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Biomedical subjects

A Iwata

Publications and source records attributed to A Iwata.

At least 19 recordsLinked to original sources
PubMed2005

Epidemiology of blood donors in Japan, positive for hepatitis B virus and hepatitis C virus by nucleic acid amplification testing.

BACKGROUND AND OBJECTIVES: The Japanese Red Cross screens seronegative blood donors by nucleic acid amplification testing (NAT) for hepatitis B, hepatitis C and human immunodeficiency virus-1 markers. NAT-positive donors thus identified seemed to have a different infectious background from serologically positive donors. The purpose of our study was to characterize this background in the hepatitis B virus (HBV) and hepatitis C virus (HCV) NAT-positive donors. MATERIALS AND METHODS: Some 328 HBV DNA-positive and 44 HCV RNA-positive donors were detected by NAT testing of seronegative blood donors. These were characterized regarding age, gender and genotype of HBV and HCV. RESULTS: Those who were HBV NAT-positive were mainly young, in particular teenage girls. In Japan, genotypes C and B have previously been dominant, but recently genotype A has increased, and genotype H was recently detected. In HBV NAT-positive donors, the rate of genotype A was high (12.2%) compared with patients in hospital (1.7-2%). Donors who were HCV NAT-positive were also young, but mostly men in their twenties. The ratio of genotype 1b to 2a or 1b to 2b in HCV NAT-positive donors differed from that of hospitalized patients in Japan. We did not find genotype 1a, which is dominant in the USA. CONCLUSIONS: The high-risk donors detected by NAT were mainly young, with a different distribution of genotypes from that of hospitalized patients, regarding both HBV and HCV. The rare HBV genotype H has been found for the first time in Japan. The findings reflect the present spread of hepatitis viruses B and C.

Adolescent↗
PubMed2001

alpha-Synuclein affects the MAPK pathway and accelerates cell death.

Insoluble alpha-synuclein accumulates in Parkinson's disease, diffuse Lewy body disease, and multiple system atrophy. However, the relationship between its accumulation and pathogenesis is still unclear. Recently, we reported that overexpression of alpha-synuclein affects Elk-1 phosphorylation in cultured cells, which is mainly performed by mitogen-activated protein kinases (MAPKs). We further examined the relationship between MAPK signaling and the effects of alpha-synuclein expression on ecdysone-inducible neuro2a cell lines and found that cells expressing alpha-synuclein had less phosphorylated MAPKs. Moreover, they showed significant cell death when the concentration of serum in the culture medium was reduced. Under normal serum conditions, the addition of the MAPK inhibitor U0126 also caused cell death in alpha-synuclein-expressing cells. Transfection of constitutively active MEK-1 resulted in MAPK phosphorylation in alpha-synuclein-expressing cells and improved cell viability even under reduced serum conditions. Thus, we conclude that alpha-synuclein regulates the MAPK pathway by reducing the amount of available active MAPK. Our findings suggest a mechanism for pathogenesis and thus offer therapeutic insight into synucleinopathies.

14-3-3 Proteins↗
PubMed2001

Molecular cloning and expression of the cDNA encoding feline granulocyte colony-stimulating factor.

Both genomic DNA and cDNA of the feline granulocyte colony-stimulating factor (G-CSF) gene were cloned from CRFK cells. Southern blot analysis showed that the haploid genome contains a single copy of the G-CSF gene. The RT-PCR analysis of several feline cell lines revealed expression of G-CSF mRNA in response to lipopolysaccharide stimulation. Sequence analysis of genomic and cDNA clones indicated that the intron-exon junction structure is conserved between the human and the feline G-CSF genes. The G-CSF coding region encodes a predicted protein of 195 amino acids including a signal sequence of 21 amino acids. The feline G-CSF amino acid sequence shares a high degree of identity with the canine (90.8%), human (87.4%), ovine (83.9%), bovine (82.8%), porcine (80.5%), murine (70.7%) and rat (66.8%) G-CSF. The feline G-CSF expressed in insect cells using recombinant baculovirus vector was biologically active as measured in a proliferation assay using NFS-60 cells and an induction assay of leukocytes in cats.

Amino Acid Sequence↗
PubMed2001

Liposome-mediated gene transfection of endothelial nitric oxide synthase reduces endothelial activation and leukocyte infiltration in transplanted hearts.

BACKGROUND: During cardiac ischemia-reperfusion injury, neutrophilic infiltration of the myocardium is mediated by adhesion molecule expression on activated coronary endothelium. Nitric oxide inhibits neutrophil adhesion to endothelium in vitro by blocking the nuclear factor (NF)-kappaB-dependent transcription of adhesion molecules. We investigated whether intraoperative gene delivery of endothelial nitric oxide synthase (eNOS) into donor hearts before transplantation would have a similar effect on an entire organ. METHODS AND RESULTS: In an allogeneic rabbit heart transplant model, liposomes complexed to the gene encoding eNOS were infused into the donor coronary circulation before transplantation. By 24 hours after transplantation, calcium-dependent nitrite production was significantly higher in eNOS-transfected donor hearts than in the 3 control groups: donor hearts transfected with empty plasmids alone, donor hearts treated with diluent only, and untransplanted native hearts. Intramyocardial neutrophil and T-lymphocyte populations were halved in eNOS-transfected hearts compared with control donor hearts (P<0.05). Moreover, the prevalence of NF-kappaB activation in microvascular endothelial cells and surrounding cardiac myocytes as well as endothelial vascular cell adhesion molecule-1 and intracellular adhesion molecule-1 expression were all significantly reduced in eNOS-transfected hearts compared with control donor hearts (P<0.01). Without immunosuppression, eNOS-transfected hearts survived longer than controls. CONCLUSIONS: Intraoperative liposome-mediated gene delivery of eNOS to donor hearts can result in early gene expression sufficient to reduce ischemia-reperfusion injury by inhibiting NF-kappaB activation, adhesion molecule expression, and the early infiltration of leukocytes, all of which may improve graft survival.

Animals↗
PubMed2001

Diffuse brain injury induces local expression of Na+/myo-inositol cotransporter in the rat brain.

We studied expression of an osmoprotective gene, sodium/myo-inositol cotransporter (SMIT) in Marmarou's animal model for human diffuse brain injury by in situ hybridization and immunohistochemistry. In rats with diffuse brain injury, transient upregulation of SMIT mRNA was exclusively observed in the lateral area of pyramidal tract in lower brainstem. The expression was induced at 1 h after injury, peaked at 24 h, and returned to almost control levels at 48 h. Upregulated expression was found mainly in small glia-like cells. By immunohistochemistry using antibodies to phosphorylated mitogen-activated protein (MAP) kinases, inductions of phosphorylated p44/42 MAP kinase were also observed after diffuse brain injury. Interestingly, the distribution patterns of induced phosphorylated p44/42 MAP kinase were completely coincident with those of upregulated SMIT mRNA after diffuse brain injury. These results suggest that diffuse brain injury induces local expression of SMIT by activation of p44/42 MAP kinase cascade. The confined SMIT induction may reflect regional differences of damage and/or cellular differences in sensitivity to neuropathological stresses caused by this injury.

Animals↗
PubMed2001

Expression of canine interferon-gamma by a recombinant vaccinia virus and its antiviral effect.

A recombinant vaccinia virus-expressing canine interferon (IFN)-gamma (vv/cIFN-gamma) was constructed. In rabbit kidney (RK13) and canine A72 cells infected with vv/cIFN-gamma, IFN activity was detected in the culture supernatants of both cell types. Canine IFN-gamma was also detected in both cell extracts by Western blot. The activity of the recombinant canine IFN-gamma in RK13 cells was higher than that in A72 cells. The vv/cIFN-gamma could not grow in A72 cells at a low multiplicity of infection, probably due to the antiviral activity of the canine IFN-gamma produced. Although exogenous IFN-gamma did not inhibit the growth of vaccinia virus, addition of anti-canine IFN-gamma serum recovered the growth of the vv/cIFN-gamma on A72 cells in a dose-dependent manner. These results suggest that the growth of vv/cIFN-gamma was inhibited by IFN-gamma produced in a paracrine and autocrine manner. In addition, the recombinant canine IFN-gamma inhibited the multiplication of canine herpesvirus, pseudorabies virus and canine adenovirus type 1 in Madin-Darby canine kidney cells. The antiviral effect of canine IFN-gamma was more effective than that of canine IFN-beta. From the present studies, we concluded the recombinant virus may be a useful suicide viral vector.

Animals↗
PubMed2001

Arthroscopic diagnosis of tibiofibular syndesmosis disruption.

PURPOSE: We have been able to diagnose tibiofibular syndesmosis injury by ankle arthroscopy, and in the present study we compare these results with the results from plain radiographs. TYPE OF STUDY: Case series. METHODS: Thirty-eight type-B (Weber system) distal fibular fractures in 38 patients were diagnosed to determine whether tibiofibular syndesmosis disruption was present. According to the Lauge-Hansen system, 16 patients had supination-external rotation fractures and 22 had pronation-abduction fractures. Standard non-weight-bearing anteroposterior radiographs and mortise radiographs were evaluated. Furthermore, ankle arthroscopy was performed on all patients. RESULTS: Tibiofibular syndesmosis disruptions were diagnosed in 16 of the 38 patients (42%) by anteroposterior radiography, 21 of 38 patients (55%) by mortise radiography, and 33 of 38 patients (87%) by ankle arthroscopy. All of the patients who were diagnosed with tibiofibular syndesmosis disruption by anteroposterior radiography and mortise radiography were also confirmed by ankle arthroscopy to have injured their tibiofibular syndesmosis. In 12 patients, ankle arthroscopy was the only method used to diagnose the tibiofibular syndesmosis disruption. CONCLUSIONS: Ankle arthroscopy excels in term of the diagnosis ratio for tibiofibular syndesmosis disruption compared with both anteroposterior and mortise radiography. Therefore, we conclude that ankle arthroscopy is necessary for the correct diagnosis of tibiofibular syndesmosis disruption.

Adolescent↗
PubMed2001

Ontogeny of the murine Ig joining chain gene and protein.

We used Northern blot analysis in order to investigate the ontogeny of the murine joining (J)-chain gene. No J-chain expression was detected in embryonic tissues, including liver, spleen and intestine, but an expression of mu heavy chain was detected in foetal liver at day 17. J-chain expression was detected in the spleen at day 9 and in the intestine at day 15 after birth. Western blot analysis was carried out in order to compare the protein levels of J and mu heavy chains in serum from day 8 to day 24 after birth, using antihuman J chain and antimouse mu chain antibodies. Although mu chain protein could be detected in serum from day 8, J-chain protein was detectable only at day 24. These results suggest that the expression of J chain is a later event than the mu chain in the mouse, which thus differs in embryogenesis from humans.

Animals↗
PubMed2001

alpha-Synuclein forms a complex with transcription factor Elk-1.

alpha-Synuclein has been identified as a component of Lewy bodies in Parkinson's disease and diffuse Lewy body disease, and glial cytoplasmic inclusions (GCIs) in multiple system atrophy (MSA). To explore the role of alpha-synuclein in the pathogenesis, we searched for molecules interacting with alpha-synuclein and discovered that GCIs are stained by anti-Elk-1 antibody. To seek the role of Elk-1 in synucleinopathies, we cotransfected alpha-synuclein and Elk-1 to cultured cells, and found small granular structure complexes where the two molecules colocalized. Moreover, alpha-synuclein and Elk-1 were co-immunoprecipitated from the cell lysates. For formation of the complex, the presence of both ETS and B-box domains of Elk-1 was required. Although there was no evidence of direct binding between alpha-synuclein and Elk-1, we discovered that alpha-synuclein and Elk-1 both bind to ERK-2, a MAP kinase. The effect of alpha-synuclein on the MAP kinase pathway was assessed using the Pathdetect system, which showed prominent attenuation of Elk-1 phosphorylation with alpha-synuclein, and especially A53T mutant. Our results suggest that alpha-synuclein reacts with the MAP kinase pathway, which might cause dysfunction of neurons and oligodendrocytes and lead to neurodegeneration in Parkinson's disease and MSA.

Amino Acid Motifs↗
PubMed2001

Vascular cell adhesion molecule-1 up-regulation and phenotypic modulation of vascular smooth muscle cells predate mononuclear infiltration in transplant arteriopathy.

OBJECTIVE: Expression of embryonic myosin heavy chain isoforms and vascular cell adhesion molecule-1 by neointimal vascular smooth muscle cells are independent indicators of atherosclerotic plaque development in both human beings and experimental animal models. We examined the chronologic change in smooth muscle cell myosin heavy chain isoforms, vascular cell adhesion molecule-1 expression, and mononuclear cell infiltration in a carotid arterial transplant model to ascertain whether similar phenotypic changes would occur in transplant arteriopathy. METHODS: Transplanted rabbit carotid arteries were examined at 7, 14, 21, and 35 days (n = 5, 7, 6, and 5, respectively). Lesion progression and the prevalence of smooth muscle cell myosin heavy chain isoforms, T-lymphocytes, macrophages, and vascular cell adhesion molecule-1 expression were evaluated immunohistochemically by computerized image analysis. RESULTS: In this carotid arterial transplant model, the intima/media area ratio increased significantly by 35 days (P =.01) as cell density decreased (P =.01), suggesting extracellular matrix elaboration. Intimal smooth muscle cells expressing embryonic phenotypes were seen as early as 7 days, a phenotypic change that predated mononuclear cell infiltration of the graft by at least 7 days. By 35 days, up to 70% of intimal smooth muscle cells expressed the embryonic phenotype, coinciding with the transition from inflammatory to chronic lesions. Although, in early lesions, vascular cell adhesion molecule-1 was identified on luminal endothelium overlying mononuclear infiltrates, in advanced lesions vascular cell adhesion molecule-1 was identified primarily on intimal vascular smooth muscle cells. CONCLUSIONS: Overall, these vascular smooth muscle cell changes mark important early events in transplant arteriopathy that may not be ameliorated by immunosuppressive regimens in routine use.

Animals↗
PubMed2001

Purification and properties of recombinant Plasmodium falciparum S-adenosyl-L-homocysteine hydrolase.

Recombinant S-adenosyl-L-homocysteine (SAH) hydrolase of the malaria parasite Plasmodium falciparum was expressed in Escherichia coli, purified to homogeneity and characterized. Comparison of the malaria parasite SAH hydrolase with that derived from the human gene indicated marked differences in kcat values. The values of both forward and reverse reactions of P. falciparum SAH hydrolase are more than 21-fold smaller than those of the human enzyme. Km values of the parasite and human SAH enzymes are 1.2 and 7.8 microM, respectively. On the other hand, IC50 values of neplanocin A, a strong inhibitor of SAH hydrolase and a growth inhibitor of P. falciparum, are 101 nM for the parasite enzyme and 47 nM for human enzyme. P. falciparum SAH hydrolase has been thought to be a target for a chemotherapeutic agent against malaria. This study may make it possible to develop a specific inhibitor for the parasite SAH hydrolase.

Adenosine↗
PubMed2001

Computed tomographic evaluation of the position of the leg for mortise radiographs.

We investigated the most advantageous internal rotation angle of the leg for mortise radiographs. One hundred and twenty-eight feet of 64 healthy volunteers with no histories of ankle or foot pathology (72 feet of 36 males, 56 feet of 28 females) were examined. The subjects had an average age of 29 years (range, 19 to 51 years), average height of 167 cm (range, 157 to 181 cm), and average foot length of 25 cm (range, 23 to 27 cm). We obtained a plain axial view at the level of the central patella and 5 mm proximal to the tibial plafond using computed tomography, and investigated the inclination angle of the distal tibiofibular joint to a horizontal plane, regarding it as a mortise angle. The mean mortise angle was 19.1 +/- 5.0 degrees. However, two peaks were observed at around 15 degrees and 20 degrees. This indicated that the mean mortise angle of the males was 21.2 +/- 4.6 degrees, and the mean mortise angle of the females was 16.4 +/- 4.1degrees, with a significant difference between the males and the females' mortise angle (P < 0.0001). There was no correlation between the mortise angle and the height (P = 0.899 in the males, and P = 0.871 in the females), nor between the mortise angle and the foot length (P = 0.359 in the males, and P = 0.512 in the females). Therefore, we concluded that the internal rotation angle of the leg for mortise radiography should be generally set up at about 20 degrees on males and 15 degrees on females.

Adult↗
PubMed2001

Comparison of the growth inhibitory effects of canine IFN-alpha, -beta and -gamma on canine cells infected with Neospora caninum tachyzoites.

The growth inhibitory effects of recombinant canine interferon alpha (IFN-alpha), beta (IFN-beta) and gamma (IFN-gamma) were examined on Madin-Darby canine kidney cells infected with Neospora caninum tachyzoites. The parasite growth was inhibited by all IFNs in a dose-dependent manner. IFN-gamma inhibited the parasite growth with greater efficacy than IFN-alpha or IFN-beta. Moreover, the effect of IFNs on N. caninum growth associated with the suppression of the host cell viability. The present study indicates IFN-alpha and -beta, besides IFN-gamma, play a crucial role for N. caninum growth in host cells.

Animals↗
PubMed2001

[Survival after oozing type cardiac rupture associated with subepicardial aneurysm evaluated by echocardiography: a case report].

A 64-year-old woman was admitted to our hospital with acute myocardial infarction. She underwent emergent percutaneous transluminal coronary angioplasty. Transthoracic echocardiography revealed mild pericardial effusion on the third day. Pericarditis or cardiac rupture were suspected, so transthoracic echocardiography was repeated serially. On the sixth day, transthoracic echocardiography showed increasing pericardial effusion and abrupt interruption of the apical myocardium of the left ventricle and intact epicardial imaging with systolic expansion. The diagnosis was oozing type cardiac rupture of a subepicardial aneurysm. Surgical treatment was successful and the accuracy of the echocardiographic diagnosis was established.

Angioplasty, Balloon, Coronary↗
PubMed2001

[Measurements of cerebral blood flow by the noninvasive microsphere method with 123I-IMP and fan beam collimator: comparison with the continuous arterial blood sampling method].

PURPOSE AND METHODS: The noninvasive microsphere (NIMS) method quantifies cerebral blood flow with N-isopropyl-p-[123I]-iodoamphetamine (123I-IMP) without blood sampling of a patient, but when a fan beam collimator is used for collecting raw data in single photon emission computed tomography (SPECT), the procedure is complicated because we have to change the collimator. In phantom and clinical studies we investigated the validity of using a fan beam collimator in collecting planar and SPECT raw data. RESULTS: The phantom study suggested that using a fan beam collimator for planar imaging was feasible because of the image magnification rate and the count rate. Mean cerebral blood flow (mCBF) values obtained by the NIMS method with the fan beam collimator were compared with mCBF simultaneously estimated from the conventional continuous arterial blood sampling (microsphere: MS) method in twenty patients with ischemic cerebral vascular diseases. There was good correlation (y = 1.033x + 8.004, r2 = 0.729, p < 0.01) in mCBF between the fan beam NIMS method and the MS method. CONCLUSION: In conclusion, this method for the measurement of CBF was acceptable for routine clinical studies.

Brain Ischemia↗
PubMed2000

Evolutionary aspects of gobioid fishes based upon a phylogenetic analysis of mitochondrial cytochrome B genes.

The Gobioidei is a large suborder in the order Perciformes and consists of more than 2000 species belonging to about 270 genera. The vast number of species and their morphological specialization adapted to diverse habits and habitats makes the classification of the gobioid fishes very difficult.A comprehensive estimation of the evolutionary scenario of all gobioid fishes using only morphological information is difficult for two major reasons: first, in addition to wide ecological diversification, there is a trend towards specialization and degeneration of morphological characters among these species; second, an appropriate outgroup of gobioid fishes has not been recognized. Based upon nucleotide sequence comparisons of gobioid mitochondrial cytochrome b genes, we established the phylogenetic relationships of their differentiation into many groups of morphological and ecological diversity. The phylogenetic trees obtained show that most species examined have diverged from each other almost simultaneously or during an extremely short period of time.

Animals↗
PubMed2000

Adeno-associated virus vector transduction of vascular smooth muscle cells in vivo.

Adeno-associated virus (AAV) vectors might offer solutions for restenosis and angiogenesis by transducing nondividing cells and providing long-term gene expression. We investigated the feasibility of vascular cell transduction by AAV vectors in an in vivo rabbit carotid artery model. Time course of gene expression, inflammatory reaction to the vector, and effects of varying viral titer, exposure time, and intraluminal pressures on gene expression were examined. Recombinant AAV vectors with an Rous sarcoma virus promoter and alkaline phosphatase reporter gene were injected intraluminally into transiently isolated carotid segments. Following transduction, gene expression increased significantly over 14 days and then remained stable to 28 days, the last time point examined. Medial vascular smooth muscle cells were the main cell type transduced even with an intact endothelial layer. Increasing the viral titer and intraluminal pressure both enhanced transduction efficiency to achieve a mean of 34 +/- 7% of the subintimal layer of smooth muscle cells expressing gene product. A mild inflammatory reaction, composed of T cells with only rare macrophages, with minimal intimal thickening was demonstrated in 40% of transduced vessels; inflammatory cells were not detected in sham-operated control arteries. These findings demonstrate that AAV is a promising vector for intravascular applications in coronary and peripheral vascular diseases.

Alkaline Phosphatase↗