Biomedical subjects

H Tsumura

Publications and source records attributed to H Tsumura.

At least 73 records · Page 4Linked to original sources

PubMed1996

Granulocyte-colony stimulating factor and stem cell factor are the crucial factors in long-term culture of human primitive hematopoietic cells supported by a murine stromal cell line.

The findings that murine marrow stromal cell line MS-5 supported the proliferation of human lineage-negative (Lin-) CD34+CD38- bone marrow cells in long-term culture have been reported. In this study, we analyzed this proliferating activity of MS-5-conditioned medium (CM) on human primitive hematopoietic cells. When Lin-CD34+CD38- cells of normal human cord blood cells were co-cultured with MS-5, colony forming cells (CFCs) were maintained over 7 weeks in vitro. Prevention of contact between MS-5 and Lin-CD34+CD38- cells by using membrane filter (0.45 micron) was negligible for this activity. This indicated that the activity of MS-5 on human primitive hematopoietic cells is a soluble factor(s) secreted from MS-5, which is not induced by the contact between MS-5 and Lin-CD34+CD38- cells. We tried to purify this soluble activity. An active material with a molecular weight of about 150 kDa, determined by gel filtration chromatography, solely supported the growth of Lin-CD34+CD38- cells and Mo7e, a human megakaryocytic cell line. This activity not only reacted with anti-mouse stem cell factor (mSCF) antibody on Western blots, but it was also neutralized in the presence of anti-mSCF antibody. Another active material with a molecular weight of about 20-30 kDa synergized with mSCF to stimulate the growth of Lin-CD34+CD38- cells but failed to do so alone, although this synergy was inhibited in the presence of soluble mouse granulocyte-colony stimulating factor (mG-CSF) receptor, which is a chimeric protein consisting of the extracellular domain of mG-CSF receptor and the Fe region of human IgG1. In addition, the latter molecule supported the growth of the G-CSF dependent cell line FD/GR3, which is a murine myeloid leukemia cell line, FDC-P2, transfected with mG-CSF receptor cDNA. Adding of anti-mSCF antibody and soluble mG-CSF receptor to the culture completely abrogated the activity of MS-5-CM. Recombinant (r) mSCF and rmG-CSF had synergistic activity on the growth of Lin-CD34+CD38- cells. These results indicated that the activity on Lin-CD34+CD38- cells included in MS-5-CM is based upon the synergistic effects of mSCF and mG-CSF.

ADP-ribosyl Cyclase↗

PubMed1995

Genomic aberrations in human hepatoblastomas detected by 2-dimensional gel analysis.

Hepatoblastoma is a frequent embryonic tumor found in children. The mechanism of its formation and development remain unknown. We analyzed genomic DNA aberrations associated with three hepatoblastomas by means of restriction landmark genomic scanning (I. Hatada et al., Proc. Natl. Acad. Sci. USA, 88:9523-9527, 1991). About 2000 NotI restriction landmarks were monitored in each of the hepatoblastomas, which revealed that the intensity of eight spots decreased in the tumor genome, suggesting allelic loss. Three of these spots were identical to those that decreased in human hepatocellular carcinomas. These results suggested that common, early events are involved in the genesis of the two types of liver tumor.

Carcinoma, Hepatocellular↗

PubMed1995

Preparation of soluble recombinant T cell receptor alpha chain by using a calmodulin fusion expression system.

We have isolated a full length T cell receptor alpha chain (TCR alpha) cDNA derived from a bee venom phospholipase A2-specific mouse suppressor T cell hybridoma. A bacterial fusion expression system was constructed using rat calmodulin as a fusion partner for production of soluble TCR alpha. In this system, calmodulin-TCR alpha fusion protein was expressed at a high level in the soluble fraction of bacterial cell lysate, and could be purified by binding of calmodulin portion of the protein to phenyl-Sepharose. Using this system, fusion proteins containing a TCR alpha peptide corresponding to the complete extracellular region, V alpha-J alpha region or C alpha extracellular region were isolated. TCR alpha peptides were then released from the fusion proteins by digestion with thrombin which recognizes a linker sequence between calmodulin portion and TCR alpha segment. Polyclonal antibodies against constant region of TCR alpha chain (C alpha) were obtained by immunization of rabbits with the recombinant C alpha peptide. ELISA for TCR protein was established by using the polyclonal antibodies and the monoclonal antibody specific for C alpha region.

Amino Acid Sequence↗

PubMed1995

Comparison of bone-implant interface shear strength of hydroxyapatite-coated and alumina-coated metal implants.

We performed a transcortical push-out test to determine the effect of surface roughness of hydroxyapatite (HA)-coated implants on bone-implant shear strength in a canine model. Hydroxyapatite- and alumina-coated SUS316L with the same surface roughness (roughness average: Ra = 5 microns) and HA-coated Ti-6A1-4V (Ra = 8.4 microns), sintered HA (Ra = 0.9 micron), and dense alumina (Ra = 1.3 microns) were inserted into the dog's femur. The interface shear strength of the dense alumina was significantly lower than that of other implants at both 4 and 12 weeks after implantation. At 4 weeks after implantation, the interface shear strength of the alumina-coated SUS316L was significantly lower than that of other implants (P < .05) except the dense alumina, but at 12 weeks, there was no significant difference between the implant types except the dense alumina. This indicates that the surface roughness of the HA coating affects the enhancement of the bone-implant interface shear strength at the early period after implantation, and that a surface roughness of several micrometers does not influence the bond strength between bone and HA. A scanning electron microscopic study indicated that in almost all cases at 12 weeks, the failure site after push-out testing was the coating-substrate interface, not the coating-bone interface. Therefore, protection of the coating-substrate interface from direct shear loading is needed.

Alloys↗

PubMed1995

Hematopoietic stem cells found in lineage-positive subsets in the bone marrow of 5-fluorouracil-treated mice.

It is known that treatment of mice with 5-fluorouracil (5-FU, 150 mg/kg) confers radioprotection. To investigate this effect, we performed bone marrow transplantation (BMT) using C57BL/6-Ly5 congenic mice treated with 5-FU five days prior to experiments. The mononuclear cells (MNC) in 5-FU-treated bone marrow (BM) were 10 times more radioprotective than those in untreated BM. Moreover, the number of BM MNC expressing c-kit on their surface from 5-FU-treated mice was markedly decreased relative to those from untreated controls. These results showed that the surface characteristics of cells that contributed to this radio-protective effect differ from those of stem cells as reported recently. BM MNC of mice treated with 5-FU were separated on the basis of expression of the lineage-specific antigens (Lin), c-kit, and Ly6A/E. When injected into lethally irradiated mice, 1,000 Lin+ and Lin-c-kit+Ly6A/E+ cells showed radioprotective effects such that 100% and 60% survived, respectively. Flow cytometric analysis 165 days after BMT showed that 88.8% and 65.1% of peripheral blood (PB) in mice transplanted with Lin+ and Lin-c-kit+Ly6A/E+ was derived from donor mice, respectively. After six months, donor-derived Lin-c-kit+Ly6A/E+ cells which showed radioprotective effects on a secondary irradiated host were detected from mice transplanted with Lin+ cells from 5-FU-treated mice. Taken together, these findings demonstrated that stem cells expressing Lin+ present in the BM of mice treated with 5-FU other than Lin-c-kit+Ly6A/E+ cells and these Lin+ cells play an important role in the recovery of myeloablative mice.

Animals↗

PubMed1995

Severity and predicted outcome of postoperative Pseudomonas aeruginosa infections.

The severity and predicted outcome of postoperative Pseudomonas aeruginosa (P. aeruginosa) infections (PPAI) was evaluated using a severity scoring system based on a simplification and modification of the APACHE II system. A total of 86 patients in whom P. aeruginosa was isolated from various sources were examined. PPAI developed in 50 patients, resulting in an overall mortality rate of 24%. An increased severity score (SS) correlated with an increased risk of developing PPAI. Thus, PPAI developed in 33% of the patients with an SS of 0-1, in 66.7% of those with an SS of 2-3, and in 100% of those with an SS of 6 or higher. Moreover, the mortality rate of the patients with an initial score of 6 or higher was 50%. The mean (+/- SD) initial severity score was 5.4 +/- 2.9 for survivors and 2.9 +/- 2.6 for nonsurvivors (P < 0.01). In the patients who subsequently died, the SS remained high throughout the clinical course despite therapy, whereas in the survivors the SS decreased progressively, reflecting a favorable clinical course. These results suggest that our severity scoring system was useful for predicting outcome and monitoring the response of PPAI to therapy.

Adult↗

PubMed1995

HLA class II genotyping by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP) in systemic lupus erythematosus patients.

Recently, genotyping of HLA class II alleles by digestion of polymerase chain reaction (PCR) amplified polymorphic DNA region with restriction endonucleases (PCR-RFLP) has been reported and claimed to be a simple and reliable technique. We tested the use of PCR-RFLP for genotyping of DQA 1, DQB 1, DPB 1 and DRB 1 alleles of 40 normal individuals and 19 patients with systemic lupus erythematosus (SLE). In agreement with previous reports, we observed an association of SLE with the DRB 1*1501-DQA 1*0102-DQB 1*0602 haplotype and noted an unreported association with DPB 1*0501. Although these observations must be confirmed with larger sample sizes, we believe that the PCR-RFLP is simpler and more practical than the sequence-specific oligonucleotides (PCR-SSO) for genotyping and can be invaluable to the more widespread molecular determination of HLA-class II specificities, including in non-specialized laboratories.

Genotype↗

PubMed1995

Three-dimensional MRI reconstructions of musculoskeletal tumors. A preliminary evaluation of 2 cases.

We have developed a new method for three-dimensional (3D) reconstruction of malignant bone and soft-tissue tumors from MRI. Data from transverse images of T2-weighted images were transferred into a personal computer. The contours of tumors, major vessels, major nerves and bones were outlined, whereupon the 3D data were processed into a 3D display, using a solid model. The determination of tumor volume both before and after preoperative adjuvant therapy was quantified. The 3D images allowed a better understanding of the relationship between the tumor and the major vessels and nerves, thereby aiding in preoperative planning. The images also provided a quantitative, reproducible measurement of the effect of adjuvant therapy.

Adolescent↗

PubMed1994

Effect of surface roughness of hydroxyapatite-coated titanium on the bone-implant interface shear strength.

We have investigated the bone-implant interface shear strength of hydroxyapatite (HA)-coated Ti-6Al-4V (HA-coating A) (roughness average, Ra = 3.4 +/- 0.5 microns) and HA-coated Ti-6Al-4V with a rougher surface (HA-coating B) (Ra = 8.4 +/- 1.8 microns). There was no significant difference between HA-coating A and HA-coating B implants with respect to the bone-implant interface shear strength as determined in push-out tests using the transcortical model in adult dogs. The bone-implant interface shear strength of bead-coated porous Ti-6Al-4V was significantly greater than that of both HA-coating A and HA-coating B implants. The failure site, as determined by scanning electron microscopy, was the coating-substrate interface, not the coating-bone interface. This indicates a need to protect the HA coating from the direct shear forces. HA coating enhances early bone growth into the porous surface of the implant. Long-term fixation should depend on bone anchoring to this porous surface. Hydroxyapatite coatings must be developed which do not obstruct the pores of the surface of the implant.

Analysis of Variance↗

PubMed1994

A girl having congenital chloride diarrhea treated with spironolactone for seven years.

We report on a girl having congenital chloride diarrhea (CCD) who has been followed for 7 years and 6 months sequentially. Dilated intestinal loops, marked enlargement of the abdominal circumference of the fetus and hydramnios were noted by ultrasound examination at 31 weeks of gestation. After delivery by cesarean section for hydramnios, she excreted profuse watery yellow green stools with marked abdominal distension. At 4 months of age, hypochloremia, hyponatremia and a high concentration of chloride in the stool were identified. She was diagnosed as having CCD. Because it was difficult to administer a large volume of potassium chloride (KCl), and sodium chloride (NaCl), we decided to administer spironolactone. After administration of spironolactone, we could generate correct serum electrolytes using less amounts of KCl. At 7 years and 6 months of age, her body size was within normal limits and her intellectual, mental and physical development had been normal. In spite of normal serum electrolytes, blood pH and the presence of chloriduria, secondary hyperaldosteronism was noted. We consider that spironolactone may be useful to decrease the amount of KCl administration in the neonatal period, but frequent measurements of renin, angiotensin and aldosterone would be necessary for adequate control in CCD cases.

Child↗

PubMed1994

A novel hypothyroid 'growth-retarded' mouse derived from Snell's dwarf mouse.

This paper describes a novel mutant mouse that has been spontaneously derived from the Snell's dwarf (DW/J) mouse. It was named the 'growth-retarded mouse' because of a characteristic growth pause followed by the delayed onset of pubertal growth. The onset of the increase in pituitary GH content that normally occurs concomitant with pubertal growth was also delayed in the growth-retarded mice. The serum concentration of thyroxine was very low in these mice from the neonatal period through adulthood, and a supplement of tri-iodothyronine was effective in shortening the growth pause and commencing the suppressed pubertal growth. Histological and immunohistochemical studies revealed that the anterior pituitary gland of the growth-retarded mouse contains clustered unusual chromophobic cells which are not reactive to various antisera against anterior pituitary hormones and the gland becomes enlarged with age. Breeding data indicated that these characteristics of the mice show an autosomal recessive inheritance and the gene responsible was designated as 'grm'. Partial linkage analysis utilizing microsatellite polymorphism demonstrated that the grm gene does not identify with the lit or hyt genes. Based on comparison of the hormonal status and growth pattern between growth-retarded, dwarf and normal mice, we have suggested the existence of a mutual interaction, possibly positive feedback regulation, between the pituitary and thyroid glands, that develops or matures the hormonal network which is responsible for rapid somatic growth and metabolic changes at puberty in mice.

Animals↗

PubMed1994

c-myc protein expression during cell cycle phases in differentiating HL-60 cells.

We examined c-myc protein expression in cell cycle phases during differentiation induction of HL-60 cells by flow cytometry using an indirect immunofluorescence method. In exponentially proliferating HL-60 cells, c-myc protein was expressed in a cell cycle dependent manner. During the differentiation induction of HL-60 cells with dimethylsulfoxide (DMSO), c-myc protein was rapidly down-regulated in the G1/0 specific phase prior to the appearance of differentiation associated markers. Our results indicate that c-myc protein functions in the G1/0 specific phase in cellular differentiation, and the rapid down-regulation of c-myc protein in G1/0 phase is closely associated with initial differentiation programs.

Cell Cycle↗

PubMed1994

The character of endogenous retrovirus in pancreatic beta-cells of NOD mice.

The character of retrovirus type C in NOD mouse pancreatic-beta-cells was investigated. First, the in vitro response of retrovirus type C to glucose stimulation was examined. When the pancreatic islets of control NON mice were observed with an electron microscopy retrovirus types C could not be detected in the beta-cells with or without glucose stimulation. Retrovirus type C particles and intracisternal type A particles (IAPs) in NOD mouse pancreatic beta-cells increased by glucose stimulation, but the increase in retrovirus type C differed from that of IAPs. The clusters of retrovirus type C were found in some beta-cells, whereas IAPs were scattered in beta-cells. Next, we investigated the expression of retrovirus type C transcript in NOD mouse pancreatic islets. From polymerase chain reaction analysis using two primers of retrovirus type C designed from a conserved U3 region, a major product was found to be endogenous polytropic retrovirus (Pmv). The subcloned PCR probe and oligonucleotide probes specific for Pmv and modified polytropic retrovirus were used for Northern blot analysis. Pancreatic islets from NOD and NON mice (control mice) contained 8.4-kb and 3.0-kb Pmv transcripts. The quantity of transcripts of Pmv in NOD mouse pancreatic beta-cells were the same as that in NON mice. The transcript level of islets was much higher than those of thymus and liver. The appearance of retrovirus type C particles in beta-cells of NOD mice may involve the mechanisms by which diabetes is generated in NOD mice.

Animals↗

PubMed1993

A study on postoperative enteritis caused by methicillin-resistant Staphylococcus aureus.

We investigated the production of staphylococcal enterotoxin (SE) with respect to coagulase types by methicillin-resistant Staphylococcus aureus (MRSA). A total of 138 strains of MRSA, which were isolated from clinical materials in the surgical ward between 1983 and 1990, were studied. Coagulase type IV strains produced SE A only, whereas coagulase type II strains were classified into four groups by SE production: SE B producing strains (32.7%), SE C producing strains (29.8%), SE B and C coproducing strains (12.5%), and SE A and C coproducing strains (25.0%). Almost all of the organisms (nine of ten) which were isolated from the feces of patients with MRSA enteritis were SE A and C coproducing strains. The coincidence in time of the prevalence of MRSA enteritis and the isolation SE A and C coproducing strains also demonstrated that these strains caused MRSA enteritis. Although SE C producing strains and SE A and C coproducing strains were simultaneously prevalent in 1990, the former tended to be sensitive while the latter tended to be resistant to minocycline. Considering the variety of antibiotic sensitivity in coagulase type II strains, it is thus considered to be of critical importance for epidemiologic purposes to further characterize isolates by SE typing.

Aged↗

PubMed1993

Mucin-producing pancreatic tumors: a study of nuclear DNA content by flow cytometry.

Nuclear DNA content in eight surgically resected mucin-producing pancreatic tumors (MPPT) consisting of two mucinous intraductal adenocarcinomas (MIDAC), two mucinous intraductal adenomas (MIDA), one mucinous cyst-adenocarcinoma (MCAC), and three mucinous cystadenomas (MCA) were measured by flow cytometry using paraffin-embedded tissue samples. The technique of Shutte was used for the preparation of paraffin-embedded tissue into single dissociated nuclei, while the method of Vindelov was used for staining the isolated nuclei with propidium iodine. Clinicopathologically, the four patients with MIDAC or MIDA were all male and had cystic lesions with a dilated pancreatic duct at the head of the pancreas, while the four patients with MCAC or MCA were all females and had cystic tumors at either the body or tail of the pancreas. All eight patients with MPPT had no metastasis to the regional lymph nodes and were all still alive without recurrence. In an analysis of nuclear DNA content, seven of eight patients had DNA diploid tumors while one patient with a MIDAC perforating the duodenum and choledochus had a DNA aneuploid tumor. Thus, these findings suggest that DNA diploid patterns in MPPT might be associated with a favorable prognosis in MPPT although some patients whose MPPT invaded the surrounding organs might have DNA aneuploid tumors.

Adenocarcinoma, Mucinous↗

PubMed1993

Bone-implant interface mechanics of in vivo bio-inert ceramics.

We have previously demonstrated that there was no significant difference between the affinity of bone to bio-inert ceramics and stainless steel in a histological study. In this study, the bone-implant interface shear strength of alumina ceramics (Al2O3), zirconia ceramics (ZrO2), stainless steel (SUS316L) and sintered hydroxyapatite (HA) were compared in 19 dogs using a transcortical push-out model of the femur 4 and 12 wk after implantation. The interface shear strength of HA was significantly greater than that of alumina ceramics, zirconia ceramics and stainless steel (P < 0.001). There was no significant difference between bio-inert ceramics and stainless steel.

Aluminum Oxide↗

PubMed1993

Biochemical and morphological alterations of macrophages and spleen cells produced by antitumor polysaccharide from Acanthopanax obovatus roots.

Acanthopanax obovatus polysaccharide (AOPS) administrated intraperitoneally (i.p.) resulted in the augmentation of phagocytosis and the chemiluminescence in peritoneal macrophages of mice. Binding of the third component of complement (C3) cleavage products to the receptors on peritoneal macrophages was enhanced by AOPS. The C3 conversion in human serum was increased by AOPS in vitro treatment. When given i.p., AOPS led to the enhancement of spleen weight, spleen cell number, and plaque-forming cells. It was also found that after in vivo administration AOPS not only enhanced the synthesis of DNA and protein but also promoted the mitogenic responses of spleen cells. The morphological observation of spleens indicated that AOPS augmented the transformation from lymphocytes to plasma cells. AOPS inhibited the growth of mouse solid Sarcoma-180 and prolonged the survival time significantly. These results suggest that AOPS is an effective biological response modifier and its antitumor activity is related to its immunopotentiating effects.

Animals↗

PubMed1993

A favourable effect of long-term alpha-interferon therapy in refractory idiopathic thrombocytopenic purpura.

We successfully treated two patients with refractory idiopathic thrombocytopenic purpura (ITP) with a weekly or monthly administration of alpha-interferon followed by short-course treatment with alpha-interferon. In both cases an increase in platelet count was brought about within 1 week after administration of alpha-interferon and the favourable response in platelet count was sustained over a few years with the weekly or monthly administration of alpha-interferon in the absence of any noticeable side-effects except for fever. So, alpha-interferon therapy seems to be beneficial in some cases of refractory ITP in terms of both remission induction and maintenance therapy.

Aged↗