availability of Micro-Tom mutant library combined with TILLING in molecular breeding of tomato fruit shelf-life     

Yoshihiro Okabe  Erika Asamizu  Tohru Ariizumi  Kenta Shirasawa  Satoshi Tabata  Hiroshi Ezura 《Breeding Science》2012,62(2):202-208

Novel mutant alleles of an ethylene receptor Solanum lycopersicum ETHYLENE RESPONSE1 (SlETR1) gene, Sletr1-1 and Sletr1-2, were isolated from the Micro-Tom mutant library by TILLING in our previous study. They displayed different levels of impaired fruit ripening phenotype, suggesting that these alleles could be a valuable breeding material for improving shelf life of tomato fruit. To conduct practical use of the Sletr1 alleles in tomato breeding, genetic complementation analysis by transformation of genes carrying each allele is required. In this study, we generated and characterized transgenic lines over-expressing Sletr1-1 and Sletr1-2. All transgenic lines displayed ethylene insensitive phenotype and ripening inhibition, indicating that Sletr1-1 and Sletr1-2 associate with the ethylene insensitive phenotype. The level of ethylene sensitivity in the seedling was different between Sletr1-1 and Sletr1-2 transgenic lines, whereas no apparent difference was observed in fruit ripening phenotype. These results suggested that it is difficult to fine-tune the extent of ripening by transgenic approach even if the weaker allele (Sletr1-2) was used. Our present and previous studies indicate that the Micro-Tom mutant library combined with TILLING could be an efficient tool for exploring genetic variations of important agronomic traits in tomato breeding.  相似文献   

Antimicrobial resistance of Staphylococcus pseudintermedius isolates from healthy dogs and dogs affected with pyoderma in Japan     

Onuma K  Tanabe T  Sato H 《Veterinary dermatology》2012,23(1):17-22, e5

Staphylococcus pseudintermedius strains were isolated from healthy dogs and dogs with pyoderma in 2000-2002 and 2009. All the isolates from dogs with pyoderma in 1999-2000 and from healthy dogs in 2000-2002 and 2009 were susceptible to cefalexin and/or other cephalosporins and oxacillin. However, 7.1-12.5 and 11.4% of S. pseudintermedius isolates from dogs with pyoderma in 2009 were resistant to cephalosporins and oxacillin, respectively. All S. pseudintermedius isolates from dogs with pyoderma in 1999-2000 and those from healthy dogs in 2000-2002 were susceptible to fluoroquinolones; however, 50% of the S. pseudintermedius strains isolated from dogs with pyoderma in 2009 and 30% of the S. pseudintermedius strains isolated from healthy dogs in 2009 were resistant to fluoroquinolones. Of the 21 oxacillin-resistant S. pseudintermedius (MRSP) isolates, 11 carried SCCmec type V and 10 carried hybrid SCCmec types II-III. Staphylococcus pseudintermedius strains that were resistant to only one of three fluoroquinolones had a mutation in the quinolone resistance determination region of grlA, whereas S. pseudintermedius strains that were resistant to two or more fluoroquinolones had mutations in the quinolone resistance determination regions of both grlA and gyrA.  相似文献   

Spontaneous Glomerular and Tubulointerstitial Lesions in Common Marmosets (Callithrix jacchus)     

Isobe K  Adachi K  Hayashi S  Ito T  Miyoshi A  Kato A  Suzuki M 《Veterinary pathology》2012,49(5):839-845

Spontaneous progressive nephropathy dominated by glomerular lesions in common marmosets has been reported. However, the histopathologic characteristics, including the relationship between glomerular and tubulointerstitial lesions, have not been described in detail. In the present study, the authors examined the histopathologic characteristics of the background renal lesions in common marmosets (3 males and 9 females, 3 to 8 years old). The severity of glomerular lesions was graded into 3 classes: grade I, no alteration; grade II, hilar/focal increase of mesangial matrix; grade III, global/diffuse increase of mesangial matrix. Tubulointerstitial lesions (tubular regeneration and hyperplasia and interstitial inflammation and fibrosis) were scored according to the area of each lesion. The renal lesions were characterized by enlargement of glomeruli, expanded mesangial area with increase of periodic acid-Schiff reaction-positive matrix, tubular regeneration and hyperplasia, and interstitial inflammation and fibrosis. Glomerular lesions progressed with increasing mesangial matrix and aging. Additionally, the tubulointerstitial lesions became exacerbated with progressing glomerular lesions. Tubular hyperplasia was divided into 4 types according to the structure of the cell layer (simple or stratified-like), the area of increased lining cells (partial or entire), cytoplasmic staining (eosinophilic or basophilic), brush border and thickness of basement membrane, and the activity of cell proliferation. In conclusion, the background renal lesions in common marmosets were characterized by glomerular lesions with increase of mesangial matrix, which progressed with aging, and secondary tubulointerstitial lesions, including tubular hyperplasia. Those lesions were thus diagnosed as progressive glomerulonephropathy in common marmosets.  相似文献   

Effects of feeding dried distillers grains with solubles (DDGS) on meat quality at the late stage of the fattening period of Holstein steers     

Toshihiro NADE  Kyouhei UCHIDA  Kouhei OMORI  Kenta MATSUBAYASHI  Nobuhiro KIMURA 《Animal Science Journal》2012,83(4):310-317

Feeding dried distillers grains with solubles (DDGS) during the late stage of the fattening period of Holstein steers was studied in regard to the influence on meat quality. Sixteen Holstein steers approximately 18 months old were used in this study. Eight animals were fed commercial concentrated feed for the entire fattening period. The other eight animals were fed 15% DDGS in the concentrated feed for 3 months before slaughtering. The moisture, ether extract and crude protein from both groups was approximately the same. The thiobarbituric acid reactive substance (TBA) value of storage for 7 days at 5°C from the animals fed DDGS showed a tendency to be lower (P = 0.059). The change in the TBA value during storage was also lower for the animals not fed DDGS (P < 0.05). There were no differences in the subcutaneous fat color between the two groups. The a*(measure of redness) and b*(measure of yellowness) of the M. longissimus from the animals fed DDGS showed a tendency to be lower (P = 0.051, 0.070). The fatty acid composition of the M. longissimus, subcutaneous and perirenal fat were not widely influenced by the feeding of DDGS. It is suggested that feeding 15% DDGS during the late stage of the fattening period for Holstein steers reduced the oxidation of the beef.  相似文献   

Segmentation of the Pathophysiological Stages of Diabetic Changes in the db/db Mouse     

Yamazaki M  Kato A  Kato C  Fujii E  Adachi K  Miyoshi A  Suzuki M 《Journal of toxicologic pathology》2009,22(2):133-137

The db/db mouse is one of the diabetes mellitus animal models and if the pathophysiological stages of diabetic changes in the mouse model could simulate the stages in human diabetes, the db/db mouse could be used to better evaluate drug candidates. Blood insulin, HbA1c levels and morphological features of pancreatic islets in db/db mice were evaluated to determine the pathophysiological stage. At 6 weeks of age, db/db mice showed the highest level of plasma insulin and lowest level of HbA1c, and histopathological examination revealed enlarged islets with a circular shape and hypertrophic islet cells. By 9 and 12 weeks of age, the plasma insulin levels had decreased to mid levels and HbA1c had increased to mid to high levels; histopathological examination at this time revealed two types of islets coexisting, enlarged circular islets and small irregular-shaped islets. By 15 and 22 weeks of age, plasma insulin had decreased further to low levels and HbA1c was at its highest level; the histopathological examination at this time revealed an increase in irregular-shaped and small islets. Based on blood insulin levels, HbA1c levels and histopathology findings in the db/db mice in this study, the clinical staging of diabetic changes were recognized. The pathophysiological stages of diabetes mellitus in this animal model were similar to the stages in humans.  相似文献   

Evaluation of a strawberry-harvesting robot in a field test     

Shigehiko Hayashi  Kenta Shigematsu  Satoshi Yamamoto  Ken Kobayashi  Yasushi Kohno  Junzo Kamata  Mitsutaka Kurita 《Biosystems Engineering》2010,105(2):160-171

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Technical note: viability and motility of vitrified/thawed primordial germ cell isolated from common carp (Cyprinus carpio) somite embryos     

Kawakami Y  Saito T  Fujimoto T  Goto-Kazeto R  Takahashi E  Adachi S  Arai K  Yamaha E 《Journal of animal science》2012,90(2):495-500

The feasibility of cryopreserving common carp (Cyprinus carpio) primordial germ cells (PGC) by vitrification of whole embryos at the 22- to 28-somite stage was investigated. Green fluorescent protein (GFP)-labeled PGC were cooled rapidly using liquid nitrogen after exposure to a pretreatment solution containing 1.5 M cryoprotectant (ethylene glycol or dimethyl sulfoxide, 30 or 50 min) and a vitrification solution containing 3 M cryoprotectant and 0.5 M sucrose (5, 10, 20, or 30 min). Embryonic cells that were pretreated for 30 min and vitrified for 20 min with ethylene glycol had the greatest rate of survival of embryonic cells (68.6%; P < 0.01), an optimal highest percentage of viable PGC (73.8 to 74.9%; P < 0.05), and no evidence of ice formation after thawing. The vitrified/thawed PGC were transplanted into blastula-stage embryos from goldfish (Carassius auratus). The PGC maintained their motility and moved to the gonadal ridge of the host embryo. Thus, the combination of vitrification and transplantation to produce germ-line chimeras is a powerful tool for the artificial production of next-generation offspring.  相似文献   

The growth,feed conversion ratio and fin damage of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> under self-feeding and hand-feeding regimes     

Kenta Suzuki  Kanta Mizusawa  Chris Noble  Mitsuo Tabata 《Fisheries Science》2008,74(4):941-943

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In vitro susceptibility and a new point mutation associated with tylosin-resistance in Japanese canine intestinal spirochetes     

Prapasarakul N  Ochi K  Adachi Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(12):1275-1280

The in vitro susceptibilities of six commonly used antimicrobial agents against 29 isolates of intestinal spirochetes isolated from dogs in Japan were examined by the agar dilution technique. In addition, the genetic basis of tylosin resistance in in vitro selected resistant mutants of two reference strains and three tylosin-susceptible field isolates obtained by three successive subcultures on blood agar containing 1 microg/ml of tylosin was investigated. Carbadox was the most active (MIC: < 0.00625) of all the antimicrobial agents. Although all the isolates were susceptible to tylosin, some were resistant to erythromycin. Tiamulin, lincomycin and dimetridazole were also very active against the isolates. All the resistant isolates did not harbor any plasmids. In vitro selected tylosin-resistant mutants of previously tylosin-susceptible isolates showed a new mutation in which their adenine at the base position equivalent to 2062 of 23S rDNA of Escherichia coli has been replaced by cytosine. These findings may both provide guidance towards the proper choice of antimicrobial agents for the treatment of canine intestinal spirochetosis, and add to the understanding of the genetic basis of tylosin resistance.  相似文献