Enzyme-linked immunosorbent assay for detection of canine chromogranin A by use of immunological cross-reactivity of rabbit anti-bovine chromogranin A antibody     

Akiyoshi H  Aoki M  Kumagai D  Saleh N  Noda K  Shimada T  Sugii S  Ohashi F 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(5):535-537

Bovine and canine chromogranin A were extracted and purified from each specie's adrenal glands. Isolated bovine 70 kDa protein showed 100% identity to bovine CgA reported previously, whereas isolated canine 68 kDa protein showed 83.3% identity to bovine CgA by the NH(2)-terminal amino acid sequence analysis. Rabbit antibody to purified bovine protein (CgA) was found to immunologically cross-reacted with purified canine protein (CgA). In sandwich ELISA with anti-bovine CgA, concentration-dependent curves were obtained ranging from 0.3 to 20 mug/ml for canine CgA. From these findings, sandwich ELISA with anti-bovine CgA is found to be useful to determine the concentration of canine CgA.  相似文献   

Expression of bone morphogenetic protein-6 and bone morphogenetic protein receptors in myoepithelial cells of canine mammary gland tumors     

Akiyoshi T  Uchida K  Tateyama S 《Veterinary pathology》2004,41(2):154-163

To study the ectopic chondrogenesis in canine mammary mixed tumors, the expression of bone morphogenetic protein-6 (BMP-6) and specific BMP receptors (BMPRs), BMPR-IA, BMPR-IB, and BMPR-II, was examined using immunohistochemical and immunoblot analysis in 39 canine mammary gland tumors. Immunohistochemically, BMP-6 and all three types of BMPRs were coexpressed in the myoepithelial cells and chondrocytes in six of eight benign mixed tumors. In complex adenomas, myoepithelial cells showed an expression pattern of BMP-6, BMPR-IA, and BMPR-II similar to those in benign mixed tumors, whereas immunoreactivity for BMPR-IB was very mild. The myoepithelial cells proliferating within the basement membrane showed more intense immunoreactivity for BMP-6 and all BMPRs as compared with those proliferating in the interstitial areas. Western blotting analysis revealed immunopositive bands at 40-45 kDa for BMP-6 in the samples from simple and complex adenomas and benign mixed tumors. The BMPR-IB-specific bands at 45 kDa were most detected in benign mixed tumors. Because among BMPRs, BMPR-IB is thought to be the major receptor for BMP-6 for primary chondrogenesis, these findings suggest that the expression of BMP and its receptors on the myoepithelial cells might play a role in the ectopic cartilage formation in canine mammary gland tumors, especially in benign mixed tumors.  相似文献   

The Effects of Frequent Electroejaculation on the Semen Characteristics of a Captive Siberian Tiger (Panthera tigris altaica)     

Daisuke FUKUI  Masashi NAGANO  Ryohei NAKAMURA  Gen BANDO  Shinichi NAKATA  Masao KOSUGE  Hideyuki SAKAMOTO  Motozumi MATSUI  Yojiro YANAGAWA  Yoshiyuki TAKAHASHI 《The Journal of reproduction and development》2013,59(5):491-495

Artificial insemination (AI) can help to avoid inbreeding and genetic degeneration for sustaining genetically healthy populations of endangered species in captivity. Collection of a sufficient quantity of viable sperm is an essential first step in the AI process. In the present study, we examined the effects of frequent electroejaculation on semen characteristics in a Siberian tiger. We collected semen in all 17 trials during 6 breeding seasons (6 years). The mean number of sperm and the percentage of motile sperm were 294.3 ± 250.2×10⁶/ejaculate and 82.4 ± 11.4%, respectively. The number of motile sperm tended to increase during frequent electroejaculation in the same breeding season. Semen collection by electroejaculation can be performed effectively up to the fourth sequential ejaculate, which contained the most sperm in the study. In conclusion, frequent collection of sperm by electroejaculation from tigers may be effective for collection of a large number of motile sperm.  相似文献   

Insulin-like Growth Factor 1 mRNA Expression in the Uterus of Streptozotocin-treated Diabetic Mice     

Yoshie MANABE  Makoto TOCHIGI  Akiyoshi MORIWAKI  Sakae TAKEUCHI  Sumio TAKAHASHI 《The Journal of reproduction and development》2013,59(4):398-404

Reproductive functions decline with the onset of diabetes in female mice. Diabetic mice have smaller uteri with an underdeveloped endometrium, suggesting diminished estrogen-induced growth. We aimed to clarify the changes in the estrous cycle and in insulin-like growth factor 1 (IGF1) expression in the uteri of streptozotocin (STZ)-treated diabetic mice, because IGF1 is one of the main growth factors involved in estrogen-induced uterine growth. ICR female mice were intraperitoneally administered STZ (10 mg/100 g BW), and blood glucose levels were determined. Mice with blood glucose levels > 200 mg/dl were classified as diabetic mice. The onset of diabetes was associated with acyclic estrous cycles. Diabetes was also induced with STZ in ovariectomized mice. Uterine Igf1 mRNA levels were reduced in ovariectomized STZ-treated diabetic mice. Estrogen is known to stimulate Igf1 mRNA expression in the uterus, but estrogen action was abolished in the uteri of STZ-treated diabetic mice. mRNA expressions of estrogen receptor α (ERα) and steroid hormone receptor coactivators (SRC-1/Ncoa1, SRC-2/Ncoa2, SRC-3/Ncoa3 and CBP/p300/Crebbp) were reduced in the uteri of ovariectomized STZ-treated diabetic mice. The present study demonstrates that diabetes induces a decline in female reproductive functions in mice. Igf1 expression in ovariectomized diabetic female mice was decreased, and decreased responsiveness to estrogen in the uteri of diabetic mice is probably associated with a reduction in ERα and steroid receptor coactivator mRNA expression.  相似文献   

Effects of omega-7 palmitoleic acids on skeletal muscle differentiation in a hyperglycemic condition     

Yayoi TOKUNAGA  Hitomi YOSHIZAKI  Akiyo TORIUMI  Ritsuko KAWAHARADA  Chisato ISHIDA  Masatoshi HORI  Akio NAKAMURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(9):1369

Maternal obesity and diabetes are known to be involved in fetal myogenesis, but the later stages of myogenesis are not well understood. In this study, we investigated the influence of a hyperglycemic environment on L6 skeletal myoblast differentiation and the function of omega-7 palmitoleic acids. Exposure to a high concentration of glucose (25 mM) in high-glucose culture medium (HG) increased the expression of myogenic genes (MyoD, Myogenin, MRF4, Myhc2x, and Myhc2a) and the synthesis of myosin. HG also activated the PI3K/AKT pathway revealed muscle cell differentiation. Furthermore, the levels of reactive oxygen species (ROS) and an inflammatory cytokine (Tnfaip3; tumor necrosis factor alpha-induced protein 3), which are crucial for the growth and differentiation of skeletal muscle, were increased by HG. Palmitoleic acids suppressed the expression levels of myogenic regulatory genes and increased the expression level of a cell proliferation-related gene (Pax3). Trans-palmitoleic acid and eicosapentaenoic acid (TPA and EPA) increased the phosphorylation level of MAPK/ERK1/2 and downregulated ROS generation and Tnfaip3 expression. In contrast, cis-palmitoleic acid inactivated MAPK/ERK1/2, leading to increased ROS generation. In conclusion, a hyperglycemic environment mediated by HG induced excessive muscle differentiation. Palmitoleic acids inhibited myoblast differentiation by downregulating muscle-specific genes. Moreover, trans-palmitoleic acids may have beneficial antioxidant and/or anti-inflammatory effects in cells.  相似文献   

Chromosome number and telomere sequence mapping of the Japanese sea cucumber Apostichopus japonicus     

Sei-Ichi Okumura  Kazuma Kimura  Mizuho Sakai  Takashi Waragaya  Suehiro Furukawa  Akiyoshi Takahashi  Kunio Yamamori 《Fisheries Science》2009,75(1):249-251

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Simultaneous steroids measurement in dogs with hyperadrenocorticism using a column-switching liquid chromatography-tandem mass spectrometry method     

Noboru SASAKI  Thandar OO  Yoshikazu YASUDA  Takahiro ICHISE  Noriyuki NAGATA  Nozomu YOKOYAMA  Kazuyoshi SASAOKA  Keitaro MORISHITA  Shouta MM NAKAYAMA  Mayumi ISHIZUKA  Kensuke NAKAMURA  Mitsuyoshi TAKIGUCHI  Yoshinori IKENAKA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(11):1634

We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2–16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.  相似文献   

α-Melanocyte-stimulating hormone directly increases the plasma calcitonin level and involves calcium metabolism in goldfish     

Ikari  Takahiro  Kobayashi  Yuki  Kitani  Yoichiro  Sekiguchi  Toshio  Endo  Masato  Kambegawa  Akira  Asahina  Kiyoshi  Hattori  Atsuhiko  Tabuchi  Yoshiaki  Amornsakun  Thumronk  Mizusawa  Kanta  Takahashi  Akiyoshi  Suzuki  Nobuo 《International Aquatic Research》2018,10(3):283-292

International Aquatic Research - The effects of α-melanocyte-stimulating hormone (α-MSH) on calcium metabolism were examined with goldfish. The scales on the left side of goldfish bodies...  相似文献   

Reference bulk density and critical degree-of-compactness for no-till crop production in subtropical highly weathered soils     

Jos Miguel Reichert  Luis Eduardo Akiyoshi Sanches Suzuki  Dalvan Jos Reinert  Rainer Horn  Inge Hkansson 《Soil & Tillage Research》2009,102(2):242

The concept of degree of compactness (DC), referred to as field bulk density (BD) as a percentage of a reference bulk density (BD_ref), was developed to characterize compactness of soil frequently disturbed, but for undisturbed soil such as under no-tillage critical degree of compactness values have not been tested. The objective of this study was to compare methods to determine BD_ref and limits of DC and BD for plant growth under no-tillage in subtropical soils. Data from the literature and other databases were used to establish relationships between BD and clay or clay plus silt content, and between DC and macroporosity and yield of crops under no-tillage in subtropical Brazil. Data of BD_ref reached by the soil Proctor test on disturbed soil samples, by uniaxial compression with loads of 200 kPa on disturbed and undisturbed soil samples, and 400, 800 and 1600 kPa on undisturbed soil samples, were used. Also, comparisons were made with critical bulk density based on the least limiting water range (BDc LLWR) and on observed root and/or yield restriction in the field (BDc Rest). Using vertical uniaxial compression with a load of 200 kPa on disturbed or undisturbed samples generates low BD_ref and high DC-values. The standard Proctor test generates higher BD_ref-values, which are similar to those in a uniaxial test with a load of 1600 kPa for soils with low clay content but lower for soils with high clay content. The BDc LLWR does not necessarily restrict root growth or crop yield under no-tillage, since field investigations led to higher BDc Rest-values. A uniaxial load greater than 800 kPa is promising to determine BD_ref for no-tillage soils. The BD_ref is highly correlated to the clay content and thus pedotransfer functions may be established to estimate the former based on the latter. Soil ecological properties are affected before compaction restricts plant growth and yield. The DC is an efficient parameter to identify soil compaction affecting crops. The effect of compaction on ecological properties must also be further considered.  相似文献   

Preventive effects of calcitriol on the development of capsular invasive carcinomas in a rat two-stage thyroid carcinogenesis model     

Kemmochi S  Fujimoto H  Woo GH  Hirose M  Nishikawa A  Mitsumori K  Shibutani M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(5):655-664

We have shown phosphoinositide 3-kinase (PI3K)/Akt signaling activation in thyroid capsular invasive carcinomas (CICs), which are highly induced by promotion with sulfadimethoxine (SDM) in a rat 2-stage thyroid carcinogenesis model. To examine the potency of calcitriol, a synthetic vitamin D3 analog, on the development or progression of CICs, male F344 rats were injected with calcitriol (0.1 μg/kg body weight) three times a week intraperitoneally, during an entire period of SDM-promotion for 13 weeks (Experiment 1) or during the last 2 weeks of a 15-week SDM-promotion (Experiment 2). Initiation with N-bis(2-hydroxypropyl)nitrosamine preceded all treatments. In Experiment 1, long-term calcitriol treatment reduced the multiplicity of CICs, while cell proliferation activity, estimated by Ki-67 cell index in the induced CICs, was unchanged with SDM-promotion alone. Considering the strong dependency of promotion with SDM during the early stages on thyroid-stimulating hormone, the reduced multiplicity in Experiment 1 may be due to the effect on an early stage of neoplastic proliferation. Although the magnitude was mild, cell proliferation activity was decreased in existing CICs after short-term calcitriol treatment in Experiment 2, which was associated with a mild decrease in cyclin-dependent kinase-2-positive cells, cytoplasmic immunolocalization of phosphorylated, inactive, Rb protein and a mild increase in nucleocytoplasmic expression of p27(kip1). Although the effect was mild at the late stage of SDM-promotion in this hypothyroidism-related thyroid carcinogenesis model, our results suggest that calcitriol targets cell proliferation via inhibition of a molecular cascade downstream of PI3K/Akt signaling, controlling G1/S transition.  相似文献