Biomedical subjects

A Nakamura

Publications and source records attributed to A Nakamura.

At least 55 records · Page 3Linked to original sources

PubMed2001

Two distinct isopentenyl diphosphate isomerases in cytosol and plastid are differentially induced by environmental stresses in tobacco.

Two distinct cDNA clones (IPI1 and IPI2) encoding IPI were isolated from Nicotiana tabacum. In situ expression of isopentenyl diphosphate isomerase-1 (IPI1)- and IPI2-green fluorescent protein fusion constructs revealed that IPI1 and IPI2 were localized in chloroplast and cytosol, respectively. The level of IPI1 mRNA was increased under high-salt and high-light stress conditions, while that of IPI2 mRNA was increased under high-salt and cold stress conditions. Both IPI transcripts were increased in an abscisic acid-independent manner. This is the first report of a cytosolic IPI. The results indicated that two distinct IPIs were differentially induced in response to stress.

Abscisic Acid↗

PubMed2001

Stereotyped stepping associated with lesions in the bilateral medial frontoparietal cortices.

The location of higher locomotion centers in humans is not well defined. The authors describe stereotyped bipedal stepping movement in a patient with meningoencephalitis with necrotic lesions localized mainly in the bilateral medial frontoparietal cortices. The patient intermittently showed stereotyped stepping in the supine position during a stuporous state. The stereotyped stepping in this patient offers clinical evidence indicating the presence of a higher locomotion center in medial frontoparietal cortex.

Cerebral Cortex↗

PubMed2001

Presence of mitochondria-type ribosomes outside mitochondria in germ plasm of Drosophila embryos.

Mitochondrially encoded large and small ribosomal RNAs (mtlrRNA and mtsrRNA) are transported out of mitochondria to polar granules, the distinctive organelles of germ plasm in Drosophila. Reduction of the extramitochondrial mtlrRNA amount leads to the failure of embryos to form the germ-line progenitors, or pole cells, suggesting that mtlrRNA, along with mtsrRNA, functions on the polar granules to specify the germ line. In this study, we provide several lines of evidence showing that there are mitochondria-type ribosomes on the polar granules during a short period before pole cell formation. Our ultrastructural analysis reveals that these ribosomes include both mitochondrial rRNAs and at least two mitochondrial ribosomal proteins (S12 and L7/L12). Furthermore, these ribosomes are integrated into well developed polysomes on the surface of polar granules. We propose that translation dependent on mitochondria-type ribosomes is an important mechanism underlying germ-line formation.

Animals↗

PubMed2001

Green tea catechins enhance tumor development in the colon without effects in the lung or thyroid after pretreatment with 1,2-Dimethylhydrazine or 2,2'-dihydroxy-di-n-propylnitrosamine in male F344 rats.

Modifying effects of green tea catechins (GTCs) on the post-initiation stage of colon, lung and thyroid carcinogenesis were examined in F344 male rats. Groups of 20 animals were given subcutaneous injections of 40 mg/kg body wt of 1,2-dimethylhydrazine twice a week for 2 weeks or oral administration of 0.1% 2,2'-dihydroxy-di-n-propylnitrosamine (DHPN) in the drinking water for 2 weeks for initiation. They then received diet containing 1 or 0.1% green tea catechin or basal diet alone for 33 weeks. Histopathological examination after final sacrifice showed that although total incidence and multiplicity of colon tumors were not significantly different from controls, values for colon adenomas were decreased while those for carcinomas and the average size of tumors were significantly increased in the 0.1% GTC group. A similar tendency was observed for the 1% GTC group. Incidences and/or multiplicity of lung hyperplasia and tumors, and thyroid lesions did not significantly vary among the DHPN-treated groups. These results indicate that GTCs do not inhibit, but rather may enhance colon carcinogenesis, while not influencing lung and thyroid carcinogenesis under the present experimental conditions.

1,2-Dimethylhydrazine↗

PubMed2001

A highly sensitive method for measurement of myosin ATPase activity by reversed-phase high-performance liquid chromatography.

A new method for measurement of myosin ATPase activity has been developed utilizing reversed-phase high-performance liquid chromatography (HPLC), which detects as low as 0.05 nmol of ADP hydrolyzed from ATP. After termination of the ATPase reaction by addition of perchloric acid, the hydrolysate ADP and substrate ATP were separated by reversed-phase HPLC. The absorbance of ADP was monitored at 259 nm, and the amount of ADP was quantified from its peak area on the chromatogram by use of the NIH Image computer software. Our method showed linearity over a wide range from 0.05 to 10 nmol of ADP per 20 microl with a coefficient of determination (r(2)) of 0.99. Myosin ATPase activities determined by the HPLC method were almost identical to those determined by the malachite green method, a widely used spectrophotometric method with range of detection from 1 to 8 nmol of phosphate. Because our method requires only a small volume of reaction solution, it will be a powerful tool for measuring ATPase activity of motor proteins, which are difficult to obtain in large amount.

Adenosine Diphosphate↗

PubMed2001

Chemical and biological evaluation of endotoxin contamination on natural rubber latex products.

Relationship between pyrogenicity and bacterial endotoxin contamination on latex products was demonstrated by chemical analysis and biological assays. In commercially available latex products' surveillance, water extracts prepared from one surgical glove and two silicone elastomer-coated Foley catheters sterilized by gamma-irradiation were obviously pyrogenic in rabbits. The induced fever was monophasic at low dose of the pyrogenic extracts and biphasic at high dose. These extracts exhibited limulus amebocyte lysate gelation activity, and induced inflammatory cytokine (interleukin-1, interleukin-6, and tumor necrosis factor-alpha) production from MM6-CA8 human monocytoid cells. These biological properties, including pyrogenicity, completely disappeared by treating the pyrogenic extracts with endotoxin-adsorbent affinity column. Limulus amebocyte lysate activity and cytokine production from MM6-CA8 cells induced by the extracts were significantly decreased by endotoxin inhibitors, an active fragment peptide of an 18-kDa cationic antimicrobial protein and a synthetic lipid A B464 analogue. Furthermore, very small amounts of 2-keto-3-deoxyoctonate and 3-hydroxy fatty acid, which are common constituents of bacterial endotoxins, were detected by gas chromatography-mass spectrometry analysis of the pyrogenic extracts. These findings clearly showed that the pyrogenicity found in these latex products originated from endotoxins contaminating the products.

Animals↗

PubMed2001

Radical fringe negatively modulates Notch signaling in postmitotic neurons of the rat brain.

Fringe was originally identified as a novel secreted signaling protein with a key role in wing formation of Drosophila. Three vertebrate fringe homologues, Radical, Lunatic and Manic fringe, were also identified, and have been shown to play major roles in neurogenesis during development. However, the expression and roles of vertebrate fringe homologues in the adult brain remain to be elucidated. We isolated the cDNA encoding rat Radical fringe (334 amino acids) from rat embryos, and found its mRNA to be most abundantly expressed in the adult rat brain by Northern blotting analysis. The localization of Radical fringe mRNA in the adult rat brain was also examined by in situ hybridization. The mRNA was abundantly expressed in most neurons, but not glial cells, throughout the brain. Notch signaling was shown to negatively modulate the stability of neurites and connections in postmitotic primary neurons. Furthermore, genetic evidence indicated that fringe modulated the Notch signaling pathway. Therefore, we examined the effects of Radical fringe on the Notch signaling pathway in primary rat neurons of the cerebral cortex using recombinant rat Radical fringe protein. Radical fringe protein significantly inhibited expression of the Notch effector Hes1 mRNA in primary neurons. These results indicated that Radical fringe functions by inhibiting Notch signaling in postmitotic neurons of the brain.

Amino Acid Sequence↗

PubMed2001

The forkhead-associated domain of NBS1 is essential for nuclear foci formation after irradiation but not essential for hRAD50[middle dot]hMRE11[middle dot]NBS1 complex DNA repair activity.

NBS1 (p95), the protein responsible for Nijmegen breakage syndrome, shows a weak homology to the yeast Xrs2 protein at the N terminus region, known as the forkhead-associated (FHA) domain and the BRCA1 C terminus domain. The protein interacts with hMRE11 to form a complex with a nuclease activity for initiation of both nonhomologous end joining and homologous recombination. Here, we show in vivo direct evidence that NBS1 recruits the hMRE11 nuclease complex into the cell nucleus and leads to the formation of foci by utilizing different functions from several domains. The amino acid sequence at 665-693 on the C terminus of NBS1, where a novel identical sequence with yeast Xrs2 protein was found, is essential for hMRE11 binding. The hMRE11-binding region is necessary for both nuclear localization of the complex and for cellular radiation resistance. On the other hand, the FHA domain regulates nuclear foci formation of the multiprotein complex in response to DNA damage but is not essential for nuclear transportation of the complex and radiation resistance. Because the FHA/BRCA1 C terminus domain is widely conserved in eukaryotic nuclear proteins related to the cell cycle, gene regulation, and DNA repair, the foci formation could be associated with many phenotypes of Nijmegen breakage syndrome other than radiation sensitivity.

Amino Acid Sequence↗

PubMed2001

Elution of bisphenol-A from hemodialyzers consisting of polycarbonate and polysulfone resins.

This study deals with bisphenol-A (BPA) analysis of the BPA-derived polymer pellets, polycarbonate (PC) and polysulfone (PS), and in the hemodialyzer casings made of PC, and the leaching of BPA from commercially available hemodialyzers into water and bovine serum, using HPLC, GC-MS, and LC-MS analyses, and NMR spectroscopy. Total contents of BPA in polymer pellets of each resin were 4.0 and 7.2 microg/g (PC) and 34.5 microg/g (PS). Amounts of BPA released from hemodialyzer PC casings lacking PS hollow-fiber were 11.7 and 13.7 ng/casing by water extraction, and 296 and 345 ng/casing by methanol extraction. On the other hand, BPA of 3.78 to 141.8 ng/module was recovered using water circulation of hemodialyzers, and 140.7 to 2,090 ng/module was detected when bovine serum was used as a circulation solvent. The elution profiles using various concentrations of ethanol/water mixtures indicated that a 17.2% (v/v) ethanol solution rather than bovine serum can be used as an extraction solvent, where a similar amount of BPA as with bovine serum circulation was eluted from the hemodialyzer. Thus, this solvent may be useful for evaluating BPA elution from hemodialyers under similar conditions to medical use.

Animals↗

PubMed2001

Studies on in vitro evaluation for the biocompatibility of various biomaterials: inhibitory activity of various kinds of polymer microspheres on metabolic cooperation.

Gap junctional intercellular communication is a function that plays an important role in maintaining cell and tissue homeostasis and in regulating cell growth, development, and differentiation. Change in this function when contacting fibroblasts with various polymer microspheres was estimated using the metabolic cooperation assay system. When the cells were in contact with the microspheres after their adhesion onto a substrate, the function did not alter. However, when they were in contact with precoated microspheres on test dishes, the function was inhibited as the quantity of microspheres increased. Moreover, the inhibition level increased as the diameters of polyethylene and polystyrene microspheres decreased. However, no inhibition was observed if precoated microspheres were composed from poly(L-lactic acid). These findings suggest that the size and the material of microspheres, and how cells recognize the microspheres, are factors affecting cell function of gap junctional intercellular communication. Therefore, estimating this function may provide valuable information about the biocompatibility of many kinds of materials even in the form of particles.

Animals↗

PubMed2001

Activation reduction in anterior temporal cortices during repeated recognition of faces of personal acquaintances.

Repeated recognition of the face of a familiar individual is known to show semantic repetition priming effect. In this study, normal subjects were repeatedly presented faces of their colleagues, and the effect of repetition on the regional cerebral blood flow change was measured using positron emission tomography. They repeated a set of three tasks: the familiar-face detection (F) task, the facial direction discrimination (D) task, and the perceptual control (C) task. During five repetitions of the F task, familiar faces were presented six times from different views in a pseudorandom order. Activation reduction through the repetition of the F tasks was observed in the bilateral anterior (anterolateral to the polar region) temporal cortices which are suggested to be involved in the access to the long-term memory concerning people. The bilateral amygdala, the hypothalamus, and the medial frontal cortices, were constantly activated during the F tasks, and considered to be associated with the behavioral significance of the presented familiar faces. Constant activation was also observed in the bilateral occipitotemporal regions and fusiform gyri and the right medial temporal regions during perception of the faces, and in the left medial temporal regions during the facial familiarity detection task, which are consistent with the results of previous functional brain imaging studies. The results have provided further information about the functional segregation of the anterior temporal regions in face recognition and long-term memory.

Adult↗

PubMed2001

A repeated 28-day oral dose toxicity study of methoxychlor in rats, based on the 'enhanced OECD test guideline 407' for screening endocrine-disrupting chemicals.

In association with the international validation project to establish an OECD Enhanced Test Guideline 407, we performed a 28-day repeated-dose toxicity study of methoxychlor, a chlorinated hydrocarbon pesticide with pro-estrogenic and anti-androgenic activities. Attention was paid to the sensitivity of certain additional parameters for detecting endocrine related effects of endocrine disrupting chemicals based on the existing TG 407. Seven-week-old Crj:CD(SD)IGS rats were allocated to one of four groups, each consisting often males and ten females, and methoxychlor was administered once daily by gavage at doses of 0 (control), 20, 100 or 500 mg/kg body weight per day. Male rats were killed on the day after the 28th administration. Female rats were killed on the day of the diestrus stage during 4 days after the 28th administration. Male rats receiving methoxychlor showed mainly atrophy of mammary acinus in the 20 mg/ kg and higher groups, together with decreases in prostate and seminal vesicle weights, and atrophy of epididymis, prostate, seminal vesicle and coagulating gland in the 100 and 500 mg/kg groups. In addition, decrease in serum testosterone level, increase in follicle-stimulating hormone level, decrease in testis and epididymis weights, atrophy of semiferous tubules and Leydig cells, decrease in the number of sperm in the caudal epididymis and their motility were observed in the 500 mg/kg group. Female rats receiving methoxychlor showed mainly abnormal estrous cycles, decrease in serum luteinizing hormone level, decrease in ovary weight, proliferation of mammary acinus, atrophy of ovary due to decrease in follicles and corpus luteum in histopathology, hypertrophy of endometrial epithelium of uterus and vagina epithelium in the 100 and 500 mg/kg groups. Among the parameters tested in the present experimental system, effects of methoxychlor on endocrine-related organs were detected with regard to serum hormone, organ weights, histopathological examination in both sexes, estrus cycle in females and sperm examination in males. Based on these results, a no-observed-adverse-effect level (NOAEL) in the present study was estimated to be below 20 mg/kg per day. In particular, the adverse effects were effectively detected in organ weights of accessory sex organs and histopathological examination.

Administration, Oral↗

PubMed2001

Evolution from pretangle neurons to neurofibrillary tangles monitored by thiazin red combined with Gallyas method and double immunofluorescence.

Double immunofluorescence for paired helical filament (PHF)-tau (AT8) and ubiquitin, enhanced by catalyzed reporter deposition amplification, was combined with thiazin red (TR), a fluorochrome, which has an affinity to fibrillary structures such as neurofibrillary tangles (NFTs). After recording these triple-fluorescent images, sections were subjected to the Gallyas silver impregnation method, so that four different staining properties could be compared on the same structure. Among pyramidal neurons quantified in the hippocampus from six cases of Alzheimer's disease, 60.3% were positive for ubiquitin, and were consistently positive for TR. TR-positive neurons (77.1%) harbored fibrillary structures in the cytoplasm and were always positive for the Gallyas stain, which stained the largest number of legions (94.5%). AT8-positive neurons without fibrillary structure were negative for TR (11.6%, pretangle neurons). Some of the pretangle neurons were positive for the Gallyas stain even without fibrillary structures. Appearance of TR stain and ubiquitin in NFTs, but not in pretangle neurons, suggests that ubiquitin is integrated into tau-positive neurons after their transformation into NFTs. Because TR-positive NFTs sometimes lacked ubiquitin-like immunoreactivity, involvement of ubiquitin may not be an early event during NFT formation. This combined method is now found useful in determining how molecules other than tau are involved during the evolution from tau-positive neurons to NFTs in various neurological disorders characterized by the deposition of tau.

Aged↗

PubMed2001

Non-expanded polyglutamine proteins in intranuclear inclusions of hereditary ataxias--triple-labeling immunofluorescence study.

Neuronal intranuclear inclusions (NIIs) found in CAG/polyglutamine-expansion disorders contain both expanded polyglutamine and the gene product without the CAG repeat. The gene product containing expanded polyglutamine has, therefore, been considered to be a major component of NIIs. In this immunohistochemical study, we showed recruitment of ataxin-2, ataxin-3 and TATA box binding protein (TBP) into NIIs of the pontine neurons of spinocerebellar ataxia type (SCA) 1, SCA2, SCA3 and dentatorubral-pallidoluysian atrophy brains. Triple-labeling immunofluorescence demonstrated colocalization of ataxin-2 and ataxin-3 in NIIs containing expanded polyglutamine, irrespective of the disease examined. These in vivo findings indicate that polyglutamine proteins recruited into NIIs are not restricted to their expanded form. Among these proteins, recruitment of ataxin-2 was least frequent in every case examined, suggesting that the rate of recruitment partly depends on the protein transported into NIIs. Because other proteins lacking polyglutamine motif were not detected in NIIs, it is suggested that the presence of polyglutamine is a prerequisite for these proteins to be recruited into nucleus and to form NIIs. Interaction between expanded and non-expanded polyglutamine may play roles during these processes.

Ataxin-3↗

PubMed2001

Different conformation of neuronal tau deposits distinguished by double immunofluorescence with AT8 and thiazin red combined with Gallyas method.

Different kinds of tau deposits were quantitatively investigated with thiazin red (TR), a fluorochrome that binds to fibrillary structures like neurofibrillary tangles (NFTs), in brains obtained at autopsy from patients with Alzheimer's disease (AD), Pick body (PB) disease, corticobasal degeneration (CBD) or diffuse NFTs with calcification (DNTC). After recording double-labeling fluorescence images with anti-paired helical filament tau (AT8) and TR, the sections were subjected to Gallyas method (GAL). This enabled three different staining properties to be compared on the identical neuron. AT8-positive neocortical neurons of AD and DNTC were fibrillary and uniformly positive for TR and GAL, consistently forming NFTs. NFTs lacking AT8 immunoreactivity (IR) were more frequent in DNTC than in AD, suggesting that evolution of NFTs is more accelerated in DNTC. Scarce TR staining in tau-positive neocortical neurons of CBD suggests their paucity of fibrillary structure. Since the affinity of TR for PB was not consistent, this may be dependent not only on the amount but also the characteristics of fibrillary structures. PBs were further characterized by the scarcity of GAL staining. This approach, which quantitatively clarifies differences between AT8-IR, TR and GAL, provides a morphological basis for further investigations of the different conformational states of tau from its deposition to fibril formation of various types.

Aged↗

PubMed2001

A new reconstructive procedure after segmental pancreatectomy: an experimental study of pancreatic end-to-end (duct-to-duct) anastomosis.

We produced experimental models of pancreatic end-to-end anastomosis, including ductal end-to-end anastomosis (with or without stent) and pancreaticojejunostomy, using mongrel dogs, with a view to evaluating reconstructive procedures after segmental pancreatectomy. We examined macroscopic findings, pancreatograms, and microangiographic and histopathological findings to determine whether pancreatic end-to-end anastomosis was as practicable as pancreaticojejunostomy. Macroscopic findings showed no suture failure in any animal in the end-to-end anastomosis group. Pancreatography revealed obstruction of the stent tube in the stent subgroup, but good patency in the no-stent subgroup. On the imaging of the microvasculature in the end-to-end anastomosis group, proliferation of neovascular vessels and formation of communicating vessels were detected. Histopathologically, no suture failure was detected, and the viability of the pancreatic end-to-end anastomosis was confirmed. From this experiment, we concluded it that it was possible to employ pancreatic end-to-end anastomosis after segmental pancreatectomy in the clinical situation.

Anastomosis, Surgical↗

PubMed2001

Preclinical study of adenoviral p53 gene therapy for esophageal cancer.

An alteration of the p53 gene function is a major factor in the development of esophageal cancer. Recently, p53 gene therapy has been applied for clinical studies in lung cancer and head and neck cancer. However, no preclinical studies have yet demonstrated an anticancer effect of adenoviral-mediated wild-type p53 gene therapy on esophageal cancer. We herein evaluated the effect of p53 adenoviral gene therapy on human esophageal squamous cell carcinoma to test the ability of clinical application. A normal esophageal epithelial cell line (EN53F) and two human esophageal cancer cell lines (ECGI-10 and T.Tn) with a p53 alteration were used. The transduction efficiency, p53 protein expression, p21 protein expression, the induction of apoptosis, and growth suppression were assessed by using the recombinant adenoviral vector Ad5CMV-p53. The transduction efficiency was 60%-80% at 100 plaque-forming units (PFU)/cell and 80%-100% at 300PFU/cell. A significant growth suppression following an Ad5CMV-p53 infection was observed in both cancer cell lines. A Western blot analysis confirmed the presence of both exogenous p53 protein expression and p21 protein induction. Apoptotic cell death was observed with TUNEL staining. T.Tn xenografts in nude mice transduced with Ad5CMV-p53 demonstrated significant growth suppression. These data suggest that Ad5CMV-p53 may thus be a potentially effective therapeutic agent for locally advanced esophageal cancer.

Adenoviruses, Human↗

PubMed2001

Glycopeptide susceptibility profiles of nosocomial multiresistant Staphylococcus haemolyticus isolates.

We investigated 48 Staphylococcus haemolyticus isolates from patients and medical staff in terms of susceptibility to and in-vitro selection for vancomycin and teicoplanin in regard to their antibiotypes. On comparison of multiresistant S. haemolyticus isolates with non-multiresistant isolates, the geometric mean minimum inhibitory concentration (MIC) of vancomycin for multiresistant S. haemolyticus was 2.9 microg/ml, and that of teicoplanin was 18.0 microg/ml, both of which values were significantly greater than the corresponding mean MICs of vancomycin (2.0 microg/ml) and teicoplanin (4.7 microg/ml) for nonmultiresistant isolates. After agar selection, the mean of the highest teicoplanin concentration of selected plates for multiresistant S. haemolyticus was 97.1 microg/ml, which was significantly higher than that for nonmultiresistant isolates (57.8 microg/ml). However, the means' of the highest vancomycin concentrations after agar selection for multiresistant and nonmulti-resistant isolates were the same, at 7.4 microg/ml, with no colonies capable of growing in 32 microg/ml of vancomycin. There was no significant difference in glycopeptide susceptibility between oxacillin-resistant and oxacillin-susceptible isolates among nonmultiresistant S. haemolyticus. The geometric mean MICs of vancomycin for oxacillin-resistant and oxacillin-susceptible isolates were 2.1 microg/ml and 1.6 microg/ml, and those of teicoplanin were 4.4 microg/ml and 5.6 microg/ml, while the means of the highest concentrations of the selected plates of vancomycin were 8.6 microg/ml and 3.3 microg/ml, and those of teicoplanin were 52.8 microg/ml and 74.7 microg/ml, respectively. Multiresistant isolates showed significantly greater mean MICs of vancomycin and teicoplanin and higher teicoplanin concentration of the selected plates than nonmultiresistant isolates, irrespective of oxacillin resistance. These results indicate that methicillin resistance may not be related to reduced susceptibility to glycopeptide in S. haemolyticus, and that a multiresistant profile is associated more with a decreasing susceptibility to glycopeptides then with resistance to oxacillin. In this study, antibiotypes showed good concordance with pulsed-field gel electrophoresis typing results, with a sufficiently high discriminatory ability index, of 0.912. We consider that primary screening with antimicrobial susceptibility testing and antibiotyping, with attention to the multiresistant profile, would be useful for monitoring nosocomial S. haemolyticus colonization and infection.

Anti-Bacterial Agents↗