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Biomedical subjects

Yukio Kato

Publications and source records attributed to Yukio Kato.

At least 73 records · Page 4Linked to original sources

Dec1 and Dec2 expression is disrupted in the suprachiasmatic nuclei of Clock mutant mice.

DEC1 and DEC2 are basic helix-loop-helix transcription factors that functionally resemble negative feedback components of the mammalian circadian clock. The genes Dec1 and Dec2 are expressed rhythmically in the rat suprachiasmatic nuclei, and Dec1 expression is stimulated by light in a time-dependent manner with the kinetics of an immediate early gene. DEC1 and DEC2 can inhibit CLOCK:BMAL1 transactivation of the clock gene Per1, suggesting that these transcription factors may help regulate circadian timing. The authors present data on the expression pattern of Dec1 and Dec2 in wild-type and homozygous Clock mutant mice. In the suprachiasmatic nuclei, the Clock mutation significantly reduces the expression of Dec1 and Dec2. Dec1 becomes arrhythmic; Dec2 remains weakly rhythmic in a 12L:12D light-dark cycle but is arrhythmic in constant darkness. A robust attenuation of the Dec1 and Dec2 signals in Clock mutant mice was detected in all brain areas examined. These data point to up-regulation of Dec1 and Dec2 by Clock in vivo.

Animals↗

Prominent expression of spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1 in LH-producing cells, LbetaT2.

The LH-producing cell line, LbetaT2, and non LH-producing cell line, alphaT3-1 cells, established from a pituitary tumor, were employed for cDNA subtraction cloning to identify genes with expression unique to LH producing cells. Several cDNAs that code for known as well as for many unidentified clones were discovered. Most clones were the spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1, the abnormality of which causes neurodegeneration and loss of cerebellar Purkinje cells. We examined whether the expression of SCA1 gene in LbetaT2 cells is related to hormone production. We also compared the expression of SCA1 with that in various other pituitary tumor derived cell lines, and confirmed the prominent expression of SCA1 in LbetaT2 cells. The effect of gonadal factor(s) for SCA1 gene expression was examined. The expression level in female rats was low and did not change during the estrus cycle, but increased significantly after ovariectomy and did not return to the normal level under low and high doses of estrogen. In the male pituitary SCA1 gene expression increased markedly after castration and was not decreased by estrogen or testosterone. The Ontogeny of SCA1 gene expression was investigated in porcine fetal and postnatal pituitaries and revealed biphasic and sexually dimorphic expression. Transient expression of SCA1 gene was observed at fetal day 50 and 65 in males and day 40 in females, followed by a decline and increased expression before birth in both genders. Thus the expression of SCA1 gene is prominent in LH-producing cells and is not under direct control of gonadal factor(s) in both genders. In addition to the variable expression of SCA1 gene during the fetus period, the present results provide a novel aspect to the understanding of Boucher-Neuhauser syndrome (Ataxia Hypogonadism Choroidal Dystrophy).

Androgens↗

Anti-microbial action against verotoxigenic Escherichia coli O157:H7 of nitric oxide derived from sodium nitrite.

The levels of verotoxin-1 and verotoxin-2 released by verotoxigenic Escherichia coli O157:H7 treated in vitro with sodium nitrite, sodium chloride and several antibiotics were evaluated. Of the three strains of E. coli O157:H7 used in this study, two strains produced both verotoxin-1 and verotoxin-2, and one strain produced only verotoxin-2. Treatment of E. coli O157:H7 with sodium nitrite (6000 mg/l, minimum inhibitory concentration) did not increase the levels of verotoxin-1 and verotoxin-2 compared with a treatment by sodium chloride or antibiotics. When the electron paramagnetic resonance spectrum of sodium nitrite-treated bacterial cells was examined at 77 K to clarify the mechanism for the anti-bacterial activity of nitric oxide derived from sodium nitrite, electron paramagnetic resonance signals with g-values of 2.035 and 2.010 were observed. These were identified as being derived from iron-nitric oxide complexes. It appears that the dinitrosyl iron complexes in the E. coli O157:H7 cells were generated from the reaction of iron-sulfur proteins (enzymes) with nitric oxide formed by the reduction of sodium nitrite. The amount of ATP was decreased by the presence of sodium nitrite in the cell suspension. These findings indicate that nitric oxide derived from sodium nitrite penetrated the cells and inactivated enzymes related to the respiratory chain.

Adenosine Triphosphate↗

Automated ribotyping, a rapid typing method for analysis of Erysipelothrix spp. strains.

Automated ribotyping classified 70 Erysipelothrix species strains, previously classified into 14 RAPD patterns and into 63 PFGE patterns, into 27 ribogroups. Twenty-three strains of the 70 analyzed and classified into 13 ribogroups were previously classified into six ribotypes by the traditional ribotyping method. Moreover, automated ribotyping differentiated seven strains that were not differentiated by PFGE. Therefore, automated ribotyping was more sensitive than RAPD and traditional ribotyping, and it might be a useful method for a rapid screening in epidemiological study of strains of this genus, and more accurate results can be obtained when this method is used together with PFGE.

Automation↗

Measurement of polynuclear aromatic hydrocarbons in canine lung after alkaline decomposition.

An alkaline decomposition method employing a KOH/alcohol solution was studied, and polynuclear aromatic hydrocarbons (PAHs) contained in particles remaining in canine lung were measured. As a result, BaA, BkF, BaP, and BghiP were found. By this method, PAHs extracted from the lungs of 32 dogs were 13.0-166.0 ng (mean, 63.0 ng) for BaA, 6.6-90.2 ng (mean, 27.4 ng) for BkF, 9.8-167.4 ng (mean 47.2 ng) for BaP, and 10.8-206.0 ng (mean, 61.8 ng) for BghiP. The results showed no correlation between the age and the concentration of PAHs in the lung, but some correlation was found between the age and the lung weight (p<0.01). There were significant correlations among the concentrations of the compounds in the lung (p<0.01). These results suggest that dogs, like humans, are affected by automobile exhaust and other common generation sources of such substances.

Age Factors↗

Enhancement of periodontal tissue regeneration by transplantation of bone marrow mesenchymal stem cells.

BACKGROUND: The use of suitable cells transplanted into periodontal osseous defects appears to be a powerful strategy to promote periodontal tissue regeneration. Mesenchymal stem cells (MSCs) isolated from bone marrow have the potential for multilineage differentiation. The purpose of this study was to examine whether auto-transplantation of MSCs into periodontal osseous defects would be useful for periodontal tissue regeneration. METHODS: Bone marrow MSCs were isolated from beagle dogs and expanded in vitro. The expanded MSCs were mixed with atelocollagen (2% type I collagen) at final concentrations of 2 x 10(6), 5 x 10(6), 1 x 10(7), or 2 x 10(7) cells/ml, and auto-transplanted into experimental Class III defects. Atelocollagen alone was implanted into the defects as a control. Periodontal tissue healing was evaluated by histological and morphometric analyses 1 month after transplantation. RESULTS: The defects were regenerated with cementum, periodontal ligament, and alveolar bone in the MSC-atelocollagen groups. Less periodontal tissue regeneration was observed in the control group compared to the MSC-atelocollagen groups. Morphometric analysis revealed that the percentage of new cementum length in the 5 x 10(6) and 2 x 10(7) cells/ml groups and the percentage of new bone area in the 2 x 10(7) cells/ml group were significantly higher than in the control group (P < 0.01). CONCLUSION: These findings suggest that auto-transplantation of bone marrow mesenchymal stem cells is a novel option for periodontal tissue regeneration.

Alveolar Bone Loss↗

Functional regions of organic cation/carnitine transporter OCTN2 (SLC22A5): roles in carnitine recognition.

The organic cation/carnitine transporter OCTN2 transports carnitine in a sodium-dependent manner, whereas it transports organic cations sodium-independently. To elucidate the functional domain in OCTN2, we constructed chimeric proteins of human OCTN2 (hOCTN2) and mouse OCTN3 (mOCTN3) and introduced mutations at several amino acids conserved among human, rat and mouse OCTN2. We found that transmembrane domains (TMD) 1-7 are responsible for organic cation transport and for sodium dependence in carnitine transport. Within TMD1-7, Q180 and Q207 of hOCTN2 are the critical amino acids for the sodium dependence, and double mutation of Q180 and Q207 resulted in minimal change in transport activity when sodium was removed from the uptake medium. We propose that sodium-dependent affinity for carnitine is dependent on sodium recognition by these critical amino acids in hOCTN2, whereas carnitine transport by OCTN2 requires functional linkage between TMD1-7 and TMD11.

Amino Acid Sequence↗

Conditioning regimen of melphalan, fludarabine and total body irradiation in unmanipulated HLA haploidentical stem cell transplantation based on feto-maternal tolerance.

Unmanipulated hematopoietic stem cell transplantation from haploidentical family donors is frequently associated with graft failure and severe graft-versus-host disease (GVHD). We employed a myeloablative conditioning regimen consisting of 125 mg/m2 of fludarabine, 140 mg/m2 of melphalan and TBI of 10 to 12 Gy for three patients. The donor in each case was a haploidentical two-loci HLA mismatch mother or son. Engraftment failure was observed in one patient. In the other two patients, engraftment was confirmed within 15 days after transplantation. Acute and chronic GVHD was observed, but was controllable in both cases. HLA mismatched transplantation based on feto-maternal tolerance may provide an alternative option for patients who do not have HLA-matched donors.

Adult↗

Recognition of dileucine-based sorting signals from HIV-1 Nef and LIMP-II by the AP-1 gamma-sigma1 and AP-3 delta-sigma3 hemicomplexes.

The sorting of transmembrane proteins to endosomes and lysosomes is mediated by signals present in the cytosolic tails of the proteins. A subset of these signals conform to the [DE]XXXL[LI] consensus motif and mediate sorting via interactions with heterotetrameric adaptor protein (AP) complexes. However, the identity of the AP subunits that recognize these signals remains controversial. We have used a yeast three-hybrid assay to demonstrate that [DE]XXXL[LI]-type signals from the human immunodeficiency virus negative factor protein and the lysosomal integral membrane protein II interact with combinations of the gamma and sigma1 subunits of AP-1 and the delta and sigma3 subunits of AP-3, but not the analogous combinations of AP-2 and AP-4 subunits. The sequence requirements for these interactions are similar to those for binding to the whole AP complexes in vitro and for function of the signals in vivo. These observations reveal a novel mode of recognition of sorting signals involving the gamma/delta and sigma subunits of AP-1 and AP-3.

Adaptor Protein Complex 3↗

Novel cisplatin-incorporated polymeric micelles can eradicate solid tumors in mice.

Polymeric micelles incorporating cisplatin (CDDP) were prepared through the polymer-metal complex formation between CDDP and poly(ethylene glycol)-poly(glutamic acid) block copolymers, and their utility as a tumor-targeted drug delivery system was investigated. CDDP-incorporated micelles (CDDP/m) had a size of 28 nm with remarkably narrow distribution. CDDP/m were very stable in distilled water even in long-time storage, but exhibited a sustained drug release accompanied with the decay of the carrier itself in physiological saline. These micelles showed remarkably prolonged blood circulation and effectively accumulated in solid tumors (Lewis lung carcinoma cells) according to the passive targeting manner (20-fold higher than free CDDP). Reduced accumulation of the micelles in normal organs provided high selectivity to the tumor. In vivo antitumor activity assay demonstrated that both free CDDP and the CDDP/m had significant antitumor activity in C 26-bearing mice compared with nontreatment (P < 0.05 for free CDDP; P < 0.01 for CDDP/m), but complete tumor regression was observed only for the treatment with CDDP/m. Four of 10 mice treated with CDDP/m (4 mg/kg; five times administration at 2-day intervals) showed complete tumor regression with no significant body weight loss, whereas free CDDP treatment at the same drug dose and regime resulted in tumor survivals and approximately 20% of body weight loss. These data suggest that CDDP/m could be a promising formulation of CDDP for the targeted therapy of solid tumors.

Adenocarcinoma↗

Anti-membrane-bound transferrin-like protein antibodies induce cell-shape change and chondrocyte differentiation in the presence or absence of concanavalin A.

Membrane-bound transferrin-like protein (MTf), a glycosylphosphatidylinositol-anchored protein, is expressed at high levels in many tumors and in several fetal and adult tissues including cartilage and the intestine, as well as in the amyloid plaques of Alzheimer's disease, although its role remains unknown. MTf is one of the major concanavalin A-binding proteins of the cell surface. In this study, we examined the effects of anti-MTf antibodies and concanavalin A on cell shape and gene expression, using cultures of chondrocytes and MTf-overexpressing ATDC5 and C3H10T1/2 cells. In cultures expressing MTf at high levels, concanavalin A induced cell-shape changes from fibroblastic to spherical cells, whereas no cell-shape changes were observed with wild-type ATDC5 or C3H10T1/2 cells expressing MTf at very low levels. The cell-shape changes were associated with enhanced proteoglycan synthesis and expression of cartilage-characteristic genes, including aggrecan and type II collagen. Some anti-MTf antibodies mimicked this action of concanavalin A, whereas other antibodies blocked the lectin action. The findings suggest that the crosslinking of MTf changes the cell shape and induces chondrogenic differentiation. MTf represents the first identification of a plant lectin receptor involved in cell-shape changes and the differentiation of animal cells.

Animals↗

Enhancement of alkaline phosphatase synthesis in pulp cells co-cultured with epithelial cells derived from lower rabbit incisors.

Dental papilla mesenchymal cells differentiate into odontoblasts through epithelial-mesenchymal interactions. However, the mechanism by which enamel epithelial cells affect the differentiation of dental mesenchymal cells remains unknown. Alkaline phosphatase (ALPase) is a marker for odontoblast-like differentiation, because odontoblasts show much higher ALPase activity than dental undifferentiated mesenchymal cells. The continuously growing rabbit incisor is a good model for the epithelial-mesenchymal interaction during odontogenesis. In the present study, we isolated and maintained rabbit incisor-derived epithelial cells and rabbit incisor pulp-derived fibroblastic cells, and examined the effect of epithelial cells on ALPase activity in fibroblastic cells. Epithelial cells were stained with anti-cytokeratin 5 and 8 antibodies and showed the expression of tuftelin mRNA. In separate cultures of epithelial cells or fibroblastic cells, ALPase activity and mRNA levels were very low, but were upregulated in co-cultures of epithelial and fibroblastic cells. Histochemical analysis found high ALPase activity in fibroblastic cells close to epithelial cells. These findings suggest that epithelial cells play an important role in promoting ALPase expression in pulp fibroblastic cells. The co-culture system developed here will be useful for examining the role of the epithelial-mesenchymal interaction during odontoblast differentiation.

Alkaline Phosphatase↗

Influence of drugs and nutrients on transporter gene expression levels in Caco-2 and LS180 intestinal epithelial cell lines.

PURPOSE: The aim of this study was to investigate the influences of various drugs and nutrients on the expression levels of intestinal drug transporters PEPT1, MDR1, MRP2 and MRP3, and drug-metabolizing enzyme CYP3A4. METHODS: Quantitative reverse transcriptase polymerase chain reaction was used to quantitate transporter and CYP3A4 mRNAs. Western blotting was used to determine protein levels of P-gp. Transport studies of P-gp were performed using cultured Caco-2 cell monolayers. RESULTS: The expression of MDR1 mRNA was increased by all-trans retinoic acid and in glucose-depleted medium, whereas little change of MRP2 and MRP3 mRNA was observed in Caco-2 cells. Substrates and inducers of P-gp or CYP3A4 tended to produce parallel changes in the expression of MDR1 and CYP3A4 mRNA in LS180 cells, whereas in Caco-2 cells no such coordinate response was observed, possibly due to the absence of the expression of steroid xenobiotic receptor (SXR) in this cell line. CONCLUSION: Several drugs and nutrients were found to affect transporter gene expression level in two human intestinal epithelial cell lines. Since SXR is involved in some expression-regulatory processes, and Caco-2 cells lack SXR, LS180 cells as well as Caco-2 cells should be used for the study of the regulation of intestinal transporters.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Immunohistochemical study of matrilin-1 in arthritic articular cartilage of the mandibular condyle.

BACKGROUND: The objective of this study was to investigate the expression of matrilin-1 in arthritic articular cartilage of the mandibular condyle by means of immunohistochemical methods. METHODS: Condylar cartilage specimens were obtained from temporomandibular joints (TMJs) of 12 patients with arthritis (osteoarthritis and internal derangement) (mean age 51.8 years; age range 28-71 years) and four patients with TMJ ankylosis (mean age 44.0 years; age range 16-64 years), diagnosed clinically and with imaging examinations. Paraffin sections were immunostained with anti-matrilin-1 antibodies. RESULTS: Matrilin-1 expression was detected in both patient groups with TMJ ankylosis and arthritis, and the level was remarkably higher in arthritic cartilage. The mean percentage of matrilin-1-producing cells to the total chondrocytes was significantly (P < 0.05) greater in the arthritic group (43.9 +/- 19.2%) than in subjects with TMJ ankylosis (28.0 +/- 8.7%). CONCLUSIONS: Articular chondrocytes in the TMJ condyle can express matrilin-1 and the expression is enhanced in arthritic cartilage, suggesting a presence of functional or adaptive remodeling in the condyle in response to degenerative changes in the TMJ structures.

Adolescent↗

Involvement of organic cation transporter 1 in the lactic acidosis caused by metformin.

Biguanides are a class of drugs widely used as oral antihyperglycemic agents for the treatment of type 2 diabetes mellitus, but they are associated with lactic acidosis, a lethal side effect. We reported previously that biguanides are good substrates of rat organic cation transporter 1 (Oct1; Slc22a1) and, using Oct1(-/-) mice, that mouse Oct1 is responsible for the hepatic uptake of a biguanide, metformin. In the present study, we investigated whether the liver is the key organ for the lactic acidosis. When mice were given metformin, the blood lactate concentration significantly increased in the wild-type mice, whereas only a slight increase was observed in Oct1(-/-) mice. The plasma concentration of metformin exhibited similar time profiles between the wild-type and Oct1(-/-) mice, suggesting that the liver is the key organ responsible for the lactic acidosis. Furthermore, the extent of the increase in blood lactate caused by three different biguanides (metformin, buformin, and phenformin) was compared with the abilities to reduce oxygen consumption in isolated rat hepatocytes. When rats were given each of these biguanides, the lactate concentration increased significantly. This effect was dose-dependent, and the EC(50) values of metformin, buformin, and phenformin were 734, 119, and 4.97 microM, respectively. All of these biguanides reduced the oxygen consumption by isolated rat hepatocytes in a concentration-dependent manner. When the concentration required to reduce the oxygen consumption to 75% of the control value (from 0.40 to 0.29 micromol/min/mg protein) was compared with the EC(50) value obtained in vivo, a clear correlation was observed among the three biguanides, suggesting that oxygen consumption in isolated rat hepatocytes can be used as an index of the incidence of lactic acidosis.

Acidosis, Lactic↗

Thyroid hormone enhances aggrecanase-2/ADAM-TS5 expression and proteoglycan degradation in growth plate cartilage.

Effects of thyroid hormone on proteoglycan degradation in various regions of cartilage were investigated. In propylthiouracil-treated rats with hypothyroidism, proteoglycan degradation in epiphyseal cartilage during endochondral ossification was markedly suppressed. However, injections of T(4) reversed this effect of propylthiouracil on proteoglycan degradation. In pig growth plate explants, T(3) also induced breakdown of proteoglycan. T(3) increased the release of aggrecan monomer and core protein from the explants into the medium. Accordingly, the level of aggrecan monomer remaining in the tissue decreased after T(3) treatment, and the monomer lost hyaluronic acid-binding capacity, suggesting that the cleavage site is in the interglobular domain. The aggrecan fragment released from the T(3)-exposed explants underwent cleavage at Glu(373)-Ala(374), the major aggrecanase-cleavage site. The stimulation of proteoglycan degradation by T(3) was less prominent in resting cartilage explants than in growth plate explants and was barely detectable in articular cartilage explants. Using rabbit growth plate chondrocyte cultures, we explored proteases that may be involved in T(3)-induced aggrecan degradation and found that T(3) enhanced the expression of aggrecanase-2/ADAM-TS5 (a disintegrin and a metalloproteinase domain with thrombospondin type I domains) mRNA, whereas we could not detect any enhancement of stromelysin, gelatinase, or collagenase activities or any aggrecanase-1/ADAM-TS4 mRNA expression. We also found that the aggrecanse-2 mRNA level, but not aggrecanase-1, increased at the hypertrophic stage during endochondral ossification. These findings suggest that aggrecanse-2/ADAM-TS5 is involved in aggrecan breakdown during endochondral ossification, and that thyroid hormone stimulates the aggrecan breakdown partly via the enhancement of aggrecanase-2/ADAM-TS5.

ADAM Proteins↗