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Biomedical subjects

M Morita

Publications and source records attributed to M Morita.

At least 127 records · Page 7Linked to original sources
PubMed2000

Peroxisomal membrane protein Pmp47 is essential in the metabolism of middle-chain fatty acid in yeast peroxisomes and Is associated with peroxisome proliferation.

Pmp47 of the methylotrophic yeast Candida boidinii belongs to a mitochondrial family of solute transporters and is localized in peroxisomal membranes. Its human homolog, Pmp34, is also known. In this study, we characterized the role of Pmp47 in fatty acid metabolism and peroxisome proliferation using the PMP47-deleted strain of C. boidinii (strain pmp47Delta). The wild-type strain grew well on a middle-chain fatty acid, laureate, as the single carbon source, and mild peroxisome proliferation was observed during its growth. The pmp47Delta strain could not grow on laureate but could grow on long-chain fatty acids including palmitate, myristate, and oleate. The levels of laureate oxidation activity in intact cells and in semi-permeabilized cells of strain pmp47Delta were lower than the respective level in the wild-type strain, although the level of laureate oxidation activity in the cell lysate and the level of lauroyl-CoA oxidation in semi-permeabilized cells of strain pmp47Delta were indistinguishable from the respective level in the wild-type strain. When lauroyl-CoA was provided in the cytosol of strain pmp47Delta through expression of Saccharomyces cerevisiae Faa2p (lauroyl-CoA synthetase) in which its peroxisome targeting signal was deleted, the growth of strain pmp47Delta on laureate was recovered to the level of growth of the wild-type strain. Laureate is converted to its CoA form in peroxisomes by the action of lauroyl-CoA synthetase. These results suggested that Pmp47 is involved in the transport of a small molecule (possibly ATP) required in the conversion of laureate to its CoA form in peroxisomes and that the absence of Pmp47 causes impairment of laureate metabolism, which results in the inability of pmp47Delta cells to grow on laureate. In addition, Pmp47 may be involved in peroxisome proliferation, because the pmp47Delta strain contained a reduced number of peroxisomes, as judged from the fluorescence analysis of cells expressing green fluorescent protein tagged with the peroxisome targeting signal 1 (GFP-AKL).

Candida↗
PubMed2000

A frozen newborn infant: froth in the air-passage after thawing.

We performed an autopsy on a frozen newborn infant who was found in a freezer at -18 degrees C. After thawing, froth emerged from the nostrils and was present in the trachea. Sometimes froth may be seen in the air-passage in cases of strangulation and drowning. In our case, however, there was neither proof of asphyxia due to strangulation nor drowning. The existence of the froth indicates that the infant was probably in a state of respiratory distress before death. Histologic findings of the lung showed that the infant did not suffer from respiratory disorders such as respiratory distress syndrome. Karyopyknosis and vacuolation of the keratinocytes, shrinkage of the hepatocytes, dilatation of the sinusoid, spaces between heart muscle fibers and deep staining of the nuclei and hemolysis were characteristic in our case. This case shows that froth persists in the internal air-passage for a long time as a result of freezing. Moreover, the froth in the air-passage, along with the findings of the lungs, demonstrates that the newborn infant was born alive.

Autopsy↗
PubMed2000

Functional reconstitution of an active recombinant human chymase from Pichia pastoris cell lysate.

We have previously reported efficient production of mature human chymase (h-chymase) using an original system of expression in Pichia pastoris (Nakakubo et al., 2000), whereby recombinant h-chymase (rh-chymase) was secreted as a mature form with the correct N-terminal amino acid sequence and was easily purified. In the course of investigation of secretory rh-chymase, we also found large amounts of chymase to be present in insoluble form in the transformant cell. Although the cellular rh-chymase had no proteolytic activity, its chymotryptic activity was restored in a reconstitution process utilizing guanidine and glutathione. As with secretory rh-chymase, efficient purification was possible by heparin affinity chromatography. The purified cellular rh-chymase showed the same mobility as secretory rh-chymase in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) before and after deglycosylation. N-terminal amino acid sequence analysis revealed that the signal peptide had been correctly removed. K(m) value (5.93 mM), as well as pH profile and inhibition profile toward protease inhibitors of reconstituted cellular rh-chymase, indicated that the rh-chymase enzymatically closely resembles native h-chymase. Furthermore, it showed a greatly restricted proteolytic activity towards Ang I, and formed Ang II without the further cleavage which is a feature of h-chymase. It was thus found that the insoluble rh-chymase stored in the cells could be solubilized and reconstituted to give the same structure as h-chymase, not only in terms of enzyme active site but also of substrate recognition site.

Amidohydrolases↗
PubMed2000

Expression of keratan sulfate at the arthroplasty surface after cup arthroplasty.

Fibrous tissue which regenerated on the acetabular arthroplasty surface was obtained from a 52-year-old woman who underwent total hip replacement after cup arthroplasty. The histological features of this newly formed fibrous tissue and expression of keratan sulfate, which is a characteristic matrix component of articular cartilage, were studied. Microscopic observation revealed that the arthroplasty surface consisted mainly of fibrous tissue which did not show metachromasia with toluidine blue staining, but there were many nodular structures communicating with the bone marrow. Immunostaining for keratan sulfate revealed clear positive staining around the cells of the nodular structures communicating with the bone marrow, while only weakly positive staining was observed in the superficial layer of the loose fibrous tissue. The present study revealed marked formation of articular cartilaginous tissue in areas having good communication with the bone marrow, which indicates that maintenance of this communication may be necessary to improve the outcome of cup arthroplasty.

Arthroplasty, Replacement, Hip↗
PubMed2000

Hyaluronan in synovial fluid of patients with loose total hip prosthesis. Comparison with hyaluronan in patients with hip osteoarthritis and idiopathic osteonecrosis of femoral head.

The concentration and molecular weight of hyaluronan (HA) in the synovial fluid of the hip joint were determined in 13 patients (aged 62.8 +/- 9.4 years) who had undergone prior total hip arthroplasty(THA), 23 patients (aged 65.0 +/- 8.2 years) with osteoarthritis of the hip joint (OA), and 13 patients (aged 40.2 +/- 2.7 years) with idiopathic osteonecrosis of the femoral head (ION). A sample of synovial fluid was obtained during revision THA because of loosening of the total hip prosthesis for the THA group, and during the first replacement surgery or osteotomy for the OA and ION groups. The concentration of HA in the synovial fluid was 0.64 +/- 0.42 mg/ml in the THA group, 1.07 +/- 0.28 mg/ml in the OA group, and 1.30 +/- 0.56 mg/ml in the ION group. The concentration of HA in the synovial fluid of the THA patients was significantly lower than that of the OA and ION patients (P = 0.0156 vs OA, P = 0.003 vs ION). The molecular weight of HA was 309 +/- 88.3 x 10(4) Da in the THA group, 377 +/- 201 x 10(4) Da in the OA group, and 240 +/- 148 x 10(4) Da in the ION group; these values do not differ significantly (P = 0.259 vs OA, P = 0.174 vs ION). Among the THA patients, there was no relation between the concentration of HA and the age of the patient, length of time since the first operation, or type of prosthesis fixation; there was also no relation between the molecular weight of HA and each of these factors. These results suggest that a pseudosynovial membrane is regenerated after THA, and that it produces HA of the same molecular weight as that in patients with OA and ION, although in smaller quantities.

Adult↗
PubMed2000

A fatal case of infantile scurvy.

We report a case of infant death due to scurvy, which is very rare in Japan. We initially had little knowledge of the disease and suspected that the bleeding in the body was caused by domestic violence. The case did not fall under the category of the battered child syndrome but the death was caused by ignorance with respect to child care. In addition the parents usually locked the child alone in a room during the day and this is probably a case of neglect.

Autopsy↗
PubMed2000

Dechlorination of hexachlorobiphenyl by using potassium-sodium alloy.

2,2',4,4',5,5'-Hexachlorobiphenyl (HCB) was dechlorinated with potassium-sodium (K-Na) alloy under an inert gas atmosphere. Solvent effect was observed in the reaction. Dechlorination yields in benzene and cyclohexane were 99.9998%, and 99.99996%, respectively. The reaction was exothermic and proceeded at room temperature. In benzene, trace amounts of polychlorinated biphenyls (PCBs) as products by stepwise dechlorination and polychlorinated quarterphenyls as product of Wurtz-Fittig reaction were detected as reaction intermediate. Reaction products were biphenyl, cyclohexylbenzene, and dicyclohexyl. In cyclohexane, there were no products of Wurtz-Fittig reaction. Dechlorination at para-position preferred to that at ortho-position, judging from analysis of PCBs as intermediates of stepwise dechlorination.

Alloys↗
PubMed2000

A new, simple method of making a spacer in interstitial brachytherapy for mobile tongue cancer.

OBJECTIVES: This article demonstrates a new method of making a spacer that increases the distance between the mandible and implanted radioactive sources in interstitial brachytherapy for patients with mobile tongue cancer. STUDY DESIGN: Fifty-three patients with mobile tongue cancer underwent interstitial brachytherapy with spacers made by this new technique. RESULTS: Our spacer is not difficult to create or to use. The spacer was made from a plastic splint by using thermoforming techniques and quick self-curing resin, which did not need waxing, wiring, or casting. The surface of the spacer, which comes in contact with the tongue, is smooth because it is covered with tissue-conditioning material. There were no complaints of pain from the patients. Osteoradionecrosis of the mandible developed in only 1 (1.9%) of these patients. CONCLUSIONS: This spacer is simple to make and prevented osteoradionecrosis.

Brachytherapy↗
PubMed2000

Photosynthetic production of microalgal biomass in a raceway system under greenhouse conditions in Sendai city.

The photosynthetic productivities of the marine microalgae Chlorophyta sp. and freshwater microalgae Chlorella sp. were investigated in a raceway system under greenhouse conditions in Sendai city. The system was constructed with a surface area of 0.986 m2 and equipped with paddle wheels (8 fins). The semicontinuous batch culture experiment was carried out for 4 months from June to October 1998 in Sendai city. The productivity of Chlorophyta sp. was ranged from 4 to 13 g.m(-2).d(-1) and the average was 8.2 g.m(-2).d(-1), which corresponded to the photosynthetic efficiency of 4.15% (PAR). The productivity of Chlorella sp. was ranged from 7 to 21 g.m(-2).d(-1) and the average was 13.2 g.m(-2).d(-1), which corresponded to the photosynthetic efficiency of 6.56% (PAR). These results indicate that stable microalgal cultivation with comparatively high photosynthetic efficiency could be obtained in a raceway system under greenhouse conditions in Sendai city located in the northern part of Japan.

Journal Article↗
PubMed2000

Molecular modeling of human serum transferrin for rationalizing the changes in its physicochemical properties induced by iron binding. Implication of the mechanism of binding to its receptor.

In order to rationalize the physicochemical properties of human serum-transferrin (STf) and the STf-receptor (TfR) recognition process, we have tried to predict the 3D structures of apo- and iron-loaded STf using a homology modeling technique to study the changes in the structural characteristics that take place upon the uptake of iron by STf in solution. The crystal structures of both forms for ovotransferrin were used as templates for the STf modeling. The modeled structure of STf gave a satisfactory interpretation for the typical physicochemical properties such that (1) STf has a negative electrophoretic mobility and its value increases with iron uptake, and (2) the radius of gyration Rg of Tf decreases with iron uptake. It was found that upon iron binding, interdomain closures take place with large movements of the NII and CII subdomains comprising the N- and C-lobes in STf through a hinge-bending motion, accompanied by the opening of the bridge region with a displacement of more than 15 A. Moreover, in view of the findings from our capillary electrophoresis experiments that the electrostatic interactions significantly contribute to a specific binding of Fe2-STf with TfR, it is inferred that the connecting (bridge) and its neighboring region associated with a surface exposure of negative charge play an important role in the STf-receptor recognition process.

Amino Acid Sequence↗
PubMed2000

Usefulness of quantitative real-time polymerase chain reaction in following up patients with Epstein-Barr virus infection after liver transplantation.

BACKGROUND: Post-transplant lymphoproliferative disease (PTLD), which is mainly induced by Epstein-Barr virus (EBV) infection, is a cause of significant morbidity and mortality for patients undergoing liver transplantation, especially when it is detected at such an advanced stage as monoclonal malignant lymphoma. METHODS: In this series, 6 of 22 liver transplant patients suffered from EBV infection. We tested quantitative DNA (Qt-DNA) by real-time polymerase chain reaction (PCR), qualitative DNA in plasma (Q1-pDNA) by PCR, and EBV-encoded mRNA 1 (EBER 1) by in situ hybridization to clarify which of them is a better marker for the early diagnosis and prediction of EBV-associated disorders. RESULTS: Four had signs or symptoms of PTLD, but 2 did not develop individualized lymphoid lesions. In all patients, both Qt-DNA and EBER 1 exceeded the cut-off level of 10(2.5) copies/microg DNA and 0.002%, respectively, at the time of diagnosis. In 2 patients, when Qt-DNA had a poor decline, EBER 1, even if it seemed to decrease after antiviral therapy, increased again after a few months and the clinical symptoms recurred. In 2 patients, Qt-DNA and EBER 1 increased again after a few months of antiviral therapy, and Q1-pDNA remained positive, whereas, in 3 patients, no reaction of EBV could be detected once Q1-pDNA became negative, even after the cessation of therapy. CONCLUSIONS: These results suggest that real-time PCR for Qt-DNA was more sensitive to the real-time activity of EBV and that Q1-pDNA could indicate when to stop antiviral therapy.

Child, Preschool↗
PubMed2000

Adverse effects of an active fragment of parathyroid hormone on rat hippocampal organotypic cultures.

Adverse effects of an active fragment of parathyroid hormone (PTH(1 - 34)), a blood Ca(2+) level-regulating hormone, were examined using rat hippocampal slices in organotypic culture. Exposure of cultured slice preparations to 0.1 microM PTH(1 - 34) for 60 min resulted in a gradual increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)); this effect was most obvious in the apical dendritic region of CA1 subfield. When PTH(1 - 34) at a lower concentration (1 nM) was added to the culture medium and its toxic effects examined using a propidium iodide intercalation method, significant toxicity was seen 3 days after exposure and increased with time. Cells in the CA1 region seemed more vulnerable to the hormone than cells in other regions. At 1 week of exposure, the toxic effects were dose-dependent over the range of 0.1 pM to 0.1 microM, the minimum effective dose being 10 pM. The adverse effects were not induced either by the inactive fragment, PTH(39 - 84), or by an active fragment of PTH-related peptide (PTHrP(1 - 34)), an intrinsic ligand of the brain PTH receptor. The PTH(1 - 34)-induced adverse effects were significantly inhibited by co-administration of 10 microM nifedipine, an L-type Ca(2+) channel blocker, but not by co-administration of blockers of the other types of Ca(2+) channel. The present study demonstrates that sustained high levels of PTH in the brain might cause degeneration of specific brain regions due to Ca(2+) overloading via activation of dihydropyridine-sensitive Ca(2+) channels, and suggests that PTH may be a risk factor for senile dementia. British Journal of Pharmacology (2000) 129, 21 - 28

Animals↗
PubMed2000

Rehabilitation of biting abilities in patients with different types of dental prostheses.

This study investigated the masticatory rehabilitation of subjects wearing different types of prostheses. Biting abilities per person (biting force, biting pressure and occlusal contact area) were assessed with a pressure detecting sheet (Prescale(R)). Five hundred and ninety volunteers were divided into four groups according to the type of posterior dentition: complete denture, removable partial denture, fixed partial denture, and full natural dentition groups. The biting forces of the fixed partial, removable partial and complete denture wearers were 80, 35 and 11% respectively, when expressed as a percentage of the subjects with a natural dentition. The complete denture wearers showed the highest biting pressure among the four groups, followed by the removable partial denture wearers. In a clinical intra-individual study, the biting abilities of 85 subjects, without (before insertion of) and with (after insertion of) renewed prostheses, were compared. No significant differences were found between biting before and immediately after insertion of the prostheses. However, the biting force and occlusal contact area increased 2 months after insertion of the prostheses. This study confirmed past clinical studies indicating an impaired masticatory function of denture wearers. The functional adaptation to new prostheses had improved at evaluation 2 months after insertion.

Adaptation, Physiological↗
PubMed2000

Molecular cloning and functional expression of triterpene synthases from pea (Pisum sativum) new alpha-amyrin-producing enzyme is a multifunctional triterpene synthase.

Ursane type triterpene is one of the most widespread triterpene aglycones found in plants, together with oleanane type, and these two types often occur together in the same plant. Pisum sativum is known to produce both types of triterpenes. Homology based PCRs with degenerate primers designed from the conserved sequences found in the known beta-amyrin synthases have resulted in cloning of two triterpene synthase cDNAs from immature seeds of P. sativum. They show high sequence identities to each other (78%) and also to the known beta-amyrin synthases (70-90%). ORFs of the full-length clones named as PSY (2277 bp, codes for 759 amino acids) and PSM (2295 bp, codes for 765 amino acids) were ligated into the yeast expression vector pYES2 under the control of GAL1 promoter. Heterologous expression in yeast revealed PSY to be a P. sativum beta-amyrin synthase. Surprisingly, however, PSM turned out to be a novel mixed amyrin synthase producing both alpha- and beta-amyrin. Several minor triterpenes were also identified as the PSM byproducts. The presence of such multifunctional triterpene synthase would account for the co-occurence of ursane and oleanane type triterpenes in plants.

Amino Acid Sequence↗
PubMed2000

Glutamate receptor subunit delta2 is highly expressed in a novel population of glial-like cells in rat pineal glands in culture.

The mammalian pineal gland uses L-glutamate as an intercellular chemical transmitter to regulate negatively melatonin synthesis. To receive glutamate signals, pinealocytes express at least three kinds of glutamate receptors: metabotropic receptor types 3 and 5 and an ionotropic receptor, GluR1. In this study, we examined whether or not the fourth class of ionotropic receptor, delta, which is known for its nondefinitive molecular function and its unique expression pattern in brain, is expressed in pineal gland. RT-PCR analyses with specific probes indicated the expression of mRNA of delta2 but not that of delta1 in pineal gland and cultured pineal cells. Western blotting analysis with polyclonal antibodies specific to the carboxyl-terminal region of the delta2 receptor recognized a single 110-kDa polypeptide of cerebellar membranes and specifically immunostained Purkinje cells. The delta2 antibodies recognized a 110-kDa polypeptide of pineal membranes and specifically immunostained huge glial-like cells with the occasional presence of several long, branching processes in a pineal cell culture. delta2 is not uniformly distributed throughout the cells and is relatively abundant at the periphery of the cell bodies and long processes, where the terminals of synaptophysin-positive processes of pinealocytes, a site for glutamate secretion, are frequently present. The delta2-positive cells constitute a very minor population among total pineal cells (approximately 0.03%). Double immunolabeling with delta2 antibodies and antibodies against marker proteins for pineal interstitial cells clearly distinguishes delta2-positive pineal cells and other known interstitial cells, including glial fibrillary acidic protein- or vimentin-positive glial-like cells. These results indicated that the delta2 glutamate receptor is expressed in a novel subpopulation of pineal glial-like cells in culture and suggest the presence of a glutamate-mediated intercellular signal transduction mechanism between pinealocytes and delta2-expressing cells. The pineal cells may provide a good experimental system for studies on the function of glutamate receptor delta2.

Amino Acid Sequence↗
PubMed2000

Development of a completely closed circuit using an air filter in a drainage circuit for minimally invasive cardiac surgery.

The completely closed circuit system is the future direction of cardiopulmonary bypass because of its compactness and superior biocompatibility. The most serious obstacle for clinical application is the sucking of air bubbles into the drainage circuit. The purpose of this study was to remove the air bubbles from the drainage circuit. Infusing 50 ml/min of air bubbles into the drainage circuit of the usual closed circuit, and infusing 50, 100, and 150 ml/min of air into the drainage circuit of a newly developed closed circuit (drainage circuit using an air filter), the number and size of air bubbles were observed at the outlet of the arterial filter. In the usual closed circuit, many air bubbles of over 40 microm were detected within 5 s at a blood flow of 4 L/min because the centrifugal pump decreased the size of the bubbles, which then passed through the oxygenator and arterial filter. Air bubbles of over 40 micro were not detected in the newly developed closed circuit within 5 min at a blood flow of 4 L/min. The removal of air mixed into the completely closed circuit was possible with a drainage circuit using an air filter that was developed. The clinical use of the completely closed circuit for minimally invasive cardiac surgery (MICS) became possible based on this development.

Air↗