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Biomedical subjects

H Ochiai

Publications and source records attributed to H Ochiai.

At least 73 records · Page 4Linked to original sources

An electron microscopic observation of the vesicular acetylcholine transporter-immunoreactive fibers in the rat dorsal raphe nucleus.

By using immunocytochemistry with an antibody directed against the vesicular acetylcholine transporter, many cholinergic neuronal processes were found to be immunopositive in the dorsal raphe nucleus. At the electron microscopic level, most of these processes were found to be axons. The immunopositive axon terminals made synapses on immunonegative dendrites and their spines whereas rare synapses were found between the immunopositive axon terminals and the immunonegative neuronal perikarya. Occasionally, the dendrites postsynaptic to an immunopositive axon terminal also received a synapse from an immunonegative axon terminal. The synapses made by the immunopositive axon terminals were usually symmetric and had a short active zone. Fewer immunostained dendrites were found, and they usually received asymmetric synapses from nonimmunostained axon terminals. The existence of cholinergic axon terminals and the synapses made by these terminals support the physiological data indicating that acetylcholine plays a role in the pain inhibition system in the dorsal raphe nucleus.

Animals↗

Synthesis of human erythropoietin in vivo in the oviduct of laying hens by localized in vivo gene transfer using electroporation.

In order to synthesize human erythropoietin protein in the oviduct of laying hens, localized in vivo gene transfer was attempted by using electroporation. In Experiment 1, transcriptional activities were compared by using four viral and cellular promoters, i.e., the 1.35-kbp long ovalbumin promoter, SV40 early promoter, Rous sarcoma virus long terminal repeat (RSV LTR), and the miw promoter, which is a hybrid of RSV LTR and chicken beta-actin promoter. These promoters were fused immediately upstream to the chloramphenicol acetyltransferase reporter gene. The results of chloramphenicol acetyltransferase activity showed that the miw promoter was the strongest, followed by SV40, RSV LTR, and the ovalbumin promoter in decreasing order. The intensity of the miw promoter was 250 times as high as that of the ovalbumin promoter. In Experiment 2, plasmid DNA containing the human erythropoietin gene, driven either by the ovalbumin promoter or the miw promoter, was transfected in vivo, and the production of human erythropoietin protein was detected by ELISA. The results indicated that the synthesis of human erythropoietin protein was attained in the chicken oviduct, and its concentration was higher when driven by the miw promoter than the ovalbumin promoter.

Animals↗

Increased cardiac angiotensin II receptors in angiotensinogen-deficient mice.

Two subtypes of angiotensin II (Ang II) receptors, type 1 (AT1-R) and type 2 (AT2-R), have been identified in the heart. However, little is known about the regulation of cardiac AT1-R and AT2-R by Ang II in vivo. Thus, we examined cardiac AT1-R and AT2-R in angiotensinogen-deficient (Atg-/-) mice that are hypotensive and lack circulating Ang II. Cardiac Ang II receptors (Ang II-R) were assessed by radioligand binding with 125I-[Sar1,Ile8]-Ang II in plasma membrane fractions. AT1-R and AT2-R were distinguished using their specific antagonists CV-11974 and PD123319, respectively. Total densities of Ang II-R and AT1-R density were significantly greater in the Atg-/- mice than Atg+/+ mice (31.1+/-2.8 versus 18.8+/-2.1, 28.7+/-3.0 versus 16.9+/-2.3 fmol/mg protein, P<.01, respectively), and AT2-R showed a slight but not significant increase in Atg-/- mice relative to Atg+/+ control animals. Kd values were not different between the two groups. In contrast to binding experiments, levels of Ang II type 1a receptor (AT1a-R) and AT2-R mRNA did not differ between Atg-/- and Atg+/+ mice. These results suggest that lack of Ang II may upregulate AT1-R through translational and/or posttranslational mechanisms in Atg-/- mice.

Angiotensin II↗

Endothelial nitric oxide synthase gene polymorphism and acute myocardial infarction.

Recently a point mutation of guanine to thymine at nucleotide position 1917 in the endothelial nitric oxide synthase (eNOS) gene has been reported to be associated with coronary artery spasm. In addition, a significant association of the 4a/b polymorphism in intron 4 of the eNOS gene with coronary artery disease has been reported. However, the implications of these polymorphisms with respect to acute myocardial infarction (AMI) remain to be established. We conducted a case-control study of 226 patients with AMI and 357 healthy gender- and age-matched control subjects. In the former group, coronary angiograms were evaluated according to angiographic criteria based on the number of diseased vessels (>/=75%) and the number of stenotic lesions (>/=50%). Homozygosity for the Glu-Asp298 polymorphism existed in 5 of 226 patients with AMI (2.2%) but not in any of the 357 control subjects (P=.0085). However, when we evaluated the coronary angiograms of 226 case patients, there was no difference in the number of diseased vessels or the number of stenotic lesions between the patients with this homozygote and those without it. By contrast, there was no evidence of a significant increase in the risk of AMI or the severity of coronary atherosclerosis among individuals with the a/a genotype of the eNOS4a/b polymorphism. Our results imply that patients who are homozygous for the Glu-Asp298 polymorphism may be genetically predisposed to AMI; however, this mutation apparently is not related to the severity of coronary atherosclerosis. Further studies are needed to confirm our results and characterize the molecular mechanisms by which eNOS is involved in susceptibility to AMI.

Case-Control Studies↗

Changes in iron and ferritin in anemic calves infected with Theileria sergenti.

Changes in iron and ferritin in calves infected with Theileria sergenti were investigated to elucidate iron metabolism in animals with extravascular hemolytic anemia. During severe anemia, serum iron was remarkably elevated while the total iron-binding capacity remained relatively unchanged or decreased slightly in the infected calves, resulting in elevated transferrin saturation. The serum ferritin concentration gradually increased with the progress of anemia. The erythrocyte ferritin content drastically increased when mean corpuscular volume was elevated. The concentration of non-heme iron and ferritin in the liver, spleen, and bone marrow of the infected calves was markedly higher than that in the respective tissues of the control animals. In particular, the liver of the anemic calves was found to contain 23 and 35 times as much non-heme iron and ferritin, respectively, as that of the non-anemic healthy cattle. The liver type (L) to heart type (H) subunit ratio of liver ferritin was significantly higher in the protozoa-infected than in the non-infected cattle. On the other hand, the L/H ratio of marrow ferritin was significantly reduced by the anemia. These results indicate that the anemic calves infected with T. sergenti apparently present symptoms of iron overload.

Anemia, Hemolytic↗

An isolated case of nephronophthisis: medullary cystic disease without typical onset.

A 16-year-old girl with renal failure was transferred to our hospital for an extensive renal examination. Computerized tomography and ultrasonography showed multiple small cysts throughout the medulla of both kidneys. Histological findings revealed tubular atrophy and dilatation and marked periglomerular fibrosis, all of which were compatible with nephronophthisis. Her development and growth were normal. A prior urinalysis, as well as her symptoms and family history were not helpful for making a diagnosis. As early diagnosis of nephronophthisis is difficult in some cases, more detailed screening is needed for children and adolescents.

Adolescent↗

Effect of bafilomycin A1 on the growth of Japanese encephalitis virus in Vero cells.

We studied the effect of bafilomycin A1 (Baf-A1), a novel and highly specific inhibitor for vacuolar-type proton (V-H+) pump, on the growth of Japanese Encephalitis virus (JEV) in Vero cells. Viral fluorescence microscopic study showed that Baf-A1 induced the complete disappearance of acidified compartments such as endosomes and lysosomes in Vero cells by the treatment with 0.1 microM Baf-A1 for 1 h at 37 degrees C. In proportion to the disappearance of acidified compartments, virus growth was inhibited when Baf-A1 was present from 1 h before infection to the end of incubation in a dose-dependent manner, or added within as early as 5 min after infection. Conversely, the virus growth was recovered in correlation with the reappearance of acidified compartments after removal of Baf-A1. These results suggest that a low pH condition, which is regulated by Baf-A1-sensitive V-H+ pumps, is essential for the early stage of JEV growth.

Acids↗

[Clinical usefulness of myocardial iodine-123-15-(p-iodophenyl)-3(R,S)-methyl-pentadecanoic acid distribution abnormality in patients with mitochondrial encephalomyopathy based on normal data file in bull's-eye polar map].

Visual interpretation of iodine-123-beta-15-(p-iodophenyl)-3(R,S)-methyl-pentadecanoic acid (123I-BMIPP) myocardial images cannot easily detect mild reduction in tracer uptake. Objective assessment of myocardial 123I-BMIPP maldistributions at rest was attempted using a bull's-eye map and its normal data file for detecting myocardial damage in patients with mitochondrial encephalomyopathy. Six patients, two with Kearns-Sayre syndrome and four with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), and 10 normal subjects were studied. Fractional myocardial uptake of 123I-BMIPP was also measured by dynamic static imaging to assess the global myocardial free fatty acid. These data were compared with the cardiothoracic ratio measured by chest radiography and left ventricular ejection fraction assessed by echocardiography. Abnormal cardiothoracic ratio and lower ejection fraction were detected in only one patient with Kearns-Sayre syndrome. Abnormal fractional myocardial uptake was detected in two patients (1.61%, 1.91%), whereas abnormal regional 123I-BMIPP uptake assessed by the bull's-eye map was detected in five patients (83%). All patients showed abnormal uptake in the anterior portion, and one showed progressive atrioventricular conduction abnormality and systolic dysfunction with extended 123I-BMIPP abnormal uptake. The results suggest that assessment based on the normal data file in a bull's-eye polar map is clinically useful for detection of myocardial damage in patients with mitochondrial encephalomyopathy.

Adult↗

[Pitfalls and their solution in MIDCAB].

Between July 1996 and September 1997, 11 patients underwent coronary artery bypass starting with left anterior small thoracotomy. All patients were evaluated with Electron Beam Computed Tomography (EBCT) preoperatively. Two patients out of 11 required conversion to sternotomy due to inadequate length of left internal mammary artery (LIMA) graft. One patient had a graft occlusion and underwent reoperation, when a graft avulsion was identified. Distance between LIMA and LAD by EBCT highly predicted needs of procedure modification to obtain additional length of graft. We demonstrated anterior pericardiopexy as an additional technique for protecting LIMA graft from abnormal tension. We conclude that it is crucial to harvest adequate length of LIMA in MIDCAB and anterior pericardiopexy may be useful to prevent a graft avulsion.

Aged↗

[Perioperative management of intracranial hemorrhage related to hematological disease].

Six patients with hematological disease complicated by intracranial hemorrhage were surgically treated in the last 2 years. In this study, in order to clarify indication for operation and perioperative management, 6 cases were classified into 2 groups. The details of each group were as follows: Group 1 was defined by the fact that the underlying hematological disease had not yet been controlled. (One case was ITP and the others were 2 AML cases). Group 2 was defined by the fact that the underlying hematological disease was well controlled. (One case was CML, one case was ATL and one case was ITP). A tendency to bleed was corrected in all patients of group 1 in the perioperative period. In the AML cases, prevention of infection was mandatory because both AML cases had been in remission, and no serious postoperative complication had occurred. The outcome of short term treatment was excellent in all but one case, in whom the recurrence of subdural hematoma caused death during the period 1 month after operation. On the other hand, no cases classified in group 2 needed specific hematological perioperative management and the short term treatment outcome was excellent. Since intracranial hemorrhage related to hematological disease has often been fatal, those patients were treated conservatively in most cases. However, from our analyses, we were able to emphasize that most intracranial hemorrhage related to hematological disease might be treated surgically and with good result, if the underlying hematological disease has entered the remission period.

Adult↗

[The significance of the cerebellar tonsillar herniation on the cause of sudden death of ruptured cerebral aneurysm].

We experienced a case with sudden unexpected death caused by rupture of an intracranial aneurysm, which was confirmed by autopsy. Depending on this case, we reported the significance of the cerebellar tonsillar herniation on the cause of sudden death of ruptured cerebral aneurysm. A 58-year-old man was admitted to us for treatment of subarachnoid hemorrhage (SAH). The CT scanning showed diffuse SAH in the whole cistern. Cerebral angiography on admission revealed an aneurysm at the bifurcation of the left middle cerebral artery in association with bleb like configuration. The aneurysmal neck was clipped on the day of admission. The postoperative course was uneventful. In the early morning of postoperative 23rd day, he was found being expired. To clarify the cause of death, an autopsy was done, disclosing diffuse SAH in association with tonsillar herniation more marked on the left. Thus, the distortion of the spinomedullary junction due to asymmetrical herniation was considered to be responsible for unexpected sudden death in this case. Examination of the major cerebral artery disclosed a ruptured anterior communicating artery aneurysm.

Aneurysm, Ruptured↗

Age and increased incidence of "forward bowing" of the iris in normal eyes.

PURPOSE AND METHODS: The incidence of forward bowing of the iris, width of the iridocorneal angle, and distance between the apex of the angle and the scleral spur were studied using high-frequency (50 MHz) ultrasound B-scan images in 90 normal eyes from Japanese volunteers. RESULTS: The incidence of forward bowing of the iris, defined as a 120-micron anterior shifting of the posterior profile of the iris, increased from 0% in subjects under 31 years old to 50% in subjects 71 years of age and older. The iridocorneal angle was significantly narrower and the distance between the apex of the angle and the scleral spur was significantly less in eyes with forward bowing. Of the risk factors (age, axial length, sex, and corneal curvature), age (p = 0.0005) was the most significant risk factor for forward bowing of the iris. CONCLUSION: Forward bowing of the iris is not uncommon in the elderly. Half of the Japanese subjects in this study 71 years of age and older had this condition, even though they had no optic nerve atrophy or elevation in intraocular pressure (IOP) at the time of examination.

Adult↗

Alteration of reticuloendothelial phagocytic function and tumor necrosis factor-alpha production after total hepatic ischemia.

BACKGROUND: This study was conducted to determine whether the duration of total hepatic ischemia influences reticuloendothelial phagocytic activity and tumor necrosis factor (TNF)-alpha production after reperfusion. METHODS: Male rats pretreated with either normal saline (NS group) or gadolinium chloride (7 mg/kg) for 2 days to inhibit Kupffer cell function (GC group) were subjected to 30, 60, or 90 min of total hepatic ischemia. RESULTS: The animals tolerated hepatic ischemia well for 30 and 60 min. Although the 7-day survival rate of the NS group decreased to 28% after 90 min of hepatic ischemia, that of the GC group improved significantly to 68% (P<0.01). In the NS group, plasma alanine transaminase and TNF-alpha levels after reperfusion increased with the length of hepatic ischemia. The phagocytic index (PI) after 60 min of reperfusion following 90 min of hepatic ischemia showed significant depression compared with the preischemic level and the value after 30 or 60 min of ischemia. The GC group had significantly lower plasma alanine transaminase and TNF-alpha levels as well as significantly less polymorphonuclear leukocyte infiltration in the liver compared with the NS group. The preischemic PI was significantly inhibited in the GC group when compared with that in the NS group, but PI in the GC group did not change significantly after reperfusion, irrespective of the ischemic time. CONCLUSIONS: This study demonstrated that warm ischemia of up to 60 min is tolerable for normal rat liver without a detrimental effect on phagocytic activity. Modulation of Kupffer cell function may have the potential to prevent reperfusion injury after hepatic ischemia, which may allow safe prolongation of the ischemic time.

Alanine Transaminase↗

Determination of simvastatin and its active metabolite in human plasma by column-switching high-performance liquid chromatography with fluorescence detection after derivatization with 1-bromoacetylpyrene.

By using a fluorescent derivatization and column-switching technique, a highly sensitive and selective high-performance liquid chromatographic (HPLC) method has been developed for the determination of simvastatin (I, beta-hydroxy-delta-lactone form) and its active hydrolyzed metabolite (II, beta,delta-dihydroxy acid form of I) in human plasma. A plasma sample spiked with internal standards was applied to a C8 solid-phase extraction column. Compounds I and II were separately extracted from plasma into two fractions. Compound I in one of the fractions was hydrolyzed to II. A fluorescent derivative was prepared by esterification of II with 1-bromoacetylpyrene in the presence of 18-crown-6 for both fractions. The pyrenacyl ester of II thus obtained was purified on a phenylboronic acid (PBA) solid-phase extraction column, and was measured by column-switching HPLC with fluorescence detection. The calibration curves for both I and II were linear in the concentration range of 0.1-10 ng/ml. The intra-day coefficients of variation were less than 11.0%, and the accuracies were between 91.7% and 117% within the concentration range for both analytes. The limits of quantification (LOQ) for both analytes were set to 0.1 ng/ml. This assay method has adequate sensitivity and selectivity to measure the concentrations of I and II in human plasma from clinical studies.

Anticholesteremic Agents↗

Stretch-induced MAP kinase activation in cardiomyocytes of angiotensinogen-deficient mice.

The renin-angiotensin system plays an important role in the hypertrophic responses in cardiac myocytes through the activation of signal transduction pathways and expression of oncogenes. In the present study, we examined mechanical stretch-induced activation of mitogen-activated protein kinases (MAP kinases) using cultured cardiac myocytes derived from neonatal angiotensinogen gene deficient mice (Agt-/-) and neonatal wild type mice (Agt+/+). Within 2 minutes of being added to cardiac myocytes, angiotensin II activated MAP kinases and the response was completely blocked by pretreatment of the cardiac myocytes with CV-11974, a selective antagonist of angiotensin II type 1 receptors. Interestingly, mechanical stretch resulted in significantly greater activation of MAP kinases in Agt-/- cardiac myocytes than in Agt+/+ cardiac myocytes. CV-11974 failed to suppress the stretch-induced activation of MAP kinases in Agt-/- cardiac myocytes while it inhibited the activation in Agt+/+ cardiac myocytes. BQ123, an endothelin type A receptor antagonist, had no effect on stretch-induced activation of MAP kinases in cardiac myocytes from either mouse strain. These results suggest that cardiac RAS is important for stretch-induced MAP kinase activation in Agt+/+ cardiac myocytes; however, angiotensin II is not indispensable for mechanical stretch-induced activation of MAP kinases in Agt-/- cardiac myocytes.

Angiotensin II↗

Ultrastructural localization of delta-1 opioid receptor in the dorsal raphe nucleus of the rat.

The ultrastructural localization of delta-1 opioid receptor in the rat dorsal raphe nucleus was studied by the preembedding avidin-biotin-peroxidase complex technique. With application of a low concentration of the first antiserum in incubation and control of short-time reaction to 3,3'-diaminobenzidine, the immunoreaction seemed to be faint at the light microscopic level. At the electron microscopic level, however, delta-1 opioid receptor immunoreaction products were found to be localized specifically on the postsynaptic membrane of dendrites, dense-cored vesicles, and the surface of the small, clear vesicles in axon terminals with strong immunoreactivity. Of the total 659 immunopositive profiles observed, up to 62.4% (411/659) were dendrites, whereas 33.8% (223/659) were axon terminals. The immunostained myelinated axons and perikarya were relatively rare, with the frequencies 0.8% (5/659) and 3.0% (20/659), respectively. Most of the immunopositive dendrites (338/411, 82.2%) were immunostained only at the postsynaptic membranes. Other immunoreactive dendrites showed their immunoreaction products also in some other contents besides the postsynaptic membranes (44/411, 10.7%) or only in those contents but not the postsynaptic membranes (25/411, 6.1%). Only four dendrites showed their immunoreactive results only at the membrane not related to synapse (4/267, 1.0%). No dendrite was found immunostained in all the contents. About half of the immunopositive axon terminals (125/223, 56.1%) were found to make synapse with nonimmunoreactive dendrites (76/223, 34.1%) or immunoreactive dendrites (49/223, 22.0%), while only one was found to make contact with immunoreactive perikarya. The present study showed that delta-1 opioid receptor in the dorsal raphe nucleus is mostly localized on postsynaptic membrane; the main function of the delta-1 receptor in the dorsal raphe nucleus is to receive signals from the opioid-containing axon terminals through synapses.

Animals↗

Inhibitory effect of ferulic acid on macrophage inflammatory protein-2 production in a murine macrophage cell line, RAW264.7.

We investigated time-related productions of certain cytokines in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, a murine macrophage cell line, by enzyme-linked immunosorbent assay. Macrophage inflammatory protein-2 (MIP-2) levels became detectable after 2 h and markedly increased over the first 8 h. Thereafter, this level remained at the same level between 10 and 16 h, and then increased again until 24 h, showing a tendency of biphasic pattern. Tumour necrosis factor (TNF)-alpha was detectable at 2 h and then increased sharply until 6 h at which it attained its peak. A low but recognizable level of interleukin (IL)-1alpha/beta was also detectable. When the inhibitory effect of ferulic acid (FA), an active component of the Rhizoma of Cimicifuga sp. used frequently as anti-inflammatory drug in Japanese Oriental medicines, was compared with that of dexamethasone (DX) on MIP-2 and TNF-alpha productions in response to LPS, both FA and DX could reduce the production of these cytokines in a dose-dependent manner. Concerning TNF-alpha, however, the inhibitory effect of FA was very weak compared with that of DX. In addition, FA as well as DX reduced MIP-2 production induced by TNF-alpha. These data suggest that MIP-2 might be induced by a direct effect of LPS and in part indirect one via initial induction of other cytokines such as TNF-alpha, leading a tendency of biphasic pattern. Comparing DX, FA is considered to be a novel and unique drug inhibiting MIP-2 production more selectively.

Animals↗