Biomedical subjects

H Osawa

Publications and source records attributed to H Osawa.

At least 127 records · Page 7Linked to original sources

PubMed1995

Soluble c-kit molecule in serum from healthy individuals and patients with haemopoietic disorders.

The proto-oncogene, c-kit, encodes a transmembrane tyrosine kinase receptor (KIT) and plays an important role in haemopoiesis. We have identified a 95 kD soluble form of KIT (S-KIT) in culture supernatant of human megakaryoblastic cell line, CMK. To study the physiological significance of S-KIT, we have established a sensitive sandwich ELISA system. Serum samples from healthy individuals contained detectable amounts of S-KIT. Next, we determined a total of 220 samples from 134 patients with haemopoietic disorders. A considerable number of patients with acute myeloid leukaemia (AML), especially those with more immature phenotypes (M0, M1 or M2) had elevated levels of serum S-KIT. Those levels decreased to the normal range after effective chemotherapy. In chronic myeloid leukaemia, patients with myeloid blastic crisis showed markedly elevated levels of serum S-KIT. In contrast, S-KIT levels decreased in cases with either acute or chronic lymphoid leukaemia. There was a tendency for patients with severe aplastic anaemia to show decreased levels, but it was not significant. In myelodysplastic syndrome, S-KIT levels appeared to vary by subsets, with higher concentration in more advanced forms of the disease. Although the functional role of S-KIT is not yet elucidated, these results suggest that the serum S-KIT levels may reflect the pathological states of various haematological disorders.

Acute Disease↗

PubMed1995

Regulation of hexokinase II and glycogen synthase mRNA, protein, and activity in human muscle.

Insulin regulates the activity of key enzymes of glucose metabolism in skeletal muscle by altering transcription or translation or by producing activity-altering modifications of preexisting enzyme molecules. Because of the small size of percutaneous muscle biopsies, these phenomena have been difficult to study in humans. This study was performed to determine how physiological hyperinsulinemia regulates the activities of hexokinase (HK), glycogen synthase (GS), and GLUT-4 in human skeletal muscle in vivo. We determined mRNA abundance, protein content, and activities for these proteins in muscle biopsies before and after a hyperinsulinemic clamp in normal subjects. HK I, HK II, GS, and GLUT-4 were expressed in muscle. HK II accounted for 80% of total HK activity and was increased by insulin from a basal value of 2.11 +/- 0.26 to 3.35 +/- 0.47 pmol.min-1.mg protein-1 (P < 0.05); HK I activity was unaffected. Insulin increased GS activity from 3.85 +/- 0.82 to 6.06 +/- 0.49 nmol.min-1.mg-1 (P < 0.01). HK II mRNA was increased 3.3 +/- 1.3-fold (P < 0.05) by insulin infusion. HK I, GS, and GLUT-4 mRNA and protein were unaffected. Because insulin infusion increased HK II but not GS mRNA, we conclude that HK II and GS may be regulated by insulin by different mechanisms in human skeletal muscle.

Adult↗

PubMed1995

Hepatitis C virus infection and membranoproliferative glomerulonephritis in Japan.

The prevalence of hepatitis C virus (HCV) infection was determined in 146 adult patients with various types of glomerulonephritis and renal diseases monitored between 1990 and 1993. Serum HCV antibody (HCV Ab) was evaluated, and positive cases were tested for HCV RNA by polymerase chain reaction. HCV infection was present in 1 (1.7%) of 58 cases of immunoglobulin A nephropathy, 0 (0%) of 14 cases of lupus nephritis, 0 (0%) of 12 cases of minimal change nephrosis, and 0 (0%) of 28 cases of other renal diseases, which is similar to the 2% prevalence observed in healthy blood donors in Japan. In contrast, HCV Ab was observed in 2 (8.3%) of 24 cases of membranous nephropathy and 6 (60%) of 10 cases of membranoproliferative glomerulonephritis (MPGN) Type I. The prevalence of HCV infection in MPGN patients was significantly higher than the frequency of HCV infection observed in the other patients with renal diseases (P < 0.001). HCV RNA was present in all cases in which HCV Ab was present. The six patients with HCV-MPGN were similar to the four patients with idiopathic MPGN with respect to age, presence of nephrotic syndrome, and renal dysfunction, but had a higher incidence of liver dysfunction, cryoglobulinemia, rheumatoid factor, and hypocomplementemia (low C3). HCV infection is present in a large percentage of patients with MPGN in Japan and clinically may differ slightly from other cases of MPGN.

Adolescent↗

PubMed1995

Postcholecystectomy syndrome mimicking angina pectoris detected by the morphine provocation test.

A 66-year old woman had had intermittent anterior chest pain and upper abdominal pain for 15 years. Angina pectoris was diagnosed at the age of 51 years, as she had typical anginal pain that was relieved by nitroglycerine, although coronary arteriography was normal and the ergonovine provocative test was negative. She had undergone cholecystectomy at the age of 38 years. Her bile duct pressure increased markedly after morphine injection and severe pain with the aforementioned distribution was produced. Postcholecystectomy syndrome due to sphincter of Oddi spasm was diagnosed and her pain was relieved by endoscopic sphincterotomy.

Aged↗

PubMed1995

Regulation of hexokinase II gene transcription and glucose phosphorylation by catecholamines, cyclic AMP, and insulin.

The hexokinases, by converting glucose to glucose-6-phosphate, help maintain the downhill gradient that results in movement of glucose into cells through the facilitative glucose transporters. GLUT4 and hexokinase (HK) II are the major transporter and hexokinase isoforms in skeletal muscle, heart, and adipose tissue, wherein insulin promotes glucose utilization. To understand whether hormones influence the contribution of phosphorylation to cellular glucose utilization, we investigated the effects that catecholamines, cyclic AMP (cAMP), and insulin have on HKII gene expression in cells representative of muscle (L6 cells) and brown (BFC-1B cells) and white (3T3-F442A cells) adipose tissues. Isoproterenol or the cAMP analog 8-chlorophenylthio-cAMP selectively increase HKII gene transcription in L6 cells, as does insulin (Printz RL, Koch S, Potter LP, O'Doherty RM, Tiesinga JJ, Moritz S, Granner DK: Hexokinase II mRNA and gene structure, regulation by insulin, and evolution. J Biol Chem 268:5209-5219, 1993), and cause a concentration- and time-dependent increase of HKII mRNA in both muscle and fat cell lines without changing HKI mRNA. Isoproterenol and insulin also increase the rate of synthesis of HKII protein and increase glucose phosphorylation and glucose utilization in L6 cells.

3T3 Cells↗

PubMed1995

[Clinical usefulness of helical-scanning CT for the evaluation of arteriosclerotic carotid lesions].

We examined arteriosclerotic carotid lesions in 76 patients using helical scanning CT (HES-CT), and evaluated the clinical usefulness of this method. A high speed slip-ring X-ray CT system was used. Scanning of the neck was performed for a 30 second period following intravenous bolus injection of non-ionic contrast medium, while couch top movement was 2.0 mm/sec. Multiplannar reconstruction images (MPR-image) and 3-dimensional surface images (3D-image) were reconstructed from the continuous raw data. MPR-images offered axial, coronal and sagittal images in which the lesion could be seen from any direction, and 3D-images that could be freely rotated were obtained by using a track ball and monitor. Eighteen cases were also evaluated by conventional angiography. Excellent HES-CT images were obtained in 73 cases, showing occlusion in 13, stenosis in 34, plaques without calcification in 15 and plaques with calcification in 74 vessels. A good correlation was obtained between HES-CT and angiogram in most cases, and in 6 cases, HES-CT was superior in the detection of stenosis, because it enabled us to observe the lesion from various directions. These results suggested that HES-CT was a minimally invasive, useful diagnostic method for the evaluation of arteriosclerotic carotid disease.

Arteriosclerosis↗

PubMed1995

TGF-beta upregulates interleukin 6 production by rat glomerular epithelial cells in vitro.

BACKGROUND: Glomerular epithelial cells (GECs) play an important role in maintaining normal glomerular permselectivity in vivo. Recent in-vitro studies have suggested that GECs are able to secrete substances which may modulate glomerular injury. Interleukin 6 (IL-6) has been shown to be a potent mediator of glomerular injury. It is also known that IL-6 could be produced by various cells. METHODS: IL-6 production by rat GECs in culture was examined in this study. IL-6 bioactivity in conditioned medium collected from cultured GECs (GEC-CM) was measured using IL-6 dependent murine hybridoma cell line, namely B9 cells. IL-6 gene expression by GECs was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). Effects of recombinant IL-6 on the proliferation of GECs and type IV collagen secretion by GECs were evaluated to examine the possible role of GECs derived IL-6. RESULTS: GEC-CM stimulated B9 cells growth in a dose dependent fashion. The mitogenic activity was inhibitable by anti-murine IL-6 antibody. De-novo synthesis of IL-6 was suggested by the demonstration of IL-6 mRNA by GECs using the RT-PCR. Secretion of IL-6 by GECs was increased by transforming growth factor beta but not by IL-1 beta. Recombinant murine IL-6 stimulated GECs growth and their type IV collagen secretion. CONCLUSIONS: These results indicate that rat GECs could produce IL-6 which may modulate glomerular inflammation and that IL-6 may function as an autocrine factor for GECs.

Animals↗

PubMed1994

Evaluation of regional cerebral blood flow changes on perifocal brain tissue SPECT before and after removal of arteriovenous malformations.

In fifteen cases of arteriovenous malformation (AVM), serial examinations of regional cerebral blood flow (rCBF) using single photon emission computed tomography (SPECT) and N-isopropyl-(123I)p-iodoamphetamine were performed. On SPECT images, the nidus was visualized as a focal rCBF defect in all cases preoperatively and seven of these cases had abnormal decreased perfusion areas in the tissues adjacent to the nidus. In five cases, the postoperative SPECT images on the day after surgery revealed an abnormal increased perfusion area adjacent to the nidus and in one case the increased perfusion was accompanied by a massive intracerebral haemorrhage detected by brain computed tomography (CT). In seven cases, postoperative SPECT images showed widespread abnormal decreased perfusion areas in the surrounding tissues and brain CT revealed either intracerebral haemorrhage or significant cerebral oedema. In the other three cases, no remarkable rCBF changes were found in comparison with the pre-operative study. There was a tendency for the preoperative abnormal decreased perfusion area adjacent to the nidus to correlate with the postoperative hyperperfusion and for the postoperative abnormal decreased perfusion area to reflect brain damage.

Adolescent↗

PubMed1994

Rat skeletal muscle hexokinase II mRNA and activity are increased by a single bout of acute exercise.

This study addresses the potential role of skeletal muscle hexokinase (HK) II in the regulation of glucose uptake and metabolism in vivo. Male rats undertook a single bout of treadmill exercise and were then killed immediately or after a predetermined recovery period. Three muscles [soleus (Sol), gastrocnemius/plantaris (Gc), and white vastus] were excised, and HK II mRNA, GLUT-4 mRNA, total HK (HK I and HK II) and heat-stable HK (predominantly HK I) activities were assessed. Three hours after the cessation of a single bout of exhaustive exercise, HK II mRNA was significantly increased in all three muscles. Ninety or thirty minutes of exercise, with a 3-h recovery, increased Gc HK II mRNA to the same extent as exhaustive exercise, but 15 min of exercise had no effect. Gc HK II mRNA continued to increase up to 8 h after the cessation of 90 min of exercise but returned to basal by 24 h postexercise. In contrast to HK II mRNA, Gc GLUT-4 mRNA was unchanged at 0, 3, 8, and 24 h after the cessation of 90 min of exercise. Total HK activity was significantly increased in Sol and Gc, 8 and 24 h after the cessation of 90 min of exercise. Heat-stable HK activity was unchanged in all three muscles. The increase in total HK activity, inferred to be an increase of HK II, may be important in the persistence of the postexercise increase in insulin action.

Animals↗

PubMed1993

Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells.

To clarify the phenotypes of various classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with CD34 and a newly developed mouse antihuman c-kit proto-oncogene product (KIT) monoclonal antibody (MoAb). We characterized three cell fractions in CD34+ cells that express KITlow and KIThigh cells in addition to KIT- cells. A clonogenic assay showed that most granulocyte-macrophage colony-forming cells (GM-CFC) were present in CD34+KIThigh populations, whereas erythroid burst-forming cells (BFU-E) were detected mainly in the CD34+KITlow population. CD34(+)-KIT- fraction contained a small number of BFU-E. Morphologic analysis showed that blast-like cells were more enriched in the CD34+KITlow fraction. KITlow cells contained CD34+CD38- cells that were considered to be very primitive progenitor cells, as determined by a replating assay. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell functions by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At week 2, more CFC recovered from the culture in the fraction initiated with a CD34+KIThigh population. However, more LTC-IC were present during weeks 5 to 9 in the CD34+KITlow population. These results indicate that primitive progenitors are more enriched in the KITlow population and that the KIThigh population contains many GM-committed progenitor cells. We also showed that anti-KIT MoAb inhibited the ability of CD34+ cells to generate CFC on the stromal layer in the LTC system. This suppressive effect was more evident in the generation of BFU-E by CD34+KITlow cells. Moreover, we confirmed that CD34+KIThigh cells emerged from CD34+KITlow cells during coculture with allogeneic stromal cells or from liquid culture in the presence of stem cell factor (SCF), interleukin-6, and erythropoietin. These results emphasize the pivotal role of the KIT and SCF interaction in hematopoiesis and indicate that KITlow cells are more primitive than KIThigh cells.

Animals↗

PubMed1993

A case of anti-nuclear antibody negative systemic lupus erythematosus associated with penile ulcer.

We report here an old male patient with anti-nuclear antibody (ANA) negative systemic lupus erythematosus (SLE) with active renal disease and penile ulcer. He revealed nephrotic syndrome, malar rash and oral ulcer. SLE was discussed, however both ANA and anti-DNA antibody were persistently negative. A penile ulcer was also observed. He died of acute respiratory distress. Autopsy findings including onion skin lesion in the spleen and haematoxylin body in the kidney resulted in the final diagnosis of SLE. To our knowledge, association of penile ulcer with SLE has not yet been reported. Therefore, the present case is thought to be extremely unusual.

Antibodies, Antinuclear↗

PubMed1993

A cancer-reactive human monoclonal antibody derived from a colonic cancer patient treated with local immunotherapy.

A human monoclonal antibody, YJ-37 (IgM) was generated through the fusion of human B lymphoblastoid cell line HO-323 with the regional lymph node lymphocytes from a colonic cancer patient who was treated with a local immunotherapy. This antibody was purified and conjugated with biotin, after which direct immunohistochemical staining was performed. The results revealed that YJ-37 selectively reacted with colonic cancer (7/19), gastric cancer (3/6), endometrial cancer (1/2) and colonic adenoma (7/13), but not with normal epithelia. Membrane immunofluorescence and FACS analysis also showed that YJ-37 bound to tumor cell surfaces. Furthermore, the chemical structure of the antigen defined by YJ-37 was analyzed by means of thin-layer chromatography immunostaining and ELISA. The results indicated that YJ-37 reacted with sialylated lacto-series carbohydrate chains, which have been reported to accumulate in cancer cells.

Adenoma↗

PubMed1993

[Immunoblastic lymphadenopathy (IBL)-like T cell lymphoma associated with granulocytopenia and autoimmune hemolytic anemia].

A 65 year-old woman was admitted in May 1990, because of fever, generalized lymphadenopathy and eruption. Laboratory examination showed granulocytopenia, polyclonal hypergammaglobulinemia, positive Coombs' test, positive cold antibody and positive anti-E erythrocyte antibody. 7 days after admission autoimmune hemolytic anemia developed. The appearance of the anti neutrophil leukocyte antibody was suspected because of the absence of the segmented neutrophils in marrow specimen. Histological findings of the biopsied lymph node and the marker study of the tumor cell disclosed IBL-like T cell lymphoma. The patient was treated with G-CSF and VEPA chemotherapy. Granulocytes increased and she showed marked symptomatic improvement with complete remission. Our case showed various clinical pictures with hemolytic anemia and granulocytopenia, which could be based on the autoimmune mechanisms.

Aged↗

PubMed1993

Expression of c-kit mRNA and protein during the differentiation of human hematopoietic progenitor cells.

To investigate how c-kit and c-kit ligand play a role in the function of hematopoietic stem cells, we determined the expression of c-kit in sorted human hematopoietic stem cells, CD34+CD33- cells and CD34+CD33+ cells. CD34+ cells constituted approximately 1% of the population of gated bone marrow cells and contained colony-forming cells. Two-color analysis by a fluorescence-activated cell sorter (FACS) revealed that about one-third to one-half of the total CD34+ cell population were positive for the CD33 antigen. To analyze the relative accumulation of c-kit mRNA in sorted cells, we used the reverse transcription-polymerase chain reaction (RT-PCR) method, followed by Southern blot analysis. There was a linear relationship between the amount of input RNA and products amplified in the range of 10(3) to 10(5) cells. Using this procedure, we carried out an analysis of c-kit mRNA expression in CD34+CD33-, CD34+CD33+, CD34-CD33+, and CD34-CD33- cells. Enhanced expression for c-kit mRNA was observed solely in CD34+CD33- cells. In contrast, flow cytometry shows that c-kit protein was expressed most abundantly in CD34+CD33+ cells. Colony-forming cells were generated on a human stromal cell layer for 5 weeks initiated with CD34+CD33- cells but not with CD34+CD33+ cells. During co-culture with stromal cells, CD34+CD33- cells differentiated into CD34+CD33+ cells. From these findings, it is concluded that CD34+CD33+ cells are direct progenies of CD34+CD33- cells. In this differentiation pathway, the expression of c-kit mRNA decreased and the c-kit protein increased.

Animals↗

PubMed1993

Deep breathing test (DBT) in predicting white coat hypertension.

We have evaluated the diagnostic value of deep breathing test (DBT) in detecting the patients with white coat hypertension on the outpatient basis. Thirty patients with untreated mild to moderate hypertension underwent 24-hour ambulatory blood pressure monitoring. Those who had a mean 24-hour blood pressure less than 135/80 mmHg were categorized as having white coat hypertension and the remainder were categorized as sustained hypertension. These two groups were compared with regard to the differences of office blood pressures before and after DBT (i.e., deep breathing 5 times for a minute in sitting position). Although the office systolic pressure significantly fell after DBT in both groups (p < 0.001 vs p < 0.05), there was no significant difference (p = 0.27) between the two groups. On the other hand, the office diastolic blood pressure significantly fell in the white coat hypertensives in contrast with no meaningful fall of it in the sustained hypertensives (p < 0.01 vs p = 0.66). At the cutoff level of -3%, -5% and -10% of the differences in office diastolic blood pressure before and after DBT, the sensitivity for the presence of white coat hypertension were 64.7%, 58.8% and 29.4%, respectively, and the specificity were 61.5%, 84.6% and 100%, respectively. These findings demonstrate that the deep breathing test is useful for identifying white coat hypertension in the outpatient clinic.

Female↗

PubMed1993

[Effective danazol therapy for a patient with Evans syndrome].

Danazol administered to a 66-year-old man with Evans syndrome, which was refractory to prednisolone therapy and pulse therapy with methylprednisolone. Danazol therapy produced an excellent and sustained improvement, increasing platelet counts and Hb concentrations. Recently, danazol therapy has been reported to be of benefit to severe idiopathic thrombocytopenic purpura (ITP) and autoimmune hemolytic anemia (AIHA). And this case marked a favorable effect also on Evans syndrome, which has both characters as ITP and as AIHA. The aim of this paper is to suggest that the administration of danazol has a positive effect on Evans syndrome and probably on other various thrombocytopenic diseases and syndromes, especially on refractory cases.

Aged↗

PubMed1992

Expression and function of adhesion molecules on human hematopoietic stem cells: CD34+ LFA-1- cells are more primitive than CD34+ LFA-1+ cells.

Advances in fluorescence-activated cell sorter technology have brought about multicolor analysis of cell phenotypes. To clarify the phenotypes of human hematopoietic stem cells (HSCs), we initially prepared novel antibodies against CD34 and labeled one of them (4A1) with allophycocyanin (APC). With this, we analyzed the phenotypes of CD34+ HSCs and showed that primitive HSCs or CD34+CD33- cells expressed adhesion molecules such as CD43, CD44, CD11a, CD11c, CD18, and leukocyte adhesion molecule (LAM-1). The more primitive hematopoietic cells or CD34+CD38- cells also expressed CD11a and CD18 with an incidence of 20% to 30%. To clarify the role of adhesion molecules in HSCs, we examined the colony forming capacity after long-term culture with allogeneic irradiated stromal layers. Among CD34+CD33- cells, CD18+ cells gave rise to colony-forming cells (CFCs) on stromal layers, but reached a maximum at week 2, after which the number of generated CFCs decreased. On the other hand, CD18- cells generated less CFCs than CD18+ cells at 2 to 3 weeks, but increased after 4 weeks of culture. When CD18 or CD11a antibody was added to a coculture system of CD34+CD33- cells with stromal layers, the number of generated CFCs decreased significantly compared with the no antibody control. Leukocyte function-associated antigen-1 (LFA-1) (CD11a/CD18) was expressed on some populations of hematopoietic cells and contributed to the proliferation by interacting with stromal cells. However, more primitive cells capable of reconstituting hematopoiesis did not express LFA-1. These data provide a rationale for the administration of anti-LFA-1 antibody after bone marrow transplantation for reducing the graft failure.

Antibodies, Monoclonal↗

PubMed1991

Combined anti-interleukin-2 receptor and low-dose cyclosporine therapy in experimental autoimmune uveoretinitis.

The effect of combination treatment with anti-interleukin-2 (IL-2)-receptor monoclonal antibody (ART18) and cyclosporine A (CsA) on the effector stage of experimental autoimmune uveoretinitis (EAU) was examined. Efferent-stage EAU was induced in Lewis rats by adoptive transfer of a T-helper cell line specific to retinal soluble antigen (SAg). Rats were treated with ART18 (0.5 mg/kg/day), low dose CsA (1.5 mg/kg/day), or a combination of both. The results were compared to groups treated with high dose CsA (10 mg/kg/day) and to sham-treated animals, with respect to clinical and histological EAU, lymphocyte proliferative responses to SAg, and the ability to transfer EAU to secondary recipients. Ten-day combination therapy with ART18 and low-dose CsA was more effective than high-dose CsA and almost completely suppressed EAU development. ART18 as sole therapy was partially effective, and was better than low dose CsA as sole therapy. Splenocytes of protected animals did not transfer EAU to secondary recipients, while splenocytes of sham-treated controls did, suggesting that the number of uveitogenic lymphocytes in the treated host was reduced by the therapy. In contrast, this therapy was completely ineffective against EAU induced by active immunization. The possible reasons for this discrepancy between the two respective models of EAU are discussed.

Animals↗