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Biomedical subjects

Y Ishizuka

Publications and source records attributed to Y Ishizuka.

At least 19 recordsLinked to original sources

Cardiovascular and respiratory actions of pituitary adenylate cyclase-activating polypeptides.

Effects of pituitary adenylate cyclase-activating polypeptide (PACAP38) and PACAP27 on the cardiovascular and respiratory systems were examined and compared to those of vasoactive intestinal polypeptide (VIP) in anesthetized beagle dogs. Intravenous PACAP27 and PACAP38 produced a decrease in mean arterial blood pressure (MBP), and an increase in both femoral arterial blood flow (ABF) and in frequency of respiration (FR) with a dose-dependent relationship between 10 and 300 pmol/kg. PACAP27 produced a dose-dependent increase in heart rate (HR) between 10 and 300 pmol/kg while PACAP38 induced tachycardia which was not dose-dependent. Administration of 300 pmol/kg PACAP38 and PACAP27 produced extreme hypertension after transient hypotension. PACAP38 produced severe bradycardia after transient tachycardia. The cardiovascular actions of PACAP38 were persistent compared to those of PACAP27. Intravenous injection of 10-300 pmol/kg VIP brought about hypotension, tachycardia and an increase in ABF and FR with a dose-dependent relationship. VIP, at 2000 pmol/kg, did not produce the biphasic response obtained by a large dose of PACAP38. The present studies demonstrate that PACAP partially possesses VIP-like cardiovascular and respiratory actions and that the C-terminal 11 amino acid residues of PACAP38 are presumably responsible for a prolongation of its actions.

Adenylyl Cyclases

Survival rate of extremely low birthweight infants and its effect on the amendment of the Eugenic Protection Act in Japan.

Because of the increasing survival rate of extremely low birthweight infants in recent years, the viability limit in the Eugenic Protection Act in Japan was amended from 24 to 22 completed weeks of gestation. The Japan Pediatric Society's survey on the outcome of infants born in 1990 focused on infants born before 24 weeks gestation and less than 500 g. The survival rates of infants born in 23, 22 and before 22 weeks gestation overall at 511 hospitals throughout Japan were 43/118 (36%), 3/36 (8%) and 0/8 (0%), respectively. Of 16 infants, none weighing less than 400 g at birth survived but 16 (12%) of 50 infants between 400 and 499 g survived. Up-to-date statistical data is essential to the amendment of the concept of viability and subsequent ethical decision-making on extremely low birthweight infants.

Birth Weight

Maturation of an NH2-terminally extended thermostable alpha-amylase in Bacillus subtilis: a possible mechanism examined by in vitro experiments.

An artificially inserted extra peptide (21 amino acid peptide) between the B. subtilis alpha-amylase signal peptide and the mature thermostable alpha-amylase was completely cleaved by B. subtilis alkaline protease in vitro. The cleavage to form a mature enzyme was observed between pH 7.5 and 10, but not between pH 6.0 and 6.5, although a similar protease activity toward Azocall was observed between pH 6.0 and 7.5. To analyze the effects of pH on the cleavage, CD spectra at pH 6, 8, and 11 of the NH2-terminally extended thermostable alpha-amylase were analyzed and the results were compared with those of the mature form of the alpha-amylase. It is suggested that the cleavage of the NH2-terminally extended peptide is controlled by the secondary and tertiary structure of the precursor enzyme. Similar cleavage of different NH2-terminally extended peptides by the alkaline protease was also found in other hybrid thermostable alpha-amylases obtained.

Amino Acid Sequence

The prevalence of types I and II feline coronavirus infections in cats.

The types of feline coronaviruses that are prevalent throughout Japan were determined by competitive enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (MAb) to feline infectious peritonitis virus (FIPV) Type II and neutralizing test using Type II FIPV as challenge virus. A total of 1,079 cat serum samples were tested by indirect fluorescent antibody (IFA) assay for FIPV Type II antigen, all 42 sample from natural cases of FIP, 138 of 647 (21.3%) from cases with some chronic diseases and 57 of 390 (14.6%) from apparently non-diseased cases were positive. Of the 42 cases with FIP, 29 (69%) and 13 (31%) were found to have infection with FIPV Types I and II, respectively. Of the cases with chronic diseases, 111 (80.4%) were shown to have infection with FIPV or FECV Type I, while 14 (10.1%) with FIPV or FECV Type II. All of the 57 apparently non-diseased cases seemed to have been infected with FIPV or FECV Type I. These results indicated that feline coronavirus Type I is more high prevalent in Japan.

Animals

Expression of rat renin in mammalian cells and its purification.

Rat renin cDNA was transfected into COS-7 and Chinese hamster ovary (CHO) cells and expressed under the control of the Simian Virus 40 early promoter. Conditioned media of the transfected cells showed renin activity only after trypsin treatment, suggesting prorenin was secreted into the medium. From the trypsinized serum-free culture of the transfected CHO cells active renin was purified to homogeneity by a simple three-step procedure. The active renin had similar specific activity, molecular weight, Km, pH optimum, and isoelectric point compared to native renin. The amino-terminal sequence was the same as that deduced from the renin cDNA. This suggests that the recombinant rat renin is similar to kidney renin in many respects, and is easily obtained by the present procedures.

Animals

[Prolactin, ovarian and adrenal steroid hormone in relation to the vaginal opening in immature female rats].

The serum prolactin levels in human girls are thought to be increased gradually throughout puberty, but some authors have reported that prolactin decreases transitorily before menarche. Prolactin has not been shown to play a significant role on the initiation of human pubertal development. However, in immature female rats, it is thought that prolactin is one of the most important hormones during sexual maturation. In experiment I, the serum concentrations of prolactin, estradiol (E2) and dehydroepiandrosterone-sulfate (DHA-S) in immature female rats were measured. Prolactin increases gradually before vaginal opening and tends to decrease after vaginal opening, while E2 increases throughout sexual maturation. No significant changes were noticed in DHA-S levels. In experiment II, sulpiride, which raises serum prolactin level, was injected daily in to 4 groups of immature female rats starting from 14, 21, 28, and 35 days of age till the day before vaginal opening. Physiological saline solution was injected daily in to age-matched controls. On the day of vaginal opening, serum levels of prolactin, E2 and DHA-S were studied, weights of bodies and internal genital organs were measured, and corpora lutea of the ovaries were inspected. Vaginal opening was accelerated in the group of rats injected with sulpiride 5mg every morning from 14 days of age (p less than 0.05). The weights of their bodies and internal organs were less than those of the control rats (p less than 0.05). Percent of ovaries which contained corpora lutea were lower than the controls. Although their serum levels of prolactin tended to be suppressed, three hormone levels were not different from those of control rats on the day of vaginal opening. However, when mean prolactin levels were depicted on the day of vaginal opening in both groups, a peak was noticed at 37 days of life, indicating that the secretion pattern of this hormone has been programmed according to the day of life. There was a close relation between prolactin and DHA-S levels (p less than 0.05). No significant relation between prolactin and E2 levels was noticed. The possible role of prolactin and DHA-S on the regulation of sexual development is suggested. To our knowledge, this is the first demonstration of a significant relation between prolactin and DHA-S in immature female rats.

Age Factors

Induction of fetal malformations after treatment of mouse embryos with methylnitrosourea at the preimplantation stages.

The effects of methylnitrosourea (MNU) on the development of preimplantation mouse embryos were investigated in this study. ICR mice were treated intraperitoneally with single doses of 10, 20, and 30 mg MNU/kg body wt on day 0, 1, 2, or 3 of pregnancy. The uterine contents were examined on day 18 of pregnancy. The fetuses were examined for external and skeletal abnormalities. No significant differences were observed in the number of implantation sites between all the MNU-treated groups and controls. MNU treatment on day 2 or 3 of pregnancy caused dose-dependent significant increases in the incidence of abnormal fetuses over the control level, while treatment on day 0 or 1 failed to cause an increase of abnormalities. Cleft palate, exencephalus, and malformed vertebrae were the most common types of abnormalities. In the embryo transfer experiments, the frequency of fetal abnormalities induced when embryos were transferred from MNU-treated females to untreated pseudopregnant females was significantly higher than that induced when embryos were transferred from untreated females to MNU-treated or untreated pseudopregnant females. The results in the present study confirm and extend the previously proposed hypothesis that the direct effects of MNU on preimplantation embryos make a significant contribution to the induction of fetal malformations.

Animals

A study on the mechanism of antibody-dependent enhancement of feline infectious peritonitis virus infection in feline macrophages by monoclonal antibodies.

Enhancement of feline infectious peritonitis virus (FIPV) infection of feline macrophages was studied using monoclonal antibodies (MAbs) to the FIPV strain 79-1146. Adherent cells recovered from the feline lung and peritoneal cavity phagocytosed fixed red blood cells, and formed Fc-mediated rosettes. Enhancement of virus infection by MAb was investigated by inoculating alveolar macrophages with a mixtures of viral suspension and MAb, and examining the cells for intracellular viral antigen by the immunofluorescence assay and the amount of infectious virus in the supernatant fluid after incubation. The replication of FIPV in macrophages was enhanced by non-neutralizing MAbs recognizing peplomer protein (S) and transmembrane protein (M) of the virus. Even among the MAbs having the ability to neutralize FIPV strain 79-1146, some reversely enhanced virus infection when they were diluted. The enhancement was suppressed by pretreatment of the MAb with protein A. The enhancement was reduced by the use of F(ab')2 fragment of MAb. These results demonstrated antibody-dependent enhancement (ADE) of FIPV infection in macrophage. The replication of FIPV 79-1146 strain in macrophages from FIPV antibody-positive cats was more enhanced than in those from antibody-negative cats.

Animals

Dopamine receptors mediate cocaine-induced temperature responses in spontaneously hypertensive and Wistar-Kyoto rats.

The involvement of dopaminergic receptors in the responses of conscious, restrained spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats to cocaine was examined using antagonists selective for DA-1 (SCH 23390) or DA-2 (sulpiride) dopamine receptors. Following pretreatment with saline vehicle, SCH 23390 (50 mg/kg, SC), sulpiride (50 mg/kg, IP) or SCH 23390 and sulpiride, cocaine was infused (1.25 mg/kg.min, IV) until death. Cocaine caused an initial pressor and tachycardiac response, which was followed by a progressively developing secondary pressor response. Combined (DA-1 and DA-2) antagonist pretreatment abolished the initial tachycardic response to cocaine. Rectal temperature during cocaine infusion increased in 38.5% of vehicle-treated SHR (designated SHRH), but decreased in the remaining SHR (SHRL) and all vehicle-treated WKY. The time-to-onset of cocaine-induced convulsions (Tc) was reduced in vehicle-treated SHRH compared to vehicle-treated SHRL and WKY. Sulpiride elevated rectal temperature in response to cocaine in SHR and WKY but reduced Tc only in SHR. SCH 23390 abolished hyperthermic responses to cocaine in SHR without altering toxicity in SHR or WKY. Combined pretreatment virtually abolished temperature responses to cocaine in SHR and WKY, but increased the Tc only in WKY. Dopamine receptors, particularly the DA-1 subtype, are involved in cocaine-induced hyperthermia.

Animals

Multi-center study of seasonal affective disorders in Japan. A preliminary report.

A multi-center study on seasonal affective disorder (SAD) was conducted from the autumn of 1988 to the spring of 1989 with the cooperation of 16 facilities in Japan. Forty-six SAD patients were identified among 1104 respondents to our advertisements in mass media, or patients seen at the outpatient clinics. Essentially similar findings to other previous reports were obtained in terms of onset age of the first episode, duration of episode, high proportion of depression in first-degree relatives and atypical vegetative symptoms. However, a nearly equal sex ratio, together with a high proportion of unipolar depression, is characteristic of the present study. Increased appetite and carbohydrate craving were predominant only in female patients, whereas hypersomnia was prominent in both sexes. Effective response to light therapy was found in 17 SAD patients. However, a controlled study on a large number of patients is required to allow final conclusions on the efficacy of light therapy in Japanese SAD patients.

Adolescent

Successful treatment of disseminated cutaneous cytomegalic inclusion disease associated with Hodgkin's disease.

A case of disseminated cutaneous cytomegalic inclusion disease associated with Hodgkin's disease is described. The patient had a diffuse eruption of pruritic, erosive erythematous nodules. Histologically, many inclusion bodies were observed in perivascular areas of the skin lesions. Immunohistologically, the inclusion bodies positively stained with both anticytomegalovirus antibody and anti-factor VIII-related antibody. On electron microscopy many virus particles and dense bodies were found in the area where inclusion bodies were observed. Treatment with high-dose intravenous acyclovir and a large amount of immunoglobulin resulted in prompt healing of the skin lesions. Subsequently, the patient's Hodgkin's disease was well controlled on chemotherapy. The patient steadily improved without relapse of skin lesions 1 year after antiviral therapy was administered.

Aged

A case of eosinophilic granuloma in the temporal bone.

The following case reports a 4-year-old boy with a solitary eosinophilic granuloma in the right temporal bone. The patient complained of an inflammatory tumor in the right external ear canal. The histopathological diagnosis made based on the first specimen of the tumor in the right ear canal was foreign body granuloma. A polyp developed again in the right ear canal with postauricular swelling. Computerized tomography of the skull at that time showed an osteolytic defect in the right temporal bone filled with soft tissue. Diagnosis was established by finding of the presence of Birbeck granules in Langerhans histiocytes by electron microscopy.

Child, Preschool

Isolation and characterization of recombinant human prorenin in Chinese hamster ovary cells.

Recombinant human prorenin (rh-prorenin) was purified from supernatants of Chinese hamster ovary (CHO) cell line transfected with the cDNA for rh-prorenin by employing a simple two-step procedure which consisted of ammonium sulfate precipitation and immunoaffinity chromatography using a monoclonal antibody specific for the profragment of human prorenin. About 100-fold purification with 35% recovery was achieved after the two steps. Purified rh-prorenin migrated as a single protein band with apparent molecular weights of 46,000-47,000 and about 50,000 on SDS-PAGE and gel filtration (HPLC), respectively, although it consisted of multiple components (pI values, 5.6-6.4) that could be resolved by isoelectric focusing (IEF). The treatment of rh-prorenin with endo-beta-N-acetylglucosaminidase converted the rather broad protein band to a sharp band on SDS-PAGE and reduced the number of multiple pI peaks on IEF. Amino-terminal sequence analysis of both the purified rh-prorenin and rh-renin revealed Leu-Pro-Thr-Asp- and Leu-Thr-Leu-Gly-, respectively, which agreed with those predicted from the base sequences of their cDNA. These data suggested that microheterogeneity of rh-prorenin is due to the carbohydrate moiety, but not to the protein moiety. Purified rh-prorenin was almost inactive, but was cleaved at the carboxyl end of a dibasic pair Lys-2-Arg-1 by trypsin and converted to active renin. However, at the early stage during trypsin activation, new intermediate forms between rh-prorenin and rh-renin were formed, suggesting multiple activation steps of rh-prorenin in addition to the one step activation.

Amino Acid Sequence

A processing enzyme for prorenin in mouse submandibular gland. Purification and characterization.

Renin is produced from an inactive precursor, prorenin, through proteolytic cleavage at paired basic amino acid residues. In this study, an enzyme which specifically cleaves mouse Ren 2 prorenin at the paired basic residues has been purified from mouse submandibular gland by CM-Toyopearl chromatography, antipain-Sepharose chromatography, and isoelectric focusing. This enzyme, named prorenin converting enzyme, consists of two polypeptide chains of 17 and 10 kDa. The enzyme has an isoelectric point of 9.5-9.8, and its pH optimum is between 7.5 and 8.5. It specifically cleaves the peptide bond on the carboxyl side of the Arg at the Lys-Arg pair of mouse Ren 2 prorenin to yield mature renin but does not cleave mouse Ren 1 and human prorenins. Studies on the effects of inhibitors indicate that this enzyme is a serine protease that differs from the enzymes processing other prohormones at paired basic amino acid residues.

Amino Acid Sequence

Restraint alters temperature responses to cocaine in spontaneously hypertensive rats.

The body temperature responses to intraperitoneal (IP) or intravenous (IV) cocaine in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats were examined under restrained and freely moving conditions. Resting values for rectal temperature (RT), in ambient temperatures of 22-24 degrees C, were significantly (p less than 0.01) higher in SHR than in WKY rats, under both restrained (39.19 +/- 0.07 vs. 38.01 +/- 0.06 degrees C) and freely moving (39.39 +/- 0.08 vs. 38.08 +/- 0.06 degrees C) conditions. Resting RT did not differ between restrained and freely moving conditions within a strain. An heterogeneity of RT response to both IV and IP cocaine was expressed by restraint in SHR. The SHR could be divided into animals which demonstrated hyperthermia to cocaine (SHRH) and those in which RT fell (SHRL) . However, cocaine produced hyperthermia in all freely moving SHR, regardless of the route of administration. The effects of IV cocaine in restrained WKY rats were similar to those in freely moving SHR, whereas IP cocaine decreased RT in all restrained WKY rats. Under conditions of restraint, divergent RT responses to cocaine were demonstrated following IV and IP administration in WKY rats, but not in SHR. These results indicate that restraint stress can significantly modify the body temperature responses to acute cocaine administration in both SHR and WKY rats.

Animals