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Biomedical subjects

A Kobayashi

Publications and source records attributed to A Kobayashi.

At least 55 records · Page 3Linked to original sources

Interaction between ATBP and DmUbc9 in the expression of the Sarcophaga lectin gene.

We previously demonstrated that (A+T)-stretch binding protein (ATBP) and Dorsal-related immunity factor (Dif) are required for the expression of the Sarcophaga lectin gene in SL-2 cells (Aozasa et al., Eur. J. Biochem. 268, 2506-2511, 2001). The present study demonstrates that DmUbc9 interacts with ATBP, and cotransfection of the DmUbc9 vector with ATBP and Dif vectors greatly enhances the expression of the luciferase reporter of the Sarcophaga lectin gene in SL-2 cells. These results suggest that sumoylation of ATBP is involved in the expression of the Sarcophaga lectin gene in this system.

Animals↗

Thermogenesis induced by intravenous infusion of hypertonic solutions in the rat.

1. Intravenous administration of 20-60 % glucose, 3.2-9.7 % NaCl or 20 % mannitol solutions (1.66 ml kg(-1)) for 5 min increased oxygen consumption in urethane-anaesthetized rats, whereas administration of physiological saline had no effect. Administration of 7.7-18.3 % urea slightly increased the oxygen consumption, but the increase was significantly smaller than that measured after the administration of other hypertonic solutions. The magnitude of the thermogenic effect correlated with the osmolality of the applied solutions. These results suggest that the thermogenesis was caused mainly by changes in osmolality rather than by a specific action of the different solute molecules. 2. Neither pretreatment with the ganglion blocker hexamethonium (20 mg kg(-1), I.P.) or the beta-adrenergic antagonist propranolol (10 mg kg(-1), I.P.), nor bilateral cervical vagotomy or bilateral adrenalectomy had any effect on the osmotically induced thermogenesis. Therefore, the autonomic nervous system and the adrenal gland were not involved in this metabolic response. 3. In response to osmotic stimulation, the temperature of the skeletal muscle increased significantly, whereas that of brown adipose tissue did not change and that of the colon and liver decreased. Accordingly, the site of osmotic thermogenesis is probably in the skeletal muscle, although osmotic stimulation was not accompanied by electromyographic activity and was not blocked by pretreatment with muscle relaxants such as dantrolene sodium or pancuronium bromide, or with the Na(+)-Cl(-) co-transport inhibitor bumetanide. 4. The increases in plasma osmolality observed after the administration of 20 % (1.3 osmol kg(-1)) glucose and 4.1 % (1.3 osmol kg(-1)) NaCl were 4.50 +/- 0.88 and 5.57 +/- 0.71 mosmol kg(-1), respectively. Since the slight increase in osmolality is well within the physiological range of changes that occur after food ingestion, diet-induced thermogenesis may have a component that is mediated by an increase in plasma osmolality, which results from the prandial increase in circulating nutrients.

Adipose Tissue, Brown↗

Superconductivity in an organic insulator at very high magnetic fields.

We investigate by electrical transport the field-induced superconducting state (FISC) in the organic conductor lambda-(BETS)2FeCl4. Below 4 K, antiferromagnetic-insulator, metallic, and eventually superconducting (FISC) ground states are observed with increasing in-plane magnetic field. The FISC state survives between 18 and 41 T and can be interpreted in terms of the Jaccarino-Peter effect, where the external magnetic field compensates the exchange field of aligned Fe3+ ions. We further argue that the Fe3+ moments are essential to stabilize the resulting singlet, two-dimensional superconducting state.

Journal Article↗

Human chromosome 19 and related regions in mouse: conservative and lineage-specific evolution.

To illuminate the function and evolutionary history of both genomes, we sequenced mouse DNA related to human chromosome 19. Comparative sequence alignments yielded confirmatory evidence for hypothetical genes and identified exons, regulatory elements, and candidate genes that were missed by other predictive methods. Chromosome-wide comparisons revealed a difference between single-copy HSA19 genes, which are overwhelmingly conserved in mouse, and genes residing in tandem familial clusters, which differ extensively in number, coding capacity, and organization between the two species. Finally, we sequenced breakpoints of all 15 evolutionary rearrangements, providing a view of the forces that drive chromosome evolution in mammals.

Animals↗

GATA DNA-binding protein expressed in mouse I-10 Leydig testicular tumor cells.

A nuclear extract of the mouse I-10 Leydig tumor cell line was analyzed by gel mobility shift assay with a combination of antibodies for various mammalian GATA proteins. Antibodies for GATA-4 caused a super-shift of the DNA-protein complex, which is formed through GATA-4 binding to an oligonucleotide with a typical GATA motif, while ones for GATA-1, GATA-2, GATA-3, and GATA-6 did not. These results indicated that I-10 cells express GATA-4 protein. Western blotting analysis of cellular proteins also demonstrated the presence of GATA-4 protein, the size of which corresponds to that of the rat orthologous protein transiently expressed in Cos-1 cells. A significant level of GATA-4 expression in I-10 cells would be advantageous for studying the roles of this protein, especially in view of gonadal function. We further examined the binding site preference of GATA-4 expressed in I-10 cells. GATA-4 showed broad sequence specificity similar to GATA-6, the order of binding core site preference being GATA > GATT > GATC, and adenine was favored on both sides of the core for strong binding. Thus the conserved zinc finger domain of GATA proteins is suggested to contribute to the binding sequence preference. GATA-4 expressed in I-10 cells was not susceptible to proteolysis coupled with cAMP signaling.

Animals↗

Magnetic-field-induced superconductivity in a two-dimensional organic conductor.

The application of a sufficiently strong magnetic field to a superconductor will, in general, destroy the superconducting state. Two mechanisms are responsible for this. The first is the Zeeman effect, which breaks apart the paired electrons if they are in a spin-singlet (but not a spin-triplet) state. The second is the so-called 'orbital' effect, whereby the vortices penetrate into the superconductors and the energy gain due to the formation of the paired electrons is lost. For the case of layered, two-dimensional superconductors, such as the high-Tc copper oxides, the orbital effect is reduced when the applied magnetic field is parallel to the conducting layers. Here we report resistance and magnetic-torque experiments on single crystals of the quasi-two-dimensional organic conductor lambda-(BETS)2FeCl4, where BETS is bis(ethylenedithio)tetraselenafulvalene. We find that for magnetic fields applied exactly parallel to the conducting layers of the crystals, superconductivity is induced for fields above 17 T at a temperature of 0.1 K. The resulting phase diagram indicates that the transition temperature increases with magnetic field, that is, the superconducting state is further stabilized with magnetic field.

Journal Article↗

Thermogenesis induced by osmotic stimulation of the intestines in the rat.

Infusion of 5-20% glucose, 1.8-3.6% NaCl, 20% methylglucose, 20% fructose, or 5-10% solutions of various amino acids (10 ml x kg(-1)) into the duodenum induced dose-dependent thermogenesis in urethane-anaesthetized rats. In contrast, infusion of 0.9% NaCl, distilled water, or safflower oil had no effect on the metabolic rate. Infusion of 7.2% urea induced a small and transient increase in the metabolic rate. These results suggested that the thermogenesis was caused mainly by changes in osmolality rather than by a specific action of the different solute molecules. The respiratory exchange ratio increased after the infusion of glucose, fructose, glycine, or serine, did not change after the infusion of NaCl, methylglucose, safflower oil, or distilled water, and decreased after infusion of arginine. Therefore, there was no relationship between substrate utilization and the occurrence of thermogenesis. Intestinal infusion of 3.6% NaCl elevated the plasma osmolality, with a plateau increase of approximately 20 mosmol x kg(-1). However, intravenous infusion of the same amount of NaCl induced a significantly smaller thermogenic response, although it elevated the plasma osmolality with a time course and magnitude similar to those obtained after the intestinal infusion. Infusion of NaCl into the hepatic portal vein or the peritoneal cavity also produced a significantly small thermogenic response. These results suggested an intestinal or mesenteric location for osmoreceptors. To test for possible stimulation of intestinal osmoreceptors after intake of a normal meal, we measured the osmolality of the intestinal contents. The osmolality of the duodeno-jejunal contents was 600-800 mosmol kg-1, whereas the plasma osmolality was 306 +/- 1 mosmol x kg(-1), which suggests that the intestinal osmoreceptors are stimulated after meals and are involved in diet-induced thermogenesis.

Amino Acids↗

High-resolution analysis of polyprenols by supercritical fluid chromatography.

A high-resolution analysis of polyprenol mixtures was achieved by supercritical fluid chromatography (SFC). The separation of polyprenols was examined on an octadecylsilane-packed column with liquid carbon dioxide as the mobile phase and ethanol as modifier. Using this chromatography system, the resolution of separation (Rs) between octadecaprenol (prenol 18) and nonadecaprenol (prenol 19) was two times higher than that using conventional reversed-phase high-performance liquid chromatography. Our SFC technique allows the advantage of baseline separation of polyprenol samples containing hydrophobic components such as terpenes or fatty acids that are unfavorable for good separation. This method is very useful for the analysis of structurally close polyprenol analogues of rubber plant metabolites.

Chromatography, Liquid↗

A novel antiferromagnetic organic superconductor kappa-(BETS)(2)FeBr(4) [where BETS = bis(ethylenedithio)tetraselenafulvalene].

The electrical and magnetic properties of kappa-(BETS)(2)FeBr(4) salt [where BETS = bis(ethylenedithio)tetraselenafulvalene] showed that this system is the first antiferromagnetic organic metal at ambient pressure (T(N) = 2.5 K). The characteristic field dependence of the magnetization at 2.0 K indicates a clear metamagnetic behavior. The small resistivity drop observed at T(N) clearly shows the existence of the interaction between pi metal electrons and localized magnetic moments of Fe(3+) ions. In addition, this system underwent a superconducting transition at 1.1 K. That is, kappa-(BETS)(2)FeBr(4) is the first antiferromagnetic organic metal exhibiting a superconducting transition below Néel temperature. The magnetic field dependence of the superconducting critical temperature indicated that the superconductivity in this system is strongly anisotropic also in the conduction plane because of the existence of the metamagnetically induced internal field based on the antiferromagnetic ordering of the Fe(3+) 3d spins in contrast to the cases of the other conventional organic superconductors. Furthermore, the specific heat measurement exhibited a lambda-type large peak of zero-field specific heat corresponding to the three-dimensional antiferromagnetic ordering of high-spin Fe(3+) ions. The lack of distinct anomaly in the C(p) vs T curve at T(c) suggests the coexistence of the superconductivity and the antiferromagnetic order below T(c).

Journal Article↗

A three-dimensional synthetic metallic crystal composed of single-component molecules.

Molecular metals normally require charge transfer between two different chemical species. We prepared crystals of [Ni(tmdt)2] (tmdt, trimethylenetetrathiafulvalenedithiolate) and carried out crystal structure analyses and resistivity measurements. The analyses and measurements revealed that these single-component molecular crystals are metallic from room temperature down to 0.6 kelvin. Ab initio molecular orbital calculations suggested that pi molecular orbitals form conduction bands. The compact molecular arrangement, intermolecular overlap integrals of the highest occupied and lowest unoccupied molecular orbitals, and tight-binding electronic band structure calculation revealed that [Ni(tmdt)2] is a three-dimensional synthetic metal composed of planar molecules.

Journal Article↗

Mutations in the TATA-binding protein, affecting transcriptional activation, show synthetic lethality with the TAF145 gene lacking the TAF N-terminal domain in Saccharomyces cerevisiae.

The general transcription factor TFIID, which is composed of the TATA box-binding protein (TBP) and a set of TBP-associated factors (TAFs), is crucial for both basal and regulated transcription by RNA polymerase II. The N-terminal small segment of yeast TAF145 (yTAF145) binds to TBP and thereby inhibits TBP function. To understand the physiological role of this inhibitory domain, which is designated as TAND (TAF N-terminal domain), we screened mutations, synthetically lethal with the TAF145 gene lacking TAND (taf145 Delta TAND), in Saccharomyces cerevisiae by exploiting a red/white colony-sectoring assay. Our screen yielded several recessive nsl (Delta TAND synthetic lethal) mutations, two of which, nsl1-1 and nsl1-2, define the same complementation group. The NSL1 gene was found to be identical to the SPT15 gene encoding TBP. Interestingly, both temperature-sensitive nsl1/spt15 alleles, which harbor the single amino acid substitutions, S118L and P65S, respectively, were defective in transcriptional activation in vivo. Several other previously characterized activation-deficient spt15 alleles also displayed synthetic lethal interactions with taf145 Delta TAND, indicating that TAND and TBP carry an overlapping but as yet unidentified function that is specifically required for transcriptional regulation.

Amino Acid Substitution↗

A lipid-coated lipase as an efficient hydrolytic catalyst in the two-phase aqueous-organic system.

A lipid-coated lipase has been known to be soluble in organic solvents and act as an efficient esterification catalyst in the dry organic solvent. It was also found to act as an efficient hydrolytic catalyst for lipophilic esters in the two-phase aqueous-organic system. Both the lipid-coated lipase and substrates are solubilized in the organic phase and the hydrolysis occurs with water molecules from the aqueous phase. Therefore, the reaction was 40-100 times faster than that of the native lipase, in which the enzyme and substrate exist separately in the aqueous and organic phase, respectively, and the reaction proceeded at the interface. The hydrolysis rates for the lipid-coated lipase were not affected by the aqueous pH and agitation speed of the two-phase. Enzymatic activity of the lipid-coated lipase was compared with that of the poly(ethylene glycol)-grafted lipase. Michaelis-Menten kinetics in the two-phase reactions was also studied.

Alcohols↗

Contribution of serotonin to liver injury following canine small-intestinal ischemia and reperfusion.

Intestinal ischemia and reperfusion (I/R) has been shown to be associated with multiple organ damages. Serotonin (5-hydroxytriptamine; 5-HT), which is synthesized in the enterochromaffin cells in the intestine and stored in platelets, is known to play an important role in platelet aggregation and vasoconstriction and may ultimately enhance such organ injuries. The purpose of this study was to investigate the association between liver damage and 5-HT levels in the liver after intestinal I/R. The entire canine small intestine, isolated on a vascular pedicle that consisted of the proximal superior mesenteric artery and superior mesenteric vein, was subjected to 4-h ischemia by clamping these vessels and the marginal arteries supplying the proximal and distal ends of the small intestine. Hepatic blood flow, liver tissue blood flow, bile flow rate, and hepatic venous ketone body ratio (HVKBR) were measured before and at the end of intestinal ischemia and at 5, 15, and 30 min, and 1 and 2 h after reperfusion. 5-HT levels in plasma of the portal vein and hepatic vein were assayed at the same intervals. Time-matched, sham-operated animals served as controls. Intestinal I/R significantly decreased the liver tissue flow, bile flow rate, and HVKBR. Compared to those in controls, 5-HT levels in the portal vein and hepatic vein were markedly increased after reperfusion. Furthermore, intravenous administration of 5-HT receptor antagonists attenuated the liver dysfunction after intestinal reperfusion. These results suggest that intestinal I/R induces continuous disturbance of hepatic microcirculation, leading to liver dysfunction, and that 5-HT may be implicated as one of the mediators of liver dysfunction after intestinal I/R.

Animals↗

Tubular aggregates observed in spindle muscle fiber of horse lumbrical muscle.

Tubular aggregates (TAs) originate from the sarcoplasmic reticulum (SR) and form polymorphic double (or single) -walled structures in cross section. TAs are involved in various human skeletal muscle disorders including periodic paralysis, congenital myasthenic syndromes, inflammatory myopathies, and malignant hyperthermias. Horse lumbrical muscle (LM) is a slender fusiform muscle that shows varying degrees of regression due to its limited activity in the limb. Double-walled TAs were found in degenerating spindle fibers and with a range of 80-116 nm (average 92 nm, n=135) for outer layer and 50-78 nm (average 59 nm, n=135) for the inner layer. TAs exhibit degradation of myofibrillar proteins, disruption of mitochondria with cristae lost, glycogen accumulation, electron-dense metabolic products, blebbing appearance of sarcolemma, and presence of various vacuoles. LM fibers also show a similarly degenerative state. The disassembly of the SR structure probably produces a large accumulation of SR proteins which remain as molecules without being further degraded and which could aggregate to form the orderly structure of TAs. We believe that TA formation may be an adaptation to store unbalanced extra proteins by forming ordered aggregates in degeneration caused by stress in cells.

Animals↗

Plasma-membrane H+-ATPases are expressed in pitchers of the carnivorous plant Nepenthes alata Blanco.

Nepenthes is a unique genus of carnivorous plants that can capture insects in trapping organs called pitchers and digest them in pitcher fluid. The pitcher fluid includes digestive enzymes and is strongly acidic. We found that the fluid pH decreased when prey accumulates in the pitcher fluid of Nepenthes alata. The pH decrease may be important for prey digestion and the absorption of prey-derived nutrients. To identify the proton pump involved in the acidification of pitcher fluid, plant proton-pump homologs were cloned and their expressions were examined. In the lower part of pitchers with natural prey, expression of one putative plasma-membrane (PM) H+-ATPase gene, NaPHA3, was considerably higher than that of the putative vacuolar H+-ATPase (subunit A) gene, NaVHA1, or the putative vacuolar H+-pyrophosphatase gene, NaV-HP1. Expression of one PM H+-ATPase gene, Na-PHA1, was detected in the head cells of digestive glands in the lower part of pitchers, where proton extrusion may occur. Involvement of the PM H+-ATPase in the acidification of pitcher fluid was also supported by experiments with proton-pump modulators; vanadate inhibited proton extrusion from the inner surface of pitchers, whereas bafilomycin A1 did not, and fusicoccin induced proton extrusion. These results strongly suggest that the PM H+-ATPase is responsible for acidification of the pitcher fluid of Nepenthes.

Amino Acid Sequence↗

Relationship between cervical curvature index (Ishihara) and cervical spine angle (C2--7).

The purpose of this study was to determine the relationship between the cervical curvature index (Ishihara) and the cervical spine angle (C2--7) from both cervical lateral radiographs and a geometrical model. The cervical lateral radiographs of 295 outpatients with no cervical symptoms were reviewed to measure the cervical curvature index (Ishihara) and the cervical spine angle (C2--7). The average cervical curvature index (Ishihara) was 10.9 (SD, 15.3) and the average cervical spine angle (C2--7) was 20.3 (SD, 14.3). A highly significant correlation (0.95; 95% confidence interval [CI], 0.94--0.96) was found between the cervical curvature index (Ishihara) and the cervical spine angle (C2--7). Their linear correlation was also validated in the geometrical model. Their correlation diminished, however, in subjects with an S-shaped cervical spine. The cervical curvature index (Ishihara) and the cervical spine angle (C2--7) were interchangeable, but this interchangeability was lost in the S-shaped cervical spines.

Adolescent↗

The occurrence of geometric polyprenol isomers in the rubber-producing plant, Eucommia ulmoides Oliver.

The chain length and geometric isomerism of polyprenols from Eucommia ulmoides Oliver were analyzed using supercritical fluid chromatography. After intensive effort to establish separation conditions for geometric isomers, a phenyl-bonded silica gel-packed column was found that cleanly separated poly-trans and -cis prenols. The presence of long-chain poly-trans prenols (>9 mers) was confirmed for the first time in plants. Trans isomers were found in the leaf, seed coat, and root, but not in the bark and seed. Poly-trans prenols in this plant may act as intermediates for trans-polyisoprene biosynthesis.

Chromatography, High Pressure Liquid↗

Receptor-mediated cell modulator delivery to hepatocyte using nanoparticles coated with carbohydrate-carrying polymers.

Cell modulators such as colchicine (CO), cytochalasin B (CY) and taxol (TX) loaded nanoparticles coated with carbohydrate-carrying polymers, as hepatocyte-specific targeting material using galactose ligands as recognition signals to asialoglycoprotein receptors were prepared by the diafiltration method. Effects of cell modulators from their loaded nanoparticles on morphology of hepatocytes were studied. Receptor-mediated endocytosis of the nanoparticles were examined by fluorescence and confocal laser microscopy. It was found that the shapes of most hepatocytes were changed for the CY-loaded, TX-loaded, or CO-loaded nanoparticles whereas their shapes were not changed in comparison with control when CY, TX, or CO were mixed with the nanoparticles. From the fluorescence and confocal laser microscopic studies, it is suggested that the nanoparticles coated with sugar-carrying polymers were internalized by the hepatocytes through the receptor-mediated mechanism.

Animals↗