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Biomedical subjects

Y Kikkawa

Publications and source records attributed to Y Kikkawa.

At least 55 records · Page 3Linked to original sources
PubMed1996

Exposure to environmental tobacco smoke results in an increased production of (+)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide in juvenile ferret lung homogenates.

Six-week-old ferrets were exposed head-only to clean air or environmental tobacco smoke (ETS) at an average particulate concentration of 38 +/- 13 mg/m3 for 2 h/d, 5 d/wk for up to 15 wk. Twenty four hours after last exposure, the ferrets were sacrificed and the metabolism of benzo[a]pyrene and (-)-7R-trans-benzo[a]pyrene-7,8-dihydrodiol was studied in lung homogenates. The results show that after ETS exposure total metabolism of benzo[a]pyrene, measured by the accumulation of hexane nonextractable radioactivity, was increased by 35% in the males and 66% in the females (p < .05), respectively, of that observed with air-exposed controls. With (-)-7R-trans-benzo[a]pyrene-7,8-dihydrodiol as substrate, the formation of both benzo[a]pyrene-r-7,t-8,9,c-10-tetrahydrotetraol and (+)-anti-benzo[a]-pyrene-7,8-dihydrodiol-9,10-epoxide-derived tetraols by lung homogenates of ETS-exposed male and female ferrets was significantly increased compared to the air-exposed controls (p < .01). DNA-bound radioactivity was significantly increased in both the males (p < .01) and females (p < .05) compared to the air-exposed ferrets.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1996

Stimulation of endothelial cell migration in culture by ladsin, a laminin-5-like cell adhesion protein.

Ladsin is a laminin-like cell-adhesive scatter factor with potent cell motility-stimulating ability and was purified from serum-free conditioned medium of a malignant human gastric adenocarcinoma cell line STKM-1. To test its possible role in tumor angiogenesis, we investigated its effect on primary culture of endothelial cells (human umbilical vein endothelial cells) and endothelial cell line ECV304 in this study. Cell adhesion and motility effects of ladsin were observed in both types of endothelial cells. In cell-attachment assay, ladsin interacted with integrin alpha 3 beta 1 that was expressed on the endothelial cell surface. In Boyden chambers, ladsin stimulated both directed and random migration of ECV304 cells. Ladsin induced repair of artificial wounds generated in ECV304 cell monolayers by stimulating cell migration. Ladsin did not affect the growth rate of ECV304 cells at a low cell density but significantly increased the saturation cell density. These results suggest that ladsin may be involved in the adhesion and migration of endothelial cells under some physiological and pathological conditions.

Cell Adhesion↗
PubMed1996

Exclusion of Sox9 as a candidate for the mouse mutant tail-short.

The Sry-related gene Sox9 has been proposed as the gene responsible for the mouse skeletal mutant Tail-short (Ts), on the basis of its expression in skeletogenic mesenchymal condensations in the mouse embryo and its chromosomal location in the region of Ts on distal Chromosome (Chr) 11. We present here detailed mapping of Ts locus relative to the Sox9, using an intersubspecific cross. Among 521 backcross progeny, 16 recombinants were detected between Sox9 and Ts, suggesting a separation of 3.5 +/- 0.01 cM, and excluding Sox9 as a candidate for Ts. A further nine recombinants were detected between Ts and the polycomb-like gene M33, suggesting that these loci are separated by 1.8 +/- 0.011 cM. Six microsatellite markers were co-localized to the Ts locus, providing reagents for positional cloning of Ts.

Animals↗
PubMed1996

Differential expression of laminin-5/ladsin subunits in human tissues and cancer cell lines and their induction by tumor promoter and growth factors.

We previously reported a new laminin variant containing laminin gamma 2 (or B2t) chain, ladsin, which exerted prominent cell-scattering, cell-adhesion, and cell-migration activities. In the present study, this laminin was further characterized, and gene expression of its three subunits in various human tissues and cancer cell lines was examined by Northern blotting. cDNA cloning of the largest subunit of ladsin and partial amino acid sequencing of its beta (or B1) subunit revealed that ladsin was identical to laminin-5 (kalinin/epiligrin/ nicein). Among various human tissues, placenta, lung, and fetal kidney expressed high levels of mRNAs for the three subunits of laminin-5 (laminin alpha 3EPA, beta 3, and gamma 2 chains). Most gastric and squamous carcinoma cell lines constitutively expressed all of the three subunit mRNAs, while other types of carcinoma cell lines expressed one or two of them. The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) and epidermal growth factor (EGF) strongly enhanced the gene expression of the three subunits, increasing 2 to 8-fold the secretion of laminin-5 from carcinoma cells into culture medium. However, TPA treatment did not increase the secretion of laminin beta 1 chain, a subunit of laminins-1, -3, and -6. The unique properties and inducibility by TPA and EGF of laminin-5 suggest that it is associated with growth and migration of cancer cells.

Blotting, Northern↗
PubMed1996

[A case of relapsing demyelinating multiple mononeuropathy with multifocal nerve enlargement].

We report a 23-year-old woman, who had a relapsing-remitting multiple mononeuropathy with multifocal nerve enlargement. The patient was characterized by asymmetrical, marked enlargement of multiple nerves in the arms (median and ulnar nerves) and neck (accessory nerve) bilaterally. Sequential nerve conduction studies revealed persistent demyelinative abnormalities, especially across the segment of nerve thickening. The MRI of the proximal arm confirmed the markedly enlarged median and ulnar nerves which showed high signal intensity on T2-weighted images and partial enhancement after administration of gadolinium. We consider that the patient had the clinical features of "multifocal pseudohypertrophic neuropathy" described by Adams et al, and that chronic demyelination and inflammation associated with impairment of the blood-nerve barrier might be underlying mechanisms.

Adult↗
PubMed1995

Analysis of genetic diversity of domestic cattle in east and Southeast Asia in terms of variations in restriction sites and sequences of mitochondrial DNA.

There are three major groups of domestic cattle in East and Southeast Asia: European cattle, Zebu cattle, and Bali cattle. Ten restriction enzymes were used to analyze restriction site variants in the mitochondrial DNA (mtDNA) in 178 individuals belonging to these three groups of cattle. The results indicate that each of the three groups has mtDNA with a specific haplotype. The sequence of the mitochondrial gene for cytochrome b in representative haplotypes of Zebu and Bali cattle was determined and was compared with that of European cattle in the literature. We calculated 51 pairwise nucleotide sequence differences between European and Zebu cattle and 91 between European and Bali cattle. Our results suggest that ancestral populations of Asiatic domestic cattle may have diverged into two lineages--Bali and European plus Zebu--more than 3 million years ago, and then the European and Zebu groups diverged more than 1 million years or so before domestication occurred.

Animals↗
PubMed1995

Metabolism of (+)-trans-benzo[a]pyrene-7,8-dihydrodiol by 3-methylcholanthrene-induced rat liver homogenates.

Using a new sensitive reverse-phase HPLC assay with on-line radioactivity detector, metabolism of (+)-trans-benzo[a]pyrene-7,8-dihydrodiol (B[a]P diol) to the ultimate carcinogen benzo[a]pyrene-7,8-diol-9,10-epoxide (B[a]PDE) was studied using 3-methylcholanthrene-induced rat liver homogenates. The results demonstrate that the stereoselectivity of B[a]PDE formation is a function of the concentration of the cellular constituents in the incubation media. At more dilute concentrations of the homogenate, the ratio of anti- to syn-B[a]PDE was the highest and decreased as the homogenate protein was increased in the incubation medium. However, there was a marked and parallel decrease of free B[a]PDE and DNA-bound radioactivity with increasing concentrations of cellular constituents in the incubation medium. The decreased DNA-bound radioactivity appears to be due to the preferential binding of B[a]PDE to glutathione and to proteins as the homogenate concentration was increased in the incubation media. These results indicate that liver homogenates, while apparently preserving the function of microsomes, present additional opportunities to study the interrelationship among cytochrome P450 monooxygenase activity, water-soluble conjugates, and binding of B[a]P diol metabolites to macromolecules in the study of benzo[a]pyrene-induced carcinogenesis.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1995

Depression of hepatic cytochrome P450 monooxygenases after chronic environmental tobacco smoke exposure of young ferrets.

Six-week-old ferrets were exposed head-only to clean air or environmental tobacco smoke (ETS) for 2 h/day, 5 days a week for a total of 8 weeks. Exposure to ETS caused a significant reduction in the levels of hepatic microsomal cytochrome P450 and P450 reductase activity in both the male and female ferrets. The content of cytochrome b5 and the activity of its reductase were significantly reduced in the hepatic microsomes of female ferrets. 7-Ethoxycoumarin O-deethylase activity and cytochrome P450 (CYP) 1A protein were markedly decreased in the hepatic microsomes of both the male and female ferrets after ETS exposure. In accord with the downregulation of P450, total metabolites formed from benzo[a]pyrene were significantly reduced in the liver homogenates of ETS-exposed animals. Similarly, sum total of free (+)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide, glutathione conjugates and DNA-bound metabolites formed from precursor (-)-7R-trans-benzo[a]pyrene-7,8-dihydrodiol showed marked reduction in both the male and female ferrets after ETS exposure in a dose-response manner. This is the first report showing downregulation of hepatic P450 and accompanying benzo[a]pyrene metabolism after tobacco smoke exposure which apparently occurred after an initial upregulation of these parameters (Rasmussen et al. (1994) FASEB J. 8, A122).

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1995

Effect of environmental tobacco smoke on the metabolism of (-)-trans-benzo[a]pyrene-7,8-dihydrodiol in juvenile ferret lung and liver.

To evaluate the effects of "environmental tobacco smoke" (ETS) on developing lungs, juvenile ferrets were exposed to ETS at an average total particulate concentration of 381 +/- 97 mg/m3 for 2 h at the breathing zone. Twenty-four hours after the exposure, the ferrets were sacrificed and the metabolism of (-)-trans-benzo[a]pyrene-7,8-dihydrodiol was studied in the lung and liver homogenates. The rate of conversion of (-)-trans-benzo[a]pyrene-7,8-dihydrodiol to the ultimate carcinogen (+)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10- epoxide was twofold higher in the liver than that observed in the lung of control ferrets. After ETS exposure, the formation of free benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide was increased by 62% in the lung (p < .01). The DNA-bound metabolites were significantly increased only in the lung, while protein-bound metabolites were significantly increased in the liver after ETS exposure. Although glutathione conjugates tended to be increased both in the lung and liver, sulfate conjugates were significantly decreased in the lung after ETS exposure (p < .05). (+)-trans-Benzo[a]pyrene-7,8-dihydrodiol was used to study the relative contributions of cytochrome P-450 and peroxyl radical-mediated formation of benzo[a]-pyrene-7,8-dihydrodiol-9,10-epoxide. Peroxyl radical- and P-450-mediated conversion of (+)-trans-benzo[a]pyrene-7,8-dihydrodiol to benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide was proportionately equal in the ferret lung, whereas in the liver the P-450-mediated pathway was predominant. After ETS exposure there was a tendency for P-450-mediated formation of benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide to increase. These results demonstrate significant differences in the metabolism of (-)-trans-benzo[a]pyrene-7,8-dihydrodiol by the lung and liver of juvenile ferrets and suggest a significant role of peroxyl radical-mediated formation of (+)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide in the lung, which may help explain discrepancy between the levels of P-450 and amounts of DNA adducts of polycyclic aromatic hydrocarbons in different organs in smokers.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1995

Hepatic cytochrome P450 enzyme imprinting in adult rat by neonatal benzo[a]pyrene administration.

The effect of neonatal exposure to benzo[a]pyrene (BaP) on the hepatic cytochrome P450 of male and female adult rats has been examined. Newborn rats (< 24 h old) were injected with a single dose of BaP (1 mg/rat, s.c.) and killed after 110 days. In both sexes, body and liver weight, microsomal protein content, and total cytochrome P450 were unchanged. Cytochrome P450 1A2 protein content and 7-ethoxyresorufin O-deethylase activity were significantly decreased (p < 0.05) in males, whereas these were unaltered in females. Male-specific cytochrome P450 2C11 of male rats was significantly increased as shown by Western blot and increased testosterone 2 alpha- and 16 alpha-hydroxylase activities by 29% (p < 0.01) and 22% (p < 0.05), respectively. Female-specific cytochrome P450 2C12 protein content was unaltered in females. In addition, the level of free hepatic glucocorticoid receptor in adult males was elevated by 35% after BaP exposure, whereas it was unchanged in adult females. These results indicate, for the first time, that neonatal BaP exposure results in gender-specific lasting effects on hepatic cytochrome P450 1A2, cytochrome P450 2C11, and glucocorticoid receptors in adult male rats, whereas these parameters are unchanged in adult female rats.

Animals↗
PubMed1994

Marked stimulation of cell adhesion and motility by ladsin, a laminin-like scatter factor.

Ladsin is a large cell-adhesive protein with potent cell-scattering activity, which was recently identified in the culture of a malignant human gastric carcinoma cell line [Miyazaki, K. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 11767-11771]. It is a heterotrimeric protein, containing a 140-kDa subunit similar or identical to the laminin B2t chain. Ladsin is similar to the keratinocyte-derived matrix proteins, "epiligrin" and "kalinin." In the present study, the cell-adhesion and cell-migration activities of ladsin were examined in comparison with those of three cell adhesion proteins, laminin, fibronectin, and vitronectin. Ladsin showed high cell-adhesion activity toward rat liver cell line BRL at concentrations 4-20-times lower than in the case of the other three proteins. In a monolayer culture, ladsin stimulated the migration of BRL cells about 2-times more strongly than the others, as compared at the minimal concentrations required for the maximal cell-adhesion activity. In Boyden chambers, ladsin stimulated both the chemotactic and chemokinetic migration of BRL cells. When the effect of anti-integrin antibodies on the adhesion of human fibrosarcoma cell line HT1080 was examined, the adhesion to ladsin was effectively inhibited by both the anti-integrin alpha 3 and beta 1 antibodies, but not the anti-integrin alpha 6 antibody, indicating that the primary receptor of ladsin is integrin alpha 3 beta 1. These results demonstrate that ladsin is a unique extracellular matrix component which may play a major role in cell migration.

Adult↗
PubMed1994

Genetic variants of ribosomal DNA and mitochondrial DNA between swamp and river buffaloes.

To clarify the genetic relationship between Swamp and River buffaloes, the restriction fragment length polymorphisms (RFLPs) of nuclear genomic ribosomal DNA (rDNA) and cytoplasmic mitochondrial DNA (mtDNA) were analysed. Blood or liver samples from 73 Swamp and three River buffaloes were collected in East and South-east Asian countries. DNA samples from cattle, goats and sheep were used for comparisons. The analysis of rDNA allowed water buffaloes, cattle, goats and sheep to be characterized by four distinct repeat-types. However, swamp and river buffaloes showed the same repeat-type. Divergence of water buffalo and cattle is considered to have occurred approximately four to six million years ago. The RFLPs for mtDNA divided water buffaloes into three haplotypes, swamp-1, swamp-2 and river types. Swamp-1 accounted for 91% of all swamp buffaloes while swamp-2 was observed only in water buffaloes from Thailand (9%). All river buffaloes were of the same haplotype. No differences were observed between swamp and river buffaloes at the rDNA level. In contrast, a few distinct differences between them were found at the mtDNA level. Therefore, mtDNA polymorphisms provide an adequate means for classifying water buffaloes into either swamp or river buffaloes.

Animals↗
PubMed1994

Neonatal hyperoxia and cytochrome P450 imprinting in adulthood.

We hypothesized that during a critical neonatal period hyperoxia may produce alterations of sex-specific cytochrome P450 isozymes in adulthood (enzyme imprinting). To test this, newborn rats were exposed to 24 or 72 h of hyperoxia (O2 > 95%) within 24 h after birth and killed at 120 d. In males, significant negative imprinting (decrease) was found in total cytochrome P450 content and male-specific CYP2C11 in the hyperoxia groups. Positive imprinting (increase) was noted in CYP1A2 and male-specific CYP3A2 in the 72-h hyperoxia group. These alterations were essentially similar when expressed on a per microsomal protein or per liver basis. In addition, the level of hepatic glucocorticoid receptor in adult male rats was elevated after neonatal hyperoxia. In females, there was a significant body and liver weight loss after hyperoxic exposure, which resulted in a negative imprinting of CYP1A2 and female-specific 2C12 in the 72-h hyperoxia group on a per liver basis, whereas the measured parameters were unaltered when expressed per microsome. In general, the changes were more marked with longer hyperoxic exposure, suggesting that more pronounced alterations may be induced with prolonged neonatal hyperoxia. Because hyperoxic exposure in premature neonates is a common clinical practice and decreased CYP2C11 in adult males is expected to result in feminization, we believe that the scope of this work should be expanded and eventually tested for its relevance in human subjects.

Animals↗
PubMed1993

Induction of cytochrome P450 1A1 and 1A2 by hyperoxia.

The effect of hyperoxia (O2 > 95%) for 48 hours on the induction of pulmonary and hepatic cytochrome P450 has been investigated in adult male rats. Northern blot analysis using six "specific" oligonucleotide probes indicated that CYP 1A1 and CYP 1A2 mRNAs in liver and CYP 1A1 mRNA in lung were significantly increased by hyperoxic exposure, whereas the major constitutive P450 mRNAs, CYP 2C11 in liver and CYP 2B1 in lung, were decreased. Since induction of CYP 1A1 has only been reported with the use of exogenously administered xenobiotics, further studies were carried out to confirm the results obtained with Northern blot analysis. cDNAs were synthesized for CYP 1A1 and 1A2 in the liver and CYP 1A1 in the lungs and amplified by reverse PCR. These results indicate that these cDNAs were amplified significantly more in the hyperoxia group than in the control animals. Futhermore, CYP 1A1 and 1A2 proteins in liver and CYP 1A1 protein in lungs as well as the corresponding monooxygenase activities were increased by hyperoxia. Hyperoxic induction of CYP 1A1 and 1A2 is the first demonstration of nonexogenous CYP 1A induction in animals and indicates the needs to pursue the changes of Ah receptor-ligand-DNA interaction in hyperoxia.

Animals↗
PubMed1993

A large cell-adhesive scatter factor secreted by human gastric carcinoma cells.

Human gastric carcinoma cell line STKM-1 secretes a large protein that induces scattering of a rat liver epithelial cell line (BRL) into disconnected individual cells in monolayer culture. This cell-scattering factor was purified from serum-free conditioned medium of STKM-1 cells and found to be composed of three disulfide-linked subunits of 140, 150, and 160 kDa. The 140-kDa peptide contains an amino acid sequence homologous to that of the laminin B2t chain. The native protein has an apparent molecular mass of > 1000 kDa and a pI of 5.0. In addition to the cell-scattering activity, the purified protein stimulates attachment of BRL cells to substrate and their migration. Similar effects have been observed toward various cell lines, including nontumorigenic epithelial, endothelial, and fibroblastic cell lines and human cancer cell lines. Similar cell-scattering activity was secreted by human squamous carcinoma and gastric carcinoma cell lines and nontumorigenic epithelial and endothelial cell lines. These results indicate that the protein, named "ladsin," is probably an extracellular matrix protein that regulates cell-cell and cell-substrate interactions and cell migration.

Animals↗
PubMed1993

Purification and characterization of a two-chain form of tissue inhibitor of metalloproteinases (TIMP) type 2 and a low molecular weight TIMP-like protein.

Multiple forms of metalloproteinase inhibitors were found in the serum-free conditioned medium of the EJ-1 human bladder carcinoma cell line by reverse zymography assay with gelatinase A as the indicator enzyme. Two novel forms of inhibitor with apparent molecular masses of 18 and 22 kDa on nonreducing SDS-polyacrylamide gel electrophoresis (PAGE), together with tissue inhibitor of metalloproteinases (TIMP) and TIMP-2, were purified from the conditioned medium by a series of chromatographic steps. Structural analysis showed that the 18-kDa inhibitor is a two-chain form of TIMP-2 (tc-TIMP-2) produced by proteolytic processing, and the 22-kDa inhibitor may be a partially glycosylated form of TIMP. The purified tc-TIMP-2 was separated into a 17-kDa peptide and a small peptide of about 2.5 kDa by reducing SDS-PAGE and into four isoforms with pI 7.6, 7.3, 7.2, and 6.8 by isoelectric focusing. tc-TIMP-2 has essentially the same inhibitory activity as TIMP-2 toward gelatinase A, collagenase, stromelysin, and matrilysin. Unlike TIMP-2, however, tc-TIMP-2 does not bind to the latent precursor fo gelatinase A. Similar two-chain forms of TIMP-2 were produced by its partial digestion with trypsin or less effectively with plasmin. These results suggest that proteolytic processing of TIMP-2 plays a role in the regulation of gelatinase A activity in the extracellular matrix.

Amino Acid Sequence↗
PubMed1993

Recombinant human granulocyte colony-stimulating factor therapy for cyclic neutropenia associated with common variable immunodeficiency.

A 14 year old boy with common variable immunodeficiency (CVID) had regularly recurring episodes of severe infections independently of the serum gamma-globulin level. Serial blood counts revealed that this patient also had cyclic neutropenia. Recently, recombinant human granulocyte colony-stimulating factor (rhG-CSF) was reported to be an effective treatment for this disease. We tried rhG-CSF therapy for this patient and a prompt increase in the neutrophil count was noted. However, the cyclic alterations and duration of the nadir of the neutrophil count were not altered, which suggested that rhG-CSF has a variable efficacy in at least some patients with cyclic neutropenia.

Adolescent↗