Biomedical subjects

N Inoue

Publications and source records attributed to N Inoue.

At least 145 records · Page 8Linked to original sources

PubMed2000

Antigens expressed in feline enteroepithelial-stages parasites of Toxoplasma gondii.

In an investigation aimed to identify Toxoplasma gondii antigens expressed in feline enteroepithelial-stages parasites, a cDNA library was constructed and fourteen positive clones were isolated by immunoscreening using sera from cats immunized with feline enteroepithelial-stages parasites. By DNA sequence homology analysis, these fourteen isolated clones were classified into four groups: hypoxanthine-guanine phosphoribosyl transferase (HGPRT) cDNA, heat shock protein 70 (HSP70) cDNA, 14-3-3 protein homologue cDNA, and cDNA encoding an unknown product. In an indirect immunofluorescence antibody test, sera from mice immunized with the recombinant protein encoded by the cDNA for HGPRT, HSP70, 14-3-3 protein, or the unknown product each showed a relatively high level of immunoreactivity with feline enteroepithelial-stages parasites.

Animals↗

PubMed2000

Distraction osteogenesis after acute limb-shortening for segmental tibial defects. Comparison of a monofocal and a bifocal technique in rabbits.

BACKGROUND: Segmental bone defects can be treated with immediate limb-shortening followed by monofocal or bifocal distraction osteogenesis. In the present study, the efficacy of monofocal distraction osteogenesis was compared with that of bifocal distraction osteogenesis in a rabbit model. METHODS: Twenty-four skeletally mature New Zealand White rabbits were divided into two equal groups: one group had monofocal distraction osteosynthesis, and the other had bifocal distraction osteosynthesis. In both groups, a one-centimeter-long segment of bone was resected from the midpart of the tibial shaft. In the monofocal reconstruction group, the limb was immediately shortened to close the segmental defect and the defect was allowed to heal for ten days. Lengthening was then begun at this site, with use of a specially designed external fixator, at a rate of 0.5 millimeter per twelve hours. In the bifocal reconstruction group, the segmental defect was closed immediately and the fragments were fixed with microplates. A subperiosteal osteotomy was performed proximal to the tibiofibular junction, and lengthening was performed at the site of the osteotomy. The animals in both groups were killed twenty days after the lengthening was completed. New-bone formation then was evaluated with use of radiographs, densitometry, biomechanical testing, and histological and histomorphometric analysis. RESULTS: Osseous consolidation occurred in all but one of the animals. Biomechanical testing demonstrated that the tibiae that had been treated with use of the simple monofocal reconstruction technique tended to have greater torsional stiffness (p = 0.14) and strength (p = 0.09). Follow-up radiographs revealed that both groups had a significant decrease in radiolucent area (p < 0.05), which occurred at essentially the same rate after lengthening. No significant differences were found between the groups with respect to new-bone mineral density, new-bone area, or the amount of callus. Thus, after resection of a diaphyseal bone segment comprising 10 percent of the original length of the tibia and acute shortening, limb reconstruction was completed successfully through distraction osteogenesis with use of either a monofocal or a bifocal technique in rabbits. CONCLUSIONS: In the present study, both monofocal and bifocal techniques of shortening and distraction osteogenesis were effective for the reconstruction of segmental bone defects. Under some conditions, the monofocal method may provide a simpler means of treating such defects. CLINICAL RELEVANCE: Damage to the soft-tissue envelope as well as venous and lymphatic stasis impose limits on the amount of limb-shortening that can be achieved with use of the monofocal method and also influence the indications for this procedure in the clinical setting.

Animals↗

PubMed2000

Lysophosphatidylcholine enhances superoxide anions production via endothelial NADH/NADPH oxidase.

Reactive oxygen species (ROS) including superoxide anions (O2(-)) play a key role in atherogenesis, and endothelial cells have the ability to generate ROS. To investigate the enzymatic sources of ROS and the effects of lysophosphatidylcholine (LPC), an atherogenic lipid, we measured ROS production in cultured bovine aortic endothelial cells (BAECs) by the lucigenin-enhanced chemiluminescence (CL) method and electron spin resonance (ESR). BAEC homogenates had the enzymatic activity of NADH/NADPH oxidase. BAECs cultured on microcarrier beads generated O2(-) under basal conditions. The inhibition of NADH/ NADPH oxidase by diphenylene iodonium (DPI) significantly attenuated O2(-) production, whereas no inhibitors of other oxidases suppressed it. Although LPC enhanced O2(-) production approximately 3.1-fold, its action was suppressed by DPI. Tyrosine kinase inhibitors significantly attenuated LPC-induced O2(-) production. ESR with DMPO demonstrated that LPC increased the formation of the DMPO-hydroxyl adduct in dose- and time-dependent manners. These data suggest that the basal production of O2(-) in endothelial cells is mainly mediated by the NADH/NADPH oxidase system and that LPC activates this oxidase to enhance O2(-) production through a tyrosine kinase-dependent pathway. The enhancement of ROS production by LPC is probably involved in its atherogenic property.

Acridines↗

PubMed2000

Comparative study of the toxic effects of gallium arsenide, indium arsenide and arsenic trioxide following intratracheal instillations to the lung of Syrian golden hamsters.

Toxic effects of gallium arsenide (GaAs), indium arsenide (InAs) and arsenic trioxide (As2O3) were studied in male Syrian golden hamsters. GaAs (7.7 mg/kg) and As2O3 (1.3 mg/kg) particles were instilled intratracheally twice a week a total of 16 times, while InAs (7.7 mg/kg) was instilled a total of 14 times. As a control, hamsters were treated with the vehicle, phosphate buffer solution. During the instillation period, the cumulative body weight gain of the InAs-, but not the GaAs- or As2O3-treated hamsters was suppressed significantly, when compared with the control group. Slight to severe inflammatory responses were observed in the lung for all treatment groups. The most severe inflammatory change, characterized by an accumulation of neutrophils and macrophages, exudation, thickness of the pleura and fibrotic proliferation was found in the InAs-treated hamsters. Extensive alveolar or bronchiolar cell hyperplasia with or without keratinizing squamous cell metaplasia was observed in almost all the InAs-treated hamsters. Furthermore, squamous cell metaplasia or squamous cell hyperplasia developed in some of the InAs-treated hamsters, but not in the GaAs- or As2O3-treated hamsters. Slight to mild lesions were found in the convoluted tubules of the kidney in both the GaAs and InAs groups. From the present study, the toxic potency of these particles was provisionally estimated to be in the following order: InAs > GaAs > As2O3, at the dosage level used in this study. Furthermore, there was evidence that InAs particles could induce pulmonary, renal or systemic toxicity, and as such, InAs particles may produce pulmonary precancerous change when instilled intratracheally into hamsters.

Animals↗

PubMed2000

Cerebral function after aortic surgery using retrograde cerebral perfusion: report of three cases.

INTRODUCTION: The neurological analysis undertaken on three cases of aortic aneurysm treated surgically using retrograde cerebral perfusion (RCP) are reported herein. This is the first detailed analysis of postoperative neurological studies in such cases. METHODS: The oxygenation state of cerebral Hb and cytochrome aa3 levels were monitored by near-infrared spectroscopy (NIR) during RCP. Postoperative neurological scoring, computed tomography (CT), magnetic resonance imaging (MRI), and monitoring of local cerebral blood flow (LCBF) quantatively by N-isopropyl-p-[123I] iodoamphetamine (IMP) single-photon emission computed tomography (SPECT) were performed for an extended follow-up period after surgery. RESULTS: The T2WI of MRI findings which consisted of many spotty high intensity areas in the striatum, cerebellum and hippocampus, may reflect the postoperative severe consciousness and mental disturbances (2-4 weeks). However, in all cases the neurological deficit gradually improved until they returned to their normal preoperative states. The improvement of LCBF correlated well with neurological recovery. CONCLUSION: The conditions of the conventional method of RCP must be improved to prevent transient severe consciousness and mental disturbances which occur after the operation in relationship with the appearance of the high intensity area on the MRI T2WI.

Adult↗

PubMed2000

[Midterm results of total aortic root replacement with pulmonary autograft (Ross operation)].

Between February 1995 and December 1999, 18 patients underwent Ross operation. Age at the operation ranged from 2 to 31 years. Diagnosis includes congenital aortic stenosis and/or regurgitation in 15, and adult aortic regurgitation in 3. In all cases autograft was implanted by the method of total aortic root replacement, associated with annuloplasty for the dilated aortic annulus in 2 and aortoventriculotomy by the Konno procedure in 3 (Ross-Konno). Right ventricular outflow tract was reconstructed by a pulmonary homograft in 12, a xenopericardial conduit in 3, or the other reconstructive procedures with autologous tissue and outflow patch in 3. There was no operative and late death. Reoperation was needed in 1 patient due to stenosis of pericardial conduit 4 years after the initial operation. Pressure gradient across implanted autograft valve was negligible (4.8 +/- 0.5 mmHg), and echocardiography revealed no aortic regurgitation in 12 cases and trivial to mild in 6, over a mean follow-up period of 23 +/- 18 months (range 2 to 60 months), signifying excellent durability of implanted autograft. Right ventricular outflow tract reconstruction with the homograft resulted in excellent mid-term performance as showing pressure gradient of 9.0 +/- 4.6 mmHg and no regurgitation in 11 of 12 cases, whereas pressure gradient was 17.9 +/- 13.1 mmHg in the patients underwent the other reconstructive procedures. We conclude that Ross procedure associated with the concomitant procedures to adjust the size discrepancy between the native aortic annulus and autograft has provided good midterm results with excellent autograft durability. And this procedure was thought to be a preferable method for children as well as young adults with congenital aortic stenosis.

Adolescent↗

PubMed2000

Hepatocyte growth factor upregulates E1AF that induces oral squamous cell carcinoma cell invasion by activating matrix metalloproteinase genes.

Hepatocyte growth factor (HGF) is thought to play a role in cell motility and invasion. Matrix metalloproteinases (MMPs) have been implicated in invasion and metastasis of tumor cells. We have previously reported that the Ets-oncogene family transcription factor E1AF positively regulates transcription of MMP genes in transient expression assays and that overexpression of the E1AF gene confers an invasive phenotype on breast cancer cells. Here we examined the effect of HGF on E1AF and MMP gene expression in terms of the invasive potential of the oral squamous cell carcinoma cell line HSC3. HGF stimulated expression of the E1AF gene. The levels of MMP-1, -3 and -9 mRNAs increased in cells treated with HGF and correlated with E1AF upregulation. In contrast, no obvious upregulation of MMP-1 and -9 mRNA was observed in ASE1AFHSC3 cells transfected with the antisense E1AF expression vector into parental HSC3 cells. The wild-type MMP-9 gene promoter was activated by endogenous E1AF in HSC3 cells, and chloramphenicol acetyltransferase (CAT) activities increased when HGF was added to transfected cells. On the other hand, CAT activity was reduced to almost two-thirds of the wild-type activity when HSC3 cells were transfected with a CAT reporter plasmid driven by a mutant MMP-9 promoter lacking the Ets-binding site, and induction of CAT activity was not observed upon addition of HGF. Analysis of organotypic raft cultures revealed that HSC3 cells invaded and degraded collagen gel actively upon addition of HGF. These results suggest that HGF induces expression of the Ets-related E1AF transcription factor gene whose product in turn activates MMP genes and leads to oral cancer cell invasion.

Adenovirus E1A Proteins↗

PubMed2000

Suggested dosage rates of melarsoprol in the treatment of mice experimentally infected with Trypanosoma brucei gambiense.

One group of BALB/c mice infected with a highly virulent strain of Trypanosoma brucei gambiense were treated intraperitoneally with three series of three injections (each injection of 10 mg/kg) of Mel-B separated by seven days of rest, while a second group was treated once by a single injection All the Mel-B treated mice in both experiments were negative for parasites when examined using either the wet blood film or buffy coat methods, but were intermittently PCR positive during the sampling period. We encourage the use of a repeat negative PCR test over a one month period in combination with corroborative clinical and parasitological investigation to be suggestive of cure in experimental animals previously infected with trypanosomosis. In view of the exorbitant costs of Mel-B and its extreme toxicity, we recommend that Mel-B be given as one course of two injections (each equivalent to 10 mg/kg) separated by 2 d of rest in experimentally infected rodent models.

Animals↗

PubMed2000

[A case treated surgically through anterior axillary thoracotomy for recurrent right spontaneous pneumothorax after repeated video-assisted thoracoscopic surgery].

A 21-year-old male who had thoracoscopic surgery for right spontaneous pneumothorax was admitted to our hospital complaining of dyspnea due to the recurrence. There was no bulla in right apex but empysematous change in visceral pleura at re-thoracoscopic surgery. Two bulla in right S6 were removal using endo-stapler. Three years after the second thoracoscopic surgery, the recurrence of right pneumothorax was recognized. Chest CT revealed a new bulla in right apex. Therefore the third operation was carried out through anterior axillary thoracotomy. A new bulla was resected with endo-stapler and pleurodesis using argon laser was also applied to prevent of recurrence of pneumothorax. The patient is now doing well without recurrence 6 months after the third operation.

Adult↗

PubMed2000

Method to remove intramedullary cement prior to mechanical testing in canine segmental replacement model.

Mechanical evaluation of fixator augmented bone healing is complicated. When the primary interest is focused on mechanical properties of the healing site, implants may need to be removed before the testing. Cement or bone ingrowth related fixation requires special techniques for successful implant removal. Five canine femurs and five specially designed intramedullary cement fixated segmental replacement prosthesis were used to show that intramedullary cement mantle could be removed safely and the procedure was repeatable without altering the mechanical properties of healing site.

Analysis of Variance↗

PubMed1999

Glutamate receptors mediate regulation of Na pump isoform activities in neurons.

Rat cerebral neurons matured in culture were stimulated with glutamate analogues, and the K+ uptake activities of Na pump isoforms were measured. Ionotropic receptor agonists, kainate, AMPA, and NMDA, increased total K+ uptake activity via activation of the alpha2/alpha3 isoforms and an inhibition of the alpha1 isoform as reported previously for glutamate. The effects of kainate or AMPA were antagonized by CNQX and those of NMDA were by APV or MK-801. In contrast, metabotropic receptor agonist ACPD had no effects on the Na pump isoform activities. Glutamate transporter substrate, PDC, was effective, but NMDA receptor antagonists abolished the effects of PDC. These results suggest that the ionotropic glutamate receptors mediate the regulation of Na pump isoform activities in neurons.

Animals↗

PubMed1999

Production of specific antibody and T helper 1-dominant cytokine elicited by dendritic cells genetically modified with an adenovirus vector.

In this study we examined how dendritic cells (DCs) transduced with an adenovirus vector encoding a model tumor antigen (beta-galactosidase; beta-gal) would influence the humoral immune response to this antigen. Mice immunized with LacZ transduced DCs by an adenovirus vector could produce more anti-beta-gal antibody, especially of IgG2a subclass, than mice immunized with DCs alone, although the amount of serum IgG antibody did not increase. Compared with mice immunized with DCs alone, splenocytes of mice immunized with LacZ transduced DCs could produce more interferon-gamma (IFN-gamma) against the beta-gal derived, H-2Ld-restricted nonapeptide (TPHPARIGL) in a dose-dependent manner. These data suggest that DCs transduced with an adenovirus vector encoding the model tumor antigen could induce the T helper 1 (Th1) dominant response against the model tumor antigen.

Adenoviridae↗

PubMed1999

Expression of NADH/NADPH oxidase p22phox in human coronary arteries.

BACKGROUND: NADH/NADPH oxidase is an important source of superoxide in the vasculature. Recently, we found that polymorphism of the gene p22phox, a critical component of this oxidase, is associated with a risk of coronary artery disease. The aim of this study was to investigate the localization of p22phox in human coronary arteries and to examine its difference in expression between nonatherosclerotic and atherosclerotic coronary arteries. METHODS AND RESULTS: Using coronary artery sections from autopsied cases (n=11), the expression of p22phox was examined by immunohistochemistry and Western blotting. In nonatherosclerotic coronary arteries, p22phox was weakly expressed, mainly in the adventitia. In atherosclerotic coronary arteries, intensive immunoreactivity was detected in neointimal and medial smooth muscle cells and infiltrating macrophages in hypercellular regions and at the shoulder region. Semiquantitative analysis and Western blotting showed that the expression of p22phox in atherosclerotic coronary arteries was more pronounced than that in nonatherosclerotic arteries. Double staining revealed p22phox expression in adventitial fibroblasts, smooth muscle cells, macrophages in the neointima and media, and endothelial cells. CONCLUSIONS: As atherosclerosis progressed, the expression of p22phox increased through the vessel wall. p22phox might participate in the pathogenesis and pathophysiology of atherosclerotic coronary disease.

Adult↗

PubMed1999

Interleukin 4 is a crucial cytokine in controlling Trypanosoma brucei gambiense infection in mice.

The role of interleukin 4 (IL-4) was studied in relation to host defense during Trypanosoma brucei gambiense IL3253 (IL3253) infection in mice. BALB/c/A-+/+ (BALB/c), BALB/c/A-nu/nu (nude) and C.B-17/Icr-scid/scid (SCID) mice were infected intraperitoneally with 5 x 10(3) bloodstream forms (BSFs) of the trypanosome. The BALB/c mice showed high resistance to IL3253 infection with sporadic parasitemia. The nude mice were also able to control IL3253 infection and experienced low, but persistent parasitemia. However, the SCID mice, which have no functional T- and B-cells, showed high susceptibility to IL3253 infection with more than 1 x 10(8) BSFs/ml. Serum IL-4 levels in the infected BALB/c mice were increased on days 12-18 post-infection (PI). In BALB/c mice depleted of CD4+ T-cells by monoclonal antibody (mAb) treatment, parasitemia was persistent, ranging from 1 x 10(4) to 1 x 10(6) BSFs/ml and was significantly higher than that of the other groups. IL-4 was not detected in the serum of CD4+ T-cells-depleted mice. On the other hand, anti-IL-4-treated IL3253-infected BALB/c mice relapsed significantly longer than the control mice (p < 0.01). These findings suggest that the CD4+ T-cells may control the levels of parasitemia in IL3253 infection through the IL-4 pathway.

Animals↗

PubMed1999

Goblet-cell-specific transcription of mouse intestinal trefoil factor gene results from collaboration of complex series of positive and negative regulatory elements.

Intestinal trefoil factor (ITF) is expressed selectively in intestinal goblet cells. Previous studies of the rat ITF gene identified one cis-regulatory element, designated the goblet-cell-response element (GCRE), present in the proximal region of the promoter. To identify additional cis-regulatory elements responsible for goblet-cell-specific expression, a DNA fragment containing 6353 bp of the 5'-flanking region of the mouse ITF gene was cloned and its promoter activity was examined extensively. In human and murine intestinal-derived cell lines (LS174T and CMT-93), the luciferase activities of a 6.3-kb construct were 5- and 2-fold greater than the smaller 1.8-kb construct, respectively. In contrast, the activity in non-intestinal cell lines (HepG2 and HeLa) was 2-4-fold lower than the smaller construct. In the region downstream from the 1.8-kb position, strong luciferase activities in LS174T and HepG2 cells were observed using a 201-bp construct. Interestingly, increased activity was almost completely suppressed in cells transfected with a 391-bp construct. Detailed analyses of this region revealed the existence of a 11-bp positive regulatory element (-181 to -170; ACCTCTTCCTG) and a 9-bp negative regulatory element (-208 to -200; ATTGACAGA) in addition to the GCRE. All three elements were well conserved among human, rat and mouse ITF gene promoters. In addition, a mutant 1.8-kb construct in which the negative regulatory region was deleted yielded the same approximate luciferase activity as a 6.3-kb construct, suggesting binding of a goblet-cell-specific silencer inhibitor (SI) between -6.3 and -1.8 kb. The SI present in goblet cells may block the silencers' binding to the pre-initiation complex and allow increased transcriptional activity driven by specific and non-specific enhancers. High-level expression of the mouse ITF gene specifically in intestinal goblet cells may be achieved through the combined effects of these regulatory elements.

Amino Acid Sequence↗

PubMed1999

Arginine kinase evolved twice: evidence that echinoderm arginine kinase originated from creatine kinase.

Arginine kinase (AK) was isolated from the longitudinal muscle of the sea cucumber Stichopus japonicus. Unlike the monomeric 40 kDa AKs from molluscs and arthropods, but like the cytoplasmic isoenzymes of vertebrate creatine kinase (CK), the Stichopus enzyme was dimeric. To explore the evolutionary origin of the dimeric AK, we determined its cDNA-derived amino acid sequence of 370 residues. A comparison of the sequence with those of other enzymes belonging to the phosphagen kinase family indicated that the entire amino acid sequence of Stichopus AK is apparently much more similar to vertebrate CKs than to all other AKs. A phylogenetic tree also strongly suggests that the Stichopus AK has evolved from CK. These results support the conclusion that AK evolved at least twice during the evolution of phosphagen kinases: first at an early stage of phosphagen kinase evolution (its descendants are molluscan and arthropod AKs) and secondly from CK later in metazoan evolution. A comparison of the amino acid sequence around the guanidino specificity (GS) region (which is a possible candidate for the guanidine substrate recognition site in the phosphagen kinase family) of the Stichopus enzyme with those of other phosphagen kinases showed that the GS region of the Stichopus enzyme was of the AK type: five amino acid deletions in the flexible loop region that might help to accommodate larger guanidine substrates in the active site. The presence of the AK-type deletions in the Stichopus AK, even though it seems that the enzyme's most immediate ancestor was probably CK, strongly suggests that the GS region has a role in substrate specificity. Stichopus AK and presumably other echinoderm AKs seem to have evolved from the CK gene; the sequence of GS region might have been replaced by the AK type via exon shuffling. The presence of an intron near the GS region in the Stichopus AK gene supports this hypothesis.

Amino Acid Sequence↗

PubMed1999

Stromelysin promoter 5A/6A polymorphism is associated with acute myocardial infarction.

BACKGROUND: Rupture of the fibrous cap of an atherosclerotic plaque is a key event that predisposes to acute myocardial infarction (AMI). Matrix metalloproteinases (MMPs) may contribute to weakening of the cap, which favors rupture. Stromelysin, a member of MMP family, is identified extensively in human coronary atherosclerotic lesions. It can degrade most of the constituents of extracellular matrix as well as activating other MMPs, which suggests that it may play an important role in plaque rupture. Recently, a common variant (5A/6A) in the promoter of the stromelysin gene has been identified. The 5A/6A polymorphism could regulate the transcription of the stromelysin gene in an allele-specific manner. METHODS AND RESULTS: To investigate the relation between the 5A/6A polymorphism in the promoter of the stromelysin gene and AMI, we conducted a case-control study of 330 AMI patients and 330 control subjects. The prevalence of the 5A/6A+5A/5A genotype was significantly more frequent in the patients with AMI than in control subjects (48.8% vs 32.7%, P<0.0001). In logistic regression models, the odds ratio of the 5A/6A+5A/5A was 2.25 (95% CI, 1.51 to 3.35). The association of 5A/6A polymorphism with AMI was statistically significant and independent of other risk factors. CONCLUSIONS: The 5A/6A polymorphism in the promoter of the stromelysin gene is a novel pathogenetic risk factor for AMI.

Alleles↗

PubMed1999

Histopathological evidence that spermatogonia are the target cells of 2-bromopropane.

To confirm the target cell of 2-bromopropane within the testis, 1355 mg/kg of 2-bromopropane was subcutaneously injected to rats for 1-5 days and the numbers of spermatogonia and spermatocytes were examined 6 h after each last injection. The number of stage I spermatogonia decreased after the first 2-bromopropane injection and the number of spermatogonia at the other stages also decreased following repetitive injection. The number of these spermatogonia decreased further by the repetition of 2-bromopropane injection. In addition, the delay in mitotic division of type B spermatogonia was frequently observed after the fifth 2-bromopropane injection. The number of stage I pachytene spermatocytes also decreased slightly after the first 2-bromopropane injection, although it did not decrease further following repetitive injection. Therefore, we concluded that spermatogonia are the target cells of 2-bromopropane in rats.

Animals↗