Establishment of a BRAF V595E-mutant canine prostate cancer cell line and the antitumor effects of MEK inhibitors against canine prostate cancer     

Masanori Kobayashi  Moe Onozawa  Shiho Watanabe  Tomokazu Nagashima  Kyoichi Tamura  Yoshiaki Kubo  Akiko Ikeda  Kazuhiko Ochiai  Masaki Michishita  Makoto Bonkobara  Masato Kobayashi  Tatsuya Hori  Eiichi Kawakami 《Veterinary and comparative oncology》2023,21(2):221-230

Canine prostate cancer (cPCa) is a malignant neoplasm with no effective therapy. The BRAF V595E mutation, corresponding to the human BRAF V600E mutation, is found frequently in cPCa. Activating BRAF mutations are recognized as oncogenic drivers, and blockade of MAPK/ERK phosphorylation may be an effective therapeutic target against BRAF-mutated tumours. The aim of this study was to establish a novel cPCa cell line and to clarify the antitumor effects of MEK inhibitors on cPCa in vitro and in vivo. We established the novel CHP-2 cPCa cell line that was derived from the prostatic tissue of a cPCa patient. Sequencing of the canine BRAF gene in two cPCa cell lines revealed the presence of the BRAF V595E mutation. MEK inhibitors (trametinib, cobimetinib and mirdametinib) strongly suppressed cell proliferation in vitro, and trametinib showed the highest efficacy against cPCa cells with minimal cytotoxicity to non-cancer COPK cells. Furthermore, we orally administered 0.3 or 1.0 mg/kg trametinib to CHP-2 xenografted mice and examined its antitumor effects in vivo. Trametinib reduced tumour volume, decreased phosphorylated ERK levels, and lowered Ki-67 expression in xenografts in a dose-dependent manner. Although no clear adverse events were observed with administration, trametinib-treated xenografts showed osteogenesis that was independent of dosage. Our results indicate that trametinib induces cell cycle arrest by inhibiting ERK activation, resulting in cPCa tumour regression in a dose-dependent manner. MEK inhibitors, in addition to BRAF inhibitors, may be a targeted agent option for cPCa with the BRAF V595E mutation.  相似文献   

A case of atypical canine lymphoma with oral mass and multiple osteolysis     

Ito T  Hisasue M  Neo S  Ishikawa T  Shida T  Watanabe T  Enomoto Y  Okuda M  Tsuchiya R  Madarame H  Ogata M  Yamada T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(9):977-980

An 8-year-old female Golden Retriever had an oral mass and lameness. Multiple osteolysis of the systemic skeleton without monoclonal gammopathy was shown on electrophoresis of serum and urine samples. Cytological and histopathological examinations of the oral mass revealed atypical polymorphic cells similar to myeloid cells, and bone marrow aspiration indicated that these abnormal cells also might have invaded the bone marrow. These cells were negative to peroxidase and non-specific esterase staining, and clonal expansion of B lymphocytes could be detected by polymerase chain reaction (PCR) assay for antigen receptor gene rearrangement. The case was diagnosed as atypical lymphoma and treated by multi-drug chemotherapy. On the 142nd day after the first admission, the case had remission and the oral mass and multiple osteolysis were improved.  相似文献   

Comparison of the growth performances of offspring produced by a pair of cloned cattle and their nuclear donor animals     

Kasai K  Sano F  Miyashita N  Watanabe S  Nagai T 《The Journal of reproduction and development》2007,53(1):135-142

In the present study, the growth performance of a calf produced by mating a somatic cell cloned dam and sire was compared with that of its full siblings produced by mating the cattle used as nuclear donors for the cloned animals. The somatic cell cloned dam and sire were derived from cultured cumulus cells and ear cells, respectively. The cloned dam was artificially inseminated with semen from the cloned sire. A female calf was produced that was reared under general group feeding conditions. The calf was subjected to a clinical examination and to hematology, serum biochemistry, and telomere length analyses; all of these tests indicated that the calf was normal. The growth characteristics (body weight and shoulder height) of the calf fell within the range of the full siblings of the same sex produced by mating the animals used as the nuclear donors of clones. These findings suggest that the same breeding performance is expected from mating a cloned dam and sire as from mating the animals used as nuclear donors for the clones.  相似文献   

Characterization of Newcastle disease virus isolated from northern pintail (Anas acuta) in Japan     

Sakai K  Sakabe G  Tani O  Watanabe Y  Jahangir A  Nakamura M  Takehara K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(12):1307-1311

A field isolate of Newcastle disease virus (NDV) isolated from northern pintail (Anas acuta) in Tohoku district, northeast Japan, was characterized. Phylogenetic analysis of the fusion protein indicated that the isolate belonged to genotype I and was closely related to isolates from the Far East corresponded to the migration route for this bird species. The isolate had the typical avirulent cleavage site of the fusion protein (112)GKQGR*L(117). In addition, pathogenicity tests indicated the isolate to have avirulent characteristics. However, the isolate has been shown to cause fusion cytopathic effects and form plaques on chicken embryo fibroblasts (CEF) in the absence of trypsin. The present results suggest that the CEF-adapted NDV, which is avirulent, is circulating among waterfowl populations.  相似文献   

Changes of cyclic AMP levels and phosphodiesterase activities in the rat ovary     

Wang Z  Shi F  Jiang YQ  Lu LZ  Wang H  Watanabe G  Taya K 《The Journal of reproduction and development》2007,53(4):717-725

Cyclic AMP (cAMP) is a second messenger that plays a critical role in follicular recruitment, development and luteinization in the mammalian ovary. The cellular level of cAMP is largely dependent on the activity of phosphodiesterase (PDE), which degrades cAMP into 5'-AMP. The present study was conducted to investigate the level of cAMP and the activity of cAMP-PDE in postnatal rats; immature rats during gonadotropin-primed follicular development, ovulation and luteinization; adult rats during normal estrous cycling; and aged rats that spontaneously developed persistent estrous (PE) by radioimmunoassay (RIA). All four rat models were confirmed by histological examination of one ovary and assayed using the other ovary by RIA. In the postnatal rats, the ovarian cAMP level was high on day 10 after birth, while ovarian cAMP-PDE activity was highest at 21 days of age. In the immature female rats, both the ovarian cAMP level and cAMP-PDE activity increased remarkably after treatment with equine chorionic gonadotropin (eCG), increased continuously 24 h after injection of human chorionic gonadotropin (hCG) for induction of ovulation and luteinization, and then declined significantly. In the adult rats during the normal estrous cycle, the ovarian cAMP levels were low on the day of estrus, and there were no significant changes in ovarian cAMP-PDE activity throughout the estrous cycle. In the PE rats, the ovarian cAMP levels were similar to those of the adult rats on the day of estrus but were lower than those on the other days of the estrous cycle; ovarian cAMP-PDE activity was lower than that in the adult rats on any day of the estrous cycle. Together, these findings indicate that the ovarian cAMP level and cAMP-PDE activity were regulated in a stage-dependent manner during ovarian follicular development, atresia and luteinization and providing evidences that cAMP and cAMP-specific PDEs are involved in these physiological processes.  相似文献   

Osteopetrosis-like disease in a cat with respiratory distress     

Fujita M  Takaishi Y  Nagae H  Watanabe N  Hasegawa D  Taniguchi A  Orima H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(6):687-690

Magnetic resonance (MR) and computed tomography (CT) were performed in an 8-year-old, spayed female cat with chronic effort respiration at the inspiration phase and stertor. Increased bone opacity in the areas of the head, neck and thorax were observed on radiography. MR images showed no signal intensity on both transverse T1WI and T2WI of the nasal cavity. CT revealed increased bone density and hypertrophy of the nasal turbinate and a narrowed nasal passage. From these results, we concluded this case had osteopetrosis-like disease, and that the respiratory distress was caused by hypertrophy of the nasal turbinate.  相似文献   

Regulation of free glutamate content in meat by dietary lysine in broilers          下载免费PDF全文

Genya Watanabe  Hiroyuki Kobayashi  Masahiro Shibata  Masatoshi Kubota  Motoni Kadowaki  Shinobu Fujimura 《Animal Science Journal》2015,86(4):435-442

Regulation of taste is important for improving meat quality and glutamate (Glu) is one of the important taste‐active components in meat. Here, the effects of dietary lysine (Lys) content on taste‐active components in meat, especially free Glu, were investigated. Fourteen‐day‐old broiler chicks (Gallus gallus) were fed on diets containing 100% or 150% of the recommended Lys content for 10 days. Concentrations of free amino acids in plasma, muscle and liver were measured. The levels of messenger RNAs (mRNAs) for enzymes related to Glu metabolism were determined in muscle and liver. The concentration of muscle metabolites was also determined. The free Glu content in muscle of chicks fed the Lys150% diet was increased by 44.0% compared with that in chicks fed the Lys100% diet (P < 0.01). The mRNA level of lysine α‐ketoglutarate reductase, which is involved in Lys degradation and Glu production, was significantly increased (P < 0.05) in the Lys150% group. Metabolome analysis showed that the Lys degradation products, muscular saccharopine, pipecolic acid and α‐aminoadipic acid, were increased in the Lys150% group. Our results suggest that free Glu content in muscle is regulated by Lys degradation. These results suggest that a short‐term feeding of high‐Lys diet could improve the taste of meat.  相似文献   

Extracellular cyclophilin A possesses chemotaxic activity in cattle     

Satoru Takanashi  Tomonori Nochi  Miku Abe  Nanami Itaya  Megumi Urakawa  Katsuyoshi Sato  Tao Zhuang  Saori Umemura  Tomohito Hayashi  Yoshio Kiku  Haruki Kitazawa  Michael T. Rose  Kouichi Watanabe  Hisashi Aso 《Veterinary research》2015,46(1)

Cyclophilin A (CyPA) was originally discovered in bovine thymocytes as a cytosolic binding protein of the immunosuppressive drug cyclosporine A. Recent studies have revealed that in mice and humans, CyPA is secreted from cells in injured or infected tissues and plays a role in recruiting inflammatory cells in those tissues. Here we found that in cattle abundant level of extracellular CyPA was observed in tissues with inflammation. To aid in investigating the role of extracellular CyPA in cattle, we generated recombinant bovine CyPA (rbCyPA) and tested its biological activity as an inflammatory mediator. When bovine peripheral blood cells were treated with rbCyPA in vitro, we observed that rbCyPA reacts with the membranous surface of granulocytes, monocytes and lymphocytes. Chemotaxis analysis showed that the granulocytes migrate toward rbCyPA and the migration is inhibited by pre-treatment with an anti-bovine CyPA antibody. These results indicate that, as for mice and humans, extracellular CyPA possesses chemotactic activity to recruit inflammatory cells (e.g., granulocytes) in cattle, and could thus be a potential therapeutic target for the treatment of inflammation.  相似文献   

Flowering ability of commercial Calibrachoa cultivars as compared to that of natural species     

Takeshi Kanaya  Hisashi Kokubun  Hitoshi Watanabe  Goro Hashimoto  Eduardo Marchesi  Laura Bullrich  Toshio Ando 《Scientia Horticulturae》2010

To evaluate and describe the current status of Calibrachoa cultivars as bedding plants, the seasonal flowering of 91 commercial cultivars were compared to those of natural species using principal components analysis. The predominant flowering of the cultivars, that is, floriferous in late spring to early summer with a tendency to reduce flower production toward winter, seems to have remained unchanged from that of natural species. However, properties desired in bedding plants, such as early flowering and continuous flowering under short-day and/or low temperature conditions, appear to have been improved in a considerable number of cultivars, although there is still room for improvement. Seasonal and total floral counts were higher in many cultivars compared to natural species. Several natural species were selected for their unique flowering ability and should be evaluated as gene resources for improving the flowering of Calibrachoa cultivars.  相似文献   

Competition as a predisposing factor of crown dieback in a secondary forest of <Emphasis Type="Italic">Betula maximowicziana</Emphasis> in Hokkaido,northern Japan     

Yasuyuki Ohno  Kiyoshi Umeki  Kazuhiko Terazawa  Michiyasu Yasaka  Ichiro Watanabe  Mika Takiya 《Journal of Forest Research》2010,15(3):161-168

We analyzed the probability that Betula maximowicziana Regel (monarch birch) would suffer crown dieback (crown-dieback probability) and the basal area growth rate (GB), which was found to be a predisposing stress factor making birch trees susceptible to crown dieback. First, we analyzed the relationship between the probability that birch trees would suffer from crown dieback in 1999 and GB from a period prior to the occurrence of crown dieback (1985–1987), using a data set of repeated measurements on 217 trees. Logistic regression analysis revealed that monarch birch had a larger crown-dieback probability when GB was low in the preceding period. Hence, there were predisposing stress factors that reduced GB and continued to affect trees for at least a decade. Next, we analyzed GB in the same period in relation to symmetrical and asymmetrical competition between trees and found that GB was reduced by symmetrical competition, suggesting that this was one of the predisposing factors for crown dieback. Based on these results, we used selected models for crown-dieback probability and GB to calculate crown-dieback probabilities for individuals with different initial basal areas and experiencing different intensities of symmetrical competition. The predicted crown-dieback probability decreased with decreasing symmetrical competition between trees. We discuss a possible process of crown dieback to death for monarch birch and the use of thinning as a method to reduce the risk of crown dieback.  相似文献