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Biomedical subjects

M Takahara

Publications and source records attributed to M Takahara.

At least 19 recordsLinked to original sources

Developmental failure of the intra-articular ligaments in mice with absence of growth differentiation factor 5.

OBJECTIVE: To show the phenotypic characteristics of the knee joints in brachypodism mice (bp mice), which carry a functional null mutation of the growth differentiation factor 5 (GDF5) gene, we investigated the adult and embryonic bp mice. METHOD: Radiographic and macroscopic examinations of the knee joint of adult bp mice were performed. A histological examination of the knee joint of bp mice from E12.5 to E18.5 was also performed. RESULTS: Radiographic and macroscopic examinations of the adult bp mice showed anterior dislocation, hypoplastic condyles, and absence of the intra-articular ligaments. Safranin O staining of knee joints of the embryonic bp mice showed severe hypoplasty of the chondroepiphyses and intra-articular ligaments at E16.5. There was no difference in the number and location of 5-bromo-2'-deoxyuridine (BrdU)-positive cells between wild-type and bp mice through E12.5 to E14.5. A terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) study showed excessive cell death of mesenchymal cells of the future knee joint in bp mice at E12.5 and E13.5. CONCLUSION: bp mice exhibit developmental failure of the condyles and intra-articular ligament of the knee joints.

Animals↗

Improved shoulder contour following forequarter amputation with an osteomyocutaneous free flap from the amputated extremity: two cases.

To cover a large soft-tissue defect and to reconstruct the shoulder contour after forequarter amputation, we used an osteomyocutaneous free flap incorporating an elbow joint from the amputated extremity in two patients. These flaps were well vascularised and reliable. They provided excellent coverage of large soft-tissue defects and they maintained shoulder contours. This procedure is useful for reconstruction after extended forequarter amputation and chest wall resection.

Adolescent↗

Wild-type p53 activates SAP expression in lymphoid cells.

SAP is an adaptor molecule with one SH2 domain and it is expressed in activated T and NK cells, where it is required for the appropriate signaling from the SLAM family of surface receptors. Deleted or mutated SAP genes that encode functionally defective protein are associated with the X-linked lymphoproliferative disease (XLP). This primary immunodeficiency is characterized by extreme sensitivity to Epstein-Barr virus (EBV) infection, dysgammaglobulinemia and a high rate of lymphoma development. The vigorous T- and B-cell proliferation that follows EBV infection and the high incidence of lymphomas (30%) in XLP patients might reflect functional defects in cell cycle and/ or apoptosis control. Our experiments show that SAP is a target of p53. In Burkitt lymphoma (BL) lines transfected with a temperatur-sensitive (ts) p53, SAP mRNA and protein expression was dependent on wild-type (wt) p53. Activation of endogenous wt p53 in BLs and lymphoblastoid cell lines led to the induction of SAP and this was inhibited by the specific p53 inhibitor pifithrin-alpha. Cell lines that carried mutant p53 did not express SAP under similar conditions. Moreover, we have shown binding of wt p53 to the promoter region of SAP by ChIP assay. Our results suggest that SAP contributes to the execution of some p53 functions.

Base Sequence↗

Expression of a Y-box protein, YB2/RYB-a, precedes protamine 2 expression during spermatogenesis in rodents.

Y-box binding proteins, a large family of proteins, are involved in a variety of functions. The present study describes the expression of YB2, a rat Y-box binding protein, and/or RYB-a, an alternatively spliced product of the YB2 gene during spermatogenesis. YB2/RYB-a is thought to be the rat orthologue of mouse Y-box protein 3 (MSY3). An antibody which recognizes YB2/RYB-a was developed and applied in an immunochemical study of rat and mouse testes. We also carried out an in-situ hybridization study and Northern blot analysis of YB2/RYB-a and protamine 2 mRNA expression. Both YB2/RYB-a mRNA and the proteins appeared in prepubertal mouse testes, prior to the expression of the mouse protamine 2 mRNA. The mRNA and protein were present at high levels in spermatocytes, decreased in round to elongated spermatids, and were absent in spermatozoa. Since the protamine 2 mRNA was present at high levels in round and elongating spermatids, the proposed function of the YB2/RYB-a protein as a translational repressor of the mRNA was supported in mouse. The level and localization of YB2/RYB-a mRNA and protein expression in the rat testis was comparable to that in mouse testis, although rat testis is known to express a very low level of protamine 2, but is also likely to affect the expression of other proteins (including protamine 1) during spermatogenesis.

Animals↗

Visualization of an FtsZ ring in chloroplasts of Lilium longiflorum leaves.

FtsZ is a bacterial division protein which forms a ring at the leading edge of the cell division site. To date, a hypothesis that the plant FtsZ forms the same structure in chloroplast division is proposed, but has not been demonstrated yet. In this study, recombinant LlFtsZ (Lilium longiflorum FtsZ) protein was produced from a previously isolated ftsZ cDNA clone [Mori and Tanaka (2000) Protoplasma 214: 57] and used to raise polyclonal anti-LlFtsZ antibodies in rabbits. In immunoblot analysis with the total protein extracted from L. longiflorum leaves, purified antibodies specifically recognized LlFtsZ whose molecular mass was approximately 43 kDa. This size corresponded to that of the recombinant LlFtsZ protein lacking N-terminal sequence, which suggests that the full-length LlFtsZ translation product has a putative N-terminal signal peptide. Moreover, fluorescent and electron microscopy revealed that the anti-LlFtsZ antibodies recognized ring structures at stromal side of the constriction point of dividing chloroplasts. Here, we show direct evidence that FtsZ ring is involved in chloroplast division.

Bacterial Proteins↗

Plastid division is driven by a complex mechanism that involves differential transition of the bacterial and eukaryotic division rings.

During plastid division, two structures have been detected at the division site in separate analyses. The plastid-dividing ring can be detected by transmission electron microscopy as two (or three) electron-dense rings: an outer ring on the cytosolic face of the outer envelope, occasionally a middle ring in the intermembrane space, and an inner ring on the stromal face of the inner envelope. The FtsZ ring, which plays a central role in bacterial division, also is involved in plastid division and is believed to have descended to plastids from cyanobacterial endosymbiosis. The relationship between the two structures is not known, although there is discussion regarding whether they are identical. Biochemical and immunocytochemical investigations, using synchronized chloroplasts of the red alga Cyanidioschyzon merolae, showed that the plastid FtsZ ring is distinct and separable from the plastid-dividing ring. The FtsZ ring localizes in stroma and faces the inner plastid-dividing ring at the far side from the inner envelope. The FtsZ ring and the inner and outer plastid-dividing rings form in that order before plastid division. The FtsZ ring disappears at the late stage of constriction before dissociation of the plastid-dividing ring, when the constriction is still in progress. Our results suggest that the FtsZ ring;-based system, which originated from a plastid ancestor, cyanobacteria, and the plastid-dividing ring;-based system, which probably originated from host eukaryotic cells, form a complex and are involved in plastid division by distinct modes.

Arabidopsis Proteins↗

Novel filaments 5 nm in diameter constitute the cytosolic ring of the plastid division apparatus.

The plastid division apparatus (called the plastid-dividing ring) has been detected in several plant and algal species at the constricted region of plastids by transmission electron microscopy. The apparatus is composed of two or three rings: an outer ring in the cytosol, an inner ring in the stroma, and a middle ring in the intermembrane space. The components of these rings are not clear. FtsZ, which forms the bacterial cytokinetic ring, has been proposed as a component of both the inner and outer rings. Here, we present the ultrastructure of the outer ring at high resolution. To visualize the outer ring by negative staining, we isolated dividing chloroplasts from a synchronized culture of a red alga, Cyanidioschyzon merolae, and lysed them with nonionic detergent Nonidet P-40. Nonidet P-40 extracted primarily stroma, thylakoids, and the inner and middle rings, leaving the envelope and outer ring largely intact. Negative staining revealed that the outer ring consists of a bundle of 5-nm filaments in which globular proteins are spaced 4.8 nm apart. Immunoblotting using an FtsZ-specific antibody failed to show immunoreactivity in the fraction containing the filament. Moreover, the filament structure and properties are unlike those of known cytoskeletal filaments. The bundle of filaments forms a very rigid structure and does not disassemble in 2 M urea. We also identified a dividing phase-specific 56-kD protein of chloroplasts as a candidate component of the ring. Our results suggest that the main architecture of the outer ring did not descend from cyanobacteria during the course of endosymbiosis but was added by the host cell early in plant evolution.

Actin Cytoskeleton↗

Hyphae and yeasts of Candida albicans differentially regulate interleukin-12 production by human blood monocytes: inhibitory role of C. albicans germination.

The role of Candida albicans yeast-to-hyphae transition in interleukin-12 (IL-12) production by monocytes was investigated. Germinating C. albicans not only failed to induce IL-12 p70 but also suppressed IL-12 production induced by heat-killed C. albicans. Comparison of the abilities of germinating C. albicans and agerminating mutants to inhibit IL-12 production showed that germination of C. albicans plays a critical role in the inhibition of IL-12 production.

Candida albicans↗

[Efficacy of tonsillectomy on psoriasis and tonsil histology].

Treatment outcomes of tonsillectomy were studied in 7 Japanese patients with psoriasis--3 males and 4 females aged 9 to 46 years (median: 23 years)--followed up 2 to 9 years after tonsillectomy. All skin lesions disappeared in 3 patients, 80% of those in 2, and no change in the remaining 2 during follow-up. Of 5 in whom skin lesions improved, 4 were females and had a history of tonsillitis making skin lesions worse. In quantitative immunohistologic analysis on tonsillar tissues by CD20 and anti-ssDNA antibodies, areas of T cell-nodules were significantly expanded, but those of the B-lymphoid follicles were smaller, and the number of apoptotic cells increased in tonsils from patients with psoriasis and PPP compared to those with recurrent tonsillitis. The area of T cell-nodules and the number of apoptosis cells were significantly larger in tonsils from 4 patients with complete recovery after tonsillectomy compared to the remaining 9 without complete recovery. This suggests that histologic evaluation may be helpful in estimating the effectiveness of tonsillectomy.

Adolescent↗

Chondro-osseous differentiation in fat tissue tumors: magnetic resonance imaging with pathological correlation.

Chondro-osseous differentiation of three benign or malignant fat tissue tumors--two chondrolipomas and a liposarcoma with cartilaginous metaplasia--was studied with magnetic resonance (MR) imaging and compared with their pathological findings. The results suggest that demarcation of cartilage tissue can be clearly defined on MR imaging when the size of the cartilaginous area is large. Myxoid matrix, degenerative fat tissue and lipodystrophic change may decrease the delineation of the cartilage tissue.

Biopsy↗

Isolation, characterization, and chromosomal mapping of an ftsZ gene from the unicellular primitive red alga Cyanidium caldarium RK-1.

The FtsZ protein is involved in eukaryote plastid division, but there is little information on its involvement in the plastid-dividing apparatus. To investigate the relationship between FtsZ and the plastid-dividing ring, the ftsZ gene was isolated from the unicellular primitive red alga Cyanidium caldarium RK-1. Comparison of several prokaryotic and eukaryotic FtsZ proteins shows that there are six highly conserved domains in the core region of FtsZ. To determine the chromosomal location of ftsZ, we first determined the electrophoretic karyotype of C. caldarium RK-1. Southern-hybridization analysis combined with CHEF revealed the chromosomes on which the ftsZ gene exist. Northern-hybridization analysis indicated that the C. caldarium RK-1 ftsZ gene is transcribed as a 1.9-kb molecule, and that the transcripts specifically accumulate just before plastid division. Phylogenetic analysis indicated that C. caldarium RK-1 and other eukaryotic ftsZ genes are the descendants of cyanobacterial ftsZ genes, supporting the current agreement that FtsZ is involved in plastid division.

Algal Proteins↗

A putative mitochondrial ftsZ gene is present in the unicellular primitive red alga Cyanidioschyzon merolae.

Two ftsZ homologues were isolated from the unicellular primitive red alga Cyanidioschyzon merolae (CmftsZ1 and CmftsZ2). Phylogenetic analysis revealed that CmftsZ1 is most closely related to the ftsZ genes of alpha-Proteobacteria, suggesting that it is a mitochondrial-type ftsZ gene, whereas CmftsZ2 is most closely related to the ftsZ genes of cyanobacteria, suggesting that it is a plastid-type ftsZ gene. Southern analysis indicates that CmftsZ1 and CmftsZ2 are both single-copy genes located on chromosome XIV in the C. merolae genome. Northern analysis revealed that both CmftsZ1 and CmftsZ2 are transcribed, and accumulate specifically before cell and organelle division. The results of Western analysis suggest that CmFtsZ1 is localized in mitochondria.

Amino Acid Sequence↗

Natural progression of osteochondritis dissecans of the humeral capitellum: initial observations.

PURPOSE: To determine the earliest findings, subsequent changes, and natural course of osteochondritis dissecans of the humeral capitellum. MATERIALS AND METHODS: Among 95 patients with osteochondritis dissecans of the humeral capitellum, 16 (mean age, 12.5 years) were selected for this retrospective study because they seemed to have early osteochondritis dissecans and had been followed up without any surgical treatment for 6 months or more (mean, 3.5 years). RESULTS: The initial imaging appearances of the 16 patients' lesions were divided into two types: localized subchondral bone flattening without fragments in seven, and nondisplaced fragments in nine. Patients with lesion flattening had younger ages and significantly shorter durations of symptoms, and most had open growth plates. In five of the seven with flattening, new bone formed over the flattened bone, and the fragments united after arm motion reduction. In contrast, patients with nondisplaced fragments at clinical presentation had longer durations of symptoms with continued arm motion, and their nondisplaced fragments failed to unite. CONCLUSION: The earliest feature of osteochondritis dissecans is subchondral bone flattening, over which new bone subsequently forms. The new bone then can unite with the underlying bone. However, if subjected to repetitive forces over a given time, unstable fragments develop. These fragments, even if not yet displaced, are unable to unite.

Adolescent↗

Sonographic assessment of osteochondritis dissecans of the humeral capitellum.

OBJECTIVE: The purpose of this study was to determine the efficacy of sonography for revealing osteochondritis dissecans of the humeral capitellum. SUBJECTS AND METHODS: Twenty-seven patients with capitellar osteochondritis dissecans (27 males; range, 11-20 years; mean age, 14 years) underwent radiography and sonography performed with a 7.5-MHz mechanical sector probe. Lesions were assessed as stable or unstable. The sonographic assessment was compared with radiographic assessment in 27 patients, MR assessment obtained in 10, and surgical findings in 15. RESULTS: Sonographic assessment agreed with radiographic assessment in 23 of the 27 patients, MR assessment in nine of the 10, and surgical findings in 14 of the 15. Sonography revealed that two lesions, which had been underestimated on radiography, were unstable. CONCLUSION: Sonography facilitates the assessment of capitellar lesions so that treatment can be optimized.

Adolescent↗

[A clinical study on pulmonary tuberculosis in younger age groups].

In 1997, the number of newly registered patients with pulmonary tuberculosis increased, compared with that in 1996, in Japan. The majority of the increase were occupied by elder patients 70 years of age or higher. But in younger group less than 30 years old, a reduction in the incidence of tuberculosis had been slowed down, until 1996. The purpose of this report is to elucidate the characteristics of these younger patients. 139 cases younger than 30 years of age, who were hospitalized in the tuberculous ward of IMCJ from April 1995 to March 1998, were investigated, and were compared with the control group (557 cases), 30-79 years old who were hospitalized during the same period. In the younger group, the proportion of women cases, discovered by health examination, foreigners, and contact with TB patients in the past was significantly higher than in the control group. But there were no difference between the both groups, concerning the proportion of those spending irregular life or living alone. The proportion of sputa smear negative cases was significantly higher in the younger group than in the control. For early diagnosis of TB among younger group, the application of bronchofiberscopy and nucleic acid diagnostic method, are encouraged.

Adolescent↗

Surgical repair of acute collateral ligament injuries in digits with the Mitek bone suture anchor.

Eighteen acute grade III collateral ligament injuries were treated by using the Mitek bone suture anchor. Seven were thumb metacarpophalangeal joint injuries, and eleven were finger proximal interphalangeal joint injuries. Seventeen patients were followed more than 12 months after surgery. All patients were able to use the digits in daily living activities within 5 weeks after surgery, and return to their original work or sports activities within 12 weeks. Pain was completely relieved in 15 patients. Loss of joint motion averaged 7 degrees. In all joints the postoperative lateral stress angle was within 10 degrees of that of the contralateral digit.

Adolescent↗

Two types of ftsZ genes isolated from the unicellular primitive red alga Galdieria sulphuraria.

FtsZ plays a crucial role in bacterial cell division, and may be involved in plastid division in eukaryotes. To investigate the evolution of the dividing apparatus from prokaryotes to eukaryotes, the ftsZ genes were isolated from the unicellular primitive red alga Galdieria sulphuraria. Two ftsZ genes (GsftsZ1 and GsftsZ2) were isolated. This suggests that duplication and divergence of the ftsZ gene occurred in an early stage of plant evolution. A comparison of the FtsZs of G. sulphuraria and other organisms shows that FtsZ is highly and universally conserved among prokaryotes, primitive eukaryotic algae, and higher plants. The GsftsZ2 gene seems to contain an intron. Southern hybridization analysis of the G. sulphuraria chromosomes separated by CHEF revealed that each ftsZ gene and its flanking region may be duplicated.

Amino Acid Sequence↗