PubMed HealthSearch

Biomedical subjects

M Umemoto

Publications and source records attributed to M Umemoto.

At least 19 recordsLinked to original sources

Bordetella bronchiseptica dermonecrotizing toxin, which activates a small GTP-binding protein rho, induces membrane organelle proliferation and caveolae formation.

We recently demonstrated that Bordetella bronchiseptica dermonecrotizing toxin (DNT) modifies a small GTP-binding protein rho and causes assembly of actin stress fibers and focal adhesions in MC3T3-E1 cells, indicating that DNT activates the function of rho protein (Horiguchi et al., 1995, J. Cell Sci. 108, 3243-3251). In this study, we examined by electron microscopy ultrastructural changes in DNT-treated MC3T3-E1 cells under conditions in which DNT elicited the above morphological changes. We found that DNT induced proliferation of cytoplasmic membrane organelles, such as Golgi apparatus, smooth and rough endoplasmic reticulum, and mitochondria, and formation of plasmalemmal caveolae. Therefore we examined also the effect of Clostridium botulinum C3 exoenzyme (C3), which has been known to ADP-ribosylate and inactivate rho protein, and found that C3 exoenzyme inhibited the development of membrane organelles in the MC3T3-E1 cells. These findings show that proliferation of membrane organelles and caveolae formation are controlled by the function of rho, which is the target of DNT action.

3T3 Cells

Expression of highly polysialylated NCAM (NCAM-H) in developing and adult chicken auditory organ.

Neural cell adhesion molecule (NCAM) is highly polysialylated (NCAM-H) in developing tissues, and recent findings suggest that NCAM-H is more essential for neural development than poorly sialylated NCAM (NCAM-L). In order to understand the precise role of NCAM-H in developing and adult inner ears, the immunohistochemical localization of NCAM-H in developing and adult chicken inner ears was examined using a monoclonal antibody which is only specific for NCAM-H. Immunoreactivity of NCAM-H was initially observed on acoustic ganglion at stage 24, when peripheral (afferent) fibers begin to emerge from the ganglion cells. At stage 38, when peripheral fibers form synapses with hair cells, NCAM-H was observed on peripheral fibers and the base of hair cells in the auditory epithelium. At stage 42, NCAM-H on nerve fibers disappeared, and only some acoustic ganglion cells were still positive for NCAM-H. This immunostain on ganglion cells was retained after birth. These data are consistent with the hypothesis that NCAM-H specifically regulates the afferent nerve fibers' growth and synaptogenesis with hair cells during inner ear development and may be associated with processing of auditory information and neuronal plasticity.

Animals

Successful hematopoietic reconstitution by transplantation of umbilical cord blood cells in a transfusion-dependent child with Diamond-Blackfan anemia.

A 4-year-old boy with Diamond-Blackfan anemia and a history of multiple transfusions underwent umbilical cord blood transplantation from his HLA-identical female sibling born by vaginal delivery at 38 weeks. The patient was prepared with busulfan, cyclophosphamide and antilymphocyte globulin. Methotrexate and cyclosporin A were given for the prophylaxis of GVHD. Regimen-related toxicity was not observed and successful engraftment occurred, including the erythroid series. No evidence of acute or chronic GVHD has been observed for 14 months after transplantation. This is the first case of successful umbilical cord blood transplantation to a patient with Diamond-Blackfan anemia.

Blood Transfusion

Conditioning with cyclophosphamide/antithymocyte globulin for allogeneic bone marrow transplantation from HLA-matched siblings in children with severe aplastic anemia.

Graft rejection has been a problem after bone marrow transplantation for patients with severe aplastic anemia (SAA). Ten children with SAA were conditioned for bone marrow transplantation from HLA-identical siblings, using cyclophosphamide (CY, 50 mg/kg) plus antithymocyte globulin (ATG, 15 mg/kg) for 4 successive days. Marrow was infused 36 h after the last dose of CY. Cyclosporin A and methotrexate were administered as graft-versus-host disease (GVHD) prophylaxis. All patients achieved durable engraftment at follow-up of 7-41+ months (mean, 25) without significant GVHD. Since investigators have used different sources, doses, and time schedules of ATG, we compared our results with other published reports. We conclude that CY/ATG conditioning is well tolerated and effective in children with SAA.

Adolescent

[Bronchogenic carcinoma located in the aortic window].

In the patients with invasion to the aortic window, we performed operation via median sternotomy combined with anteroaxillar thoracotomy. In such patients with T4 invasion, conventional pneumonectomy could not be performed because of the extensive invasion near the main pulmonary artery trunk. In these patients in this study, complete resection of the involved pulmonary artery could be performed using a vascular clamp without CP bypass. Operative technique was as follows: first, the pericardium was opened and taping of the aorta was applied. When the uninvolved part of the intrapericardial pulmonary artery was long enough to cut, we could use a stapling device, but the stapling device could not be used in many cases because the length of the uninvolved segment was too short to cut the left pulmonary artery. In order to carry out complete resection, it was necessary to clamp the central part of the main pulmonary artery diagonally from the left lower side to the right upper side. The pulmonary arterial stump was closed with continuous 4-0 monofilament mattress and over and over suture. We recommend an aggressive surgical approach for the tumor with invasion to the aortic window, because the prognosis is dismal in nonresected locally advanced lung cancer.

Aorta, Thoracic

Juvenile systemic granulomatosis manifesting as premature aging syndrome and renal failure.

We describe a case of juvenile systemic granulomatosis in a 22-year-old woman. The rash consisted of purple papules and first appeared at the age of one year. She had persistent symmetrical painless boggy tenosynovitis with minimal roentgenographic changes and chronic granulomatous symptoms. Uveitis resulted in visual impairment. She also had granulomatous changes in her vessels. Renal impairment developed; however, neither renal artery stenosis nor hypercalcemia was found. Clinical features included the development of premature aging with alopecia, which differed from the previously reported progeria syndrome. Poikiloderma may cause a prematurely aged appearance. Our report expands the clinical spectrum of systemic granulomatosis to include the development of premature aging with alopecia.

Adult

[Relapse after video-assisted thoracoscopic surgery for spontaneous pneumothorax].

Postoperative relapse was noted in 8 (14.8%) of 54 patients who underwent video-assisted thoracoscopic surgery for spontaneous pneumothorax followed for more than one year. Five of the eight patients with relapse were among the 15 initial-stage cases. Concerning treatment, only drainage was performed in 1 patient, and thoracotomy was performed in 5 patients. Findings on additional surgery consisted of missed cystic lesions, in 2 patients, a small opening adjacent to a suture caused by an autosuture device in 1 patient, and a cyst around a suture in patients. There was no missed cystic lesion after the 15th case. In subsequent cases, there were changes at the periphery of a suture. In patients with other diseases, there were no changes observed when an autosuture device was used under thoracoscopic guidance. Therefore, the changes may be associated with hypertonus of the emphysematous lung at the periphery of the site where an autosuture device was used. Thoracoscopic surgery for spontaneous pneumothorax using an autosuture device has limitations such as the presence of a dead angle, insufficient evaluation of emphysematous changes in the collapse lung, a large volume of resected tissue, and hypertonus of the peripheral lung. These limitations may contribute to a high recurrence rate of spontaneous pneumothorax. To reduce the recurrence rate, conditions and problems in cases showing relapse should be clarified before selecting treatment including pleurodesis in individual cases.

Adolescent

Subcellular distribution and phosphorylation of the nuclear localization signal binding protein, NBP60.

We previously purified a nuclear localization signal binding protein, NBP60, from rat liver (1993, J. Biochem. 113, 308-313). In this study, the subcellular localization of NBP60 was examined using anti-NBP60. Most NBP60 was found to be localized in the nuclear envelope fraction of rat liver obtained on cell fractionation followed by immunoblotting. Staining of the nuclei of cultured cells by the antibody was observed on immunofluorescence microscopy. NBP60 was widely detected in rat nuclear fractions prepared from other tissues and also in nuclei of cultured cells derived from other species. It was shown by immunoelectron microscopy that most NBP60 is present in the nuclear envelope and at least some of that is present on nuclear pore complexes. Although NBP60 was localized in the nuclear envelope in interphase cells, it diffused into the cytoplasm in the mitotic phase. The purified NBP60 was highly phosphorylated by a cdc2 mitotic kinase, whereas nuclear pore proteins p144, p62, p60, and p54 were not phosphorylated by the kinase directly. NBP60 was also phosphorylated by protein kinase A, calmodulin-dependent protein kinase II, and casein kinase II. The phosphorylation of NBP60 by cdc2 kinase and/or the other kinases may be related to the change in the protein's location during the mitotic phase.

Animals

Localization of bone morphogenetic protein-4 messenger RNA in developing mouse cochlea.

Bone morphogenetic protein-4 (BMP-4) is a cytokine that belongs to the TGF-beta superfamily. It has been implicated that it plays an important role during embryogenesis including epithelial-mesenchymal interactions and mesenchymal cells condensation. To investigate whether BMP-4 is involved in the embryogenesis of the inner ear, we performed in situ hybridization at various stages of the developing inner ear of mice. BMP-4 mRNA was detected only in the developmental stage. Its initial expression was detected in part of the otic vesicle at 9 days post-coitum (PC). As development proceeded, the hair cells of the cochlear duct were morphologically distinguishable. and BMP-4 expressing cells were found in Claudius cell region at 16 days PC. At this stage, the signal was most intense during development and the structure notably changed under the expressing cells. Mesenchymal cells were assembled and condensed underneath the BMP-4-expressing cells. The basilar membrane, with a rich extracellular matrix and elasticity, would be produced in the area between the Claudius' cells and condensed mesenchymal cells. Previous reports support the notion that BMP-4 expressed in the epithelium causes mesenchymal cell condensation and induces a structure with elasticity. In the developing cochlea. BMP-4 may be involved in the condensation and differentiation of the mesenchymal cells as well as basilar membrane formation. BMP-4 might be an essential factor for the normal development of the basilar membrane.

Animals

Adrenal tumor producing 11-deoxycorticosterone, 18-hydroxy-11-deoxycorticosterone and aldosterone.

A case of adrenal tumor producing 11-deoxycorticosterone, 18-hydroxy-11-deoxycorticosterone and aldosterone is reported. A 55-year-old woman had hypertension, hypokalemia, low plasma renin activity and an adrenal tumor. The plasma level of aldosterone was normal, and the levels of 11-deoxycorticosterone and 18-hydroxy-11-deoxycorticosterone were extremely high. After the tumor removal, the plasma level of aldosterone decreased and plasma levels of 11-deoxycorticosterone and 18-hydroxy-11-deoxycorticosterone were normalized. The tumor was benign adenoma and the production of steroid hormones was under control of adrenocorticotropic hormone. The enzyme activity of 21-hydroxylation in the tumor was elevated and that of 11 beta-hydroxylation was decreased compared with the adjacent tissue.

18-Hydroxydesoxycorticosterone

The Ca(2+)-standardization of Japan only may cause problems for the importation of foreign analyzers.

A serum based reference material for Ca2+ measurement has been established in Japan. cCa2+ in the reference material was assigned with a IFCC reference cell. Some improvements of the IFCC reference cell are described. A few non-Japanese Ca2+ analyzers have given very inaccurate results with the reference material. The Japanese authorities can therefore be expected to hamper their importation in the future. A world-wide external quality assessment scheme with the Japanese reference material is suggested to further identify and solve the problems.

Calcium

Accuracy evaluation and hematocrit effect of commercial ionized calcium analyzers using Japanese serum reference material.

The serum reference material for concentration measurement of ionized calcium in blood has been established by our research group in Japan. The reference method used is the reference standard cell (RSC:IFCC/WGSE, Covington-Umemoto Cell) system. Here we present the results of the evaluation of accuracy using the serum reference material and the hematocrit effect of the nine commercial ionized calcium analyzers being used in routine measurements by Japanese round robin test program. The ionized calcium concentration and pH of two concentration levels of the reference material were analyzed in triplicate measurements according to the standard procedures designated by manufacturers. The effect of hematocrit in ionized calcium concentration measurement was tested using plasma and whole blood with hematocrit values of 35%, 50% and 65%, respectively. The range of bias of ionized calcium values were -0.14 approximately +0.09 mmol/L, -0.12 approximately +0.09 mmol/L for ionized calcium, respectively. The effect of hematocrit in ionized calcium concentration measurement was strongly observed in GEM Premier, CAI-101 and GASTAT-2 analyzers.

Calcium

[Case of intralobar pulmonary sequestration with increased serum sialyl Lewis X-i (SLX)].

A 37-year-old male whose chief complaint was fever and back pain was admitted to our hospital because of a mass shadow in the left lower P3ng field found in X-ray examination. The level of serum SLX on admission increased to 1,338 U/ml. He was diagnosed intralobar pulmonary sequestration by angiography. The left lower lobe was resected 2 months after second admission. His general condition was steady, and he was discharged 18 days after the operation because the level of serum SLX decreased to 49.9 U/ml.

Adult

[A case of right postpneumonectomy empyema treated by open window thoracostomy, thoracoplasty, pedicled omental flap and musculocutaneous flaps].

A case of right postpneumonectomy empyema treated by open window thoracostomy, thoracoplasty, pedicled omental flap and musculocutaneous flaps is reported. After open window thoracostomy, pedicled omental flap and musculocutaneous flaps transplantation utilizing techniques used in plastic surgery were performed, and right postneumonctomy empyema was successfully treated. In this study, open window thoracostomy was effective against depuration of the empyema cavity and omental and musculocutaneous flaps against the infection control and plombage of the cavity. Furthermore, to maximize the effect of musculocutaneous flaps and to use them effectively, it is thought that techniques utilized in plastic surgery are effective and necessary.

Adenocarcinoma

Targeted disruption of the NF-IL6 gene discloses its essential role in bacteria killing and tumor cytotoxicity by macrophages.

To investigate the role of NF-IL6 in vivo, we have generated NF-IL6 (-/-) mice by gene targeting. NF-IL6 (-/-) mice were highly susceptible to infection by Listeria monocytogenes. Electron microscopic observation revealed the escape of a larger number of pathogens from the phagosome to the cytoplasm in activated macrophages from NF-IL6 (-/-) mice. Furthermore, the tumor cytotoxicity of macrophages from NF-IL6 (-/-) mice was severely impaired. However, cytokines involved in macrophage activation, such as TNF and IFN gamma, were induced normally in NF-IL6 (-/-) mice. Nitric oxide (NO) formation was induced to a similar extent in macrophages from both wild-type and NF-IL6 (-/-) mice. These results demonstrate the crucial role of NF-IL6 in macrophage bactericidal and tumoricidal activities as well as the existence of a NO-independent mechanism of these activities. We also demonstrate that NF-IL6 is essential for the induction of G-CSF in macrophages and fibroblasts.

Animals

Hair cell regeneration in the chick inner ear following acoustic trauma: ultrastructural and immunohistochemical studies.

The regeneration of hair cells in the chick inner ear following acoustic trauma was examined using transmission electron microscopy. In addition, the localization of proliferation cell nuclear antigen (PCNA) and basic fibroblast growth factor (b-FGF) was demonstrated immunohistochemically. The auditory sensory epithelium of the normal chick consists of short and tall hair cells and supporting cells. Immediately after noise exposure to a 1500-Hz pure tone at a sound pressure level of 120 decibels for 48 h, all the short hair cells disappeared in the middle region of the auditory epithelium. Twelve hours to 1 day after exposure, mitotic cells, binucleate cells and PCNA-positive supporting cells were observed, and b-FGF immunoreactivity was shown in the supporting cells and glial cells near the habenula perforata. Spindle-shaped hair cells with immature stereocilia and a kinocilium appeared 3 days after exposure; these cells had synaptic connections with the newly developed nerve endings. The spindle-shaped hair cell is considered to be a transitional cell in the lineage of the supporting cell to the mature short hair cell. These results indicate that, after acoustic trauma, the supporting cells divide and differentiate into new short hair cells via spindle-shaped hair cells. Furthermore, it is suggested that b-FGF is related to the proliferation of the supporting cells and the extension of the nerve fibers.

Acoustic Stimulation

Endocytosis and transepithelial transport of endolymph in the endolymphatic sac.

The fate of cationized ferritin (CF) injected into the endolymphatic space of the endolymphatic sac was observed by transmission electron microscopy. At 10 min after the injection, CF particles bound to the apical plasma membrane of epithelial cells of the sac and were then endocytosed with coated pits. However, they never passed through the junctional complexes between the epithelial cells. At 30 min after the injection, the CF particles were transferred to endosomes and lysosomes by small vesicles of 100-150 nm in diameter. CF particles were also found in small vesicles close to Golgi cisternae and in multivesicular bodies. Acid phosphatase positive lysosomes were found close to endosomes containing CF particles. In addition, a small fraction of the small vesicles containing CF particles became inserted into the basolateral plasma membrane. At 60 min after the injection, many CF particles were found in acid phosphatase positive secondary lysosomes. These observations suggest that endocytosis of endolymph is actively performed by the epithelial cells of the sac, and transepithelial vesicular transport across the epithelial cells occurs.

Acid Phosphatase