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Biomedical subjects

M Shino

Publications and source records attributed to M Shino.

At least 19 recordsLinked to original sources
PubMed2001

Serological survey of parapoxvirus infection in wild ruminants in Japan in 1996-9.

The prevalence of parapoxvirus infection was examined in free-ranging wild ruminants in Japan, Japanese serow (Capricornis crispus) and Japanese deer (Cervus nippon centralis), in 1996-9. We collected a total of 151 serum samples from 101 Japanese serows and 50 Japanese deer and tested for antibodies against parapoxvirus by an enzyme-linked immunosorbent assay and an agar gel immunodiffusion test. Overall seroprevalences among Japanese serows were 5/25 (20.0%) in 1996, 4/14 (28.6%) in 1997, 5/32 (15.6%) in 1998 and 2/30 (6.7%) in 1999, respectively. The seroprevalence increased with age but was not affected by sex. No antibodies were detected from any of 50 serum samples taken from Japanese deer. Our results in this study suggest that parapoxvirus infection is widespread among the population of Japanese serows, however, Japanese deer appear to be still free of the disease.

Age Factors↗
PubMed2001

Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva.

Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purfied swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-Il) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 +/- 0.017 mg/mg total protein.

Animals↗
PubMed2001

Generation of live rat offspring by intrauterine insemination with epididymal spermatozoa cryopreserved at -196 degrees C.

This study reports the development of a reliable method for cryopreservation of rat epididymal spermatozoa and the production of live young by artificial insemination using these cryopreserved spermatozoa. The motility and membrane integrity of rat spermatozoa were investigated after spermatozoa had been subjected to physical stress and frozen with various concentrations of glycerol (0, 3 and 6%) either in the presence or absence of Equex Stem as cryoprotective agents. The ability of cryopreserved spermatozoa to generate normal offspring by intrauterine insemination was also evaluated. Rat spermatozoa that had been centrifuged at 700 g for 5 min showed a significant decrease in motility compared with non-centrifuged spermatozoa. In addition, after centrifugation three times the percentage of membrane-intact spermatozoa decreased to approximately 0%. The percentage of membrane-intact spermatozoa was significantly higher (P < 0.01) in semen samples that had been frozen in medium without glycerol than in samples frozen in medium with 3% glycerol. Although the addition of 0.7% Equex Stem to medium without glycerol or with 3% glycerol did not influence rates of sperm motility after freezing and thawing, the percentage of membrane-intact spermatozoa was improved by the presence of 0.7% Equex (P < 0.05). Therefore, rat spermatozoa were handled gently to avoid physical stress and were frozen in medium containing 23% egg yolk, 8% lactose monohydrate and 0.7% Equex Stem, at pH 7.4 adjusted with 10% Tris(hydroxymethyl)aminomethane solution. Thirteen female rats were inseminated into the oviductal end of both uterine horns with frozen-thawed spermatozoa. Forty-one normal live offspring were obtained from nine of the inseminated females. These results indicate that frozen-thawed rat spermatozoa can generate normal offspring. To our knowledge, this procedure is the first successful production of offspring using spermatozoa cryopreserved in liquid nitrogen.

Animals↗
PubMed1999

Developmental competence, after transfer to recipients, of porcine oocytes matured, fertilized, and cultured in vitro.

The objective of this study was to evaluate the developmental ability of early porcine embryos produced in vitro and transferred to recipient gilts. Porcine cumulus-oocyte complexes were matured in modified North Carolina State University-37 solution for 44-46 h (in vitro maturation, IVM). In vitro fertilization (IVF) was performed with frozen-thawed epididymal spermatozoa. Inseminated oocytes were cultured in vitro (IVC) for 0, 24, or 48 h in modified NCSU-37 solution. Embryos were surgically transferred to the oviducts of recipients in which estrus had been synchronized with eCG and hCG. On the 29th day post-IVF, the uteri of some recipients were surgically examined for pregnancy; then pregnant females were hysterectomized in order to examine number and weight of the fetuses. Developmental rates to fetuses for IVM/IVF oocytes cultured for 24 and 48 h were significantly lower (p < 0.05, 1.7% and 2.0%, respectively) than that of IVM/IVF oocytes without IVC (6.7%). However, the weights of fetuses (1.0-1.2 g) did not differ among the experimental groups. The other recipients were examined for pregnancy using an ultrasound pregnancy detector, and pregnant females were allowed to go to term. Healthy piglets were delivered by some recipients to which embryos cultured for 0 or 24 h had been transferred; however, no farrow was obtained from embryos cultured for 48 h before the transfer. The results indicate that the viability of in vitro-produced porcine embryos is decreased by IVC after IVF; however, these embryos have competence to develop to term. An improved IVC system of porcine IVM/IVF oocytes is needed to generate advances in this field.

Animals↗
PubMed1995

Acinic cell tumour of the maxillary sinus: an unusual case initially diagnosed as parotid cancer.

An unusual case of acinic cell tumour of the maxillary sinus is presented. The patient, a 41-year-old male who had undergone incomplete excision of the tumour in the left parotid region previously, was referred to our department for further treatment. The initial pathological diagnosis was adenocarcinoma of the parotid gland. CT-scan not only revealed tumours in the left pre-auricular and upper neck region, but also an enhanced mass in the left maxillary sinus. Although there were neither nasal symptoms nor destruction of the maxillary bone, aspiration biopsy of the maxillary sinus revealed class V. Total maxillectomy, radiotherapy and systemic chemotherapy were performed just after total parotidectomy and radical neck dissection at the left side. The clinical and histopathological findings are discussed.

Adenocarcinoma↗
PubMed1993

[Determination of ubiquinones in biological material by high performance liquid chromatography].

Quantitative high performance liquid chromatographic (HPLC) methods are described for the determination of reduced, oxidized, and total ubiquinone (UQred, UQox, and total UQ, respectively) in biological material. UQ was extracted into n-hexane from an aqueous homogenate after precipitation of protein by addition of alcohol. The n-hexane extract was evaporated to dryness under a N2 atmosphere at 30 degrees C. In order to determine the total UQ, UQred was converted into the corresponding oxidized form (UQox) with ferric chloride. UQox was separated on a reversed-phase column and detected by its ultraviolet absorption (UV275n m). For the simultaneous assay of UQred and UQox, the n-hexane extract was injected onto a HPLC having a UV detector and an electrochemical detector (ECD) coupled in series. UQox was detected by its UV275n m, and UQred by ECD. The HPLC methods described here were applied satisfactorily to the determination of UQred, UQox, and total UQ in human and animal tissues.

Animals↗
PubMed1991

Pharmacokinetics of a novel dopamine DA1 receptor agonist, E4101, and an improvement of its bioavailability in dogs.

Pharmacokinetic parameters of E4101,(-)-(3R,4R)-3-(2-chloro-3-hydroxyphenyl)- 4-(3,4-dihydroxyphenyl)-pyrrolidine hydrochloride, in plasma were determined in conscious beagles. E4101 was extracted from plasma using a Bond Elute C18 cartridge and determined by HPLC connected to a reverse phase ODS column and an electrochemical detector. After an i.v. administration of 1 mg/kg, the plasma level of E4101 decreased according to the three compartment model. The half lives of the alpha, beta and gamma phases were 1.3, 8.1 and 27 min, respectively. The mean residence time (MRT) and total clearance were 14.5 min and 78 ml/min/kg, respectively. When 3 mg/kg of E4101 were administered orally under fasting, E4101 in plasma reached the peak (Cmax: 22.4 ng/ml) 45 min after administration. The bioavailability was 3.6%. When 3 mg/kg of E4101 in combination with 15 mg/kg of ascorbic acid was orally administered in capsule, the area under concentration curve (AUC) was increased 7-fold compared with that in a matched control study. A combination with 15 mg/kg of tartaric acid, citric acid, L-aspartic acid or L-cysteine also increased the AUC, byt they were less effective than ascorbic acid. When 10 mg/kg of E4101 in combination with 25 mg/kg of ascorbic acid as a sustained-release dosage form using oily somisolid matrix orally administered to non-fasted beagles, the duration of effective plasma concentration (2 ng/ml plasma in i.d. administration) was 8.4 +/- 1.0 hr (mean +/- SE, n-4). The Cmax was 16.2 +/- 2.5 ng/ml.

Animals↗
PubMed1989

Production and characterization of monoclonal antibodies against particulate guanylate cyclase in porcine kidney.

Three monoclonal antibodies (Ig G1 type) to particulate guanylate cyclase from porcine kidney cortex have been produced by fusing spleen cells from immunized BALB/c mouse with P3X63 myeloma cells. The antibodies were detected by their ability to bind immobilized antigen and by immunoprecipitation of enzyme activity. After subcloning by limiting dilution, hybridomas were injected intraperitoneally into mice to produce ascitic fluid. The antibodies recognized a 180,000 dalton protein in Lubrol-PX extract of porcine kidney cortex membrane, and when immobilized on Sepharose 4B, they co-precipitated both [125I]human atrial natriuretic peptide (ANP)-receptor complex and guanylate cyclase activity. The antibodies caused a greater increase in generation of cGMP than that of ANP.

Animals↗
PubMed1982

Digital processing methods for structural analysis of an electron micrograph.

Digital processing method using the scanning-densitometer with microcomputer provides a useful description system. It draws attention to the usefulness of local lattice averaging in clarifying images of periodic structures in crystals as well as crystalloids and para crystalline arrays in biological specimens. Digital processing of lattice images in crystal structures from the selected diffraction spots is successfully developed to the structural analysis of evaporated gold, graphitized carbon and human tooth enamel of hydroxyapatite. Furthermore, known microbody crystalloids in enzyme and icosahedral capsids in virus are confirmed and clarified by the procedure of selecting diffraction spectra transformed from the information taken with high resolution electron micrographs. The processed results can provide significant improvements in structural analysis of an electron micrograph.

Crystallography↗
PubMed1980

Maintenance of gonadotrophs in pituitary autografts under the kidney capsules of female rats given sex hormones or LRH.

Female Sprague-Dawley rats were hypophysectomized and the anterior pituitary gland was immediately placed under the kidney capsule. For 1 week after surgery, groups of pituitary autograft-bearing animals were treated with twice-daily injections of estradiol 17 beta (E), progesterone (P), estradiol 17 beta and progesterone (EP), or luteinizing hormone-releasing hormone (LRH). Within 2--4 hours following the last injection, the pituitary grafts were removed and placed into organ culture. They were maintained in culture with or without added LRH (10(-7) M) for 1 hour at 37 degrees C. The culture media were then frozen for later radioimmunoassay of FSH and LH. The tissues were kept in culture for an additional 24 hours, at which time they were fixed and prepared for immunocytochemistry or electron microscopy. Results showed that treatment of the animals with E, EP, or LRH enhanced the release of FSH and LH into the culture media, and that the release of these hormones was increased further by acute incubation with LRH. The ultrastructure of the gonadotrophs was well maintained by treating the animals with E or the combination of E and P or with LRH. Graft tissue from animals treated with LRH, which was incubated subsequently for 24 hours with LRH, showed the best maintenance of gonadotroph morphology. This experimental procedure should be useful for obtaining gonadotrophs for use in establishing gonadotroph cell lines.

Animals↗