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Biomedical subjects

N Inoue

Publications and source records attributed to N Inoue.

At least 163 records · Page 9Linked to original sources

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↗

[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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

Induction of human leukocyte antigen (HLA)-A2-restricted and MAGE-3-gene-derived peptide-specific cytolytic T lymphocytes using cultured dendritic cells from an HLA-A2 esophageal cancer patient.

BACKGROUND AND OBJECTIVES: Using peripheral blood mononuclear cells (PBMCs) from a 10-year survivor with established human leukocyte antigen (HLA)-A2(+) and MAGE-3(+) esophageal cancer cell line (KYSE-170), we examined the induction of HLA-A2-restricted and MAGE-3-gene-derived peptide (FLWGPRALV, amino acids 271-279)-specific cytolytic T lymphocytes (CTLs). METHODS: Autologous dendritic cells (DCs) cultured with granulocyte-macrophage colony stimulating factor and interleukin-4 were used as antigen presenting cells. PBMCs were stimulated by peptide-pulsed DCs in vitro. RESULTS: PBMC cocultured with FLWGPRALV-pulsed DCs could induce the relevant peptide-specific CTLs, which had tumor necrosis factor production and specific cytotoxicity against relevant peptide-pulsed autologous DCs (34%, effector:target ratio = 40:1). Moreover, they showed specific cytotoxicity against the autologous esophageal cancer cell line KYSE-170 (17%, effector:target ratio = 40:1). CONCLUSIONS: These results suggest that FLWGPRALV-pulsed cultured DCs would be a potent candidate for peptide vaccine against HLA-A2(+) and MAGE-3(+) esophageal cancer.

Antigen-Presenting Cells↗

alpha(1)-adrenoceptor-induced trafficking of aquaporin-5 to the apical plasma membrane of rat parotid cells.

Incubation of rat parotid tissue with 10 microM epinephrine resulted in a transient and marked trafficking of aquaporin-5 (AQP5) from intracellular membranes to the apical plasma membrane (APM) that was maximal at 1 min. This effect of epinephrine was mimicked by phenylephrine, but not by clonidine, dobutamine, or salbutamol, and it was inhibited by phentolamine, but not by propranolol. Furthermore, the epinephrine-induced trafficking of AQP5 was inhibited by phospholipase C inhibitor U73122 as well as dantrolene and TMB-8, both of which inhibit the release of Ca(2+) from intracellular stores. Cytochalasin D and tubulozole-C also inhibited this action of epinephrine. These results indicate that epinephrine, acting at alpha(1)-adrenoceptors, induces the trafficking of AQP5 to the APM by triggering the release of Ca(2+) from intracellular stores through inositol 1,4,5-trisphosphate and ryanodine receptors. In addition, the potent involvement of the cytoskeleton was shown in the epinephrine-induced trafficking of AQP5.

Adrenergic alpha-Agonists↗

Effect of 12-O-tetradecanoyl-phorbol ester and incisional wounding on neuropsin mRNA and its protein expression in murine skin.

The expression of neuropsin mRNA in vivo in mouse skin was examined by in situ hybridization and Northern blotting under stimulated conditions. Two kinds of epidermal stimuli, a topical application of a chemical tumor promoter and incisional wounding, were used. A single topical application of 12-O-tetradecanoyl-phorbol 13-acetate induced epidermal hyperplasia and simultaneously induced an extensive increase in neuropsin mRNA in the suprabasal cells. A full-thickness skin incision also induced a profound increase in neuropsin mRNA in the suprabasal cells surrounding the wound but not in actively proliferating basal cells. The increases in neuropsin mRNA occurred rather late and were limited to the site of drug application or around the incision. Interestingly, neuropsin mRNA was not expressed in the epithelial tongue migrating toward the wound during re-epithelialization. Thus, neuropsin might participate in accelerated epidermal differentiation rather than in the proliferation or migration of keratinocytes in the wound.

Animals↗

Metabolic activities of partially degenerated hypertrophic chondrocytes: gene expression of hyaluronan synthases.

Ultrastructural aspects of hypertrophic chondrocytes in hamster and mouse epiphysial cartilage were examined in relation to their metabolic activities. With the hypertrophic change, cytoplasmic vacuolization proceeded leaving the partially intact endoplasmic reticulum (ER). In the hypertrophic cells, cytoplasmic hyaluronan was stained with the biotinylated hyaluronan-binding region (b-HABR) of aggrecan, and mRNAs of hyaluronan synthase (Has 1, Has 2 and Has 3) were detected by in situ hybridization. When the epiphysial cartilage was cultured in the presence of 35S, 3H-GlcNAc, 3H-proline or 14C-palmitic acid, vacuolated late hypertrophic chondrocytes were labeled with these radioactive precursors. The evidence indicates that late hypertrophic chondrocytes are metabolically active, which appears to be essential for the enlargement of chondrocytes.

Animals↗

Increased susceptibility to Toxoplasma gondii infection in SAG-1 transgenic mice.

SAG-1, one of the major surface proteins of Toxoplasma gondii, has been reported to play an important role in immune and pathogenic mechanisms of the parasites but its exact function is still unclear. We investigated the time courses of T. gondii infection in B6C3F1 transgenic mice carrying the SAG-1 gene. SAG-1 transgenic mice were infected intraperitoneally with a high virulent RH strain or a low virulent Beverley strain of T. gondii. When infected with RH strain tachyzoites, no significant differences in time courses of survivals between SAG-1 transgenic and wild-type mice were observed. Both groups succumbed to an acute infection within 8 days after infection. However, a lower survival rate (20%) was observed in SAG-1 transgenic mice than in wild-type (80%), when infected with Beverley strain cysts. This result indicates that SAG-1 transgenic mice are more susceptible to T. gondii infection as compared with their wild-type counterpart. ELISA using recombinant SAG-1 protein indicates that SAG-1 transgenic mice do not produce antibodies to the SAG-1 molecule. These findings may provide a critical tool for analysing the molecular mechanisms of pathogenesis and host immune responses during toxoplasmosis.

Animals↗

New bone formation in an osteoblastic tumor model is increased by endothelin-1 overexpression and decreased by endothelin A receptor blockade.

OBJECTIVES: The osteoblastic response of bone to metastatic prostate cancer is both characteristic and enigmatic. The potent vasoconstrictor endothelin-1 (ET-1), produced by prostate cancer, has been identified as a potential factor in new bone formation. METHODS: Using a novel method to quantitate new bone formation induced by the WISH tumor, we examined the effects of ET-1 overexpression and endothelin receptor antagonists on the osteoblastic response. RESULTS: WISH, a human tumor cell line derived from amnion, produces ET-1 mRNA and protein and induces abundant new bone formation and splenomegaly in vivo. Stable transfection of WISH with an ET-1 overexpression cDNA construct produced clones that secreted 18-fold more bioactive ET-1 than vector-only controls. After 14 days of growth in the lower leg of nu/nu mice, ET-1 overexpressing tumors produced significantly more new bone than vector-only controls. Conversely, areas of new bone formation were significantly less in animals treated with a selective endothelin A (ET(A)) receptor antagonist A127722. CONCLUSIONS: The activity of ET-1 in this osteoblastic model provides a unique target for therapy.

Animals↗

Asn-linked sugar chain structures of recombinant human thrombopoietin produced in Chinese hamster ovary cells.

Human thrombopoietin (TPO) that regulates the numbers of megakaryocytes and platelets is a heavily N- and O-glycosylated glycoprotein hormone with partial homology to human erythropoietin (EPO). We prepared recombinant human TPO produced in Chinese hamster ovary (CHO) cells and analyzed the sugar chain structures quantitatively using 2-aminobenzamide labeling, sequential glycosidase digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). We found bi-, tri- and tetraantennary complex-type sugar chains with one or two N-acetyllactosamine repeats, which are common to recombinant human EPO produced in CHO cells. On the other hand, there were triantennary sugar chains with one or two N-acetyllactosamine repeats that were specific to the recombinant human TPO, and their distributions of branch structures were also different. These results suggested that proximal protein structure should determine the branch structure of Asn-linked sugar chains in addition to the glycosyltransferases subset.

Animals↗