Biomedical subjects

T Hara

Publications and source records attributed to T Hara.

At least 649 records · Page 36Linked to original sources

PubMed1994

c-fos induction in the subnucleus caudalis following noxious mechanical stimulation of the oral mucous membrane.

c-fos expression in response to noxious mechanical stimulation of the peripheral receptive field was used to study the somatotopic representation of the oral mucous membrane and trigeminal cutaneous receptive fields in the trigeminal subnucleus caudalis and the cervical spinal dorsal horn. Fos-neurons (neurons with c-fos protein-like immunoreactivity) following stimulation of the cutaneous receptive field were found in laminae I and II of the first and second cervical segments of the spinal cord ipsilateral to the stimulation and exhibited a clear somatotopic segregation; i.e., the fos-neurons responding to receptive fields in the mandibular, maxillary, and ophthalmic divisions were arranged in a mediolateral sequence. On the other hand, fos-neurons responding to the oral mucous membrane were found in the similar laminae of the rostral pole of subnucleus caudalis bilaterally. They were all found in the dorsalmost part of the subnucleus without segregation; e.g., topographic distributions of fos-neurons resulting from the mandibular and maxillary gingival stimulations were indistinguishable. The central mechanism of trigeminal nociception is discussed with respect to the difference between intra- and extraoral receptive fields.

Animals↗

PubMed1994

Development of an ELISA assay for soluble CD35 (C3b/C4b receptor): high levels of soluble CD35 in LE-positive patients with hematological malignancies.

Malignant cells usually lack CD35 (complement receptor type 1, C3b/C4b receptor), a differentiation surface antigen. We measured soluble forms of CD35 in the plasma of normal subjects and patients with various malignant diseases. A microassay for the determination of CD35 was established based on a sandwich enzyme immunoassay using 2 monoclonal antibodies that recognize different epitopes. Soluble CD35 was not detected in any plasma samples from normal subjects or from patients with a variety of solid cancers: i.e., levels were below 20 ng/ml. On the other hand, 3 of 70 patients with hematological malignancies showed high levels of plasma CD35. The molecular mass of the soluble form was about 200 kDa, which is similar in size to membrane forms of CD35. Although the clinical conditions differed in these patients, they had high transaminase titers and detectable autoantibody. Complement titers (CH50) and the levels of membrane complement regulatory proteins were within the normal range in these patients. Although the mechanism by which it is produced remains unknown, soluble CD35 is present in significant amounts in association with immunological disorders secondary to hematological malignancies.

Enzyme-Linked Immunosorbent Assay↗

PubMed1994

Complications associated with endocapsular balloon implantation in rabbit eyes.

After phacoemulsifying the lens in six rabbit eyes through a 1.3 mm anterior capsule opening, we implanted a silicone balloon in the capsular bag and filled it with liquid silicone. The eyes were enucleated nine to 21 months postoperatively, embedded in paraffin, stained with toluidine blue, and examined by stereoscopic and light microscopy. We looked at the postoperative complications primarily. All balloons were securely fixated within the capsule. Surface wrinkles were observed in two balloons. The injected silicone remained transparent. None of the lens capsules was completely transparent, although all remained relatively clear. Retrocorneal membranes were produced in one eye in which the silicone leaked.

Accommodation, Ocular↗

PubMed1994

Mutations in transcript isoforms of the neurofibromatosis 2 gene in multiple human tumour types.

The neurofibromatosis 2 gene (NF2) has recently been isolated and predicted to encode a novel protein related to the moesin-ezrin-radixin family of cytoskeleton-associated proteins. Here we describe a novel isoform of the NF2 transcript that shows differential tissue expression and encodes a modified C terminus of the predicted protein. Mutations affecting both isoforms of the NF2 transcript were detected in multiple tumour types including melanoma and breast carcinoma. These findings provide evidence that alterations in the NF2 transcript occur not only in the hereditary brain neoplasms typically associated with NF2, but also as somatic mutations in their sporadic counterparts and in seemingly unrelated tumour types. The NF2 gene may thus constitute a tumour suppressor gene of more general importance in tumorigenesis.

Alternative Splicing↗

PubMed1994

Regulation of IL-3 receptor expression: evidence for a post-transcriptional mechanism that dominantly suppresses the expression of beta subunits.

The IL-3 receptor (IL-3R) is composed of alpha and beta subunits. Two homologous beta subunits of IL-3R are present in the mouse: AIC2A is the IL-3 specific beta subunit, and AIC2B is the common beta subunit shared by IL-3, IL-5 and granulocyte macrophage colony stimulating (GM-CSF) factor receptors. Both beta subunits form functionally indistinguishable high-affinity IL-3Rs with the same IL-3R specific alpha subunit (IL-3R alpha or SUT-1). Cell surface expression of the alpha and beta subunits of IL-3R was found to be diminished in an IL-3 non-responsive variant (MC/9.IL-4) derived from an IL-3-dependent mast cell line, MC/9. This IL-3R-defective phenotype was dominant based on cell fusion experiments. Moreover, regulatory mechanisms of the alpha and beta subunits are distinct since cell hybrids between MC/9.IL-4 and a CTLL-2 transfectant (CTLL/AS) expressing AIC2A and IL-3R alpha showed a significantly reduced expression of the AIC2A mRNA, while the IL-3R alpha expression was unchanged. Since transcription of both AIC2A and IL-3R alpha cDNAs in the CTLL/AS was driven by an artificial promoter, SR alpha, and nuclear run-off assays showed similar transcriptional rates of the AIC2A gene in both CTLL/AS and the cell hybrids between MC/9.IL-4 and CTLL/AS, the dominant suppression of the beta subunits is post-transcriptional and sequence-specific. A target sequence of the negative regulator must be present within 2756 bases of AIC2A mRNA, which is transcribed from the transfected cDNA in CTLL/AS cells. Similar dominant suppression of the beta subunit expression was also found in a B cell line WEHI231. As the negative regulator suppresses expression of the beta subunits of IL-3, IL-5 and GM-CSF receptors, it has the potential to eliminate all three high-affinity receptors simultaneously.

Animals↗

PubMed1994

In-vitro evidence for the regulation of 17,20-lyase activity by cytochrome b5 in adrenocortical adenomas from patients with Cushing's syndrome.

OBJECTIVE: The electron transfer system molecules, NADPH-cytochrome P450 reductase (Red) and cytochrome b5 (b5) are known to increase the relative activity of 17,20-lyase to 17 alpha-hydroxylase in vitro. Consistent with this hypothesis, we have reported recently that adrenocortical adenomas from patients with Cushing's syndrome that produced exceptionally high concentrations of androgens also contained more b5 mRNA as well as greater 17,20-lyase activity than adenomas that produced low concentrations of androgens. This finding was suggestive but inconclusive in linking b5 functionally to this difference in adenoma 17,20-lyase activity. In the present study, we have extended this finding by examining the effect of b5 on microsomal 17,20-lyase activity using an antibody against cytochrome b5. DESIGN: Biochemical quantitation of the content of b5 and Red activity in the microsomal fraction of the tumours and determination of 17,20-lyase activity in the microsomes in the presence or absence of an antibody against b5. PATIENTS: Seven patients with a clinical diagnosis of Cushing's syndrome secondary to benign adrenocortical adenoma were studied. MEASUREMENTS: The microsomal activities of 17 alpha-hydroxylase, 17,20-lyase and 3 beta-hydroxysteroid dehydrogenase were measured by in-vitro enzyme assay with thin layer chromatography. Microsomal Red activity was assayed by measuring NADPH-dependent cytochrome c reduction reaction. Cytochrome b5 concentration was determined by spectrophotometric analysis. RESULTS: An in-vitro enzyme assay of microsomal fractions from the adenoma showed that the 17,20-lyase activities of two adenomas that produced high concentrations of adrenal androgen were threefold greater than those of five other adenomas that produced low concentrations of androgens. Cytochrome b5 concentrations were greater in the two adenomas with high 17,20-lyase activity than in the other adenomas, while no significant difference in Red activity was observed among all the adenomas. The increased 17,20-lyase activity in the two adenomas was partially but significantly antagonized by an antibody against b5. CONCLUSIONS: These results suggest that differences in tissue b5 are functionally associated with differences in 17,20-lyase activity in adrenocortical adenomas in Cushing's syndrome, resulting in a dissociated secretion of cortisol and androgens in some patients.

Adenoma↗

PubMed1994

An experimental study on histopathological changes in the tissue covered with denture base without occlusal pressure.

In order to clarify the basic histopathological effect of a denture prosthesis on the denture supporting tissues, a new experimental appliance was developed. The denture base was designed to make contact with palatal mucosa of the experimental animal without any mechanical pressure and transmit no occlusal force to the mucosa. The objectives were: (i) non-pressure contact of denture base with palatal mucosa; and (ii) lack of physiological stimulation for palatal tissue by tongue and/or food. The results of the study revealed that covering the palatal mucosa with a denture base without any mechanical pressure and with the maintenance of a clean palate and denture base, did not cause any inflammatory change and bone resorption in the denture supporting tissues, except for a slight transient proliferative reaction in the epithelial ridges.

Animals↗

PubMed1994

Myxoid leiomyoma of the vulva mimicking aggressive angiomyxoma.

A case of vulvar leiomyoma with extensive myxoid change in a 40 year old female is described. The tumor had a unique connection with a non-degenerative leiomyoma that compressed the rectum and the bladder. Scattered smooth muscle cells in a loose myxoid stroma were immunoreactive for desmin. Fibroblast-like spindle cells were immunoreactive for vimentin but not for desmin. The initial, although incorrect, pathological diagnosis of the tumor was aggressive angiomyxoma based on the similarity in both clinical and pathological aspects with this more invasive tumor. Myxoid vulvar leiomyoma should also be differentiated from angiomyofibroblastoma. The key to the differential diagnosis is the presence of interlacing smooth muscle cells and an awareness of tendency toward myxoid change in vulvar leiomyomas.

Adult↗

PubMed1994

Severe aplastic anemia in a patient with primary immunodeficiency.

This paper reports the case of a 6 year old boy with primary immunodeficiency disease, whose marrow cells showed dyshematopoietic features, that was subsequently transformed to severe aplastic anemia. He was first diagnosed as having congenital immunodeficiency comprising deficiency of immunoglobulin A (IgA), IgG2 and IgG4, depressed mitogen responses, lymphopenia with inverted CD4/CD8 ratio and an increased proportion of T-cell receptor gamma/delta-bearing cells. Cytogenetic study of the peripheral blood showed a normal karyotype of 46,XY, but that of the marked hypoplastic marrow demonstrated one cell with monosomy 7 and another with trisomy 8 in the 20 cells examined.

Anemia, Aplastic↗

PubMed1994

A variant form of hypobetalipoproteinaemia associated with ataxia, hearing loss and retinitis pigmentosa.

A six-year-old Japanese boy had ataxia, mental retardation, peripheral neuropathy, proximal myopathy, hearing loss, retinitis pigmentosa and deficiencies in apolipoprotein AI, B, CII and CIII. His clinical features except for hearing loss resembled those of abetalipoproteinaemia or symptomatic hypobetalipoproteinaemia, but his apolipoprotein abnormalities were distinct from these disorders. He had apolipoprotein B-100 with a normal molecular weight. Although most of his neurological manifestations were compatible with those of vitamin E deficiency, their early onset and the presence of hearing loss was unusual for that condition. There has been slight deterioration of ataxia during two years follow-up despite high-dose vitamin E supplementation. Other abnormalities in lipid metabolism might be associated with the neurological damage in this case.

Ataxia↗

PubMed1994

Mechanism of abnormal production of adrenal androgens in patients with adrenocortical adenomas and carcinomas.

The production of adrenal androgens can be modulated by the activities of steroidogenic enzymes and by the electron transfer system, NADPH-cytochrome P450 reductase (Red) and cytochrome b5 (b5), both of which have been shown to increase 17,20-lyase activity in vitro. To clarify the mechanism of diminished secretion of adrenal androgens in patients with adrenocortical adenoma and Cushing's syndrome and of excess secretion in patients with adrenocortical carcinoma, we investigated the enzymatic activities of cytochrome P45017 alpha, 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), and Red as well as the content of b5 in five adenomas, three carcinomas, and two normal adrenal glands. An in vitro enzyme assay using a microsomal fraction of the tissues indicated that all the tumors had almost the same degree of 17 alpha-hydroxylase activities as the normal adrenals. However, the relative activity ratio of 17,20-lyase to 17 alpha-hydroxylase of the three adenomas was markedly diminished, and 3 beta-HSD activity was apparently lower in the three carcinomas. The messenger RNA concentrations of P45017 alpha were similar in all tumors, whereas those of 3 beta-HSD were markedly lower in the carcinomas than in other tissues. Both the content of b5 and the activity of Red were significantly lower in the adenomas. These results suggest that low concentrations of adrenal androgens in patients with adrenocortical adenomas are mainly due to low 17,20-lyase activity, which may be explained in part by a lower content of b5 and Red. In addition, high concentrations of adrenal androgens in patients with adrenocortical carcinoma are mainly due to the diminished activity of 3 beta-HSD.

3-Hydroxysteroid Dehydrogenases↗

PubMed1994

No mutation in cytochrome P450 side chain cleavage in a patient with congenital lipoid adrenal hyperplasia.

Molecular basis of lipoid adrenal hyperplasia (lipoid CAH) in a Japanese patient was investigated. A 46XY Japanese female patient was clinically diagnosed as having lipoid CAH based on her clinical history of adrenal crisis at birth and the low basal concentrations of cortisol, aldosterone, adrenal androgens and testosterone in serum. In vitro studies of testicular mitochondrial enzymes confirmed a specific impairment of cholesterol side chain cleavage (SCC) activity. However, in spite of the virtual reduction of SCC activity, the amounts of immunodetectable P450scc, adrenodoxin reductase, and adrenodoxin in testicular mitochondria were almost same as those of normal testis. Furthermore, the size of each protein was similar to that of normal testis. Enzymatic amplification of the complementary DNA encoding P450scc from the patient's testis RNA and its nucleotide analysis by direct sequencing revealed no mutation. These results indicate that defective P450scc is not the lesion in this patient, confirming a previous report showing no P450scc mutations in patients with lipoid CAH. The exact lesion causing lipoid CAH in this patient is currently unknown.

Adolescent↗

PubMed1994

Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues.

Ad4BP, a zinc finger DNA-binding protein, was identified as a transcription factor regulating steroidogenic P-450 genes in a cAMP-dependent manner. Immunochemical and immunohistochemical studies with steroidogenic tissues, adrenal, ovary, and testis, were performed using the antiserum to Ad4BP. Ad4BP was expressed to the same extent in the three zones of the adrenal cortex. Immunohistochemical examination of ovarian follicle and corpus luteum showed the expression of Ad4BP. The granulosa and thecal cells, the two distinct types of the steroidogenic cells in the follicle, gave Ad4BP signals, which were stronger than in the latter cells than in the former. Immunoblot analyses of mature and regressed corpora lutea indicated a parallel expression of Ad4BP and side-chain cleavage P-450, and both proteins significantly decreased in the regressed tissues. Leydig cells surrounding seminiferous tubules gave clear immunostaining signals for Ad4BP. ELP, a mammalian counterpart of Drosophila FTZ-F1 detected in EC cells, and are isoforms transcribed from the same gene. The Ad4BP and ELP forms recognize same nucleotide sequences. Reverse transcriptase-polymerase chain reaction with specific primers for ELP revealed that steroidogenic tissues contained ELP as well as Ad4BP. The effects of the two proteins on the transcription of the CYP11B gene were compared using the expression vectors of Ad4BP and ELP. ELP did not activate transcription and showed a weak inhibitor effect on the Ad4BP-dependent transactivation of the CYP11B gene promoter when transfected simultaneously. A gel shift analysis using in vitro synthesized Ad4BP and ELP revealed that the binding activity of ELP is significantly weaker than that of Ad4BP.

Adrenal Cortex↗

PubMed1994

EFFECTS OF DILUTED NATURAL WATER AND ALTERED IONIC ENVIRONMENTS ON GUSTATORY RESPONSES IN RAINBOW TROUT (ONCORHYNCHUS MYKISS)

1. The effects of adaptation to diluted natural water (NW) and various salt solutions on the gustatory responses recorded from the palatine nerve in rainbow trout (Oncorhynchus mykiss) were studied. 2. The magnitude of the response to 1 mmol l-1 l-proline (l-Pro) decreased when the perfusing NW was diluted with artificial fresh water (AFW) that maintained concentrations of major cations. AFW suppressed the responses to l-Pro by about 70 %. 3. The responses to 1 mmol l-1 l-Pro, 0.1 mmol l-1 quinineHCl (Q-HCl) and 10 nmol l-1 taurolithocholic acid (TLCA) were eliminated or reduced (to <10 %) by adapting the palate to distilled water (DW). The addition of 0.1100 mmol l-1 salts (NaCl, KCl, CaCl2, MgCl2) and choline chloride restored the gustatory responses to about 50 % of those in NW. The addition of salts to NW had no effect on the gustatory responses. 4. The gustatory responses to 5 % CO2 were similarly reduced when the palate was adapted to solutions that contained no NW (DW, AFW, 10 mmol l-1 NaCl in DW). However, the reduction was independent of salt concentration, suggesting a different transduction mechanism for CO2. 4. Tetrodotoxin (1 µmol l-1) had no effect on the gustatory responses to l-Pro. 5. We conclude that NW is required and that cations alone are not sufficient to support maximal gustatory responses. The results suggest that an unknown substance(s) contained in NW plays an essential role in gustatory reception and that permeation of cations through the apical membrane of gustatory cells is not involved in gustatory transduction in rainbow trout.

Journal Article↗

PubMed1994

Urinary excretion levels of hydroxylysine glycosides in osteoporotic patients.

The urinary excretion levels of hydroxylysine glycosides [Hyl-Gs; O-beta-D-Galactopyranosylhydroxylysine (GH) and 2-O-alpha-D-glucopyranosyl-O-beta-D-galactopyranosylhydroxylysine (GGH)], provide a new index of collagen metabolism. The determination of urinary hydroxylysine glycoside excretion levels was applied to 20 osteoporotic patients and 208 healthy control subjects (69 females and 139 males) in order to evaluate the conditions of bone collagen metabolism. Urinary Hyl-Gs were analyzed by the method of pre-column fluorescent derivatized HPLC. Compared to the age and sex-adjusted control, the urinary excretion levels of hydroxylysine glycosides, especially GH, in osteoporotic patients were significantly higher than in the age and sex-adjusted control, and the urinary GGH/GH ratio was lower. Sixteen of the 20 patients whose Hyl-Gs levels we followed exhibited significantly lower values after 200 d compared to the initial levels. These results suggest that clinical therapeutics affected the bone resorption and, therefore, demonstrated the usefulness of determining urinary excretion levels of Hyl-Gs for evaluating the conditions of bone metabolism.

Adult↗

PubMed1994

Determination of regional myocardial blood flow with 13N-ammonia positron emission tomography during low-grade exercise for evaluating coronary artery stenosis.

We measured regional myocardial blood flow (RMBF) by positron emission tomography (PET) with 13N-ammonia at rest and during low-grade exercise using a bicycle ergometer fixed at 25 watts for 6.5 min. The ischemic area with severe coronary artery stenosis was evaluated in terms of quantitative RMBF. The study was performed in 46 subjects consisting of 19 patients with old myocardial infarction, 20 with effort angina pectoris, and 7 normal individuals. RMBF was calculated using the radioactivity in myocardial tissue measured by PET and the radioactivity in arterial blood. The PET data were compared with coronary arteriographic findings and ECG findings. When the analysis was restricted to the anterior wall of the myocardium, it was found that exercise caused RMBF to increase by 44.9% (n = 28) in normal area, to increase by 23.3% (n = 37) in severely stenotic area (stenosis > or = 90%), and to decrease by 17.4% (n = 20) in infarcted area. When we considered only the infarcted area of the entire heart, exercise caused RMBF to decrease by 26.6% (n = 30) in the area of angiographically positive collaterals, and to increase by 0.8% (n = 31) in the area of negative collaterals. Thus, we were able to quantitate the degree of ischemia in myocardium that was associated with severe coronary artery stenosis. By application of low-grade exercise, RMBF in normal myocardium increased, while RMBF in myocardium perfused by collaterals decreased.

Adult↗

PubMed1994

Using H2(15)O and C15O in noninvasive pulmonary measurements.

Regional pulmonary blood flow (PBF), regional pulmonary extravascular water (PEW), and regional pulmonary blood volume (PBV) were measured quantitatively with positron emission tomography (PET) using H2(15)O and C15O in normal human subjects and patients before and after thoracotomy. The method was based on the Kety's model and a modification of the Mintun's method of measuring regional PBF with H2(15)O. The method consisted of intravenous injection of H2(15)O and dynamic PET scanning of the lung field and the right heart blood pool. Another scan following C15O inhalation was performed to visualize the blood pool, and the influence of the blood pool radioactivity to the H2(15)O image was subtracted. A mathematic model was applied to the data analysis, and the least-square fitting procedure was used with a computer for obtaining the optimal parameters. The mean values of the measured PBF, PEW, and PBV in two normal human subjects were 67.2 +/- 23 ml/min/100 ml, 17 +/- 5 ml/100 ml, and 19 +/- 6 ml/100 ml, respectively. The PBF, PEW, and PBV increased in the direction of gravity in the transverse cross-section image of the lung. Using this method, we studied the condition of patients who underwent lung surgery with thoracotomy. The PEW increased and the PBF decreased in the patients after surgery. This condition lasted for a few days, and the patients then recovered. The well-known phenomenon of pulmonary edema occurring after thoracotomy, which is usually recognized by symptoms and radiologically, was thus confirmed quantitatively by our method.

Aged↗

PubMed1994

Adsorption of salivary proteins to the surface of oral streptococcal cells.

Oral tissues, especially tooth surfaces, are covered with a layer of salivary proteins. Oral bacterial cells that adsorb to salivary components accumulated on the tooth surface are, as a rule, covered with the same components, especially proteins. Thus, it is possible that the salivary proteins covering the bacterial cells are related to the adhesion of bacteria to oral tissues. The aim of this study was to clarify the mechanisms of adsorption of salivary proteins to the surface of Streptococcus sanguis, S. mitis and S. salivarius using an adsorption assay with salivary proteins labeled with tritiated formaldehyde. The results showed that salivary proteins adsorbed more to S. salivarius than to S. mitis, and least to S. sanguis. It was evident that hydrophobic bonding was involved in the adsorption of salivary proteins to the bacterial cells tested. The amount of salivary proteins adsorbed to S. mitis and S. salivarius was decreased by the presence of phosphate, that to S. sanguis was increased by the presence of a divalent cation such as Ca2+, and that to all bacteria tested was inhibited in different ways by the presence of sugars. The amount of salivary proteins adsorbed to S. sanguis and S. salivarius was reduced effectively by pretreatment of the cells with trypsin, chymotrypsin and papain. In the case of S. mitis, the amount of adsorbed salivary proteins was decreased by pretreatment of the cells with chymotrypsin only, and was increased by pretreatment with lipase. These results indicate that there are different mechanisms of adsorption of salivary protein to the cell surfaces of oral streptococci.

Acetylglucosamine↗