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Biomedical subjects

K Yazaki

Publications and source records attributed to K Yazaki.

At least 37 records · Page 2Linked to original sources
PubMed1998

Characterization of homo-oligomeric complexes of alpha and beta chaperonin subunits from the acidothermophilic archaeon, Sulfolobus sp. strain 7.

The chaperonin from the acidothermophilic archaeon, Sulfolobus sp. Strain 7, is composed of two kinds of subunits designated as Scp alpha and Scp beta. In this study, we characterized the recombinant Scp alpha and Scp beta, which were separately expressed in Escherichia coli. Both of them were able to assemble to homo-oligomeric double-ring complexes, similar to subunits of group II chaperonins from Thermoplasma acidophilum and Thermococcus strain KS-1. Both complexes have no or at most trace ATPase activities. However, they could arrest spontaneous refolding of chemically denatured enzyme in the same way as the purified Sulfolobus chaperonin. We found that they dissociated in the presence of 15% ethanol to monomers, which spontaneously assembled to oligomers when concentrated in the absence of ethanol. Both the reconstituted homo-oligomers were unstable, and easily dissociated to monomers. Further structural and functional characterization is necessary to elucidate if these homo-oligomers exist and if so, their function in vivo.

3-Isopropylmalate Dehydrogenase↗
PubMed1998

Production of macrocyclic ellagitannin oligomers by Oenothera laciniata callus cultures.

Callus cultures of Oenothera laciniata grown on LS agar medium supplemented with IAA and kinetin produced large amounts of the macrocyclic ellagitannin dimer, oenothein B, and a trimer, oenothein A, accompanied with related monomeric hydrolysable tannins. The content of the main compound oenothein B (65 mg/g dry wt) in calli cultured on modified LS medium containing 10 mM NH4+ and 5 mM NO3- was nearly two times higher than that in intact leaves.

Biopolymers↗
PubMed1998

Mitochondrial dysfunction of human muscle in chronic alcoholism detected by using 31P-magnetic resonance spectroscopy and near-infrared light absorption.

We previously examined the effect of alcohol on muscle energy metabolism in chronic alcoholics by using 31P-magnetic resonance spectroscopy. Measurements of intracellular pH and PCr index [PCr/(PCr + Pi)] during resting, hand grasping, and recovery in the left flexor digitorum superficialis muscle of alcoholics with neurological signs showed a marked decrease and delayed recovery of pH, but rapid recovery of PCr index indicating that the muscle produces lactate during and after exercise to maintain the ATP level. To clarify the reason for this preference for anaerobic metabolism, we conducted simultaneous measurements of the muscle blood supply during and after exercise by using the near-infrared light method and energy metabolism by using 31P-magnetic resonance spectroscopy. In alcoholics with neurological signs, we observed a significant increase of oxyhemoglobin after exercise with a slight increase of total hemoglobin. In healthy volunteers and chronic alcoholics without neurological signs, such an overshoot of oxyhemoglobin was not observed. We conclude that chronic alcoholics with neurological signs have an abnormality of aerobic metabolism owing to muscle mitochondrial dysfunction.

Acid-Base Equilibrium↗
PubMed1998

[Effects of a short-term pulmonary rehabilitation program on patients with chronic respiratory failure due to pulmonary emphysema].

To evaluate the effects of a short-term pulmonary rehabilitation program on dyspnea, exercise capacity, and lung function, 15 patients with chronic respiratory failure due to pulmonary emphysema were enrolled in such a program for 3 weeks as inpatients. The program consisted of pursed lip breathing, diaphragmatic breathing, respiratory muscle stretch gymnastics, and walking with synchronized breathing. Dyspnea as measured with a visual analogue scale at the end of a 6-minute walk before and after the program (49.7 +/- 4.0% to 24.2 +/- 3.8%) decreased significantly (p < 0.01). As a measure of functional exercise capacity, the 6-minute walking distance (226.9 +/- 32.4 m to 292.1 +/- 35.8 m) increased significantly (p < 0.01). As an indicator of maximal exercise capacity, endurance time on an incremental treadmill test did not improve. Spirometric data did not change during the study. Total lung capacity (TLC) (8.44 +/- 0.70 L to 7.58 +/- 0.74 L) and residual volume (RV) (5.13 +/- 0.53 L to 4.28 +/- 0.59 L) decreased significantly (p < 0.01). The findings suggest that this program relieves dyspnea, increases functional exercise capacity, and decreases TLC and RV on patients with chronic respiratory failure due to pulmonary emphysema.

Aged↗
PubMed1997

Structural and functional characterization of homo-oligomeric complexes of alpha and beta chaperonin subunits from the hyperthermophilic archaeum Thermococcus strain KS-1.

To elucidate the function of group II chaperonin, the gene for the chaperonin from the hyperthermophilic archaeum Thermococcus strain KS-1 was cloned and sequenced. Two distinct genes coding for chaperonin subunits, designated alpha and beta, were obtained, and their deduced amino acid sequences are highly homologous to those of group II chaperonins from other sources. The alpha and beta subunits were individually expressed in Escherichia coli. Both of the recombinant subunits assemble to constitute the homo-oligomeric double-ring complexes, which are prone to form large aggregates. The alpha aggregate is dissociated into the typical chaperonin ring complex by incubation in buffer containing 15% (v/v) methanol, while the beta aggregate cannot be dissociated. At high temperature, both of the recombinant complexes have weak ATPase activities. They are able to arrest refolding of a chemically denatured thermophilic enzyme in the absence of ATP, and refolding is resumed when ATP is supplemented. These results suggest that homo-oligomeric complexes of the archaeal chaperonin have activity.

3-Isopropylmalate Dehydrogenase↗
PubMed1997

Envelope protein of parasitic wasp symbiont virus, polydnavirus, protects the wasp eggs from cellular immune reactions by the host insect.

Cotesia kariyai polydnavirus (CkPDV) virions are present in the oviducts of C. kariyai wasp and are injected with eggs into the hemocoel of the host armyworm Pseudaletia separata larvae during parasitization. Evidence that the presence of polydnavirus particles on the surface of the wasp eggs may be essential for prevention of cellular immune reactions by the host hemocytes was obtained by isolating an immunoevasive factor from CkPDV virions. The purified proteinaceous factor protects foreign materials from adhesion and encapsulation by hemocytes of the host P. separata larvae but not by those of common cutworm Spodoptera litura larvae which is an incompatible host for the C. kariyai wasp. Purification procedures consisted of extraction with ethanol/trifluoroacetic acid and reverse-phase high performance liquid chromatography. A factor with a molecular mass of approximately 50 kDa is demonstrated to be present on the envelope of CkPDV virion by immunoelectronmicroscopic observations. Furthermore, immunoreactive proteins are found in plasma of the armyworm larvae but not in the common cutworm larvae, indicating that only the natural host of C. kariyai wasp shares a similar epitope with CkPDV. The sequence of 23 amino acid residues at the amino terminus of the factor was determined to be Ile-Ser-Val-Glu-Asn-Val-Xaa-Thr-Thr-Gly-Ile-Phe-Leu-Asp-Ser-Gly-Glu-Xaa- Val- Pro-Tyr-Ala-Thr-Lys-Pro.

Animals↗
PubMed1997

Nerve growth factor promotes giant-cell transformation of mouse trophoblast cells in vitro.

We examined the effects of NGF on the TGC (trophoblastic giant cell) transformation of the ectoplacental cone (EPC) cells derived from the mouse pregnant uteri on day 7.5 p.c. The addition of NGF strongly accelerated the rate of the TGC transformation of EPC cells in a dose-dependent manner. The promoting action of NGF was abolished by the addition of anti-NGF MAb. Histochemical staining with succinyl wheat germ agglutinin (s-WGA), an in situ marker for the TGCs on day 10.5 p.c., revealed that NGF induced the expression of s-WGA-binding glycoproteins in TGCs in vitro. RNA analysis revealed that NGF mRNA was expressed in the pregnant uterus on day 7.5 p.c., mainly in the decidua, but it could not be detected in the EPC. p75NGFR mRNA was expressed in the EPCs, whereas TrkA mRNA was not detected in the placental tissues throughout day 7.5 to 10.5 p.c. We therefore conclude that maternally derived NGF may play a role in mouse placentation by promoting the giant-cell transformation of trophoblast cells through p75NGFR.

Animals↗
PubMed1997

Transmission electron and atomic force microscopic observation of polysomes on carbon-coated grids prepared by surface spreading.

A simple modification of surface spreading that enables easy examination of subcellular structures by both transmission electron microscopy (TEM) and atomic force microscopy (AFM) was developed in the present study. Specimens were adsorbed onto carbon-coated electron microscopy (EM) grids with parlodion backing for comparison of images obtained using TEM and AFM. Fine structures of ribosomes and polysomes were observed in this study. Washing the mounted specimen with detergent such as 0.05% Photo-Flo 200 before conducting negative staining or metal shadowing brought clear visualization of ribosomes and polysomes. The same preparation could apply to AFM imaging without any additional treatment.

Animals↗
PubMed1997

cDNA cloning and gene expression of phenylalanine ammonia-lyase in Lithospermum erythrorhizon.

Two cDNA clones (LEPAL-1 and LEPAL-2) encoding phenylalanine ammonia-lyase were isolated from cell suspension cultures of Lithospermum erythrorhizon. Northern kinetic studies showed that LEPAL-1 mRNA contents markedly increased one day after inoculation of the cells into fresh medium, then decreased to the steady-state level. The course of mRNA accumulation paralleled that of PAL enzyme activity. The rapid induction of PAL activity seems to reflect the induction of dihydroechinofuran biosynthesis, while shikonin was produced at the steady-state level of PAL activity. The course of LEPAL-2 mRNA accumulation seemed to be similar to, but much lower than that of LEPAL-1. In the intact plant, both genes are expressed mainly in the root, the organ in which shikonin is exclusively produced and accumulated. Genomic Southern blot analyses showed that both genes are present in the genome as single copies.

Amino Acid Sequence↗
PubMed1996

cDNA cloning, characterization, and functional expression of 4S-(-)-limonene synthase from Perilla frutescens.

A molecular biological study on limonene synthase that catalyzes the cyclization of geranyldiphosphate to yield the olefin 4(S)-limonene, an intermediate in the biosynthesis of a monoterpenoid, perillaldehyde, in Perilla frutescens Britton has been carried out. We isolated and characterized 10 cDNAs homologous to spearmint limonene synthase cDNA from an expression library constructed from cotyledons of a Perilla strain homozygous for two pairs of dominant genes, G and H, which are responsible for the formation of 4(S)-limonene. Two of these cDNA clones were functionally expressed in Escherichia coli, yielding enzymes which were catalytically active in generating 4(S)-limonene from geranyldiphosphate. The longest open reading frame in the representative cDNA clone PFLC1 consisted of 1812 nucleotides corresponding to 603 amino acids. Its identity to the ORFs of spearmint limonene synthase, tobacco 5-epi-aristolochene synthase, and castor bean casbene synthase were 65, 35, and 30%, respectively. Genomic Southern blot analyses of various genotypes (GGHH, GGhh, ggHH, and gghh) of P. frutescens suggested that more than one copy of the PFLC1 DNA exists in strains having the HH genotype. In contrast, no PFLC1 DNA sequences were found in the genomes of strains with the hh genotype that are incapable of producing cyclohexanoid monoterpenes for lack of limonene synthase activity. Northern blot analysis, using a PFLC1 3'-flanking region as a hybridization probe, showed that PFLC1 mRNA accumulated in all the aerial parts of the GGHH plants, particularly in the leaves. In the ggHH plants, on the other hand, PFLC1 mRNA was detected only in minute amounts in the stem and calyx.

Amino Acid Sequence↗
PubMed1996

Identification of two Sox17 messenger RNA isoforms, with and without the high mobility group box region, and their differential expression in mouse spermatogenesis.

The different mRNA isoforms of the mouse Sox17 gene were isolated from adult mouse testis cDNAs. One form (referred to as form Sox17) encodes an Sry-related protein of 419 amino acids containing a single high mobility group box near the NH2-terminus, while the other form (referred to as form t-Sox17) shows a unique mRNA isoform of the Sox17 gene with a partial deletion of the HMG box region. Analysis of genomic DNA revealed that these two isoforms were produced at least by alternative splicing of the exon corresponding to the 5' untranslated region and NH2-terminal 102 amino acids. RNA analyses in the testis revealed that form Sox17 began at the pachytene spermatocyte stage and was highly accumulated in round spermatids. Protein analyses revealed that t-Sox17 isoforms, as well as Sox17 isoforms, were translated into the protein products in the testis, although the amount of t-Sox17 products is lower in comparison to the high accumulation of t-Sox17 mRNA. By the electrophoretic mobility-shift assay and the random selection assay using recombinant Sox17 and t-Sox17 proteins, Sox17 protein is a DNA-binding protein with a similar sequence specificity to Sry and the other members of Sox family proteins, while t-Sox17 shows no apparent DNA-binding activity. Moreover, by a cotransfection experiment using a luciferase reporter gene, Sox17 could stimulate transcription through its binding site, but t-Sox17 had little effect on reporter gene expression. Thus, these findings suggest that Sox17 may function as a transcriptional activator in the premeiotic germ cells, and that a splicing switch into t-Sox17 may lead to the loss of its function in the postmeiotic germ cells.

Amino Acid Sequence↗
PubMed1996

Reinitiation of spermatogonial mitotic differentiation in inactive old BDF1 mouse seminiferous tubules transplanted to W/Wv mouse testis.

The seminiferous epithelia of old mice (33 mo of age) are composed of spermatogonia and Sertoli cells. Histochemical examination using the anti-c-kit monoclonal antibody demonstrated that the number of differentiating type A spermatogonia decreases with age. To elucidate the differential activity of old mouse spermatogonia, we transplanted extremely thin seminiferous epithelia of old BDF, mice into W/Wv mouse testes and examined whether or not they could reinitiate differentiation. Artificially cryptorchid mice were used as the control. At 2 wk after transplantation, spermatocytes and round spermatids were detected in transplanted seminiferous tubules of the control, whereas the most advanced spermatogenic cells in those of old mice were spermatocytes. At 4 wk after transplantation, although elongated spermatids were detected in transplanted tubules of the control, haploid cells (spermatids) were still undetectable in those derived from old mice. Thus, meiosis was never restored, although spermatogonia of old mice can reinitiate differentiation into spermatocytes under suitable testicular conditions. Since it has been reported in several mammalian species that age-related changes in the testicular microenvironment lead to the gerontal cessation of spermatogenesis, the present results suggest that both a defective extratubular environment and a defective intratubular environment may cause the cessation of spermatogenesis in old BDF, mice.

Aging↗
PubMed1996

Mitosis-specific phosphorylation of smooth muscle regulatory light chain of myosin II at Ser-1 and/or -2 and Thr-9 in sea urchin egg extract.

We analyzed the kinase activities capable of phosphorylating the regulatory light chain of myosin-II (MRLC) from chicken gizzard in unfertilized and fertilized sea urchin egg extracts. Total kinase activity phosphorylating MRLC in vitro did not fluctuate throughout the first cell cycle. Phosphopeptide mapping analysis showed that MRLC was phosphorylated at two different sites corresponding to myosin light chain purified from chicken gizzard (MLCK) and protein kinase C (PKC) phosphorylation sites, namely MLCK and PKC sites, respectively. The activity of the kinase(s) responsible for phosphorylation of MRLC at PKC sites showed a significant increase at metaphase. Phosphoamino acid analysis revealed that this increase in MRLC phosphorylation was due to phosphorylation at serine residue (Ser-1 and/or Ser-2) and a threonine residue (Thr-9). This increase in phosphorylation at PKC sites is occurred concomitantly with an increase in histone H1 kinase activity. In contrast, MRLC phosphorylation at MLCK sites showed no significant changes during the first cell cycle. Butyrolactone I, a selective inhibitor of p34cdc2 kinase, inhibited the activity of the kinase(s) responsible for phosphorylation of MRLC at PKC sites at metaphase. These results suggest that the increase in MRLC phosphorylation at PKC sites (Ser-1 and/or -2, and Thr-9) at metaphase may be induced by p34cdc2 kinase. Thus, p34cdc2 kinase may be involved in the regulation of MRLC phosphorylation during cell division.

4-Butyrolactone↗
PubMed1996

Treatment of essential and parkinsonian tremor with nipradilol.

Nipradilol is a new type of beta-blocker which possesses nitroglycerin-like vasodilating action in addition to beta-blocking action. We investigated the efficacy and safety of nipradilol for treating tremor in 20 patients with essential tremor (ET group) and 20 patients with Parkinson's disease (PD group). All patients received nipradilol (6 mg per day) for more than 8 weeks. Improvement of tremor appeared within 2 or 4 weeks after the start of nipradilol therapy, and the efficacy rate, defined as "moderately effective" or over, was 42.5% in all 40 patients, while that defined as "slightly effective" or over was 87.5%. The efficacy rate tended to be higher in the ET group compared with the PD group. Mean blood pressure was significantly decreased from the 4th week after the start of treatment and heart rate was significantly reduced from the 2nd week of treatment. Laboratory examination showed no significant changes.

Adrenergic beta-Antagonists↗
PubMed1996

Effect of chronic alcohol intake on energy metabolism in human muscle.

We investigated the effect of alcohol on muscle energy metabolism by using 31P magnetic resonance spectroscopy in 12 chronic alcoholics [6 with neurological signs and symptoms (such as cerebellar ataxia or diplopia) and 6 without neurological signs or symptoms], compared with five healthy subjects who also received acute alcohol loading. Intracellular pH and phosphocreatine (PCr) index [PCr/ (PCr + Pi)] were measured during rest, exercise, and recovery in the left flexor digitorum superficialis muscle. In healthy subjects, acute alcohol loading did not influence the changes of muscle pH and PCr index. Alcoholics with neurological signs showed marked decreases in muscle intracellular pH and PCr index during exercise and a marked delay of pH recovery after exercise. There was no delay of PCr index recovery. Alcoholics without neurological signs showed slight decreases in intracellular pH and PCr index, but rapid recovery of both intracellular pH and PCr index was observed. Marked decrease and delayed recovery in pH, but rapid recovery of PCr index, indicate that the muscle of patients with neurological signs produced lactate during and after exercise to maintain the ATP level, which implies that anaerobic metabolism is favored over aerobic metabolism in these patients.

Adult↗
PubMed1995

Effects of extracellular matrix on differentiation of mouse fetal gonads in the absence of mesonephros in vitro.

The influence of mesonephric tissues and the extracellular matrix on mouse gonadal differentiation was examined in vitro. Gonadal ridges, with or without the adjacent mesonephric region, were removed from mouse embryos on day 12 post coitum (p.c.), and cultured in the presence or absence of reconstituted basement membrane (matrigel) for 5 days. Culturing control undifferentiated testes with mesonephric tissues induced normal testicular differentiation. When testes without mesonephric tissues were cultured in the absence of matrigel, testicular cord formation was not observed in the explants. Sertoli cells were irregularly arranged in the testicular parenchyma, and no continuous basal lamina was formed around the Sertoli cells. However, when testes without mesonephric tissues were embedded in matrigel and cultured for 5 days, the Sertoli cells were organized into testicular cord-like structures. The Sertoli cells positioned at the base of the cord-like structures were closely connected to the matrigel at their basal surface, and showed a polarized distribution of vimentin filaments in their basal cytoplasm. Leydig cells, on the other hand, were differentiated in all testicular explants. In all ovarian explants, germ cells normally entered meiotic prophase. Therefore, these findings indicate that the extracellular matrix permits testicular differentiation in the absence of the mesonephros, and that removal of mesonephric tissues leads to developmental failure of cord formation because the components of the extracellular matrix around pre-Sertoli cells are incomplete.

Animals↗