[Subacute spongiform encephalopathy].
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Animals↗
Biomedical subjects
T Mizutani
Publications and source records attributed to T Mizutani.
Single-step reverse transcriptase-polymerase chain reaction for detection of Borna disease virus RNA in vitro and in vivo.
There are few copies of Borna disease virus (BDV) genome in peripheral blood mononuclear cells and no reliable standard reverse transcriptase-polymerase chain reaction (RT-PCR) method for the detection of BDV RNA, which is both highly sensitive and free of contamination. Single-step RT-PCR, in which both reverse transcription and amplification by Taq DNA polymerase work efficiently in a single buffer, was applied to detect the p24 region of BDV RNA in vitro and in vivo. Using in vitro synthesized RNA, it was demonstrated that at least 100 copies of BDV RNA could be detected and the sensitivity and specificity were nearly equal to those obtained by RT-nested PCR. We could detect BDV RNA from more than 1 pg of cellular RNA obtained from BDV-persistently infected MDCK cells. Furthermore, this method was successfully performed on brain specimens obtained from a BDV-infected rat at 11 weeks post-inoculation. This single-step RT-PCR method will be convenient for detecting limited amounts of BDV RNA in various cells and tissue samples.
Animals↗
[Physiological atrophy and pathological atrophy: morphological aspect in brain aging].
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Adolescent↗
Inhibitory effect of intestinal bacteria on spontaneous multiple polyps in the small intestine of gnotobiotic BALB/c mice.
The incidence of multiple polyps and number of polyps per mouse were significantly lower in conventionalized (CVz), and chloroform-resistant bacteria (CRB)- or fusiform bacteria (FB)-associated mice than in germfree (GF) BALB/c mice. The concentration of fecal fatty acids was also higher in mice associated with either CRB or FB than in GF mice. The incidence of multiple polyps and number of polyps per mouse were significantly correlated with the concentration of fatty acids in the feces of CVz and CRB-GB mice. This study demonstrated that inhibition of multiple polyps in the small intestine of BALB/c mice was affected by the production of intestinal fatty acids.
Animals↗
[A surgically treated case of giant pseudoaneurysm arising as a complication following carotid endarterectomy].
We reported a case of giant pseudoaneurysm (PA) originating at the left extracranial internal carotid artery after carotid endarterectomy (CEA). A 60-year-old man underwent CEA for severe stenosis of the left internal carotid artery. A day after the operation, a left cervical painful and pulsatile mass beneath the operative scar was noticed. Soon after that, the patient showed dyspnea, and intratracheal intubation was carried out. 6 days after CEA, a painful swelling reoccurred. A left carotid angiogram visualized a 5 x 5 cm sized PA in the region of the left carotid bifurcation and demonstrated patency of the two major carotid branches. After a balloon catheter had been inserted and inflated at the carotid bifurcation, the patient underwent a second operation. The bleeding from arteriotomy was well controlled by the inflated balloon catheter at the carotid bifurcation. The continuous SURGILENE suture appeared to be disrupted and a saccular PA was found to have arisen from a defect on the lateral surface of the carotid bifurcation. The suture was completely removed and arteriotomy was closed with a continuous SURGILENE suture again and some single sutures were added. A PA arising as a complication following CEA is rare, but the possibility of PA after CEA should be considered in any patient presenting with a painful or/and pulsatile mass in the neck at any time interval following the operation. Angiography at the early post-op stage is necessary to rule out the various complications after CEA. To control the bleeding, the balloon catheter was thought to be very useful.
Aneurysm, False↗
[Intraoperative video-assisted resection of left atrial myxoma through the right parasternal approach].
A 58-year-old man admitted with atypical chest pain, and an echocardiogram showed a tumor in the left atrium. The tumor widely originated from the interatrial septum and the superior-anterior inner surface of the left atrium did not disturb a mitral valve motion. A right coronary angiogram demonstrated feeding arteries of the tumor. The right parasternal skin incision was made and the tumor was completely removed through the interatrial incision under the video-assisted fiberscopic system. Nd-YAG laser was exposed on the tumor-resected surface for laser coagulation resulting in tumor necrosis. Pathological examination revealed myxoma with rich neovascularization. On the 12th day the patient discharged without complications.
Cardiac Surgical Procedures↗
[Aseptic meningitis induced by high-dose gamma-globulin in an adult woman with antiphospholipid syndrome and chronic idiopathic thrombocytopenic purpura].
It is well known that the administration of high-dose gamma-globulin concentrate is effective in alleviating thrombocytopenia in patients with idiopathic thrombocytopenic purpura (ITP). However, treatment can sometimes induce aseptic meningitis. A 25-year-old Japanese woman with antiphospholipid syndrome and ITP was conditioned with high-dose gamma-globulin concentrate prior to splenectomy. Three days after the initial gamma-globulin course, she experienced severe headache, vomiting, and high-grade fever. Cerebrospinal fluid examination yielded a diagnosis of aseptic meningitis. The patient's clinical symptoms and abnormal cerebrospinal fluid findings disappeared immediately after the discontinuation of gamma-globulin therapy. Cases of aseptic meningitis induced by high-dose gamma-globulin therapy are uncommon in the literature, and most involve children. Moreover, to our knowledge, only 1 Japanese adult ITP case of gamma-globulin-induced aseptic meningitis has been reported to date. Aseptic meningitis may be one of the important adverse effects of the administration of high-dose gamma-globulin concentrate to pediatric as well as adult ITP patients.
Adult↗
[Familial parkinsonism and dementia with ballooned neurons, argyrophilic neuronal inclusions, atypical neurofibrillary tangles, tau-negative astrocytic fibrillary tangles, and Lewy bodies].
We reported a new type of familial Parkinson's disease (familial PD) previously. After that, we examined this family by both the immunohistochemical staining using anti-alpha-synuclein antibody and the analysis of alpha-synuclein gene. We reported these results, and briefly reviewed both familial PD and "familial frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17)". Immunohistochemical staining with anti-alpha-synuclein antibody revealed that the argyrophilic neuronal inclusions and atypical barely tau-positive neurofibrillary tangles were strongly immunoreactive, whereas the ballooned neurons and astrocytic fibrillary tangles were unreactive. DNA analysis of the leukocytes obtained from one live patient of our family did not show any mutations in the entire exons of alpha-synuclein gene. Our results indicated that our family had familial diffuse Lewy body disease with atypical features and without alpha-synuclein gene abnormalities. Features of familial PD included 1) autosomal dominant inheritance, 2) not uncommon atypical clinical features, 3) variable symptomatology and dopa-responsiveness, and 4) low incidence of alpha-synuclein gene abnormalities. Our familial PD showed similar features, and neuropathological findings of our patients also resembled FTPD-17 in the presence of frequent ballooned neurons and neurofibrillary tangles in the cerebral cortex, but were different in the presence of Lewy bodies and paucity of tau pathology.
Humans↗
Inhibition of polyposis in the small intestine of BALB/c mice by intestinal bacteria.
The incidence of polyposis and the number of polyps per mouse were significantly lower in conventionalized (CVz) mice than in germ-free (GF) mice. There was no significant difference in the average number of polyps between GF and gnotobiotic (GB) mice monoassociated with the various strains of intestinal bacteria. However, the incidence of polyposis and the number of polyps per mouse were significantly lower for mice associated with either chloroform-resistant bacteria (CRB) or fusiform bacteria (FB) than for GF mice. This study demonstrated that polyposis was suppressed by FB and CRB in the small intestine of BALB/c mice.
Animals↗
Changes in the expression of protein kinase C (PKC), phospholipases C (PLC) and D (PLD) isoforms in spleen, brain and kidney of the aged rat: RT-PCR and Western blot analysis.
The age-dependent changes of expression of protein kinase C (PKC), phospholipase C (PLC) and phospholipase D (PLD) isozymes were analyzed in spleen, brain and kidney of young-adult (12-16 week-old) and aged (82-88 week-old) rats. The activities of spleen cPKC and nPKC were significantly decreased by nearly 35 and 30% in aged rats compared to those of young adults, respectively (P < 0.05). The level of PKC beta1 was significantly decreased in aged rats as assessed by RT-PCR and Western blot analyses. In aged rat brain where the activity of cPKC was significantly decreased by nearly 25% (P < 0.05), PKC alpha and beta1 isozymes were significantly down-regulated. In kidney, the level of PKC beta2 was decreased. In spleen the both mRNA and protein levels of PLC beta2 and gamma2 were significantly down-regulated in aged rat (P < 0.05). PLC beta1 was also significantly lower in aged rat brain (P < 0.05) as assessed by RT-PCR and Western blotting. Moreover, PLC beta1 was significantly down-regulated in both mRNA and protein levels in aged rat kidney (P < 0.05). In contrast, the tissues examined, the expressions of PLD isozymes (PLD1a, 1b and 2) were rather stable in the course of aging. These results indicate that mRNAs of PLD isozymes were rather stable but that particular PKC and PLC isozymes were down-regulated in different tissues during aging, suggesting age-dependent decline of specific PKC and PLC isozymes in organs which may, at least in part, be implicated in tissue dysfunction with aging.
Aging↗
Oxidative DNA damage and apoptosis induced by metabolites of butylated hydroxytoluene.
DNA damage by metabolites of a food additive, butylated hydroxytoluene (BHT), was investigated as a potential mechanism of carcinogenicity. The mechanism of DNA damage by 2,6-di-tert-butyl-p-benzoquinone (BHT-quinone), 2,6-di-tert-butyl-4-hydroperoxyl-4-methyl-2,5-cyclohexadienone (BHT-OOH), and 3,5-di-tert-butyl-4-hydroxybenzaldehyde (BHT-CHO) in the presence of metal ions was investigated by using 32P-labeled DNA fragments obtained from the c-Ha-ras-1 proto-oncogene and the p53 tumor suppressor gene. BHT-OOH caused DNA damage in the presence of Cu(II), whereas BHT-quinone and BHT-CHO did not. However, BHT-quinone did induce DNA damage in the presence of NADH and Cu(II). Bathocuproine inhibited Cu(II)-mediated DNA damage, indicating the participation of Cu(I) in the process. Catalase also inhibited DNA damage induced by BHT-quinone, but not that induced by BHT-OOH. The DNA cleavage pattern observed with BHT-quinone plus NADH was different from that seen with BHT-OOH. With BHT-quinone plus NADH, piperidine-labile sites could be generated at nucleotides other than adenine residue. BHT-OOH caused cleavage specifically at guanine residues. Pulsed field gel electrophoresis showed that BHT-OOH and BHT-quinone induced DNA strand breaks in cultured cells, whereas BHT-CHO did not. Both BHT-quinone and BHT-OOH induced internucleosomal DNA fragmentation, which is the characteristic of apoptosis. Furthermore, flow cytometry analysis revealed an increase of peroxides in cultured cells treated with BHT-OOH or BHT-quinone. These results suggest that BHT-OOH participates in oxidative DNA damage directly, whereas BHT-quinone causes DNA damage through H2O2 generation, which leads to internucleosomal DNA fragmentation.
Apoptosis↗
A G.U base pair in the eukaryotic selenocysteine tRNA is important for interaction with SePF, the putative selenocysteine-specific elongation factor.
In Escherichia coli, selenocysteine biosynthesis and incorporation into selenoproteins requires the action of four gene products, including the specialized selenocysteine tRNA(Sec) and elongation factor SELB, different from the universal EF-Tu. In this regard, the situation is less clear in eukaryotes, but we previously reported the existence of SePF, a putative SELB homologue. The secondary structure of the tRNA(Sec) differs slightly in eukaryotes, due to a change in the lengths of several stems. Two non-Watson-Crick base pairs, G5a x U67b and U6 x U67, reside in the acceptor stem and are conserved in the course of evolution. Since it has already been reported that changing them to Watson-Crick base pairs did not affect the serylation or selenylation levels of tRNA(Sec), we asked whether these non-Watson-Crick base pairs are required for the interaction with SePF. To this end, tRNA(Sec) variants carrying Watson-Crick changes at these positions were tested for their ability to maintain the interaction with SePF. In these assays, the tRNA(Sec)-SePF interaction was determined by the protective action it confers against hydrolysis of the amino acid ester bond, under basic conditions. All the changes introduced at U6 x U67 did not significantly affect the interaction. Interestingly, however, the G5a x U67b to G5a-C67b substitution was sufficient, by itself, to lead to unprotection of the ester bond. Therefore, our finding strongly suggests that SePF is unable to interact with a tRNA(Sec) mutant version carrying a Watson-Crick G5a-C67b instead of the wild-type G5a x U67b base pair, establishing that G5a x U67b constitutes a structural determinant for SePF interaction.
Animals↗
Reversible hypothyroidism with corticotropin releasing hormone deficiency.
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Aged↗
X-ray absorption of bromonaphthalene dissolved in supercritical fluid xenon.
The results of Br K-edge X-ray absorption measurements of 1-bromonaphthalene dissolved in supercritical fluid Xe are reported. As the pressure of Xe confined in a high-pressure cell is increased, the absorption spectrum of bromonaphthalene gradually appears, showing that Xe in the supercritical fluid state solvates the bromonaphthalene molecule. The spectrum of the dissolved sample shows a remarkable difference from that of the pure liquid sample in the near-edge region of the X-ray absorption spectrum, reflecting the interaction of the solute material with solvent Xe. Ab initio full multiple-scattering calculations can reproduce the spectral features qualitatively. A large value of the Debye-Waller factor must be introduced to give the best fit, corresponding to a large degree of spatial disorder and violent thermal vibration of the Xe atoms around the Br atom.
Journal Article↗
Molecular cloning and functional expression of chicken luteinizing hormone receptor.
A complementary DNA for chicken luteinizing hormone (LH) receptor containing the entire coding region was isolated from chicken F1 granulosa cell cDNA library. Nucleotide sequence analysis revealed that there are characteristic GC-rich regions around the N-terminal part. Chicken LH receptor consists of a 19-residue signal peptide, a 366-residue extracellular domain, a 267-residue region containing seven transmembrane segments, and a 76-residue cytoplasmic C-terminal tail. The deduced amino acid sequence of the chicken LH receptor shares 67%, 69%, and 69% identity with the human, rat and porcine LH receptor sequences, respectively, and 51% with chicken FSH receptor. However, an insertion of about 30 amino acid residues is found in chicken LH receptor in the extracellular domain about 44 amino acid residues upstream of the first transmembrane segment. In addition, alternative splicing seems likely to occur at the point where the insertion starts (nucleotide position 933), resulting in the truncated forms of chicken LH receptor with only the extracellular domain. Northern blot analysis revealed the presence of multiple transcripts of LH receptor, a major 3.0-kb and minor 7-kb and 1.5-kb bands, in chicken F1 to F3 granulosa cells. The full length chicken LH receptor cDNA was transiently expressed in COS-7 cells and the transfected cells displayed a concentration-dependent increase in cAMP production when exposed to varying concentrations of chicken LH. This clearly indicates that the cloned cDNA encodes a functional chicken LH receptor protein.
Amino Acid Sequence↗
Intramuscular injection of plasmid DNA expressing mRNA7 coding the nucleocapsid protein of JHMV partially protected mice against acute infection with JHMV.
We constructed a plasmid expressing mRNA7 coding the nucleocapsid (N) protein of JHM strain of mouse hepatitis virus (JHMV) under the control of Rous sarcoma virus LTR, referred to as pRSV-mRNA7. When C57BL/6 mice injected intramuscularly (i.m.) with control plasmid DNA which contained no viral sequence were infected with JHMV, necrotic figures of neural cells and diffuse immersion of lymphatic cells in the pedunculus cerebri were observed. In the hypothalamus, vascular cuffing consisting of lymphatic cells was observed. In contrast, no histological change was observed throughout these areas of the brains in the JHMV-infected mice after injection with pRSV-mRNA7. These results showed that the injection with plasmid DNA expressing mRNA7 of JHMV partially protected mice against acute infection with JHMV in the brain. The plasmid DNA was i.m. injected into mice and the cytolytic activity of spleen cells from the mice was assessed by 51Cr-release assay. The spleen cells from the mice administrated with pRSV-mRNA7 showed a significant level of cytolytic activities against the transfected cells expressing the viral N protein even though the spleen cells were not cocultivated with stimulator cells. When the spleen cells from administrated mice with pRSV-mRNA7 were cocultivated with stimulator cells, higher cytolytic activity was observed against the transfected cells, compared with the activity without stimulation.
Acute Disease↗
Muscular dystrophy: centronucleation may reflect a compensatory activation of defective myonuclei.
Muscular dystrophy has long been believed to be characterized by degeneration and abortive regeneration of muscle fibers (the muscle degeneration theory), but unfortunately its pathogenesis is still unclear and an effective treatment has yet to be developed. As a challenge to the theory, we have proposed an alternative muscle-defective-growth theory and a further bone muscle growth imbalance hypothesis supposing possible defects in bone-growth-dependent muscle growth based on our findings in hereditary dystrophic dy mice (C57BL/6J dy/dy). This review presents some new insights into the pathogenesis of the disease along with our hypothesis, focusing on the physiological meaning of centronucleation, one of the major pathological changes commonly observed in dystrophic muscles of man and experimental animals.
Animals↗
Familial parkinsonism and dementia with ballooned neurons, argyrophilic neuronal inclusions, atypical neurofibrillary tangles, tau-negative astrocytic fibrillary tangles, and Lewy bodies.
We report four patients with a new type of familial parkinsonism and dementia consisting of an autosomal dominant inheritance, dopa-responsive parkinsonism, severe dementia, variable myoclonus and autonomic disturbances. Autopsy of two patients revealed symmetrical cerebral atrophy with fronto-temporal dominant distribution, and marked depigmentation in the substantia nigra and locus ceruleus. Neuronal loss and gliosis were observed in the deep cerebral cortex and amygdala as well as in the areas vulnerable to Parkinson's disease. In the cerebral cortex, swollen neurons with frequent granulovacuolar changes were observed, consisting of ballooned neurons and those with argyrophilic intracytoplasmic inclusions, in addition to neuropil threads. Atypical neurofibrillary tangles, which barely stained with tau antibodies, were numerous in the upper cortical layers, consisting of 15-nm straight tubules. In addition, tau-negative astrocytic fibrillary tangles were also frequent. Electron microscopically, the ballooned neurons and argyrophilic neuronal inclusions contained filamentous structures coated with fuzzy electron-dense deposits. The inclusions showed immunohistochemical features different from those of cortical Lewy bodies and Pick bodies. Occasional Lewy bodies were present in the brain stem lesions of both patients. In two of our patients, the pathology in the brain stem was similar to that of Parkinson's disease, whereas their cerebral pathology was unusual and has not been reported previously.
Adult↗