T Inaba

Osaka Prefecture University, Sakai, Ōsaka, Japan

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Publications (107)184.5 Total impact

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    ABSTRACT: Materials used for the past 30 years as immunoadjuvants induce suboptimal antitumor immune responses and often cause undesirable local inflammation. Some bacterial lipopeptides that act as Toll-like receptor (TLR) 2 ligands activate immune cells as immunoadjuvants and induce antitumor effects. Here, we developed a new dendritic cell (DC)-targeting lipopeptide, h11c (P2C-ATPEDNGRSFS), which uses the CD11c-binding sequence of intracellular adhesion molecule-1 to selectively and efficiently activate DCs but not other immune cells. Although the h11c lipopeptide activated DCs similarly to an artificial lipopeptide, P2C-SKKKK (P2CSK4), via TLR2 in vitro, h11c induced more effective tumor inhibition than P2CSK4 at low doses in vivo with tumor antigens. Even without tumor antigens, h11c lipopeptide significantly inhibited tumor growth and induced tumor-specific cytotoxic T cells. P2CSK4 was retained subcutaneously at the vaccination site and induced severe local inflammation in in vivo experiments. In contrast, h11c was not retained at the vaccination site and was transported into the tumor within 24 h. The recruitment of DCs into the tumor was induced by h11c more effectively, while P2CSK4 induced the accumulation of neutrophils leading to severe inflammation at the vaccination site. Because CD11b+ cells, but not CD11c+ cells, produced neutrophil chemotactic factors such as macrophage inflammatory protein (MIP)-2 in response to stimulation with TLR2 ligands, the DC-targeting lipopeptide h11c induced less MIP-2 production by splenocytes than P2CSK4. In this study, we succeeded in developing a novel immunoadjuvant, h11c, which effectively induces antitumor activity without adverse effects such as local inflammation via the selective activation of DCs. © 2014 Wiley Periodicals, Inc.
    International Journal of Cancer 05/2014; · 6.20 Impact Factor
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    ABSTRACT: To investigate whether pNF-H is a prognostic biomarker of spinal cord injury (SCI) in paraplegic dogs with thoracolumbar intervertebral disc herniation (IVDH). Prospective, case-control clinical study ANIMALS: Dogs (n = 60) with SCI from IVDH and 6 healthy dogs. Serum from 60 thoracolumbar IVDH dogs (Grade 4: 22 dogs; Grade 5: 38 dogs) collected 1-3 days after injury, and 6 control dogs, was analyzed using enzyme-linked immunosorbent assay (ELISA) against a phosphorylated form of the high-molecular-weight neurofilament subunit NF-H (pNF-H). Serum pNF-H levels were compared between different IVDH grades and their prognostic value was investigated. pNF-H levels were significantly greater in Grade 5 than Grade 4 dogs. There were significant differences in pNF-H levels between dogs that regained voluntarily ambulation and those that did not. All 8 dogs that had high pNF-H levels 1-3 days after injury did not regain the ability to walk after surgery. Serum pNF-H levels might be a biomarker for predicting prognosis of canine SCI.
    Veterinary Surgery 01/2014; · 1.24 Impact Factor
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    ABSTRACT: The existence of Th1 responses in a tumor microenvironment elicits a better prognosis for the patients. Transfection of Th1 polarizing cytokines, such as IFNγ, into tumor cells is an effective way to set up an appropriate microenvironment. Using a novel type synthetic vector composed of polyamidoamine dendrons, we transfected canine IFNγ gene into canine tumor cell lines, and examined direct and indirect effects of dendritic cells (DCs) against tumor growth in vitro. A cloned canine IFNγ gene expressed functional protein that induces maturation of DCs. When the canine IFNγ gene was transfected into canine tumor cell lines using the synthetic vector, most cells secreted canine IFNγ. Secretion of IFNγ reduced with time, but was maintained for 48 hours. DCs incubated with the IFNγ-transfected tumor cells exhibited greater suppressive activity and induced significantly higher cytotoxic activity against the tumor cells, relative to those incubated with untransfected tumor cells and comparable dose of IFNγ. Successful transfection of IFNγ by the synthetic vector efficiently enhanced the anti-tumor immune function of DCs, and sets up a suitable microenvironment for improvement in tumor therapy.
    Veterinary Immunology and Immunopathology 01/2014; · 1.88 Impact Factor
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    ABSTRACT: CD40 ligand (CD40L) expressed by activated T cells is shown to induce maturation of immature dendritic cells (DCs) and this maturation is a vital part in DC based tumor immunotherapy. We constructed an expression vector by cloning the extracellular domain of canine CD40L fused to the signal sequence of canine IL-12p40. When PBMCs were incubated with canine granulocyte-macrophage (GM) -CSF and IL-4, expression of CD86 was significantly elevated, but the majority of cells displayed the morphology of immature DCs. Following addition of the expressed canine soluble CD40L (csCD40L) to the DC-inducing culture, the cell morphology shifted to that of mature DCs, and expression of CD80, CD86, MHC class II and CD1a was significantly enhanced. This morphological change and enhancement of expression was observed even when the csCD40L was present only in the second half period of the culture. Furthermore, the csCD40L caused a significant increase in IL-12 production from DCs. These results show that the csCD40L significantly promotes the maturation and activation of canine monocyte derived DCs.
    Veterinary Immunology and Immunopathology 09/2013; · 1.88 Impact Factor
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    ABSTRACT: This study examined two female offspring of a somatic cell cloned Holstein cow that had reproduction problems and milk production performance issues. The two offspring heifers, which showed healthy appearances and normal reproductive characteristics, calved on two separate occasions. The mean milk yields of the heifers in the first lactation period were 9,037 kg and 7,228 kg. The relative mean milk yields of these cows were 111.2% and 88.9%, respectively, when compared with that of the control group. No particular clinical abnormalities were revealed in milk yields and milk composition rate [e.g., fat, protein and solids-not-fat (SNF)], and reproductive characteristics of the offspring of the somatic cell cloned Holstein cow suggested that the cloned offspring had normal milk production.
    Journal of Reproduction and Development 08/2013; · 1.76 Impact Factor
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    ABSTRACT: Feeding rumen bypass polyunsaturated fatty acids (PUFA) affects the reproduction of Japanese Black cows, though its influence on superovulatory response in donor cows and conception in recipient cattle has not been well studied. Here we investigated the effects of PUFA to Japanese Black cows on blood biochemistry, the numbers of ova and embryos or transferable embryos, and pregnancy rate following embryo transfer (ET) to recipient Holstein heifers. PUFA (40% linoleic acid) was fed at 300 g/day in the experimental group since the last estrus day until day of artificial insemination for superovulatory treatment. Blood was collected on the first day of follicle-stimulating hormone administration. Total cholesterol level was significantly higher in the 15 to 19 days feeding group (117.4 mg/dl) than in the control group (95.0 mg/dl). The numbers of ova and embryos or transferable embryos were significantly higher in the 15 to 19 days feeding group than in the control group. The numbers of transferable embryos in the 15 to 19 days feeding group was significantly higher than in the 10 to 14 days feeding group. The pregnancy rate at Day 60 was significantly higher in the experimental group (66.7 and 57.1%) than in the control group (51.1 and 44.0%) after transfer of fresh and frozen-thawed embryos, respectively. In conclusion, the numbers of ova and embryos or transferable embryos after superovulatory treatment increased and pregnancy rate after ET was higher in Japanese Black cows fed PUFA than in the control group.
    Journal of Veterinary Medical Science 07/2013; · 0.88 Impact Factor
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    ABSTRACT: Effect of Insulin-like Peptide 3 on Testosterone Secretion and cAMP Release in Mouse Leydig Cells Indunil Pathirana1,2,*, Masahiro Takahashi2, Shingo Hatoya2, Toshio Inaba2, Hiromichi Tamada2 and Noritoshi Kawate2 Insulin-like peptide 3 (INSL3) has recently been identified as an exclusive product of testicular Leydig cells. The role of INSL3 during fetal and neonatal period is well elucidated, but very little is known about the role of INSL3 in developing and adult males. Recently, it was shown that INSL3 is potentially involved in the prevention of germ cell apoptosis and in proliferation of osteoblasts in adults. However, regardless of the localization of its receptor (RXFP2) in Leydig cells of several mammalian species, i.e., humans, mice and dogs, the effects of INSL3 on endocrine function Leydig cells are totally unknown.The objectives of the present study were to examine the effects of INSL3 on: (1) testosterone secretion; and (2) cAMP release in mouse Leydig cells. Purified Leydig cells were isolated from testicular interstitial cells obtained from 8-week-old male mice, using a 3-step discontinuous gradient (specific gravities: 1.05, 1.06 and 1.08) of Percoll. Cells were then plated in the presence or absence of mouse, human, canine or bovine INSL3 (0–100 ng/ml) for 18 h in multiwell-plates (96 wells) in different cell densities (2500, 5000, 10,000 or 20,000 cells per well). The effects of bovine INSL3 (100 ng/ml) on testosterone secretion by Leydig cells were examined in the presence or absence of an adenylate cyclase inhibitor, SQ 22536 (1 μM) or INSL3 antagonist (bovine and human; 100 ng/ml). Testosterone and cAMP in spent medium were measured by enzyme immunoassay. All INSL3 species stimulated (P<0.0001) the testosterone secretion in Leydig cells, and the maximum stimulation (nearly twofold; P<0.05) was observed with 100 ng/ml bovine INSL3 at the lowest Leydig cell density (2500 cells per well). Moreover, bovine INSL3 (100 ng/ml) stimulated (P<0.0001) the cAMP production from Leydig cells maximally at 1 h, and remained elevated (P<0.0001) even at 18 h. SQ 22536 and INSL3 antagonists (bovine and human) reduced (P<0.0001) INSL3-stimulated testosterone secretion from Leydig cells. In conclusion, the observed stimulatory effects of INSL3 on testosterone secretion in Leydig cells are exerted via the activation of cAMP, suggesting a new autocrine function of INSL3 in males. Keywords: autocrine, cAMP, INSL3, Leydig cell, Testis, Testosterone
    Conference on Japan Sri Lanka Collaborative Research; 03/2013
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    ABSTRACT: Thrombocytopenia (TTP) is a blood disease common to canines and human beings. Currently there is no valid therapy for this disease except blood transfusion. In this study, we report the generation of canine induced pluripotent stem cells (ciPSCs) from canine embryonic fibroblasts, and a novel protocol for creating mature megakaryocytes (MKs) and functional platelets from ciPSCs. The ciPSCs were generated using lentiviral vectors, and differentiated into MKs and platelets on OP9 stromal cells supplemented with growth factors. Our ciPSCs presented in a tightly domed shape and showed expression of a critical pluripotency marker, REX1 and normal karyotype. Additionally, ciPSCs differentiated into cells derived from three germ layers via the formation of an embryoid body. The MKs derived from ciPSCs had hyperploidy and transformed into proplatelets. The proplatelets released platelets early on that expressed specific megakaryocyte and platelet marker CD41/61. Interestingly, these platelets, when activated with adenosine diphosphate (ADP) or thrombin, bound to fibrinogen. Moreover, electron microscopy showed that the platelets had the same ultrastructure as peripheral platelets. Thus, we have demonstrated for the first time the generation of ciPSCs that are capable of differentiating into megakaryocytes and release functional platelets in vitro. Our system for differentiating ciPSCs into MKs and platelets promises a critical therapy for canine TTP and appears to be extensible in principle to resolve human TTP.
    Stem cells and development 02/2013; · 4.15 Impact Factor
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    ABSTRACT: Objective-To compare methods for harvesting canine bone marrow stromal cells (BMSCs) and determine the biological properties of canine BMSCs at successive passages in vitro. Sample-BMSCs collected from the femurs of 9 Beagles. Procedures-A fibroblast assay was performed to compare 2 methods for harvesting BMSCs: the aspiration and perfusion method. Flow cytometric analysis was performed to evaluate the cell surface markers. Changes in proliferative activity were analyzed by examining radioactivity of hydrogen 3-thymidine. Cell senescence was studied via senescence-associated β-galactosidase staining, and differentiation properties (osteogenesis and adipogenesis) were estimated in association with passage. Results-The aspiration method yielded significantly more fibroblasts than the perfusion method. The cells harvested by both methods gave positive results for CD44 and CD90 and negative results for CD34 and CD45. After induction, the cells had osteogenic and adipogenic phenotypes. The biological properties of BMSCs harvested by the aspiration method were estimated in association with passage. With increasing number of passages, the proliferative activity was reduced and the proportion of cells with senescence-associated β-galactosidase staining was increased. The capacity of differentiation was reduced at passage 3. Conclusions and Clinical Relevance-The aspiration method was superior for collection of BMSCs. In early passages, canine BMSCs had the proliferative activity and potential of osteogenic and adipogenic differentiation, but this decreased with increased number of passages. Consideration of passage will be important to the success of any strategy that seeks to regenerate tissue though the use of BMSCs.
    American Journal of Veterinary Research 11/2012; 73(11):1832-40. · 1.35 Impact Factor
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    ABSTRACT: Testicular Leydig cells secrete insulin-like peptide 3 (INSL3) and express its receptor, RXFP2. However, the effects of INSL3 on endocrine function of Leydig cells are unknown. The present study examines the effects of INSL3 on mouse Leydig cells taking testosterone and cAMP secretions as endpoints. Leydig cells were isolated from testicular interstitial cells obtained from 8-week-old male mice. Cells were then plated in the presence or absence of mouse, human, canine or bovine INSL3 (0-100 ng/ml) for 18 h in multiwell-plates (96 wells) in different cell densities (2500, 5000, 10,000 or 20,000 cells per well). The effects of bovine INSL3 (100 ng/ml) on testosterone secretion by Leydig cells were examined in the presence or absence of, an adenylate cyclase inhibitor, SQ 22536 (1μM) or INSL3 antagonist (bovine and human; 100 ng/ml). Testosterone and cAMP in spent medium were measured by enzyme immunoassay. All INSL3 species tested significantly stimulated the testosterone secretion in Leydig cells, and the maximum stimulation was observed with 100 ng/ml bovine INSL3 at the lowest Leydig cell density (2500 cells per well). Moreover, bovine INSL3 (100 ng/ml) significantly stimulated the cAMP production from Leydig cells maximally at 1h, and remained significantly elevated even at 18 h. SQ 22536 and INSL3 antagonists (bovine and human) significantly reduced INSL3-stimulated testosterone secretion from Leydig cells. Taken together, stimulatory effects of INSL3 on testosterone secretion in Leydig cells are exerted via the activation of cAMP, suggesting a new autocrine function of INSL3 in males.
    Regulatory Peptides 07/2012; 178(1-3):102-6. · 2.06 Impact Factor
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    ABSTRACT: This study was undertaken to develop a simple and practical method to control the time of ovulation in cynomolgus monkeys. Diets containing a synthetic gestagen, levonorgestrel (LNG) were given daily to normally cycling female monkeys for 2 weeks, and plasma concentrations of estradiol-17β and progesterone were determined by EIA in order to estimate the time of ovulation. Doses of LNG (0, 3.2, 8, 20, 50, or 125 µg) were given from Day 2 (Day 0 = the first day of menstruation) through Day 15. The numbers of days from the last administration of LNG to the estimated ovulation in the groups treated with LNG at 20 µg and above were significantly greater than those in the controls, and the values in the group treated with LNG at 50 µg were within a narrow range. In a second experiment, LNG was administered at 50 µg in different phases of the menstrual cycle (Days 9-22, 16-29, and 23-36), and the results indicated that ovulation occurred more than 12 days after the last administration in all monkeys, and the number of days from the last administration of LNG to the estimated ovulation in the group treated on Days 16-29 (luteal phase) was significantly greater than that in the group treated on Days 23-36. These results indicate that daily provision of a diet containing 50 µg LNG could be applicable for delaying ovulation, and suggest that the total level of (exogenous and endogenous) progestins is critical for determining the length of ovulation delay in cynomolgus monkeys.
    Journal of Veterinary Medical Science 07/2012; · 0.88 Impact Factor
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    ABSTRACT: This study examined factors involved in the patency of uterine cervices in the bitch with pyometra. The uterine cervices were obtained from the bitches with pyometra at the time of ovariohysterectomy. Cervical patency was measured by inserting the stainless steel rods with different diameter into cervical canals. Collagen concentration and collagenase activity (for type I collagen) in the tissue were determined and the number of neutrophils, which contain the enzymes related to collagen metabolism, and morphological changes in collagenous fibers were studied by histological examination. Levels of mRNA expressions for hormonal factors, estrogen receptor-α (ER-α), progesterone receptor (PR), relaxin (Rlx) and an attractant of neutrophils, interleukin-8 (IL-8), were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). In the statistical analysis, the cervical patency positively correlated with the collagenase activity, and negative correlation was found between the cervical patency and collagen concentration. Histological examination indicated distinct positive correlation between the cervical patency and the number of neutrophils in the cervical stroma and that the collagenous fiber in the uterine cervix became thinner and degraded with increase of the cervical patency. Although there was no relationship between the cervical patency and the level of mRNA for ER-α, PR or Rlx, IL-8 mRNA level has significant positive correlation with the cervical patency and the number of neutrophils in the cervical stroma. These results suggest that the increased number of neutrophils in the uterine cervix, which could be related to the local expression of IL-8, may be involved in collagen degradation and connective tissue remodeling to increase cervical patency in the bitch with pyometra.
    Research in Veterinary Science 06/2012; 93(3):1203-10. · 1.77 Impact Factor
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    ABSTRACT: To assess the feasibility and safety of transplantation of autologous bone marrow stromal cell (BMSC) in dogs with acute spinal cord injury (SCI). An open-label single-arm trial. Dogs (n = 7) with severe SCI from T6 to L5, caused by vertebral fracture and luxation. Decompressive and stabilization surgery was performed on dogs with severe SCI caused by vertebral fracture and luxation. Autologous BMSCs were obtained from each dog's femur, cultured, and then injected into the lesion in the acute stage. Adverse events and motor and sensory function were observed for >1 year after SCI. Follow-up was 29-62 months after SCI. No complications (eg, infection, neuropathic pain, worsening of neurologic function) were observed. Two dogs walked without support, but none of the 7 dogs had any change in sensory function. Autologous BMSC transplantation is feasible and safe in dogs with acute SCI. Further studies are needed to determine the efficacy of this therapy.
    Veterinary Surgery 04/2012; 41(4):437-42. · 1.24 Impact Factor
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    ABSTRACT: Progressive ataxia and paralysis in three Miniature Dachshunds were found to be caused by idiopathic sterile pyogranulomatous inflammation of epidural fat between T5 and L4. All dogs were managed by hemilaminectomy and removal of epidural compressive material. Surgical findings and histopathological evaluation were necessary to diagnose epidural pyogranulomatous inflammation. A dog did not regain motor and sensor function after the surgery. Two dogs had exhibited improved neurological function after the surgery, but they recurred. Oral cyclosporine treatment was useful for their long remission. Idiopathic sterile pyogranulomatous inflammation of epidural fat can be considered to be a cause of thoracolumbar myelopathy in dogs.
    Journal of Veterinary Medical Science 04/2012; 74(8):1071-4. · 0.88 Impact Factor
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    ABSTRACT: The objectives were to: (1) develop a time-resolved fluorescence immunoassay (TRFIA) to measure insulin-like peptide 3 (INSL3) in canine plasma; (2) investigate changes of plasma concentrations of INSL3 and testosterone with age in normal male dogs; and (3) compare hormonal concentrations among cryptorchid, normal, and castrated dogs to evaluate endocrine function of the Leydig cell component in retained testes. Blood samples were taken from normal male dogs from prepubertal age to advanced age (4 mo to 14 y, n = 89), and from unilateral cryptorchid (n = 31), bilateral cryptorchid (n = 7), and castrated dogs (n = 3). Canine plasma INSL3 was measured with a newly developed TRFIA. The minimum detection limit of the INSL3 assay was 0.02 ng/ml and the detection range was 0.02 to 20 ng/ml. Plasma INSL3 concentrations increased (P < 0.05) from prepubertal age (4-6 mo) to pubertal age (6-12 mo), and then declined (P < 0.05) from pubertal age to post-pubertal age (1-5 y), reaching a plateau. Plasma testosterone concentrations increased (P < 0.0001) dramatically from prepubertal to pubertal ages, and then seemed to plateau. Concentrations of both INSL3 and testosterone were lower (P < 0.0001 for each) in bilateral cryptorchid dogs than in normal and unilateral cryptorchid dogs. The INSL3 (range: 0.05-0.43 ng/ml) and testosterone (range: 0.10-0.94 ng/ml) concentrations were readily detected in bilateral cryptorchids, but not in castrated dogs (INSL3 < 0.02 ng/ml; testosterone < 0.04 ng/ml). In conclusion, plasma INSL3 concentrations in male dogs measured by a newly developed TRFIA had a transient surge at a pubertal age, whereas testosterone did not. Lower plasma concentrations of INSL3 and testosterone in bilateral cryptorchid dogs suggest impaired endocrine functions of Leydig cell component in paired retained testes. Therefore, peripheral plasma INSL3 and testosterone concentrations have potential diagnostic value in predicting the presence of bilaterally retained testes in male dogs.
    Theriogenology 02/2012; 77(3):550-7. · 2.08 Impact Factor
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    ABSTRACT: The objectives were to: (1) develop an enzyme immunoassay (EIA) for insulin-like peptide 3 (INSL3) or relaxin-like factor (RLF) in bovine plasma; (2) investigate changes of plasma INSL3 concentrations from birth to pubertal age of beef bulls; and (3) compare changes in plasma concentrations of INSL3, testosterone, and LH. Plasma samples were collected from beef bull calves (n = 15) at birth (0 d) and at 28, 56, and 84 d after birth. Furthermore, in beef bulls around pubertal age (n = 26; age range 3 to 22 mo), plasma samples were collected at 1 to 4 mo intervals. Plasma INSL3 concentrations increased (P < 0.05) from 0 to 28, 28 to 56, and from 56 to 84 d of age. Plasma testosterone concentrations increased (P < 0.001) from 0 to 28 d, and from 28 to 56 d, but did not change from 56 to 84 d. For bulls around pubertal age, plasma INSL3 concentrations did not change from the prepubertal phase (3 to 6 mo) to the early pubertal phase (6 to 12 mo), but increased (P < 0.05) from the early to late pubertal phases (12 to 18 mo), and from the late pubertal to postpubertal phases (18 to 22 mo). Plasma testosterone concentrations increased from the prepubertal to early pubertal phases (P < 0.001), but did not change thereafter. Plasma LH concentrations did not change from 0 d to 84 d, but decreased (P < 0.001) from prepubertal to early pubertal phase, with no significant change thereafter. Plasma INSL3 concentrations increased during the first 3 mo of life and throughout the pubertal age in beef bulls. There were similar dynamic patterns for INSL3 and testosterone during the first 3 mo of life, but patterns subsequently diverged in bulls around pubertal ages.
    Theriogenology 08/2011; 76(9):1632-8. · 2.08 Impact Factor
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    ABSTRACT: To evaluate effects of transplantation of bone marrow stromal cells (BMSCs) into the CSF for the treatment of chronic spinal cord injury in dogs that had not responded by 1 month after decompressive surgery. 23 dogs. Dogs with paraplegia and loss of nociception in the pelvic limbs for at least 1 month after decompressive surgery were assigned to transplantation or control groups. Dogs in the transplantation group received BMSCs injected into the CSF 1 to 3 months after decompressive surgery. Dogs in the control group did not receive additional treatments. Improvements in gait, proprioceptive positioning, and nociception were evaluated by use of the Texas Spinal Cord Injury Scale for ≥ 6 months after BMSC transplantation. 6 of 10 dogs in the transplantation group regained the ability to walk, whereas only 2 of 13 dogs in the control group regained the ability to walk. Scores for the Texas Spinal Cord Injury Scale in the transplantation group were significantly higher than scores in the control group at the endpoint of the study (6 months after BMSC transplantation or after decompressive surgery for the transplantation and control groups, respectively). Only 1 dog (transplantation group) recovered nociception. All dogs from both groups had fecal and urinary incontinence. No complications were observed in relation to BMSC transplantation. Injection of BMSCs into the CSF caused no complications and could have beneficial effects on pelvic limb locomotion in dogs with chronic spinal cord injuries.
    American Journal of Veterinary Research 08/2011; 72(8):1118-23. · 1.35 Impact Factor
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    ABSTRACT: The objective was to determine the effects of estradiol-17β, monobutyl phthalate (MBP) and mono-(2-ethylhexyl) phthalate (MEHP) on testosterone and insulin-like peptide 3 (INSL3) secretions in cultured testicular interstitial cells isolated (enzymatic dispersion) from scrotal and retained testes of small-breed dogs. Suspension cultures were treated with estradiol-17β (0, 10, and 100 ng/mL), MBP (0, 0.8, and 8 mmol/L) or MEHP (0, 0.2, and 0.8 mmol/L) for 18 h, in the presence or absence of 0.1 IU/mL hCG. Testosterone (both basal and hCG-induced) and INSL3 (basal) concentrations were measured in spent medium. Effects of estradiol-17β, MBP, and MEHP on testosterone and INSL3 secretions were not affected (P > 0.15) by cell source (scrotal versus retained testis); therefore, data were combined and analyzed, and outcomes reported as percentage relative to the control. In testicular interstitial cells, basal testosterone secretion was increased (P < 0.01) by 100 ng/mL estradiol-17β (130.2 ± 10.6% of control). Among phthalates, 0.2 and 0.8 mmol/L MEHP stimulated (P < 0.01) basal testosterone secretion (135.5 ± 8.3% and 154.6 ± 12.9%, respectively). However, hCG-induced testosterone secretion was inhibited (P < 0.01) by 8 mmol/L MBP (67.7 ± 6.0%), and tended to be inhibited (P = 0.056) by 0.8 mmol/L MEHP (84.5 ± 5.6%). Basal INSL3 secretion was inhibited (P < 0.01) by 8 mmol/L MBP (73.6 ± 6.8%) and 0.8 mmol/L MEHP (76.9 ± 11.3%). In conclusion, we inferred that estradiol-17β and certain phthalate monoesters had direct effects on secretions of testosterone and INSL3 in canine testicular interstitial cells, with no significant difference between scrotal and retained testes.
    Theriogenology 07/2011; 76(7):1227-33. · 2.08 Impact Factor
  • The Veterinary record. 06/2011; 168(25):669.
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    ABSTRACT: Differences in the distribution of single nucleotide polymorphisms (SNPs) and haplotypes in the estrogen receptor α gene (ESR1) were examined in Miniature Dachshunds (n = 48), Chihuahuas (n = 20) and Toy Poodles (n = 18). Five DNA fragments located in the 40-kb region at the 3' end of ESR1 were amplified by polymerase chain reaction and were directly sequenced. We compared allele, genotype and estimated haplotype frequencies at each SNP in the 3' end of ESR1 for these three breeds of small dog. The frequency of the major allele and the genotype frequency of the major allele homozygotes, were significantly higher in Toy Poodles for five SNPs (SNP #5, #14-17) than in Miniature Dachshunds, and significantly higher in Toy Poodles than Chihuahuas for three SNPs (SNP #15-17). A common haplotype block was identified in an approximately 20-kb region encompassing four SNPs (SNPs # 14-17). The frequencies of the most abundant estimated haplotype (GTTG) and GTTG homozygotes were significantly higher in Toy Poodles than in the other two breeds. These results imply that homozygosity for the allele, genotype and haplotype distribution within the block at the 3' end of ESR1 is greater in Toy Poodles than in Miniature Dachshunds and Chihuahuas.
    Animal Science Journal 06/2011; 82(3):390-5. · 1.04 Impact Factor

Publication Stats

578 Citations
184.50 Total Impact Points

Institutions

  • 1988–2014
    • Osaka Prefecture University
      • Graduate School of Life and Environmental Sciences
      Sakai, Ōsaka, Japan
    • Kanagawa University
      Yokohama, Kanagawa, Japan
    • University of Washington Seattle
      • Department of Obstetrics and Gynecology
      Seattle, WA, United States
  • 2012
    • Nara Hospital
      Ikuma, Nara, Japan
    • Shin Nippon Biomedical Laboratories, Ltd.
      Kagosima, Kagoshima, Japan
  • 1997
    • Yamaguchi University
      • Faculty of Agriculture
      Yamaguchi-shi, Yamaguchi-ken, Japan
  • 1988–1989
    • Meiji University
      Edo, Tōkyō, Japan