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Biomedical subjects

H Kondo

Publications and source records attributed to H Kondo.

At least 289 records · Page 16Linked to original sources
PubMed1999

Incidence of abnormalities in laboratory tests found in surveillance of adults over 40 years of age.

National health check-up systems have been used for 5 years in Japan for adults who are over 40 years of age. As part of a national project, Osaka prefecture is also conducting a program for health check-up testing and cancer screening for this age group. This surveillance revealed that incidence of obesity, hypertension, cholesterolemia, albuminuria, or abnormal ECG was high. Analysis of surveillance results should contribute to understanding the present status and recent trends in diseases in the aged. With continuation of this surveillance for a number of years, trends in life-style related diseases in Japan should be detectable.

Adult↗
PubMed1999

Current status of liver function tests in Japan.

Inter-laboratory variations in data obtained from surveillance in Japan were studied. The items evaluated were related to liver function and were as follows: total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (gamma-GT), cholinesterase (CHE), lactate dehydrogenase (LD), alkaline phosphatase (ALP) and hepatitis markers. Inter-laboratory coefficients of variations for bilirubin, AST and ALT were acceptable, being less than 10%. but higher variations were found for thle other enzyme assays. Detection of hepatitis markers was acceptable. However. even for parameters with lower inter-laboratory variation, differences in obtained values among different reagents or methods still existed. Thus, standardization will be needed for laboratory data in Japan, and this will contribute to international standardization in laboratory medicine in the future.

Biomarkers↗
PubMed1998

Rac1 is required for the formation of three germ layers during gastrulation.

The Rac1, a member of the Rho family proteins, regulates actin organization of cytoskeleton and cell adhesion. We used genetic analysis to elucidate the role of Rac1 in mouse embryonic development. The rac1 deficient embryos showed numerous cell deaths in the space between the embryonic ectoderm and endoderm at the primitive streak stage. Investigation of the primary epiblast culture isolated from rac1 deficient embryos indicated that Rac1 is involved in lamellipodia formation, cell adhesion and cell migration in vivo. These results suggest that Rac1-mediated cell adhesion is essential for the formation of three germ layers during gastrulation.

Animals↗
PubMed1998

Alz-50/Gallyas-positive lysosome-like intraneuronal granules in Alzheimer's disease and control brains.

The Gallyas-Braak silver impregnation method revealed neurons containing well-defined intraneuronal granules in both Alzheimer's disease and normal control brains. The granules were immunostained prominently with the Alz-50 antibody and, to a lesser degree, with the tau-2 antibody, but not with other anti-tau antibodies examined. The areas of distribution of granule-containing neurons detected by the Gallyas-Braak method appeared to overlap with the reported main sites of subcortical distribution of neurofibrillary tangles. They, however, were not observed in the cerebral cortex, including the hippocampal region. The Alz-50 immunoreactive granules showed ultrastructural features similar to those of lysosomes or lipofuscin. These findings suggest that denatured tau might be degraded in lysosomes.

Aged↗
PubMed1998

Identification of a novel mutation of the CPO gene in a Japanese hereditary coproporphyria family.

Hereditary coproporphyria (HCP) is an autosomal dominant disease characterized by a deficiency of coproporphyrinogen oxidase (CPO) caused by a mutation in the CPO gene. Only 11 mutations of the gene have been reported in HCP patients. We report another mutation in a Japanese family. Polymerase chain reaction-single strand conformational polymorphism and direct sequence analyses demonstrated a C to T substitution in exon 1 of the CPO gene at nucleotide position 85, which lies in the putative presequence for targeting to mitochondria. This mutation changes the codon for glutamine to a termination codon at amino acid position 29. MaeI restriction analysis showed two other carriers in the family. The C-T mutation is located within a recently proposed putative alternative translation initiation codon (TIC-1), supporting that TIC-1 is the real TIC rather than TIC-2.

Adult↗
PubMed1998

Localization of alpha-tocopherol transfer protein in rat brain.

Vitamin E (alpha-tocopherol) is a fat-soluble antioxidant that is transported by plasma lipoproteins in the body. Alpha-tocopherol transfer protein (alpha-TTP), which was identified as a product of the causative gene for familial isolated vitamin E (FIVE) deficiency, is a cytosolic liver protein which plays an important role in the efficient circulation of plasma vitamin E in the body. In the present study, we detected the message for alpha-TTP at low levels in some rat tissues including brain, spleen, lung and kidney. In the brain, the alpha-TTP transcript was detected predominantly in the cerebellar cortex, as revealed by in situ hybridization histochemistry. In the cerebellar cortex, clusters of the hybridization signal were aligned with the Purkinje cell layer, although the signal was not detected in the Purkinje cells, but in the small cells around the Purkinje cells. This distribution pattern strongly suggests that the message for alpha-TTP is expressed in the Bergmann glial cells. Combined with the previous observation that there was severe Purkinje cell loss in patients with FIVE deficiency, the present data suggest that vitamin E is supplied to the Purkinje cells from the surrounding Bergmann glial cells with the help of alpha-TTP.

Animals↗
PubMed1998

The p47 co-factor regulates the ATPase activity of the membrane fusion protein, p97.

The highly conserved ATPase p97, a member of the AAA-ATPases, is found in a complex with its co-factor p47 in rat liver cytosol. Previously it had been shown that p97-mediated reassembly of Golgi cisternae from mitotic Golgi fragments requires p47 which mediates the binding of p97 to a Golgi t-SNARE (soluble N-ethylmaleimide-sensitive factor attachment factor receptor), syntaxin 5. Here we show that it also suppresses the ATPase activity of p97 by up to 85% in a dose-dependent and saturable manner suggesting that it has other roles in the membrane fusion cycle.

Adenosine Triphosphatases↗
PubMed1998

Studies on selectin blockers. 7. Structure-activity relationships of sialyl Lewis X mimetics based on modified Ser-Glu dipeptides.

We have previously found that heterochiral fucodipeptides, L-Ser-D-Glu (3a) and D-Ser-L-Glu (3b), exhibited up to 20-100 times more potency than a sialyl Lewis X (sLeX, 1) and a 3'-sulfated Lewis X analogue (2) toward E-selectin binding and have also proposed, from molecular dynamics calculation, that their strong activities would depend on a possible formation of the type II and/or type II' beta-turn of compounds 3a,b (Tsukida, T.; Hiramatsu, Y.; Tsujishita, H.; Kiyoi, T.; Yoshida, M.; Kurokawa, K.; Moriyama, H.; Ohmoto, H.; Wada, Y.; Saito, T.; Kondo, H. J. Med. Chem. 1997, 40, 3534-3541). To clarify our hypothesis, we synthesized several analogues of compounds 3a,b and investigated their structure-activity relationships. As a result, it was indicated that the type II and/or type II' beta-turn conformation would be a comparatively tight form and would play important roles in favorable binding to E-selectin. These findings indicate that sLeX mimetics with type II and type II' beta-turn dipeptides could be a useful methodology for the design of an active selectin blocker.

Dipeptides↗
PubMed1998

A stereoselective alpha-fucosylation reaction using 1-hydroxy 2,3,4-tri-O-benzyl-L-fucose donor for the practical synthesis of selectin blocker.

A 1-hydroxy 2,3,4-tri-O-benzyl-L-fucose donor affords a high stereoselectivity of glycosylation in the presence of TMSOTf and is a very useful substrate for the preparation of an alpha-L-fucosyl dipeptide in a stereoselective manner. The donor will be a key component in the preparation of an attractively biological selectin blocker 1.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1998

Highly efficient recovery of functional single-chain Fv fragments from inclusion bodies overexpressed in Escherichia coli by controlled introduction of oxidizing reagent--application to a human single-chain Fv fragment.

An improved and efficient refolding system for a single-chain antibody fragment (scFv) from inclusion bodies expressed in Escherichia coli was developed. Stepwise removal of denaturing reagent and controlled addition of oxidizing reagent were found to be the most effective conditions to achieve for almost complete recovery of functional monomeric scFv from inclusion bodies. Adding L-arginine to the refolding solution also increased the yield of refolded functional scFv. The single-chain Fv fragments of both a mouse anti-lysozyme monoclonal antibody, HyHEL10, and a human monoclonal antibody against the D antigen of the Rh blood group, D10, in solubilized inclusion bodies could be refolded under these conditions with yields of up to 95%. The refolding procedures developed in this study will contribute to providing a stable supply of large amounts of human single-chain Fv fragments.

Animals↗
PubMed1998

Localization of mRNAs for CDP-diacylglycerol synthase and phosphatidylinositol synthase in the brain and retina of developing and adult rats.

CDP-diacylglycerol (CDP-DAG) synthase (CDS) is known as one of the key enzymes in the lipid synthesis including phosphoinositides (PIs) production. Phosphatidylinositol (PtdIns) synthase (PIS) catalyzes a formation of PtdIns from CDP-DAG in the PI cycle which produces several second messengers. We compared the gene expression for a presumably PI cycle specific-CDS molecule and a PIS using Northern blot analysis and in situ hybridization histochemistry in the central nervous system (CNS) and retina of developing and mature rats. Whereas no significant expression for CDS was detected during the prenatal stage in any CNS regions, PIS mRNA had already expressed on the prenatal day 15 throughout the neuroaxis including the spinal cord. During the postnatal stages, the gene expression for both CDS and PIS was detected widely in the gray matters throughout the entire brain. The expression for CDS was at higher levels in the olfactory mitral cells, the occipital cortex, the subiculum and hippocampal CA1 pyramidal cells, and the cerebellar Purkinje cells. On the other hand, the expression for PIS was at high levels in the olfactory mitral cells, the cerebral cortex, the hippocampal and dentate neuronal layer and the cerebellar Purkinje and granule cells. No significant expression for CDS or PIS was detected in the ventricular germinal zone, the cerebellar external granular layers, the mature ependyma or entire white matters. The expression for CDS and PIS decreased slightly throughout the CNS on P49. The significance of the parallel and discrepant expression patterns in terms of relative intensity between the two enzyme molecules was discussed in relation to the membrane turnover and signal transduction.

Animals↗
PubMed1998

Molecular characterization and tissue distribution of a new organic anion transporter subtype (oatp3) that transports thyroid hormones and taurocholate and comparison with oatp2.

Two complementary DNAs for the organic anion transporter subtypes oatp2 and oatp3, which transport thyroid hormones as well as taurocholate, were isolated from a rat retina cDNA library. The sequence of oatp2 is identical to that recently reported (Noé, B., Hagenbuch, B., Stieger, B., and Meier, P. J. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 10346-10350), whereas the sequence of oatp3 is novel. oatp3 consists of 670 amino acid residues and exhibits a structural architecture common to the organic anion transporter family, possessing the 12 putative membrane-spanning segments. Oocytes injected with oatp2 and oatp3 cRNAs showed taurocholate uptake in a saturable manner. The oatp2 and oatp3 cRNA-injected oocytes also showed significant uptake of both thyroxine and triiodothyronine. Northern blot and in situ analyses showed that the oatp2 mRNA was widely expressed in neuronal cells of the central nervous system, especially in the hippocampus, cerebellum, and choroid plexus as well as in the retina and liver. The oatp3 mRNA was highly expressed in the kidney and moderately abundant in the retina. This suggests that oatp2 and oatp3 are multifunctional transporters involved in the transport of thyroid hormones in the brain, retina, liver, and kidney.

Amino Acid Sequence↗
PubMed1998

Hammerhead ribozymes against gamma-glutamylcysteine synthetase mRNA down-regulate intracellular glutathione concentration of mouse islet cells.

gamma-Glutamylcysteine synthetase (gamma-GCS) is a key enzyme in glutathione synthesis and is thought to play a significant role in intracellular detoxification systems. To specifically suppress gamma-GCS gene expression, we constructed two different hammerhead ribozymes against gamma-GCS mRNA transcripts. Two cleavage sites were targeted as follows: site 1 for anti-gamma-GCS ribozyme (H), a GUU triplet located from +348 to +350 of the gamma-GCS heavy chain, and site 2 for anti-gamma-GCS ribozyme (L), a GUU triplet located from +235 to +237 of the gamma-GCS light chain. The anti-gamma-GCS ribozymes effectively cleaved gamma-GCS mRNA in a cell-free system. When transfected into a Min-6 mouse islet cell line, these anti-gamma-GCS ribozymes not only suppressed gamma-GCS gene expression, but also reduced intracellular glutathione concentration. These results suggest that the ribozyme-mediated down-regulation of gamma-GCS gene expression may be useful for analyzing the glutathione-associated cellular defense systems of pancreatic islet cells.

Animals↗
PubMed1998

Studies on selectin blockers. 6. Discovery of homologous fucose sugar unit necessary for E-selectin binding.

We describe a mimic of the sugar unit of the E-selectin ligand, sialyl Lewis X (sLeX). Carbohydrates are entering the realm of rational drug design, aided by the growing understanding of the structure-function relationships. We investigated a new methodology of preparing sLeX mimetics and developed a potent E-selectin blocker characterized by beta-turn dipeptides. Another characteristic point of this E-selectin blocker is that the six-membered fucose ring was replaced with a five-membered fucose ring. Interestingly, it was found that the five-membered fucose ring could also bind to a calcium ion on the E-selectin, which could be an important role of the six-membered fucose ring. Especially, the L-Ser-D-Glu and D-Ser-L-Glu derivatives 3a,b showed 65-90-fold more potent inhibitory activities than the sulfated LeX analogue 1. In addition, molecular dynamics (MD) studies indicated that the 2- and 3-OH groups of the six-membered fucose ring, which were necessary for the calcium binding, overlapped well with the 2- and 3-OH groups of the five-membered fucose ring. These new findings could be useful for the design of new types of selectin blockers.

Carbohydrate Sequence↗
PubMed1998

2-Arachidonoylglycerol, an endogenous cannabinoid receptor agonist: identification as one of the major species of monoacylglycerols in various rat tissues, and evidence for its generation through CA2+-dependent and -independent mechanisms.

The molecular species compositions of monoacylglycerols obtained from various rat tissues were examined by reverse-phase high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analyses. We confirmed that 2-arachidonoylglycerol, an endogenous cannabinoid receptor agonist, is one of the most abundant molecular species of monoacylglycerols in the brain. Substantial amounts of 2-arachidonoylglycerol were also found in the liver, spleen, lung and kidney, but the levels were considerably lower than that in the brain. We found that a small amount of 2-arachidonoylglycerol was generated in a brain homogenate during incubation in the absence of Ca2+. Importantly, the generation of 2-arachidonoylglycerol was markedly augmented in the presence of Ca2+, suggesting that Ca2+ plays a key role in regulation of the generation of 2-arachidonoylglycerol in this tissue.

Animals↗
PubMed1998

Intracellular Ca2+ store-operated influx of Ca2+ through TRP-R, a rat homolog of TRP, expressed in Xenopus oocytes.

To elucidate whether rat transient receptor potential (TRP-R), a rat TRP4 homolog, functions as a store-operated Ca2+ channel (SOC), we have measured the Ca2+ entry after thapsigargin treatment in Xenopus oocytes injected with mRNA for TRP-R. While non-injected oocytes elicited an SOC response, significantly larger responses were observed in the oocytes expressing TRP-R. The oocyte-native SOC response was inhibited by injection of antisense oligodeoxyribonucleotide for mammalian TRP1. When Ca2+ concentration-SOC response curve was examined, the EC50 value was much smaller in oocytes expressing TRP-R than that of non-injected oocytes. These results suggest that TRP-R functions as SOC having higher sensitivity to external Ca2+ than amphibian TRP1 channel.

Animals↗
PubMed1998

Cytological compartmentalization in the staggerer cerebellum, as revealed by calbindin immunohistochemistry for Purkinje cells.

The staggerer mouse carries a deletion in a gene encoding the nuclear hormone receptor RORalpha, which leads to severe impairments in phenotypic differentiation of cerebellar Purkinje cells. We previously found parasagittal compartments in the mature staggerer cerebellum, as defined by different transcription levels of Purkinje cell-specific molecules including calbindin. In the present study, we developed a hightiter anti-calbindin antibody to examine morphological features of the staggerer Purkinje cells. Immunohistochemistry for calbindin revealed compartmentalized Purkinje cell populations with different cell sizes, alignments, cell densities, and dendritic arborization, as well as different immunoreactivities, corresponding to the "transcriptional" compartments. Based on these immunohistochemical and cytological characteristics, the rostral cerebellum was clearly subdivided into three to seven parasagittal zones (Zones I-VII). Purkinje cells in Zones I and III were associated with the strongest calbindin immunoreactivities and exhibited morphological features reminiscent of the wild-type cells, i.e., large flask-shaped cell bodies, monolayer alignment, and arborized dendrites. Purkinje cells in Zone V were also labeled strongly, but they were small in cell size, ectopic and possessed long unbranched dendrites. On the other hand, Purkinje cells in Zones II, IV, and VI were very low in calbindin immunoreactivity and marked by small cell size, ectopia, poorly-developed dendrites and low cell density. Considering that this unique cytological compartmentalization emerges as the result of RORalpha gene mutation, it is suggested that normal cytodifferentiation of Purkinje cells is governed by both RORalpha-dependent and -independent mechanisms, and further that the latter mechanism might exert unevenly along the mediolateral cerebellar axis.

Animals↗