PubMed HealthSearch

Biomedical subjects

H Ohtani

Publications and source records attributed to H Ohtani.

At least 19 recordsLinked to original sources

Photoreaction of N560 intermediate in the photocycle of bacteriorhodopsin.

Sophisticated measurements were made on the nanosecond time-resolved absorbance change of the purple membrane of Halobacterium halobium under cw background light irradiation (440-800 nm, 11-441 mW/cm2). A red-shifted transient species R660 (KN, Q) was found in alkaline conditions (pH > 9.3). Background light intensity effect shows that (i) R660 is photochemically formed from N560 intermediate which is accumulated under background light irradiation because of the elongated lifetime in alkaline suspension, and that (ii) the slow decaying M412 is not photochemically formed from N560 but from bR568.

Bacteriorhodopsins

Time-resolved fluorometry of purple membrane of Halobacterium halobium. O640 and an O-like red-shifted intermediate Q.

The photocycle of the light-adapted purple membrane was studied with a time-resolved fluorometry apparatus: fluorescence of the sample suspension (greater than 660 nm) was pumped with a 633-nm cw laser and the temporal change induced by a 532-nm pulsed laser was measured with a photon-counting-type transient recorder. The formation and the decay of the O640 intermediate were clearly observed in the pH region between 4.0 and 11.4. A photochemical cycle of N560 was apparently driven in alkaline suspension (pH greater than 9.3). An O-like fluorescent intermediate Q appears and decays with time constants of less than 0.1 ms and 1.7 +/- 0.2 ms, respectively.

Bacteriorhodopsins

Changes in plasma thrombomodulin antigen in rabbit developing endotoxin-induced disseminated intravascular coagulation and the effect of heparin.

Soluble thrombomodulin (TM) antigen level was 1.64 +/- 0.64 microgram/ml (n = 18, mean +/- S.D.) in plasma of normal male rabbits as measured by enzyme immunoassay, and the antigen consisted of subspecies of 94, 83 and 51 kd. When disseminated intravascular coagulation (DIC) was induced by intravenous infusion of endotoxin into rabbits, the TM antigen level in plasma was elevated to about 1.5 times of the control value, and an increase in the 83 kd subspecies as well as the appearance of new subspecies of 76 and 48 kd was observed concomitantly with disappearance of the 94 kd subspecies in plasma. Elevation of the antigen level and disappearance of the 94 kd subspecies caused by infusion of endotoxin were reduced by simultaneous infusion of heparin. Addition of leukocytes stimulated with endotoxin plus FMLP to cultured endothelial cells induced release of TM antigen to the medium accompanying cell injury as measured by 51Cr release, which was prevented by treatment with heparin. It was suggested that the increase in plasma TM antigen level in parallel with the generation of DIC reflected endothelial injury of rabbits, and that the elevation of TM antigen and the endothelial cell injury were prevented by heparin treatment.

Animals

A general transcription initiation factor, human transcription factor IID, overexpressed in human lung and breast carcinoma and rapidly induced with serum stimulation.

A general transcription factor IID which binds to the TATA box promoter element on RNA polymerase II genes regulates and initiates eukaryotic mRNA synthesis. A quantitative polymerase chain reaction procedure was developed and the human transcription factor IID (hTFIID) transcript was measured in normal human tissues, lung carcinomas, lung carcinoma cell lines, and breast carcinomas. In some normal tissues such as liver, fetal lung, and placenta, relatively low to moderate levels of hTFIID mRNA were detected. In contrast, hTFIID transcript was highly expressed in nearly all solid lung carcinomas and cell lines including both small cell lung cancer and non-small cell lung cancer. hTFIID mRNA was present to a greater extent in small cell lung cancer than non-small cell lung cancer in solid tumors and cell lines. In solid carcinomas of breast, overexpression of hTFIID was also detected. A serum induction study using a serum-starved small cell lung cancer cell line, Lu134BS, indicated hTFIID transcription to be rapidly induced at 15 min following stimulation and its response essentially similar to that of protooncogene, c-fos. These results indicate the involvement of the expression of the general transcription factor hTFIID in lung and breast carcinoma, such as being associated with poor differentiation and high mitotic activity.

Base Sequence

Light and electron microscopic immunolocalization of endothelial leucocyte adhesion molecule-1 in inflammatory bowel disease. Morphological evidence of active synthesis and secretion into vascular lumen.

Endothelial leucocyte adhesion molecule-1 (ELAM-1) is a rapidly inducible adhesion molecule for neutrophils in vascular endothelial cells. We investigated its immunohistochemical localization in 17 cases of inflammatory bowel disease. ELAM-1 was preferentially expressed in venules in actively inflamed mucosa and granulation tissue. Most capillaries were negative for ELAM-1. In areas with mild inflammation its expression diminished markedly and in normal mucosa of the colon and small intestine its expression was sparse. Electron microscopically, venules in active inflammation had swollen endothelial cells with well-developed rough endoplasmic reticulum. Immunoelectron microscopy revealed ELAM-1 localization along the luminal plasma membrane and in rough endoplasmic reticulum and round granules, findings suggestive of active production in endothelial cells. Furthermore, exocytosis of immunoreactive substance into the lumen was confirmed. Our study suggests that venules in actively inflamed area play an important role in eliciting and/or maintaining acute inflammatory processes by active permeation of neutrophils from the blood stream into the tissue, and that ELAM-1 may be a secretory protein as well as a transmembrane receptor protein.

Adult

Clinical evaluation of cardiac function by ambulatory ventricular scintigraphic monitoring (VEST): validation and study of the effects of nitroglycerin and nifedipine in patients with and without coronary artery disease.

Global left ventricular function and ECGs were continuously monitored by radionuclide ambulatory ventricular function monitoring (VEST) and validated against multigated blood pool analysis (MUGA) and left ventriculography in 26 subjects (study 1). Ejection fraction by VEST (Y) showed good correlation with Y = 5.5 +/- 0.79 X (r = 0.91), Y = 1.7 +/- 0.86 X' (r = 0.91), and Y = 11.6 + 0.68 X" (r = 0.82) to sitting and supine MUGA and left ventriculography, respectively. In study 2 left ventricular function and ECGs were evaluated at rest and during exercise without any drug (control), with nitroglycerin, and with nifedipine in 21 patients with coronary disease (group I) and six normal subjects (group II). In group I abnormal ejection fraction responses (exercise increase less than or equal to 6%) during the control exercise period were found in 15 patients (71%), ST segment abnormalities in seven (33%), and chest pain in four (18%). Control exercise increased end-diastolic volume (100 to 112 +/- 8%) and end-systolic volume (53 +/- 15% to 63 +/- 22%) and decreased the ejection fraction (47 +/- 15% to 43 +/- 21%). The ejection fraction during exercise increased after nitroglycerin (50 +/- 22%) or nifedipine (54 +/- 21%) (p less than 0.05). In group II the ejection fraction was unchanged between rest and exercise with or without nitroglycerin or nifedipine. Thus combined radionuclide and ECG monitoring by VEST could detect changes in left ventricular function at rest and during exercise over a prolonged period and demonstrated that nitroglycerin and nifedipine improved cardiac function in the ischemic setting with an increased ejection fraction in the upright position.

Coronary Disease

L-tryptophan alleviates fatty liver and modifies hepatic microsomal mixed function oxidase in laying hens.

1. Three experiments were conducted to study the effect of dietary L-tryptophan supplementation (250-1000 ppm) on lipid accumulation, an occurrence of hemorrhages and microsomal mixed function oxidase in the liver of laying hens. 2. Dietary L-tryptophan supplementation resulted in significant decreases in hepatic lipids, in particular triglyceride, and occurrence of hemorrhage in laying hens. 3. Hepatic lipid accumulation by estrogen injection in starved-refed growing chicks decreased as dietary tryptophan content increased. 4. Supplementation of L-tryptophan at 1000 mg/kg diet enhanced alanine aminotransferase activity in the hepatic tissue and at 500 mg/kg diet, increased cytochrome b5, a component of the mixed function oxidase, in the hepatic microsomes. 5. These results demonstrate that L-tryptophan alleviates fatty liver in laying hens and modifies microsomal mixed function oxidase in the liver.

Animals

Globotriaosyl ceramide glycolipid in seminoma: its clinicopathological importance in differentiation from testicular malignant lymphoma.

Glycolipids were biochemically extracted from 14 specimens of seminoma, 2 of testicular malignant lymphoma (both of which were difficult to differentiate from seminoma with a high mitotic index) and 4 of normal testicle. The pattern of their expression was compared. Marked accumulation of globotriaosyl ceramide was observed in seminoma but it was present in a small amount in testicular malignant lymphoma. Differentiation between seminoma and malignant lymphoma is sometimes difficult by histopathological findings but it is considered to be greatly facilitated by examination of the pattern of glycolipid expression.

Antigens, Differentiation, B-Lymphocyte

De novo expression of aromatase in gastric carcinoma. Light and electron microscopic immunohistochemical and immunoblot study.

We have performed immunohistochemical and immunochemical studies of steroidogenic enzymes involved in estrogen biosynthesis in 30 cases of gastric carcinoma in order to investigate possible in situ production of estradiol (E 2) in carcinoma cells. Positive incidence of immunoreactivity for E 2, testosterone (T), cholesterol side-chain cleavage enzyme (P-450 scc) and aromatase (P-450 arom) were 17/30 (56.7%), 11/30 (36.7%), 3/30 (10.0%) and 23/30 (76.7%), respectively on light microscopy. Estrogen receptor (ER) immunoreactivity was not observed in any of the 30 cases examined. Normal gastric mucosa was negative for P-450 arom and P-450 scc. Examination of serial sections revealed that immunoreactivity of E 2 and P-450 arom were located in the same cells of carcinomatous glands. Immunoelectron microscopy demonstrated that E 2 and P-450 arom were located along the membrane and cisternae of smooth endoplasmic reticulum (sER). Western blot analysis showed one major band of 55 kDa of P-450 arom in the gastric carcinoma tissues examined. Retrospective analysis of immunohistochemistry of E 2 in 108 cases of gastric carcinoma revealed that E 2 positive carcinoma cases were likely to demonstrate better survival rate than negative cases. These results above strongly suggest that E 2 is produced by de novo expressed aromatase in gastric carcinoma cells and is possibly involved in the biology of gastric carcinoma cells.

Aromatase

Glomeruloid structures as vascular reaction in human gastrointestinal carcinoma.

Malignant tumors induce angiogenesis and modulation of microvasculature. Based on histologic and immunohistochemical analysis of human surgical material, we describe here the occurrence of glomeruloid structures in gastrointestinal carcinomas, and compare them with the microvasculature in inflammatory granulation tissue. The glomeruloid structures were composed of clusters of mutually fused capillaries with prominent swelling of endothelial cells and pericytes. They were thought to be specific for glioblastoma of the brain. The glomeruloid structures were observed juxtaposed to carcinoma nests in one-third of gastric carcinoma of intestinal type and colorectal carcinoma in the area of invasive growth beyond the muscularis mucosae. They were not observed in gastric carcinoma of diffuse type, intramucosal carcinoma, or inflammatory granulation tissue. The glomeruloid structures can be regarded as an extreme example of endothelial hyperplastic changes observed in cancer stroma. Our results suggested that glomeruloid structures can occur in carcinomas as vascular reaction, a mechanism different from that in inflammatory granulation tissues.

Actins

[A case of Trichosporon beigelii peritonitis in CAPD].

Continuous ambulatory peritoneal dialysis (CAPD) was introduced to Japan ten years ago and was established as the treatment for end-stage renal disease along with HD. Although the incidence of peritonitis in CAPD has decreased by educating the patients and parents and the improvement of various devises of CAPD, peritonitis is still one of the major complications of CAPD. Fungus is a rare pathogen for peritonitis in CAPD, but it must be considered as a causative agent in cases of intractable peritonitis. This report describes the first case of Trichosporon beigelii (T. beigelii) peritonitis in CAPD in Japan. A nine year old boy with chronic renal failure due to bilateral vesicoureteral reflux was given CAPD treatment four years prior to admission. This patient had been admitted to our hospital frequently because of recurrent bacterial peritonitis. The peritonitis in CAPD was usually treated by changing the peritoneal fluid and antibiotic treatment. In this case T. beigelii was proved to be a pathogen of peritonitis by culture of CAPD fluid and also serum antibody titers. T. beigelii infection was successfully eradicated from the peritoneal cavity by administration of MCZ and by the removal of peritoneal catheter. The patient was switched from CAPD to HD. In the case of intractable peritonitis in CAPD, rare fungal pathogens such as T. beigelii must be considered as a causative agent.

Child

Human lung mast cells and pulmonary macrophages produce tumor necrosis factor-alpha in sensitized lung tissue after IgE receptor triggering.

Tumor necrosis factor (TNF) is considered to play a key role in the pathogenesis of allergic disorders. We examined TNF production in human lung fragments after IgE receptor triggering at mRNA and protein levels. IgE receptor triggering was performed by sensitizing lung fragments with monoclonal human IgE and then exposing them to anti-human IgE antibody. Cytotoxic activity against L929 cells appeared in the culture supernatant of lung fragments 2 h after IgE receptor triggering and increased for up to 4 h. This cytotoxic activity was completely neutralized by anti-human TNF antibody. Northern blot analysis demonstrated that 1.8-kb TNF mRNA transcripts in sensitized lung fragments were expressed as early as 1 h after IgE receptor triggering and continued up to 4 h. Immunohistochemical analysis revealed TNF localization in tissue mast cells, alveolar macrophages, tissue macrophages, and bronchial epithelial cells. Double staining with anti-TNF antibody and alcian blue clearly identified that lung mast cells are one of the TNF-positive cell types in the pulmonary tissue. With immunoelectron microscopy, TNF immunoreactivity was detected in the rough endoplasmic reticulum and the perinuclear spaces in tissue macrophages, and in the cytosol and the perinuclear spaces in bronchial epithelial cells. In addition, IgE was detected on the cell surface of mast cells, tissue macrophages, and alveolar macrophages. These results suggest that TNF is released from mast cells and pulmonary macrophages through IgE receptor triggering and may play a key role in the allergic reaction in human airway.

Animals

Identification of type I collagen-producing cells in human gastrointestinal carcinomas by non-radioactive in situ hybridization and immunoelectron microscopy.

Invasive growth of cancer cells induces desmoplastic reaction as one of the host reactions. It has been a matter of controversy whether stromal collagen is produced by cancer cells or stromal fibroblasts. In the present study, we investigated the cellular origin of Type I collagen in human gastrointestinal carcinomas by in situ hybridization and immunoelectron microscopy. In situ hybridization technique with digoxigenin-labeled RNA probes revealed that spindle-shaped fibroblasts in the stromal area were abundantly positive for transcripts of pro alpha 1(I) collagen in intestinal-type adenocarcinoma. Gland-forming carcinoma cells were negative. In diffuse-type carcinoma of the stomach, spindle-shaped or stellate fibroblasts were positive, whereas dissociated, oval carcinoma cells were negative. A precise determination of cell type was done by immunoelectron microscopy. Intracellular immunoreactivity for Type I collagen was observed in rough endoplasmic reticulum of fibroblasts (including myofibroblasts) in the stromal area. No definite reactivity was obtained in cancer cells by either in situ hybridization or immunoelectron microscopy. Our results indicated that stromal Type I collagen is produced by stromal fibroblasts, which are activated by cancer invasion.

Adenocarcinoma

Immunohistochemical identification of transforming growth factor-beta and its binding protein in human gastrointestinal carcinoma.

We attempted to clarify the tissue localization of TGF-beta and latent TGF-beta binding protein (LTBP) in human gastrointestinal carcinomas by immunohistochemistry. The immunoreactivity for TGF-beta was observed in both carcinoma cells and stromal cells, particularly in diffuse-type gastric carcinoma. The immunoreactivity for LTBP was observed only in stromal cells. These results suggest that both carcinoma cells and stromal cells may produce TGF-beta, and that only stromal cells may produce LTBP in gastrointestinal carcinoma.

Gastrointestinal Neoplasms

Heterogeneity of monoclonal immunoglobulins with antistreptolysin-O activity detected in the cases of essential monoclonal gammopathy and multiple myeloma.

Two different types of monoclonal human immunoglobulins (M-components) with antistreptolysin-O (ASO) activity were investigated. The M-component FM with essential monoclonal gammopathy revealed to have an ASO activity, demonstrated not only by streptolysin-O neutralizing assay according to Ranz-Randall's method, but also by passive agglutination assays and precipitation on agar. The ASO activity was shown to reside in the Feb. These findings suggest that the M-component FM have a true antibody activity. On the other hand, ASO activity of M-component TT with multiple myeloma was detected only by streptolysin-O neutralizing assay, but the passive agglutinating assays and precipitation on agar showed no positive results. It has not been fully confirmed if the M-component TT behaves as a true antibody activity. Heterogeneity of the M-components with ASO activity was discussed.

Aged

[Detection of cytomegalovirus in urine specimens from renal transplant recipients using polymerase chain reaction].

Human cytomegalovirus (CMV) in this study, a polymerase chain reaction (PCR) assay was developed to detect human cytomegalovirus (CMV) from urine specimens taken from renal transplant patients and one congenital CMV infection. Twenty eight urine specimens of 20 renal transplant patients were analyzed by the PCR. CMV infection was detected in 11 specimens (8 recipients). The incidence (40.0%) of PCR positive in the renal transplant patients was highest among the other conventional methods including fluorescent antibody study, anti-IgM EIA and complement fixation test. In two cases, CMV infection could be identified by PCR before antibody development. The PCR procedure is more sensitive and rapid than conventional methods for the diagnosis of CMV. Rapid diagnosis of CMV infection using PCR may be useful in clinical diagnosis and have therapeutic value.

Adult

Viable myocardium identified by reinjection thallium-201 imaging: comparison with regional wall motion and metabolic activity on FDG-PET.

Reinjection thallium-201 scans were performed in 68 patients with coronary artery disease after the routine stress-delayed scans for more accurate identification of new fill-in. Following the stress and 3 hour delayed thallium-201 SPECT scans, 40 MBq (1.1 mCi) was injected at rest, and 10 min later, the reinjection SPECT scan was obtained. To determine whether the reinjection method can aid in identifying ischemic but viable myocardium, the thallium-201 findings were compared with regional wall motion on radionuclide ventriculography in 61 patients and with metabolic activity on positron emission tomography (PET) using F-18 fluorodeoxyglucose (FDG) in 18 patients. The reinjection scan identified new fill-in which had not been shown on the stress-delayed scans in 6 of the 22 patients (27%) or in 29 of the 105 segments (28%). Regional wall motion was preserved more in the segments that exhibited new fill-in after reinjection (wall motion score = 1.64 +/- 1.29) than in those without new fill-in (score = 2.72 +/- 1.04) (p < 0.01). In the comparative study with FDG-PET, persistent FDG uptake was observed in all segments with new fill-in (20/20 segments: 100%); whereas, it was seen in only 7 of the 28 segments (25%) without new fill-in after reinjection (p < 0.05). We concluded that the segments having new fill-in after reinjection may represent ischemic but viable myocardium. Thus, the reinjection thallium-201 scan should be performed to identify ischemic myocardium which occasionally cannot be detected by the routine stress-delayed thallium-201 scans.

Adult

Comparison of technetium-99m-glucarate and thallium-201 for the identification of acute myocardial infarction in rats.

The scintigraphic identification of acute severe ischemic myocardial injury requires a marker that localizes rapidly and specifically in zones of damaged myocardium. Technetium-99m-glucarate, a six-carbon dicarboxylic acid, which behaves in vivo somewhat like fructose, was recently described as a marker of severe acute ischemic injury with necrosis. This study was performed to determine the interval between the onset of myocardial ischemia and initial uptake and the duration of a positive scan in experimental animals. Serial injections and images were recorded over 10 days following ligation of the left anterior descending coronary artery of the rat. The distribution of 99mTc-glucarate was compared to that of regional myocardial perfusion monitored with 201TI. The findings on radionuclide imaging were compared to histologic changes in the myocardium. Sequential pinhole images of both radionuclides were collected at 3 hr, 24 hr, 72 hr and 7-10 days following ligation. Ten rats had normal 201TI distributions, no uptake of glucarate and no evidence of infarction by TTC staining at autopsy. Twenty-one rats had either 201TI lesions or evidence of infarction at autopsy. In 17 of these rats, significant acute 99mTc-glucarate uptake was noted, decreasing at 24 hr, and was not seen at 72 hr or 7-10 days. The extent of perfusion abnormality was greatest at 3 hr in most animals; the lesion decreased in four (33%), increased in one (8%) and remained stable in the remainder. These data suggest that 99mTc-glucarate may be a useful marker of acute myocardial injury.

Animals