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Biomedical subjects

J Mori

Publications and source records attributed to J Mori.

At least 19 recordsLinked to original sources

Morphology and antigenicity of recombinant B19 parvovirus capsids expressed in transfected COS-7 cells.

COS-7 cells transfected with parvovirus B19-simian virus 40 (SV40) hybrid vectors have previously been shown to express B19 structural proteins. In this study the morphology and antigenicity of B19 proteins expressed in these cells were investigated. At 84 h after transfection, approximately 10% of the COS-7 cells expressed B19 antigen, and the yield was equivalent to 2 x 10(3) to 2 x 10(5) B19 particles/transfected cell. The B19 proteins self-assembled into capsids that were morphologically and antigenically similar to native B19 virions, and could substitute for native antigen in a B19 IgM assay. Recombinant capsids lacking the recently described 11 kDa protein also resembled native virions.

Animals

Fetal survival in ovariectomized uterus-incised rats.

This study was undertaken to investigate whether the actions of ovarian hormones not related to the uterine physical state are essential for fetal survival during late pregnancy in rats. Bilateral ovariectomy and longitudinal incision in one uterine horn, which enabled exposure of the fetuses without detaching the placentas, were performed in each rat on day 14, 15, 16, 17, or 18 of pregnancy, and fetal survival was examined on day 21 or 3 days after the operation. Although almost none of the fetuses were maintained in the intact uterine horn, the fetal survival rate in the incised uterine horn increased linearly as the operation was performed during the later stage of pregnancy. In control animals, which received sham ovariectomy and the unilateral uterine incision, fetal survival rates were high in the intact and incised uterine horns. In the rats given bilateral ovariectomy and unilateral uterine incision on day 14 of pregnancy, progesterone substitution maintained fetuses in the intact and incised uterine horns examined on day 17. The results suggested that the actions of ovarian progesterone not related to the uterine physical state become less essential for fetal survival as pregnancy progresses to the late stage in rats.

Animals

Polymerase chain reaction and sequencing for typing rhinovirus RNA.

Primers were designed and tested for their ability to distinguish rhinoviruses from enteroviruses. A primer set derived from the 5'-UTR/VP coding region junction was able to amplify all the rhinovirus serotypes tested. Enteroviruses were either not amplified by these primer pairs or produced a band of larger size that could easily be discriminated from the rhinovirus-specific product. In contrast, primers embedded in the 5'-UTR region alone were able to amplify both rhinovirus and enterovirus RNA. It is shown that rhinoviruses could be specifically typed by sequencing the amplicon derived from the 5'-UTR set. The sequences of the 5'-UTR region of ten previously unsequenced rhinoviruses were derived. The sequences obtained cluster into two groups: 1B, 41, 15, 30, 63, 31, 56, and 44; and 17, 69, and 70. Amplicons from serotypes 17,69, and 70 also group by sequence with the equivalent region of HRV14 from the genetic database, while the others group with 2 and 89.

Animals

Structure of genes that encode isozymes of aspartate aminotransferase in Panicum miliaceum L., a C4 plant.

The cytosolic and mitochondrial isozymes of aspartate aminotransferase (AspAT) function in the C4 photosynthetic cycle in NAD-malic enzyme-type C4 plants and are expressed at high levels in mesophyll cells and bundle sheath cells, respectively. We constructed a genomic library from Panicum miliaceum, a NAD-malic enzyme-type C4 plant, and cloned the genes for these isozymes. The sequence of the cloned gene for cytosolic AspAT spans 7800 bp and consists of 12 exons. The sequence of the cloned gene for mitochondrial AspAT spans 9000 bp and consists of 10 exons. The results of primer-extension analysis suggest that transcription may be initiated from multiple adjacent sites. Both genes have significant GC-rich regions around the site of initiation of transcription, and these regions showed no CpG suppression. The 5'- flanking regions of both genes include several short sequences similar to the regulatory elements found in other genes for components of the photosynthetic machinery. In particular, the cytosolic AspAT gene contains sequences similar to nuclear protein-binding sites in other mesophyll-expressed C4 photosynthetic genes and the mitochondrial AspAT gene contains elements for light-sensitive and constitutive expression of a bundle sheath-expressed gene. The results of Southern analysis indicated that there are at least two genes that encode each isozyme in the genome of P. miliaceum. A comparison of intron-insertion positions between AspAT genes of plants and animals revealed that several introns are located at identical positions. On the basis of a phylogenetic tree among AspATs and tyrosine aminotransferase, we have shown that the introns of aminotransferase genes antedate the divergence of eubacteria, archaebacteria, and eukaryotes.

Animals

Developmental changes in testicular luteinising hormone receptors and androgens in the dog.

The changes with age in the levels of testicular luteinising hormone (LH) receptors in dogs, were correlated with plasma androgen concentrations and the effects of stimulation with human chorionic gonadotrophin (hCG). The presence of high-affinity binding sites for LH was demonstrated in the testes of all the animals studied, and the apparent affinity of the LH receptors did not change significantly with age. The total number of LH receptor sites per testis increased continuously with age and the growth of the testis. The major rise in the number of LH receptors occurred between six and 10 months of age, and coincided with the phase of rapid testicular growth. The concentrations of androstenedione, testosterone and 5 alpha-dihydrotesterone in plasma increased gradually until six months of age, and then increased rapidly with the increase in the number of testicular LH receptors. An increase in the concentrations of androgens in plasma was observed in response to stimulation with hCG after the number of testicular LH receptors began to increase progressively. The increase in the sensitivity of the dogs' gonads to LH could be responsible for the marked increase in the secretion of androgens which occurs during puberty in the male dog.

Androgens

Secretion of progesterone and 20 alpha-dihydroprogesterone during pregnancy in goats.

The relationships between the secretion of progesterone and 20 alpha-dihydroprogesterone and the sites of their production in pregnant goats were investigated. The progesterone concentration in the peripheral plasma increased after the goats mated, and was high from day 10 to day 140 of pregnancy. The 20 alpha-dihydroprogesterone concentration increased gradually from day 0 to day 140 after mating. The progesterone concentration before parturition decreased rapidly on day 1 prepartum, but the 20 alpha-dihydroprogesterone concentration remained high until the day of parturition and then began to fall. The ratio of progesterone to 20 alpha-dihydroprogesterone decreased and fell to less than one on day 1 prepartum. The plasma concentrations of progesterone in ovarian vein and of 20 alpha-dihydroprogesterone in the ovarian vein, umbilical vein, and umbilical artery were higher than those in the jugular vein. The ratio of progesterone to 20 alpha-dihydroprogesterone in placental tissue was less than one. These results suggest that in pregnant goats the main production site is the ovary for progesterone and the ovary and feto-placental unit for 20 alpha-dihydroprogesterone, and 20 alpha-dihydroprogesterone may take help to regulate progesterone production within the ovary and the feto-placental unit.

20-alpha-Dihydroprogesterone

Secretion of progesterone and 20 alpha-dihydroprogesterone during the estrous cycle in goats.

The secretion of progesterone and 20 alpha-dihydroprogesterone (20 alpha-DHP) during the estrous cycle and the effects of prostaglandin F2 alpha (PGF2 alpha) and 20 alpha-DHP on the secretion of these two steroids during the midluteal phase in goats were studied. The mean progesterone concentration in the peripheral plasma was extremely low (< 0.01 ng/mL) on the day of estrus. The concentration increased to a maximum (7.80 ng/mL) on about day 10 after estrus, and decreased rapidly during the last 3 days of the cycle. The plasma concentration of 20 alpha-DHP also was low (0.86 ng/mL) on the day of estrus, but increased gradually after estrus and decreased gradually in the last 5 days of the cycle (to 3.22 ng/mL). The ratio of progesterone to 20 alpha-DHP was > 1 after day 2 of estrus, but decreased and fell to < 1 during the last 3 days of the cycle. An injection of 3 mg of PGF2 alpha decreased the secretion of progesterone within 30 min, but affected the secretion of 20 alpha-DHP little until 2 h. The ratio of progesterone to 20 alpha-DHP fell to < 1 after 24 h. Three of the five goats given such an injection came into estrus within 2 days, and the other two came into estrus within 3 days. An injection of 15 mg of 20 alpha-DHP did not affect progesterone secretion or the estrous cycle length. These results indicate that in the goat after estrus, there is much 20 alpha-DHP in the peripheral plasma. Progesterone may be catabolized to the biologically inactive steroid, 20 alpha-DHP, during luteolysis.

20-alpha-Dihydroprogesterone

Epidermal growth factor--induced implantation and decidualization in the rat.

Effects of epidermal growth factor (EGF) on blastocyst implantation and experimentally-induced decidualization were examined in the rat. Intraluminal injection of EGF into each uterine horn induced implantation in the ovariectomized progesterone-treated delayed implanting rat in a dose-dependent manner. This induction was inhibited by indomethacin, an inhibitor of prostaglandin synthesis. Intraluminal injection of EGF on day 5 of pseudopregnancy elicited a greater decidual response when compared to the vehicle-injected contralateral uterine horn. These results suggest that EGF may play important roles in the process of implantation and decidualization in the rat.

Animals

Effects of single and repeated administration of prostaglandin F2 alpha on secretion of testosterone by male rats.

The effects of single and repeated administration of prostaglandin (PG) F2 alpha on the secretion of testosterone and the effect of PGF2 alpha on the action of human chorionic gonadotropin (hCG) were investigated in adult male rats. The concentrations of plasma and testicular testosterone followed by a single injection of 100 micrograms of PGF2 alpha into each testis were similar to controls given a single injection of 0.05 ml of vehicle into one testis. When 1 or 10 IU of hCG were injected into the jugular vein simultaneously with the intratesticular injection of PGF2 alpha, concentrations of testosterone increased in response to dose; again, the effect was similar in controls. When 100 micrograms of PGF2 alpha was injected into each testis daily for 5 days, plasma testosterone concentration decreased below that in the controls, but the LH concentration was unchanged. When 1 or 10 IU of hCG were injected into the jugular vein the day after the fifth injection of PGF2 alpha, concentrations of plasma and testicular testosterone increased in response to dose; the effect was similar in controls. These results indicate that repeated stimulation by PGF2 alpha directly inhibits testosterone production in the rat testis without mediation by LH, and that PGF2 alpha does not affect the action of an LH-like substance.

Animals

Effect of oxytocin and indomethacin on the estrous cycle of goats.

The effect of oxytocin and indomethacin on luteal function during the estrous cycle in goats was investigated. Daily subcutaneous administration of oxytocin given on days 12-15 of the estrous cycle had no effect on length of the cycle or concentrations of progesterone and estradiol-17 beta. Oxytocin given on days 3-6 shortened the cycle and decreased concentrations of progesterone, but increased the estradiol-17 beta to levels similar to seen at estrus. These changes caused by oxytocin were prevented by indomethacin, a prostaglandin synthetase inhibitor, although indomethacin given alone did not affect luteal function. These results suggest that luteal function affected by prostaglandin in the early luteal phase of goats may be different from that in cattle.

Animals

Effect of nilvadipine on the development of neurological deficits in stroke-prone spontaneously hypertensive rats.

BACKGROUND AND PURPOSE: Several types of antihypertensive drugs have been reported to protect stroke-prone spontaneously hypertensive rats from stroke. However, the clinical relevance remains unclear. This study was performed to investigate the effect of nilvadipine, a calcium channel blocker, on the development of neurological deficits in stroke-prone spontaneously hypertensive rats. In addition, plasma levels of nilvadipine were measured to determine the clinical relevance. METHODS: Salt-loaded stroke-prone spontaneously hypertensive rats were orally administered nilvadipine mixed with a powder diet (0.01% and 0.03%, wt/wt). Non-salt-loaded rats were maintained on tap water. Chronological changes in neurological deficit scores and systolic blood pressure were recorded. After 6 weeks of medication, measurement of plasma levels of nilvadipine, serum biochemical analysis, and pathological observation of both the brain and the kidney were performed. RESULTS: In the salt-loaded control group, both severe hypertension and neurological deficit developed, and the final survival rate was 30%. Systolic blood pressure decreased significantly in the high-dose nilvadipine-treated group but not in the low-dose nilvadipine-treated group. However, the development of neurological deficit was almost completely inhibited in both nilvadipine-treated groups that had no deaths (P < .01). The mean plasma levels of nilvadipine in the low-dose group and in the high-dose group at the time of death were 0.21 ng/mL and 0.61 ng/mL, respectively. CONCLUSIONS: Nilvadipine inhibited the development of neurological deficit in stroke-prone spontaneously hypertensive rats at plasma concentrations lower than that in clinical use. Thus, nilvadipine might prevent cerebral vascular disorders at doses routinely used for essential hypertension.

Animals

Optimal regional anesthesia for circumcision.

Dorsal penile nerve block (DPNB) is a useful procedure for analgesia in circumcision. It has minor complications and a reported failure rate of from 4% to 6.7%. To evaluate the intraoperative value of additional anesthesia of the perineal nerves--a branch of the pudendal nerve--during circumcision, we conducted a prospective randomized double-blind study on 250 adults. The postoperative period was not studied. The subpubic space technique of DPNB was used. Patients received DPNB (Group I) or DPNB with an additional ventral injection (Group II) for perineal nerve analgesia. Seventeen patients (13.6%) from Group I suffered pain. Of these, nine (7.2%) had discomfort and mild pain but no supplemental analgesia was needed. In the remaining eight patients, however, it was necessary to add local analgesia. This represents a total failure rate of 6.4%. On the other hand, only six patients (4%) in Group II had a mild diffused discomfort with no need for additional local anesthesia (P < 0.01). The average operating time was 12.4 +/- 2.7 min (range 9-22 min) in Group I and 10.7 +/- 1.6 min (range 8-15 min) in Group II (P < 0.001). We think that perineal nerves play an important part in innervation of the penis and must be anesthetized during the penile block.

Adult

Vasorelaxant mechanism of the new vasodilator, FK409.

To define the vasorelaxation mechanism of FK409, we examined the effect of the compound on vascular tension and cyclic nucleotide levels in isolated rat thoracic aorta contracted with norepinephrine, and on activities of guanylate cyclase and cyclic GMP phosphodiesterase prepared from rat or rabbit thoracic aorta. FK409 (1 x 10(-9) to 1 x 10(-6) M), like nitroglycerin (1 x 10(-9) to 1 x 10(-6) M), produced a potent vasorelaxant effect associated with an increase in cyclic GMP content of the tissue. There was no change in cyclic AMP levels. The vasorelaxant effect of FK409 was independent of the integrity of the endothelium, and was unaffected by L-NG-monomethylarginine (0.1 mM) or oxyhemoglobin (1 microM). On the other hand, FK409 (3.2 x 10(-7) M) activated soluble guanylate cyclase, and the activating effect was completely inhibited by oxyhemoglobin (10 nM). Cyclic GMP phosphodiesterase was unaffected by FK409 (1 x 10(-7) to 1 x 10(-5) M). Furthermore, in rat aortic soluble fraction FK409 (3 mM) was found to liberate nitric oxide (NO) which was evaluated spectrophotometrically after diazotization of sulfanilic acid and coupling with N-(1-naphthyl)-ethylenediamine. The liberation occurred even in the absence of L-cysteine (5 mM), in contrast to the case with nitroglycerin (3 mM). These results suggest that the vasorelaxant effect of FK409 is associated with an increase in intracellular cyclic GMP, and that the cyclic GMP accumulation is due to activation of soluble guanylate cyclase. The enzyme activation is probably due to NO released from the compound molecule in the vascular smooth muscle cells.

3',5'-Cyclic-GMP Phosphodiesterases

Detection of HIV-1 in serum, using reverse transcription and the polymerase chain reaction (RT-PCR).

We have used a simple method for detecting HIV-1 in the serum of infected individuals using the polymerase chain reaction (PCR). This is useful if only serum, or other specimens which would not be expected to harbour proviral DNA, is available for diagnostic testing. Viral RNA present in serum is bound to silica particles in the presence of a high concentration of guanidinium thiocyanate (GuSCN) which denatures any proteins present, specifically ribonucleases. After washing the RNA/silica pellet, the RNA is eluted in water and reverse transcribed using random primers and Moloney murine leukaemia virus reverse transcriptase in the presence of a modified PCR buffer. The resultant cDNA is amplified using nested PCR and the products of amplification are detected by gel electrophoresis and ethidium bromide staining. The identity of bands on the gel is confirmed using a digoxigenin-labelled oligomer probe. The method is a general one applicable to amplification of any RNA species.

Base Sequence

Effect of FK409, a novel nitric oxide donor, on acute experimental myocardial ischemia.

The anti-ischemic heart effect of (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]- 5-nitro-3-hexenamide (FK409), a novel nitric oxide donor, was studied in dog and rat preparations in vivo and in vitro. In anesthetized dogs with partially occluded coronary artery that were subjected to atrial pacing at a constant blood pressure, FK409 (1-100 micrograms/kg, i.v.) suppressed the ST-segment elevation on epicardial electrocardiograms. Glyceryl trinitrate (GTN; 10, 32 micrograms/kg) or dipyridamole (1000 micrograms/kg) failed to suppress the ST-segment elevation, although continuous i.v. infusion of GTN (32, 100 micrograms/kg/min) was effective. FK409 also suppressed the ST-segment elevation induced by methacholine in anesthetized rats by both i.v. (10, 100 micrograms/kg) and intraduodenal (i.d., 100, 1000 micrograms/kg) injections, while GTN (100 micrograms/kg, i.v.; 1000 micrograms/kg, i.d.) was effective only by the i.v. route. FK409 (0.32 microgram/kg/min, i.v.) and GTN (10 micrograms/kg/min) increased the blood flows of the endomyocardium (ENDO) and the epicardium (EPI) and the flow ratio of ENDO/EPI in the ischemic zone in anesthetized dogs with occluded coronary artery. Furthermore, in isolated dog vascular preparations, FK409 (4.6 x 10(-10)-4.6 x 10(-7) M) had a greater vasorelaxing effect on the large coronary artery [2.0-2.5-mm outer diameter (od)] than on the small coronary artery (0.3-0.5-mm od) or the saphenous artery. The results suggest that FK409 protects against acute experimental myocardial ischemia through relaxation of the large conductive coronary artery, and may be a useful oral drug for the treatment of angina pectoris.

Animals

Multiple mediators and mechanisms are involved in the adaptive cytoprotection provided by certain mild irritants.

We investigated the participation of prostaglandins (PG) and nitric oxide (NO) in adaptive cytoprotection using 0.6 N HCl-induced gastric lesions in the rat stomach. Indomethacin reversed the protective effect of 0.2 N HCl more strongly than that of 0.35 N HCl, both of which markedly inhibited HCl ulcer. NG-Nitro-L-arginine (L-NNA) did not affect the protective effect afforded by either 0.2 N HCl or 0.35 N HCl. Combined pretreatment with indomethacin and L-NNA did not diminish the protective action induced by 0.35 N HCl, but almost completely abolished the indomethacin-resistant protection afforded by 0.1 N NaOH. Acid mild irritant increased the gastric fluid volume concentration-dependently, whereas alkaline mild irritant had little or no effect on the volume. These results suggest that: 1) The mediators involved in adaptive cytoprotection afforded by 0.1 N NaOH may be fully ascribed to PG and NO; 2) PG is a major mediator in the protection induced by 0.2 N HCl; 3) In the case of 0.35 N HCl, the mediators remain to be determined since increased gastric fluid volume could contribute to the protection through dilution. These findings thus may indicate that multiple mediators and mechanisms are implicated in adaptive cytoprotection.

Animals

Nerve growth factor (NGF) and its receptor in rat olfactory epithelium.

Nerve Growth Factor (NGF) is a bioactive protein that plays an important role in the genesis, differentiation, growth, maintenance and aging of the central and peripheral nervous systems. The purpose of this study was to explore the significance of NGF in the turnover, regeneration and maintenance of the olfactory epithelium. We tried to detect NGF and NGF-receptor in the olfactory epithelium of young and mature rats with an immunohistochemical technique. Immunoreactivity to NGF and NGF-receptor was detected in all specimens of olfactory epithelium. Immunoreactivity to NGF was positive in the olfactory nerve cells, especially in the cytoplasm and dendrites, in the basal cell layer, and on fibers piercing the olfactory epithelium. Immunoreactivity to NGF-receptor was positive in the basal cell layer and on fibers piercing the olfactory epithelium. These results suggest that NGF plays a role in the maintenance of olfactory nerve cells, and in the differentiation from the basal cell to the mature olfactory nerve cell, in both the young and the mature rat. In addition, the differences in the density of immunoreactive cells between different parts of the olfactory epithelium support the concept of an active and a quiescent zone in olfactory epithelium turnover.

Aging

[Clinical studies of S-1108, a new oral cephem, in pediatric patients].

Clinical studies of S-1108, a new oral cephem, in pediatric patients were conducted and results are summarized below. 1) Clinical effects of S-1108 against 18 cases of bacterial infections were excellent in 5 cases, good in 10 cases, fair in 1 case and poor in 2 cases, thus the clinical efficacy rates was 83.3%. 2) Bacteriological effects were evaluated in 16 strains. The elimination rate was 100%. 3) No adverse effects nor abnormal laboratory test results were observed in any of the cases.

Administration, Oral