Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
66 巻 , 11 号
November
選択された号の論文の30件中1~30を表示しています
Anatomy
FULL PAPER
  • Kazumi TANIGUCHI
    2004 年 66 巻 11 号 p. 1311-1314
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    The expression of T1R3, a taste receptor essential for the perception of sweetness and umami-taste, was examined by immunohistochemistry to determine whether and where it may be localized in the liver and pancreas. In the liver, both immunopositive and immunonegative reactions were detected; bile ducts and intercalated portions of the bile ductules were immunopositive to T1R3, while arterioles and venules were immunonegative in interlobular connective tissue. In the hepatic lobule, all other cells including liver cells (hepatocytes) and bile capillaries were immunonegative. In the pancreas, all endocrine portions of the pancreas were immunonegative to T1R3. Within the exocrine portions, immunopositive reactions were detected in excretory duct cells, intercalated cells, and centroacinar cells. In contrast, acinar cells were immunonegative, as were vessels, lymph capillaries, nerve fibers, and connective tissue cells in the exocrine portions. The restricted localization of T1R3 in the duct cells of the liver and pancreas in the present study may indicate that T1R3 is involved in monitoring changes in the makeup of bile and pancreatic juices in the hepatic and pancreatic duct systems.
  • Nobuaki NAKAMUTA, Shigeru KOBAYASHI
    2004 年 66 巻 11 号 p. 1365-1370
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Proteins encoded by p63 gene a have structural similarity with tumor suppressor p53, and were thought to induce cell cycle arrest and apoptosis during development. The p63 proteins are also expressed in the basal cells of many epithelial tissues in the adult, and supposed to play important roles in maintaining the epidermal stem cells. Previously, we reported the p63 expression in the testis of mouse embryos, suggesting their involvement in the growth arrest and apoptosis of testicular germ cells (Nakamuta and Kobayashi, J. Vet. Med. Sci. 65:853-856). In this study, we investigated the timing of this p63 expression in the germ cells during migration and colonization to the gonads. Immunohistochemical analysis of mice from embryonic day (E) 7.5 to E12.5 demonstrated that p63 positive reactivity was seen as early as E8.5 when the founder cells of germ cells, primordial germ cells (PGCs), were located in the hind gut epithelium, but PGCs were negative for p63 at E7.5 when they first appeared. p63 is expressed as six isoforms, resulting from alternative splicing at C-terminus and by the use of two promoters that generate variations at N-terminal end. RT-PCR analyses suggested that different types of p63 mRNAs were likely to be expressed in PGCs during development. These results imply that p63 may be involved in the regulation of PGC development by controlling the gene expression required for their migration and colonization to the gonads.
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Bacteriology
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  • Eiki SASAKI, Ro OSAWA, Yosuke NISHITANI, Robert A. WHILEY
    2004 年 66 巻 11 号 p. 1467-1470
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    A total of 23 Streptococcus gallolyticus strains, consisting of 12 strains from feces of healthy animals and 11 from clinical cases of human or cow mastitis milk, were examined genealogically. Four strains of S. bovis "biotype II/1" and 3 strains of S. equinus, the closely related organisms to S. gallolyticus, were also analyzed for outgroup comparison. Neither the amplified ribosomal DNA restriction analysis (ARDRA) nor the randomly amplified polymorphic DNA (RAPD) analysis that had been designed to recognize S. gallolyticus strains virulent in pigeons could differentiate clinical strains from the others of S. gallolyticus. No correspondence between the DNA profile in either analysis and the host animal species was detected.
Biochemistry
FULL PAPER
  • Yoshiaki YAMANO, Masanobu YOSHIOKA, Yoshihisa TODA, Yuichi OSHIDA, Shi ...
    2004 年 66 巻 11 号 p. 1323-1327
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    We investigated the effects of electrical foot shock stress on the melanocortin signaling cascade and the hypothalamus-pituitary-adrenal (HPA) system by observing levels of mRNA expression of corticotropin releasing factor (CRF), pro-opiomelanocortin (POMC), and melanocortin receptor subtype 4 (MC4R) in the rat amygdala and hypothalamus. When rats were exposed to electrical shock for 0.5 hr or 1 hr, plasma ACTH and corticosterone concentrations increased, indicating stress. The rats were then sacrificed to obtain RNA preparations from the brain tissue. In the amygdala, the expression of MC4R and POMC mRNA as well as CRF mRNA was significantly increased by electrical foot shock stress. In the hypothalamus, MC4R and POMC mRNA increased, but CRF mRNA remained unchanged. The duration of increased gene expression of MC4R and POMC in the amygdala was more sustained than in the hypothalamus. These results have provided the first evidence that exposure to stress increases expression of the MC4R system in the amygdala and hypothalamus.
  • Takuya IMAMURA, Nanami MIYAUCHI-SENDA, Satoshi TANAKA, Kunio SHIOTA
    2004 年 66 巻 11 号 p. 1387-1393
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    In normal tissues, methylation of CpG islands is generally accepted to be limited to the inactive X-chromosome and imprinting clusters. Gene Sphk1 has shown complex organization, indicated by multiple alternative splicing and tissue-dependent DNA methylation within the limited area (T-DMR) of the CpG island in the rat. Comparisons among human, mouse and rat SPHK1/Sphk1 genomic DNA revealed five coding exons and association of a CpG island at the 5' end in common. We also found two novel subtypes, for a total of eight mRNA subtypes generated through selective usage of untranslated first exons. A 38-bp region at the 5'-end of T-DMR is highly conserved. This restricted area is specifically hypomethylated in the brain. Here, we examine the complex genetic/epigenetic features of the SPHK1/Sphk1 CpG island, and suggest that the T-DMR is the core target for tissue-dependent CpG island methylation.
Clinical Pathology
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  • Mahmoud Rushdi ABD ELLAH, Kazuhiro NISHIMORI, Masanobu GORYO, Keiji OK ...
    2004 年 66 巻 11 号 p. 1421-1422
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    A total of 60 cattle were examined for the presence of pathological liver lesions. The liver lesions were classified as glycogen degeneration, liver abscess, sawdust liver and fatty degeneration. The value of serum adenosine deaminase (ADA) activity was investigated as a pilot study for diagnosing liver diseases in cattle. Serum ADA activity was significantly higher in cases with glycogen degeneration (9.8 ± 3.8 U/l) , liver abscess (10.4 ± 3.2 U/l), sawdust liver (11.5 ± 7.3U/l) and fatty degeneration (20.8 ± 7.7 U/l) than in the controls. The results indicate that ADA activity increases with the degree of hepatocellular damage. We concluded that serum ADA activity may be of value in bovine liver disease diagnosis.
Immunology
FULL PAPER
  • Hidekazu NAGAYA, Toshimichi KANAYA, Hiroki KAKI, Yoneko TOBITA, Masash ...
    2004 年 66 巻 11 号 p. 1395-1401
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    We developed a procedure for the large-scale purification of bovine interferon-τ (boIFN-τ) by means of a silkworm-baculovirus gene expression system. Recombinant boIFN-τ (rboIFN-τ) was efficiently produced in the silkworm infected with boIFN-τ cDNA recombinant baculovirus and accumulated in the haemolymph. To establish a purification method suitable for mass production, we tried three crude purification methods, namely, an acidification and neutralization treatment (ANT), silica gel column chromatography (SGCC), and Blue sepharose column chromatography (BSCC) with a combination of Q-sepharose (QSC) and chelating sepharose column chromatographies (CSCC). As a result, the acidification and neutralization treatment was found to be the most efficient and cost effective. With this combination, we obtained 91% pure products. To confirm the applicability of the procedure for mass production, we inoculated 100 silkworms with the recombinant virus, and recovered about 4.55 mg (1.26 × 108U/mg) of 91% pure rboIFN-τ by means of a combination of the ANT, followed by QSC and CSCC.
Internal Medicine
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Laboratory Animal Science
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  • Akinobu OKADA, Takao SHISHIDO, Kenji NAKANO, Kyoko ITO, Kotaro TAMURA, ...
    2004 年 66 巻 11 号 p. 1441-1445
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    In order to evaluate the physiological roles of the testicular endothelin (Edn) signaling via Edn receptor subtype-A (Ednra) in mammals, the localization of Ednra was investigated by in situ hybridization and immunohistochemistry in the testis of rats, dogs, and monkeys. For in situ hybridization, a rat Ednra RNA probe which is highly homologous to the subcloned canine and monkey Ednra (88.7% and 87.9% identical, respectively) was used. Both Ednra mRNA and protein were detected in interstitial cells and cells in the basal compartment of the seminiferous tubules, mainly Sertoli cells, as well as spermatogonia and some early spermatocytes, but not spermatids. The localization pattern of Ednra was exhibited in a same manner among species, indicating that the physiological role of Edn signaling throughout Ednra was maintained in the mammalian testis.
Parasitology
FULL PAPER
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Surgery
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Theriogenology
FULL PAPER
  • Naoki TAKENOUCHI, Kazunaga OSHIMA, Kazuhiro SHIMADA, Masayoshi TAKAHAS ...
    2004 年 66 巻 11 号 p. 1315-1321
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    The development of a sensitive enzymeimmunoassay (EIA) for the determination of estrone (E1) and estradiol-17β (E2 β) in bovine plasma is described. The assay is a homologous double-antibody EIA with E2β 17hemisuccinate (HS) as hapten for the immunoreactive reagent. The antiserum was raised against E2β 17HS bovine serum albumin conjugate in the rabbit, and E2β 17HS-horseradish peroxidase was used as steroid-enzyme conjugate. Each estrogen EIA was distinguished only by using the each working standard and sample for the EIA. Bovine plasma E1 and E2β were extracted and purified before EIA. The antiserum was used at 1:1,750,000 dilutions for EIA. Estrone and E2β showed high cross-reactivity with the antiserum (E1: 350.7%, E2β:100%). The sensitivities were <0.03 pg/well for E1 and <0.12 pg/well for E2β. Recovery rates of E1 and E2β added to bovine blood plasma were 94.5% and 93.9%, respectively. The precision for EIA of estrogens was below 9.7%. The profiles of either estrogen as determined by EIA corresponded closely well with follicle dynamics in the cow during the estrous cycles and with placental function in pregnant animals. In conclusion, our new EIA can be applied with sufficient sensitivities, recovery and precision for the routine analysis of E1 and E2β concentrations in bovine plasma.
  • Tsukasa OKANO, Tetsuma MURASE, Makoto ASANO, Toshio TSUBOTA
    2004 年 66 巻 11 号 p. 1359-1364
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    This study re-evaluated a protocol for cryopreservation of canine semen. Semen from 4 beagle dogs was pooled, concentrated by centrifugation and adjusted to increasing sperm concentrations by adding back seminal plasma. The prepared or original semen was diluted with an extender (Egg yolk-Tris-citrate-glucose) and cooled to 4°C (cooling), followed by a second dilution with the same extender including glycerol, equilibrated at 4°C (equilibration), then stored in liquid nitrogen. The semen was diluted for frozen samples having a fixed sperm concentration with increasing dilution rates or for those having the reverse combinations. Various dilution rates of 2.5-10 folds or sperm concentrations of 0.25-2.5 × 108/ml had no significant effect on post-thaw sperm characteristics. When cooling was done for different times (0-26 hr) with glycerol equilibration for 1 hr, post-thaw characteristics were better at 2 and 3 hr of cooling, while various times for equilibration (0-4 hr) with cooling for 3 hr had no effect. These results suggest that different dilution rates and sperm concentrations within the ranges tested may not affect the post-thaw sperm characterisitics and that sufficient time for cooling may be essential but a specific equilibration time may not necessarily be required.
  • Tsukasa OKANO, Tetsuma MURASE, Toshio TSUBOTA
    2004 年 66 巻 11 号 p. 1371-1376
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    The Japanese black bear (Ursus thibetanus japonicus) is endangered for extinction in some areas of Japan, and semen collection and cryopreservation are an important means to preserve genetic resources. The aim of this study was to characterize and cryopreserve semen of free-ranging Japanese black bears. Semen was collected by electroejaculation procedure from 4 free-ranging Japanese black bears at the capture point in the field. Ejaculates containing motile sperm were recovered from all of the animals and ejaculate volume, total sperm count, % motility (percentage of motile spermatozoa), % viability (percentage of spermatozoa that excluded eosin) and % abnormal morphology (range (mean)) were 0.65-2.20 (1.51) ml, 99-1082 (490) × 106, 5-100 (31), 42-97 (66) and 20-87 (53), respectively. Three of the 4 ejaculates were diluted with an egg yolk-TRIS-citrate-glucose extender and cryopreserved in liquid nitrogen. Motile spermatozoa were observed after freezing and thawing in all cases. This study showed that electroejaculation would be a useful method for collecting semen from free-ranging Japanese black bears and that at least motile spermatozoa would be obtained by freezing the thus collected electroejaculates.
  • Yukikazu ISHIKAWA, Ken NAKADA, Katsuro HAGIWARA, Rikio KIRISAWA, Hiros ...
    2004 年 66 巻 11 号 p. 1403-1408
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Reproductive diseases after parturition are a serious problem in dairy cattle. It is important to predict postpartum reproductive diseases early and to develop prophylaxis. The objectives of this study were to demonstrate changes in the peripheral blood concentration of interleukin-6 (IL-6) before parturition, which was mainly produced by T helper 2 type (Th2) cells, and to investigate a correlation between the IL-6 concentration and the occurrence of the postpartum retained placenta, endometritis and/or follicular cyst in dairy cattle. Twenty-seven Holstein-Friesian cows were used for this study. Thirteen had no clinical disease, 8 had retained placenta, 4 were diagnosed with endometritis by vaginal inspection, and 2 were diagnosed with follicular cyst by rectal palpation at 1 and 2 months after parturition. Blood samples were collected 60 days pre- and post-partum. They used for IL-6, progesterone (P4) and estradiol-17β (E2) concentration determination. This study showed that the IL-6 concentration prepartum was higher than postpartum. Low levels of IL-6 and P4 in peripheral blood prepartum tended to affect retained placenta and a high level of IL-6 prepartum tended to affect endometritis. These results indicate that measurement of change in the IL-6 concentration during pregnancy is one useful tool for predicting crisis in postpartum reproductive diseases in dairy cattle.
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Toxicology
FULL PAPER
  • Jin-Wook JUNG, Joon-Suk PARK, Jae-Woong HWANG, Kyung-Sun KANG, Yong-So ...
    2004 年 66 巻 11 号 p. 1329-1333
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    The recent DNA microarray technology enables us to understand a large number of gene expression profiling. The technology has potential possibility to comprehend mechanism of multiple genes were related to compounds which have toxicity in biological system. So, the toxicogenomics through this technology may be very powerful for understanding the effect of unknown toxic mechanisms in biological system. We have studied that the effect of compounds related to hepatotoxin in vivo system using DNA microarray and classified chemicals which have been well characterized. We have studied three compounds; 2 peroxisome proliferators: Clofibrate (ethyl-p-chlorophenoxyisobutyrate), gemfibrozil (5-2[2,5-dimethyl-phenoxy]2-2-dimethyl-pentanonic), and an antiepileptic drug: phenytoin (5,5-diphenylhydantoin). Male Sprague-Dawely VAF+ albino rats of 5-6 weeks old were treated with each compound for 24 hr and 2 weeks. 4.8 K cDNA microarray in house has been used for gene expression profiling. We found that the clustering of gene expression had similarity like as the toxic phenotype of compounds.
  • Wan-Seon LEE, Gyoum-Jung LEE, Gyoung-Jae LEE, Chan-Dong YEO, Jong-Soo ...
    2004 年 66 巻 11 号 p. 1335-1338
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Toxicogenomics is now emerging as one of the most important genomic application because the toxicity test based on gene expression profiles is expected to be more precise and efficient than current histopathological approaches in a pre-clinical phase. One of the challenging issues in toxicogenomics is the construction of intelligent database management system which can deal with heterogeneous and complex data from many different experimental and information sources. TEST(Toxicogenomics for Efficient Safety Test) database is especially focused on the connectivity of heterogeneous data and the intelligent query system which enable users to obtain relevant useful information from the complex data sets. The database deals with four kinds of information; compound, histopathology, gene expression, and annotation information. Currently, TEST database maintains toxicogenomics information for 16 compounds, 45 microarrays, 190 animal experiments, and customized 4.8 K rat clone set. Our presented system is expected to be a good information source for studying of toxicology mechanism in the genome-wide level and can also be applied to the designing toxicity test chip.
  • Gyoung-Jae LEE, Wan-Seon LEE, Ki-Seon JEON, Chan-Hwi UM, Yang-Suk KIM, ...
    2004 年 66 巻 11 号 p. 1339-1345
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Toxicogenomics, the subdiscipline that merges genomics with toxicology, hold the promise to contributing toward the goal of elucidating mechanism by studying genomic profiling related with various drugs. The application of gene expression profiling technology to examine multiple genes and signaling pathways promises a significant advance in understanding the toxic mechanisms of various drugs and prediction of new drug candidate. Toxicogenomics is emerging field combining genomics and bioinformatics to identify and characterize mechanisms of toxicity of drug and various compounds. The principal hypothesis underlying on this field is that chemical-specific pattern of altered gene expression is related with each chemicals properties, especially toxicological property, and it will be revealed using high-density microarray analysis of sample from exposed organisms. So, in this study we compare the gene expression pattern of two anticancer drugs paclitaxel and orally absorbable paclitaxel, using the cDNA microarray. And from the result of this study, it is possible to provide the new possibility for genome-wide insight into mechanism of their anticancer activity and toxicological phenotype.
  • Eun-Yong JUNG, Beom-Jun LEE, Young Won YUN, Jong-Koo KANG, In-Jeoung B ...
    2004 年 66 巻 11 号 p. 1347-1354
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Genistein, a soybean-originated isoflavone, is widely consumed by humans for putative beneficial health effects but its estrogenic activity may adversely affect the development of male reproductive system. Twenty one-day-old ICR mice weaned from dams fed with a soybean-based diet throughout gestation and lactation were exposed by gavage to genistein (2.5 mg/kg b.w./day) or 17β-estradiol (7.5 μg/kg b.w./day) for five weeks. Corn oil was used as a negative control. The animals were fed with a casein-based AIN-76A diet throughout the experimental periods. There were no significant differences in body and organ weights of mice among experimental groups. No significant differences in sperm counts and sperm motile characteristics were found between control and genistein groups. Treatment of 17β-estradiol caused a significant decrease in prostate weight and epididymal sperm counts compared to the control (p<0.05). The levels of phospholipid hydroxide glutathione peroxidase in the testis and prostate of mice exposed to genistein or 17β-estradiol were significantly higher than that of the control mice (p<0.05). 17β-estradiol treatment caused degeneration and apoptosis of germ cells in the testis, depletion and degeneration in the epididymal epithelium, and hyperplasia of mucosal fold region in the prostate of mice. Genistein treatment did not cause any lesion in the testis, epididymis, and prostate. These results suggest that dietary uptake of genistein during juvenile period may not affect male reproductive development and functions.
  • Zein SHABAN, Samir El-SHAZLY, Shawky ABDELHADY, Ibrahim FATTOUH, Kaamp ...
    2004 年 66 巻 11 号 p. 1377-1386
    発行日: 2004年
    公開日: 2004/12/07
    ジャーナル フリー
    Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates a spectrum of toxic and biological effects of 2,3,7,8-tetrachloro dibenzo-p-dioxin (TCDD) and related compounds. Peroxisome proliferator activated receptor alpha (PPARα) is a nuclear receptor involved in the maintenance of lipid and glucose homeostasis. In this study we hypothesized that one of the possible mechanisms for the effect of TCDD and its related chemicals on fat metabolism could be through down regulation of PPARα functions. We treated Wistar rats with an AhR ligand, Sudan III (S.III), and/or PPARα ligand, Clofibric Acid (CA), for 3 days. We analysed the expression of one of the PPARα-target gene products, CYP4A protein and its mRNA. We also tested HepG2 cells with the afore-mentioned treatments and evaluated their effects on PPARα and RXRα protein. Treatment of Wistar rats with S.III was found to down regulates CYP4A protein expression and reduced its induction with CA. It also decreased mRNA expressions of CYP4A1, CYP4A2, CYP4A3 and PPARα. In HepG2 cells, PPARα and RXRα protein expression was decreased by S.III treatment in a dose dependent manner. Our results suggest that AhR has an inhibitory effect on PPARα function and a new pathway by which AhR ligands could disturb lipid metabolism.
Virology
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