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Biomedical subjects

K Kajiwara

Publications and source records attributed to K Kajiwara.

At least 73 records · Page 4Linked to original sources

[Estimation of myocardial viability and clinical significance of reverse redistribution in resting technetium-99m sestamibi myocardial single photon emission computed tomography in patients with acute myocardial infarction].

The clinical significance of reverse redistribution of technetium-99m sestamibi (MIBI) was investigated in 36 patients with acute myocardial infarction and angiographically confirmed single-vessel disease, but without previous infarction using resting MIBI myocardial single photon emission computed tomography (SPECT) and exercise-reinjection thallium-201 (Tl) myocardial SPECT. MIBI myocardial SPECT was performed 90 min and 300 min after injection of MIBI 370 MBq at rest. Four hours after exercise Tl imaging was completed, reinjection imaging was obtained. Wall motion abnormalities on left ventriculograms were analyzed at the onset of infarction and 1 month later. The severity scores on the MIBI early image, MIBI delayed image and Tl reinjection image were 98 +/- 18, 170 +/- 22 and 90 +/- 18, respectively. The reverse redistribution of MIBI was marked in acute infarction. A significant correlation of severity score was found between the MIBI early image and Tl reinjection image (r = 0.89). In 18 patients with significant stenosis of an infarct-related artery, there was a significant correlation between the degree of reverse redistribution and that of Tl redistribution (r = 0.826). A good correlation was found between the severity score on the MIBI early image and wall motion abnormality at 1 month after infarction (r = 0.816). There was a significant correlation between the degree of reverse redistribution and wall motion improvement (r = 0.782). Despite stenosis of the infarct-related artery, the wall motion abnormality was less in 22 patients with marked reverse redistribution (defect score on the MIBI delayed image was double that on the early image) than the other 14 patients. In conclusion, the MIBI early image may reflect myocardial viability and the reverse redistribution of MIBI was observed frequently in patients with acute myocardial infarction. Marked reverse redistribution was observed in patients with preserved left ventricular function. Because of the close correlation of reverse redistribution with Tl redistribution and wall motion improvement, reverse redistribution of MIBI is considered to occur in areas at risk for acute myocardial infarction.

Adult↗

[Elucidation of the mechanism of retinal degeneration and regeneration and the prospects for its clinical application].

In order to obtain the basic knowledge necessary to develop therapeutical intervention for blindness due to the damaged retina and optic nerve, the mechanism of retinal degeneration and regeneration in an amphibian model, Cynops pyrrhogaster, was studied. In the retinal degenerative process following enucleation and reimplantation of the eye ball, evidence was found for active cell death of neural retinal cells. As the degeneration proceeded, Musashi, an ribonucleic acid (RNA)-binding protein, started its expression in the daughter cells of proliferating retinal pigment epithelium (RPE) cells, messenger RNA (mRNA) expression of proneural genes with basic helix-loop-helix motif was then detected in the newly developing retina. These results suggest that transdifferentiation of RPE cells to neural retina involves at least partial cascade, if not entirely, of neural induction from uncommitted ectodermal tissue. Search for genes that are required for transdifferentiation of RPE cells to neural retinal cells, in addition to those mentioned above, will provide the basic knowledge for successful retinal transplantation and retinal regeneration in higher vertebrates.

Amphibians↗

Proliferative potentials of glioma cells and vascular components determined with monoclonal antibody MIB-1.

Proliferative activity in 78 glioma specimens was assessed immunohistochemically by determining proliferating index of tumor cells (PTC-PI) and endothelial cells (PEC-PI) using the MIB-1 monoclonal antibody. The PTC-PI of anaplastic astrocytoma (9.0 +/- 5.8: mean +/- standard deviation) was significantly higher than that of astrocytoma (1.2 +/- 0.4, < 0.01), and lower than that of glioblastoma multiforme (12.0 +/- 5.6, < 0.05). We then compared PTC-PI values, patients' age and extent of tumor resection with the prognosis of patients with malignant glioma (both glioblastoma and anaplastic astrocytoma). Kaplan-Meier survival rate analysis demonstrated higher survival rates in patients with less than 8.0% of PTC-PI at 5 and 10 years (p < 0.05). The mean age of patients who survived more than a year was lower than that of patients who died within a year (53.0 y.o., vs. 59.7 y.o., p < 0.01). Total or subtotal resection of the tumor was more often performed in the former than latter patients (51% vs. 21%, p < 0.01). These results suggested PTC-PI provides useful information which may allow better assessment of the biological behavior and clinical prognosis of glioma, in addition to histological grading, patients' age and extent of tumor resection. While the average PEC-PI value (3.3) was lower than that of PTC-PI (7.0), there was a significantly close relationship between PTC- and PEC values (p < 0.01), providing an impetus to develop novel therapies directed toward suppression of microvascular regeneration.

Adult↗

Conformation of cyclic and linear (1 --> 2)-beta-D-glucans in aqueous solution.

The conformations of cyclic (1 --> 2)-beta-D-glucan chains having degree of polymerization (dp) 17 to 24 were characterized by means of small-angle X-ray scattering and Monte Carlo simulation. The results indicate that cyclic (1 --> 2)-beta-D-glucan chains adopt the shape of a doughnut-like ring with a thickness of about 10 A for all the samples. The diameter of the annulus for the cyclic glucan having dp 21 is estimated to be only about 4-5 A. Two linear (1 --> 2)-beta-D-glucans possessing dp 19 and 21 prepared by acid hydrolysis of a cyclic glucan and subsequent fractionation showed different scattering profiles from those obtained for cyclic glucans having the corresponding dp. Although the Monte Carlo simulation does not completely reproduce the scattering profiles observed by small-angle X-ray scattering, linear (1 --> 2)-beta-D-glucans seem to possess a characteristic cylindrical shape with cross-sectional diameters of 11.8 and 13.2 A for linear glucans of dp 19 and 21, respectively.

Chromatography, High Pressure Liquid↗

Evidence for a role of phosphatidylinositol 3-kinase in IL-4-induced germline C epsilon transcription.

Association of interleukin-4 receptor (IL-4R) with phosphatidylinositol 3-kinase (PI3-kinase) has been demonstrated as the proximal event of IL-4 signaling. We investigated the role of this enzyme in the IL-4 signaling pathway in a human Burkitt lymphoma B cell line, DND39, that expresses germline C epsilon transcripts in response to IL-4. Stimulation of DND39 cells with IL-4 resulted in an accumulation of PI-3-monophosphate as well as a decrease of PI-4,5-bisphosphate, which were abrogated by wortmannin, a potent inhibitor of PI3-kinase. Activation of PI3-kinase was further confirmed by the finding that IL-4 caused an increase in PI3-kinase activity coimmunoprecipitated with anti-IL-4R and with anti-JAK3 kinase antibodies. As a possible downstream event of PI3-kinase activation, the translocation of a zeta isoform of protein kinase C (PKC) from the cytosol to the membrane fraction was observed after IL-4 stimulation, and wortmannin also suppressed this translocation. Moreover, IL-4-induced expression of germline C epsilon transcription was inhibited not only by wortmannin, but also by a PKC inhibitor, K252a. These results suggest that the signaling pathway involving PI3-kinase and PKC zeta plays an important role in induction of germline C epsilon transcription in DND39 cells by IL-4.

Amino Acid Sequence↗

Cloning of SEZ-12 encoding seizure-related and membrane-bound adhesion protein.

SEZ-12 is one of the seizure-related cDNAs which was isolated by differential hybridization from primary cultured neurons from the mouse cerebral cortex with or without pentylenetetrazol (PTZ). SEZ-12 expression is transiently down-regulated in the mouse brain by injection of PTZ. To characterize SEZ-12, isolation of full-length cDNA and nucleotide sequence analysis were performed. The deduced amino acid sequence of SEZ-12 revealed that it encodes membrane-bound C-type lectin and has a significant homology to that of human cDNA, DGCR2 and IDD, which were cloned from a balanced translocation breakpoint associated with the DiGeorge syndrome. The isolated cDNA was about 4 kb in length and the message was expressed ubiquitously in various organs with low-abundance. Previously, we also cloned a transmembrane protein which is probably involved in cell-cell interaction by the differential hybridization technique. These findings suggest that transmembrane signaling in neuronal cells may have an important role in PTZ-induced seizure.

Amino Acid Sequence↗

Molecular characterization of seizure-related genes isolated by differential screening.

To isolate seizure-related genes, we applied differential screening technique to the cDNA library which constructed from primary cultured cerebral cortical cells treated with pentylenetetrazol (PTZ). Northern blotting analysis of mRNA levels in the cerebra after systemic administration of PTZ confirmed the results of differential screening procedure: 6 clones showed increased mRNA level and 3 clones showed decreased expression with PTZ. Interestingly, 4 genes which were isolated by this technique were related to intracellular calcium action. They were cytosolic phospholipase A2, 78 kDa glucose regulated protein, SEZ-15 which has an EF hand motif and PTZ-17 that causes calcium current in Xenopus oocyte with PTZ application. These data and our previous results suggest that intracellular calcium may play an important role for seizure-related pathophysiological changes in neuronal cells.

Amino Acid Sequence↗

Alanine protects liver from injury caused by F-galactosamine and CCl4.

The liver is the main organ involved in amino acid metabolism, and it utilizes glucogenic amino acids as substrates for glucose or adenosine triphosphate (ATP), but this process is impaired in clinical and experimental liver diseases. In this study, we administered high doses of amino acids in rats or cultured hepatocytes with experimental models of liver injury to examine whether such supplementation could attenuate liver damage. We found that the addition of alanine reduced enzyme leakage from primary cultured rat hepatocytes treated with D-galactosamine (D-gal), while other amino acids did not. A significant decrease of lactate dehydrogenase (LDH) leakage was observed when cells were cultured with >6 from mmol/L alanine. Alanine also reduced enzyme leakage from normal hepatocytes that were not treated with D-gal. In D-gal-treated rats, constant infusion of a high dose of alanine significantly reduced the plasma transaminase and total bilirubin levels when compared with infusion of an amino acid mixture. Bolus administration of alanine significantly prevented the elevation of plasma transaminase levels and histological liver damage in CCl4-treated rats, while fructose-1,6 bisphosphate (FDP) had little effect. Alanine might promote the restoration of damaged liver in hepatotoxicant-treated rats, because significant effect was found after the elevation in plasma transaminase levels. Alanine also prevented the decrease of cellular ATP caused by D-gal and appeared to promote ATP production in primary cultured rat hepatocytes. These results indicate that alanine reduces experimental liver damage by a direct effect on hepatocytes.

Adenosine Triphosphate↗

Atypical MRI findings in treatment-related leukoencephalopathy: case report.

A patient with acute myeloblastic leukaemia showed atypical findings on MRI following combination therapy including intrathecal methotrexate and radiation. MRI findings not previously been reported are ring as well as patchy enhancement, marked mass effect and lesions extending to the putamen and corpus callosum.

Adult↗

In vitro and in vivo analysis of human leukocyte binding by the antitumor polysaccharide, lentinan.

Lentinan (LNT), a (1-->3)-beta-D-glucan with (1--6)-beta-D-glucopyranoside branches, has marked antitumor effects in syngeneic and autochtonous hosts. Clinically, LNT has proved effective with chemotherapeutic agents in patients with recurrent gastric and colorectal cancer. However, the mechanism that triggers subsequent immunologic reactions remains obscure. We hypothesized that LNT must first bind to the host cells. Accordingly, we analyzed LNT binding to host cells in healthy volunteers after incubating their cells under a variety of conditions as well as intravenously injecting LNT then subjecting them to flow cytometry and immunofluorescent staining using monoclonal antibody (anti-LNT mAb). LNT bound to monocytes and neutrophils, but not to lymphocytes in vitro. The most avid LNT binding was to monocytes. The percentage of LNT-bound monocytes after 60 min incubation at 4 degrees C was greater than that at 37 degrees C. The binding of LNT to monocytes was inhibited slightly by anti-iC3b receptor (anti-CR3) mAb, strongly by anti-C3b receptor (anti-CR1) mAb, and completely by anti-CR1 and anti-CR3 mAb together. The percentages of LNT-binding monocytes in the peripheral blood increased significantly 3 and 4 after 2 mg LNT-injection and returned to low levels 5 h later. However, no increase in LNT-binding neutrophils and lymphocytes was observed. We concluded that binding of LNT to human monocytes may initiate the influence of this compound on the immune system and differ between individuals. Its binding site may be similar to the C3b receptor.

Adult↗

Involvement of nuclear factor-kappa B activation in IgE synthesis in human B cells.

Nuclear factor-kappa B (NF-kappa B) is a transcription factor that binds to the consensus DNA sequence in the cis-acting elements of various genes. Although NF-kappa B activates the expression of many genes involved in immune and inflammatory responses, little is known about the role of NF-kappa B activation in the induction of IgE synthesis in human B cells. Therefore we first examined the participation of NF-kappa B in germline C epsilon transcription in a human Burkitt lymphoma B cell line, DND39. Stimulation of DND39 cells with IL-4 or anti-CD40 monoclonal antibody (mAb) activated phosphatidylinositol 3-kinase and subsequently induced nuclear expression of NF-kappa B, which was identified by electrophoretic mobility shift assays. n-Acetyl-L-cysteine (NAC), a potent antioxidant, blocked NF-kappa B activation caused by IL-4 and by anti-CD40 mAb. Although inhibition of IL-4-driven germline C epsilon transcription by NAC was not sufficient, the agent remarkably diminished anti-CD40 mAb-mediated up-regulation of germline C epsilon transcription. Second, we studied the effect of NAC on IgE synthesis in human normal B cells costimulated with IL-4 and anti-CD40 mAb. NAC was effective in inhibiting mature C epsilon transcription and IgE synthesis in the T cell-independent culture system. However, NAC did not significantly affect the spontaneous production of IgE by atopic B cells. These results indicate that NF-kappa B activity is commonly inducible in DND39 cells by IL-4 and anti-CD40 mAb and suggest that NF-kappa B sensitive to NAC may play a role in regulating IgE synthesis in B cells.

B-Lymphocytes↗

Immune responses to adenovirus vectors in the nervous system.

Non-replicating adenovirus vectors are being developed as vehicles for gene transfer into cells of the nervous system. An important requirement for successful gene transfer is the absence of deleterious cytotoxic or inflammatory side effects of the delivery system. Despite offering relatively stable reporter gene expression, currently available adenovirus vectors also elicit immune responses in the brain, both at the site of vector delivery and at synaptically linked distant sites. However, although an anti-viral T-lymphocyte response eliminates the vector and damages local tissue in many peripheral organs, the immune response to adenovirus in the brain is less effective and enables the vector to persist. Nevertheless, in this persistent state the adenovirus vector remains a potential target for a destructive immune response that can also cause local demyelination. The development of strategies to minimize this damaging immune response, through either vector modification or immunomodulation, will be crucial for the future success of genetic therapies in the brain.

Adenoviridae↗

Effect of alanine on D-galactosamine-induced acute liver failure in rats.

The effect of high-dose alanine on survival and liver function in rats with acute liver failure caused by a lethal dose of D-galactosamine (D-gal) was studied. Greater than 90% of control animals died within 5 days after D-gal injection, but alanine significantly decreased mortality, even when treatment was started at 12 hours after D-gal injection. Alanyl-glutamine had a slight effect, but glucose produced no improvement. There was marked elevation of the plasma aspartate transaminase (AST) level, prolongation of the prothrombin time, and a decrease of the arterial ketone body ratio (AKBR) and hepatic adenosine triphosphate (ATP) content within 12 hours after D-gal injection. The AKBR decreased in parallel with the decrease of the hepatic ATP content. These parameters were significantly improved in alanine-treated rats at 48 hours after the induction of liver damage, which was just before control rats began to die. The hepatic ATP content was significantly greater in alanine-treated rats than in the other rats (including normal controls), but glucose pretreatment had no effect. It was also found that the liver labeling index of partially hepatectomized rats was significantly elevated by alanine administration at 3 hours before measurement. In conclusion, alanine is effective for the treatment of experimental acute liver failure, probably caused by promotion of ATP synthesis. Ala may be a good candidate for clinical application because of its preventive effect on hepatocyte necrosis and its promotive effect on liver regeneration.

Adenosine Triphosphate↗

Cloning and characterization of seizure-related gene, SEZ-6.

SEZ-6 is a brain-specific cDNA. Its expression is increased by convulsant drug and it encodes membrane protein with five copies of short consensus repeat (SCR; complement C3b/C4b binding site) and two repeated sequences which are partially similar to CUB domain (complement Clr/s-like repeat). In this study, we analyzed the gene product of SEZ-6 by antibody raised against SEZ-6 peptide. In vitro translation and immunoblot revealed that SEZ-6 protein is modified post-translationally and expressed in the cerebrum and the cerebellum. To analyze the SEZ-6 expression in detail, we searched for other types of SEZ-6 cDNA by the polymerase chain reaction. Two additional types of cDNAs were isolated and sequence analysis showed that one cDNA encodes membrane protein with a different C-terminal region and the other encodes secreted protein with two SCRs and one CUB-like domain.

Amino Acid Sequence↗

Cloning and characterization of pentylenetetrazol-related cDNA, PTZ-17.

cDNAs related to pentylenetetrazol-induced bursting activity in neurons were screened by a differential hybridization method using normal and pentylenetetrazol-treated primary cultured neurons from the cerebral cortex of mice. Twenty clones of candidate cDNA with expression increased or decreased by treatment with pentylenetetrazol were obtained. One of them, PTZ-17, was sequenced. Injection of PTZ-17 derived RNA into Xenopus oocytes showed a large calcium inward current with extracellular application of pentylenetetrazol.

Animals↗

Immunohistochemical localization and biological significance of the phylogenically conserved thymus-brain antigen (UB-13 antigen) in skate, rat and human.

A monoclonal antibody (UB-13) originally raised against the brain of the skate (Raja kenojei, a cartilaginous-fish) was found to react with lymphoid and brain tissues from many species when examined immunohistochemically. In rat and human thymus, UB-13 antigen was observed to be closely associated with reticular tissue in the medulla and cortex. Interestingly, a few or several thymocytes were encircled by the UB-13-reactive reticular tissue. At 14 days gestation, rat thymus consisted mainly of reticular epithelial tissue, after which strong thymocyte production started. At this stage, some of the reticular tissue was heavily stained with UB-13. In the thymus tissues of the irradiated and recovering rats, where reduction and massive reproduction of thymocytes were observed, extensive UB-13 antigen expression localized on the reticular epithelial tissue, an observation which may support the thymocyte re-population. These findings suggest that the antigen recognized by UB-13 may be important for thymocyte proliferation and maturation. UB-13 antigen was found in the fibrous structure of the molecular and granular layer of the human cerebellum. Some glial cells were also stained strongly with UB-13 in the human cerebellar or cerebral grey and white matter. In rat, glial cells, especially astroglias, and the endothelial structure of blood vessels were stained strongly with UB-13. These findings suggest that UB-13 may be a useful monoclonal antibody for analysis of brain-lymphoid antigen in many species.

Adult↗

Cloning and expression of SEZ-6, a brain-specific and seizure-related cDNA.

To clarify the molecular mechanism of neuronal bursting activity of seizures, we have constructed a cDNA library from mouse cerebrum cortex-derived cells treated with pentylentetrazole (PTZ), one of the convulsant drugs. Using a differential screening technique, several cDNA clones whose expressions change with PTZ-treatment were obtained. Among these clones, SEZ-6 was characterized by increased expression with PTZ. Detailed northern analysis showed that expression of SEZ-6 was limited to the brain and increased by the administration of PTZ not only in in vitro cultured cells but also in vivo. Analysis of SEZ-6 cDNA revealed multiple motifs, including typical signal sequence, threonine-rich domain, five copies of short consensus repeats (SCRs) or sushi domain (complement C3b/C4b binding site), two repeated sequences which were partially similar to the CUB domain or complement C1r/s-like repeat, one transmembrane domain and a short cytoplasmic segment in the C-terminal region. Although many proteins with multiple SCRs or CUB domains other than complement-related proteins have been found, this is the first report about a brain-specific cDNA which encodes membrane protein with both SCRs and CUB domain-like segments. Based on these findings, it is evident that SEZ-6 encodes a novel type of protein which may be related to seizure.

Animals↗

Solution X-ray scattering study of reconstitution process of tobacco mosaic virus particle using low-temperature quenching.

The reconstitution process of tobacco mosaic virus (TMV) was investigated by the solution X-ray scattering measurements with the synchrotron radiation source using low-temperature quenching. TMV assembly in an aqueous solution is completely stopped below 5 degrees C. The TMV assembly was traced by the small-angle X-ray scattering (SAXS) measurements at 5 degrees C on a series of solutions prepared by low-temperature quenching after incubation either at 15, 20 or 25 degrees C for an appropriate interval between 0 and 60 min. The SAXS results were analyzed by the Guinier plot, the Kratky plot and the distance distribution function. In order to account the time course of SAXS profiles in terms of the elongation of TMV assembly, a model calculation was performed to simulate the Guinier plot, the Kratky plot and the distance distribution function by applying Glatter's multibody method using models that were constituted of the spheres representing a column of piled two-layer disks of TMV-protein. The three simulated functions thus obtained support the conclusion derived from the three functions calculated from the experimental results that the incubation of the RNA and protein of TMV began to reconstitute TMV instantly after mixing, proceeded steeply to a long rod, and then extended asymptotic to the full length of the TMV particle. This process is in good agreement with that obtained from electron microscopic studies.

Journal Article↗