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Biomedical subjects

J Hata

Publications and source records attributed to J Hata.

At least 91 records · Page 5Linked to original sources
PubMed1999

Photoautotrophic cultures of the host and transformed cells of Marchantia polymorpha under controlled incident light intensity.

Photoautotrophic cultures of the host and transformed cells of the liverwort, Marchantia polymorpha, were examined. In cultures in flat glass flasks under various light intensities, it was found that the growth rates of both the cells increased with increase in light intensity in the range of 0 to 25 W/m2, but further increase in light intensity caused photoinhibition of the growth of the cells. Cultures of both the types of cells under light-controlled conditions using an externally illuminated bioreactor were carried out taking into consideration the inhibition of cell growth by excessive light and the light intensity distributions in the cell suspensions. In these cultures, 2.1 (transformed cells) and 3.3 (host cells) kg dry cell weight per m3 were harvested at culture times of 9.0 and 10 d, respectively. These values were larger than those obtained in cultures of the respective cells at a fixed incident light intensity of 25 W/m2.

Journal Article↗
PubMed1999

Takayasu's aortitis with dissection in systemic lupus erythematosus.

A forty-seven-year-old Japanese woman under treatment for systemic lupus erythematosus (SLE), complained of severe back pain. Chest X-ray and MRI showed an aneurysmal dilatation of the ascending aorta. Subsequently an aortic replacement was performed. Microscopically, the resected aorta showed Takayasu's aortitis with chronic dissection. Both aortitis and dissection are rare events in SLE patients. To our knowledge, this is the first report of Takayasu's aortitis with dissection in a patient with SLE.

Aortic Dissection↗
PubMed1999

Development and validation of an ultrasonographic activity index of Crohn's disease.

OBJECTIVES: We developed and validated an ultrasonographic index of intestinal inflammatory activity for patients with Crohn's disease. METHODS: Fifty-five patients with Crohn's disease were examined by transabdominal ultrasonography. The pathological findings were classified into three types (A-C) on the basis of wall thickness and wall stratification. To calculate the index, we divided the intestine into eight segments, and the scores for each segment were summed to calculate the index (ultrasonographic activity index of Crohn's disease) as follows: 1 point for type A lesions, [wall thickness (mm) -2] x 2 for type B lesions, and [wall thickness (mm) -2] x 4 for type C lesions. Endoscopic or barium contrast findings were also scored in a similar fashion, with the following parametric scores: 10 for cobblestoning, 5 for longitudinal ulcers, 3 for aphthoid ulcers, and 1 for chronic inflammatory changes. RESULTS: A strong correlation (r2 = 0.62, P<0.01) was found between the ultrasound index and the endoscopic/radiological score, while weak correlations were found between the endoscopic/radiological score and the Crohn's disease activity index or biological indices of inflammation. CONCLUSIONS: Our results show that the ultrasonographic activity index of Crohn's disease can be of value in the ongoing assessment and treatment of patients.

Adolescent↗
PubMed1999

Axonal and perikaryal involvement in chronic inflammatory demyelinating polyneuropathy.

OBJECTIVES: To assess the extent of loss of myelinated nerve fibres and spinal motor neuron loss in chronic inflammatory demyelinating polyneuropathy (CIDP), a clinicopathological study was conducted on biopsied sural nerves and necropsied spinal cords from patients with CIDP. METHODS: The myelinated fibre pathology of 71 biopsied sural nerves and motor neuron pathology of nine necropsied spinal cords at L4 levels in patients with CIDP were quantitatively and immunohistochemically assessed. RESULTS: Myelinated nerve fibre density was significantly diminished to 65.4% of the control values (p <0.0001), correlating inversely with the extent of segmental demyelination and remyelination (r = -0.43, p < 0.0005) and duration of illness (r = -0.31, p < 0.01). Numbers of large spinal motor neurons in CIDP were variably but significantly diminished (range from 46.0 to 97.6% of the age matched control value (p < 0.005)), and reactive astrogliosis was evident in the ventral horn in CIDP. The frequency of ventral horn neurons exhibiting central chromatolysis and the accumulation of phosphorylated high molecular weight neurofilament protein was significantly higher in CIDP than in controls (p<0.01 and p<0.05). CONCLUSIONS: The loss of nerve axons and spinal motor neurons is common in CIDP, and extensive in some cases. These neuronal and axonal losses may influence the functional prognosis in CIDP.

Adolescent↗
PubMed1999

Cardiomyocytes can be generated from marrow stromal cells in vitro.

We have isolated a cardiomyogenic cell line (CMG) from murine bone marrow stromal cells. Stromal cells were immortalized, treated with 5-azacytidine, and spontaneously beating cells were repeatedly screened. The cells showed a fibroblast-like morphology, but the morphology changed after 5-azacytidine treatment in approximately 30% of the cells; they connected with adjoining cells after one week, formed myotube-like structures, began spontaneously beating after two weeks, and beat synchronously after three weeks. They expressed atrial natriuretic peptide and brain natriuretic peptide and were stained with anti-myosin, anti-desmin, and anti-actinin antibodies. Electron microscopy revealed a cardiomyocyte-like ultrastructure, including typical sarcomeres, a centrally positioned nucleus, and atrial granules. These cells had several types of action potentials, such as sinus node-like and ventricular cell-like action potentials. All cells had a long action potential duration or plateau, a relatively shallow resting membrane potential, and a pacemaker-like late diastolic slow depolarization. Analysis of the isoform of contractile protein genes, such as myosin heavy chain, myosin light chain, and alpha-actin, indicated that their muscle phenotype was similar to that of fetal ventricular cardiomyocytes. These cells expressed Nkx2.5/Csx, GATA4, TEF-1, and MEF-2C mRNA before 5-azacytidine treatment and expressed MEF-2A and MEF-2D after treatment. This new cell line provides a powerful model for the study of cardiomyocyte differentiation.

Action Potentials↗
PubMed1999

An HIV autopsy--characterization of zidovudine-resistant subtype E HIV-1 from autopsy tissue suggests the route of infection and an alternative protocol of therapy.

This CPC concerns a 47-year-old male patient with acquired immunodeficiency syndrome (AIDS). The patient became symptomatic when he developed Pneumocystis carinii pneumonia, but recovered sufficiently to be treated as an outpatient. Two years after falling ill, he developed septic shock and died within a short time. During this period, he failed to respond to HIV drugs, and there was no improvement in his immunodeficient status. The HIV retrieved from the patient's organs at autopsy was found to be type E and to have acquired resistance to Zidovudine. It was also possible to determine the route of infection. HIV treatment guidelines are continuously being revised on the basis of HIV research and the development of new treatment plans, and at the present time, when no definitive method of treatment has yet been established, it is essential for the clinician to keep abreast of the latest information. Since HIV patients are compromised hosts, it is important to diagnose and treat other infectious complications, not only complications unique to AIDS, and we have briefly described the latest HIV therapy.

Acquired Immunodeficiency Syndrome↗
PubMed1998

Hypoplasia of pancreatic islets in transgenic mice expressing activin receptor mutants.

Activin, a member of the TGF-beta superfamily, regulates the growth and differentiation of a variety of cell types. Based on the expression of activin in pancreatic rudiments of rat embryos and stimulation of insulin secretion from adult rat pancreatic islets by activin, activin is implicated in the development and function of islets. To examine the significance of activin signaling in the fetal and postnatal development of islets, transgenic mice expressing a dominant negative form of activin receptor (dn-ActR) or a constitutively active form of activin receptor (ActR-T206D) in islets were generated together with the transgenic mice expressing intact activin receptor (intact ActR) as a negative control. Transgenic mice with both dn-ActR and ActR-T206D showed lower survival rates, smaller islet area, and lower insulin content in the whole pancreas with impaired glucose tolerance when compared with transgenic mice with intact ActR or littermates, but they showed the same alpha cell/beta cell ratios as their littermates. In addition to islet hypoplasia, the insulin response to glucose was severely impaired in dn-ActR transgenic mice. It is suggested that a precisely regulated intensity of activin signaling is necessary for the normal development of islets at the stage before differentiation into alpha and beta cells, and that activin plays a role in the postnatal functional maturation of islet beta cells.

Activin Receptors↗
PubMed1998

Up-regulated expression of murine Mcl1/EAT, a bcl-2 related gene, in the early stage of differentiation of murine embryonal carcinoma cells and embryonic stem cells.

We have cloned a murine homologue of the human Mcl1/EAT gene, a Bcl-2 related gene. Sequence analysis revealed that murine Mcl1/EAT (mMcl1/EAT) has three Bcl-2 homology domains, two PEST sequences, and immediate response boxes (IRB). The presence of IRB indicates that mMcl1/EAT is an immediate-early gene. mMcl1/EAT increases dramatically with exposure to retinoic acid in murine embryonal carcinoma cell lines (F9 and PCC3) as well as embryonic stem cells, both of which are models of early embryogenesis.

Amino Acid Sequence↗
PubMed1998

Isolation and expression of a human SRY-related cDNA hSOX20.

SOX is a family of genes related to the testis-determining gene, SRY. We have isolated and sequenced an hSOX20 cDNA from a cell line of human embryonic carcinoma. This cDNA contains an open reading frame (ORF) encoding 233 amino acids. The protein encompasses an SRY-type HMG box exhibiting strong homologies to those of mouse Sox15 and Sox16. Various adult and fetal tissues were tested for hSOX20 mRNA by Northern analysis. Its expression is restricted to the fetal testis and the size of the transcript is 1.5 knt. Electrophoretic mobility shift assay indicated that recombinant hSOX20 polypeptide is capable of binding to AACAAT sequence.

Adult↗
PubMed1998

Rapid diagnosis of red sea bream iridovirus infection using the polymerase chain reaction.

A simple and sensitive polymerase chain reaction (PCR) based assay is described for detection of the red sea bream iridovirus (RSIV) in infected fish. The assay involves amplification of a portion of the ribonucleotide reductase small subunit (RNRS) gene of the virus from DNA isolated from the spleen. The system was tested on red sea bream following an experimental infection. In our infection model, disease signs first became apparent 5 to 6 d post-infection, and mortality commenced at Day 6 and reached 90% by Day 9. No amplified product was detected from fish at 1 or 2 d post-infection, but 3 of 5 fish tested positive at Day 3, and all fish tested positive at Days 5 and 8. Thus, infection could be detected prior to the appearance of overt symptoms. This PCR method should be of considerable value for aquaculture to detect RSIV infection.

Animals↗
PubMed1998

Interleukin 7 transgenic mice develop chronic colitis with decreased interleukin 7 protein accumulation in the colonic mucosa.

We have demonstrated that intestinal epithelial cells produce interleukin 7 (IL-7), and IL-7 serves as a potent regulatory factor for proliferation of intestinal mucosal lymphocytes expressing functional IL-7 receptor. To clarify the mechanism by which locally produced IL-7 regulates the mucosal lymphocytes, we investigated IL-7 transgenic mice. Here we report that transgenic mice expressing murine IL-7 cDNA driver by the SRalpha promoter developed chronic colitis in concert with the expression of SRalpha/IL-7 transgene in the colonic mucosa. IL-7 transgenic but not littermate mice developed chronic colitis at 4-12 wk of age, with histopathological similarity to ulcerative colitis in humans. Southern blot hybridization and competitive PCR demonstrated that the expression of IL-7 messenger RNA was increased in the colonic mucosal lymphocytes but not in the colonic epithelial cells. IL-7 protein accumulation was decreased in the goblet cell-depleted colonic epithelium in the transgenic mice. Immunohistochemical and cytokine production analysis showed that lymphoid infiltrates in the lamina propria were dominated by T helper cell type 1 CD4+ T cells. Flow cytometric analysis demonstrated that CD4+ intraepithelial T cells were increased, but T cell receptor gamma/delta T cells and CD8alpha/alpha cells were not increased in the area of chronic inflammation. Increased IL-7 receptor expression in mucosal lymphocytes was demonstrated in the transgenic mice. These findings suggest that chronic inflammation in the colonic mucosa may be mediated by dysregulation of colonic epithelial cell-derived IL-7, and this murine model of chronic colitis may contribute to the understanding of the pathogenesis of human inflammatory bowel disease.

Animals↗
PubMed1998

Gene expression of noncollagenous bone matrix proteins in the limb joints and intervertebral disks of the twy mouse.

The twy (tiptoe walking Yoshimura) mouse is an autosomal recessive mutant manifesting multiple osteochondral lesions characterized by pathologic calcium deposition. To elucidate the pathophysiology of the limb joint lesions and the intervertebral disk lesions of the twy mouse, we assessed the mRNA expression of noncollagenous bone matrix proteins such as osteocalcin, osteonectin, osteopontin, and matrix Gla protein (MGP) by in situ hybridization, but only expression of MGP was observed in association with the pathologic calcium deposits in twy mice. Mild degeneration and abnormal growth of the cartilage in contact with the joint capsule was observed at 5 weeks in the articular cartilage of the ankle joint of the twy mouse, and MGP gene expression was observed at the same time. Simultaneous growth of synovial membrane cells and relatively undifferentiated articular cartilage cells in the knee joint, and of cartilage-like cells near the insertion of the cruciate ligament was observed in the twy mouse, and MGP gene expression was found to be present at the same time. Hypertrophy of abnormally proliferated chondrocyte-like cells, which are different from fibrocartilaginous cells of the annulus fibrosus, was observed in the intervertebral disks of the twy mouse at 3 weeks of age, and MGP gene expression was noted at the same time. These findings suggest that abnormal expression of MGP plays a major role in the pathologic calcification of the twy mouse.

Animals↗
PubMed1998

Abnormalities of basal cell keratin in epidermolysis bullosa simplex do not affect the expression patterns of suprabasal keratins and cornified cell envelope proteins.

Basal keratins, suprabasal keratins, filaggrin, and cornified cell envelope (CCE) precursor proteins are expressed during the differentiation of epidermal keratinocytes. These molecules are coordinately expressed during epidermal differentiation. The present study investigated the expression patterns of keratins and CCE precursor proteins in 15 patients with epidermolysis bullosa simplex (EBS), which is caused by mutations in the genes that encode for the basal keratins, keratins 5 and 14. The patterns of expression of keratins 5, 14, 1 and 10, filaggrin, and of the three major CCE precursor proteins, involucrin, loricrin and small proline-rich proteins 1 and 2 (SPRs), were studied immunohistochemically and by electron microscopy. In 14 of the 15 patients with EBS, the distribution pattern of keratins was not altered. In one neonate with EBS, basal cell keratins were expressed in the suprabasal layers. Ultrastructurally, numerous clumped tonofilaments were observed in the basal and suprabasal cells. In all cases, findings were positive for filaggrin in the granular cells, with positivity for involucrin in the upper spinous and granular cells. The upper spinous cells and granular cells were positive for SPRs 1 and 2, and loricrin was expressed in granular cells. Ultrastructurally, no marked abnormality was observed in the suprabasal layers such as a decrease in, or agglutination of, keratin filaments, except in one neonate. A CCE about 15 nm thick was formed normally in the cell membrane of cornified cells. The patterns of distributions of basal cell keratins, suprabasal keratins, filaggrin, and CCE precursor proteins, as well as the ultrastructural findings, resembled those of normal skin. Thus, the abnormality in basal cell keratins in patients with EBS did not appear to alter the patterns of expression of the keratins and CCE precursor proteins.

Adolescent↗
PubMed1998

Evaluation of the colorectal wall in normal subjects and patients with ulcerative colitis using an ultrasonic catheter probe.

BACKGROUND: Ulcerative colitis is usually evaluated by barium enema and colonoscopy, methods of imaging that are limited to the mucosal surface. Endoscopic ultrasonography (EUS) is the best modality for the evaluation of transmural changes in the bowel wall. We therefore evaluated the colorectal wall in normal control subjects and patients with ulcerative colitis using an ultrasonic catheter probe. METHODS: Endoscopic ultrasound with a catheter probe was performed on 36 normal control subjects (36 examinations) and 72 patients (111 examinations) with documented ulcerative colitis. RESULTS: In normal control subjects, the total wall, mucosa, submucosa, and muscularis propria were significantly thicker in the rectum than in the colon. There was no significant difference in the thickness of the colon and rectum among different age groups or between men and women. In ulcerative colitis, we classified the boundary of each layer into three patterns (smooth, irregular, and blurred) and then classified the wall into six types. In Matts grade 2 and 3 ulcerative colitis lesions, the total wall and each layer were significantly thicker than lower grade lesions and normal control subjects. For grade 1 and most of the grade 2 lesions, the boundary of each layer was smooth. In some of the grade 3 lesions, the mucosa-submucosa and submucosa-muscularis propria layer borders were abnormal. In all grade 4 cases, the mucosa-submucosa boundary was blurred. There was some correlation between the Matts grade and EUS findings except for Matts grade 3 lesions which had various EUS patterns. CONCLUSION: Endoscopic ultrasound with a catheter probe is a useful modality for the transmural assessment of the colorectal wall and, when used in conjunction with the many clinical and endoscopic parameters currently available, may contribute to the diagnosis and treatment of ulcerative colitis.

Adult↗
PubMed1998

Establishment of a retinoic acid-resistant human acute promyelocytic leukaemia (APL) model in human granulocyte-macrophage colony-stimulating factor (hGM-CSF) transgenic severe combined immunodeficiency (SCID) mice.

To understand the mechanisms and identify novel approaches to overcoming retinoic acid (RA) resistance in acute promyelocytic leukaemia (APL), we established the first human RA-resistant APL model in severe combined immunodeficiency (SCID) mice. UF-1 cells, an RA-resistant APL cell line established in our laboratory, were transplanted into human granulocyte-macrophage colony-stimulating factor (GM-CSF)-producing SCID (hGMTg SCID) mice and inoculated cells formed subcutaneous tumours in all hGMTg SCID mice, but not in the non-transgenic control SCID mice. Single-cell suspensions (UF-1/GMTg SCID cells) were similar in morphological, immunological, cytogenetic and molecular genetic features to parental UF-1 cells. All-trans RA did not change the morphological features of cells or their expression of CD11b. RA did not alter the growth curve of cells as determined by MTT assay, suggesting that UF-1/GMTg SCID cells are resistant to RA. These results demonstrate that this is the first RA-resistant APL animal model that may be useful for investigating the biology of this myeloid leukaemia in vivo, as well as for evaluating novel therapeutic approaches including patients with RA-resistant APL.

Animals↗