Biomedical subjects

Takeshi Kitano

Publications and source records attributed to Takeshi Kitano.

15 recordsLinked to original sources

PubMed2006

Stereoselective reduction of 4-benzoylpyridine in the heart of vertebrates.

The stereoselectivity in the reduction of 4-benzoylpyridine (4-BP) was examined in the cytosolic fractions from the heart of 9 vertebrates (pig, rabbit, guinea pig, rat, mouse, chicken, soft-shelled turtle, frog and flounder). 4-BP was stereoselectively reduced to S(-)-alpha-phenyl-4-pyridylmethanol [S(-)-PPOL] in the cytosolic fractions from the heart of pig, rabbit and guinea pig. However, of mammalian heart cytsol tested, only rat heart cytosol had little ability to reduce stereoselectively 4-BP. In an attempt to elucidate this reason, amino acid sequence of rat heart carbonyl reductase (RatHCR) was deduced from the cloned cDNA and compared with that of pig heart carbonyl reductase (PigHCR), which shows a high stereoselectivity in the reduction of 4-BP to S(-)-PPOL. RatHCR showed a high identity with PigHCR in amino acid sequence. Furthermore, recombinant RatHCR was confirmed to reduce stereoselectively 4-BP to S(-)-PPOL with a high optical purity comparable to recombinant PigHCR. It is possible that in the cytosolic fraction from the heart of rat, constitutive reductase other than RatHCR counteracts the stereoselective reduction of 4-BP to S(-)-PPOL, by catalyzing the reduction of 4-BP to the R(+)-enantiomer.

Alcohol Oxidoreductases↗

PubMed2006

Transactivation activity of thyroid hormone receptors in fish (Conger myriaster) in response to thyroid hormones.

We previously isolated cDNAs encoding conger eel (Conger myriaster) thyroid hormone (TH) receptors (TRs). In the present study, we investigated the transactivation activities of conger eel TRs treated with THs (3,3',5-triiodo-l-thyronine [T3], l-thyroxine [T4], and 3,3',5'-triiodo-l-thyronine [rT3]), or ligands and activators of other nuclear receptors. Following transient transfection into the Japanese eel (Anguilla japonica) hepatocyte cell line Hepa-E1, the conger eel TRs (cTRs) showed TH-dependent activation of transcription from a TH-responsive promoter. However, no transactivation activity of any of the four cTRs was observed with ligands or activators of other nuclear receptors. Although T3 activation for cTRs is stronger than other THs (T3>T4>rT3), the transactivation sensitivity was different from the activity of cTRs with THs, respectively. Therefore, we conclude that cTRs can act in concert with THs in fish metamorphosis.

Animals↗

PubMed2005

Organochlorine contaminants in human adipose tissues from China: mass balance approach for estimating historical Chinese exposure to DDTs.

Concentrations of persistent organochlorines (OCs), such as DDTs, hexachlorocyclohexanes (HCHs), hexachlorobenzene (HCB), polychlorinated biphenyls (PCBs), and chlordane compounds (CHLs) were determined in 34 human adipose tissues collected from Guizhou Province, southern China, during 2002. DDT was the predominant contaminant among OCs analyzed; concentrations ranged from 420 to 20 000 ng/g on a lipid wt basis (average +/- sd: 5700 +/- 4100 ng/g). Concentrations of DDTs and the ratio of p,p'-DDT/ sigmaDDT in humans in China were significantly higher than those reported for developed countries. DDT levels in humans in Guizhou Province were comparable to those from Shanghai City, implying the presence of significant sources of DDTs in inland and coastal areas in China. Age-dependent accumulation of HCH concentration was found in this study, possibly because of the considerable reduction in average dairy intake (ADI) of HCHs by Chinese during the recent two decades. On the basis of the information of the ADI rates, half-lives, and body burdens of DDTs, the magnitude of historical DDT exposures by Chinese was estimated. This suggested that approximately 80% of DDTs deposited in Chinese adipose tissues was accumulated before the 1990s. The monitoring of OC levels and epidemiological studies are needed in China to understand the status of contamination and the risks to humans.

Adipose Tissue↗

PubMed2005

Concentrations and compositions of organochlorine contaminants in sediments, soils, crustaceans, fishes and birds collected from Lake Tai, Hangzhou Bay and Shanghai city region, China.

Contamination by persistent organochlorines (OCs), such as DDTs, hexachlorocyclohexane isomers (HCHs), chlordane compounds (CHLs), hexachlorobenzene (HCB) and polychlorinated biphenyls (PCBs) were examined in sediments, soils, fishes, crustaceans, birds, and aquaculture feed from Lake Tai, Hangzhou Bay, and in the vicinity of Shanghai city in China during 2000 and 2001. OCs were detected in all samples analyzed, and DDT and its metabolites were the predominant contaminants in most sediments, soils and biota. Concentrations of p,p'-DDT and ratio of p,p'-DDT to SigmaDDTs were significantly higher in marine fishes than those in freshwater fishes. While the use of DDTs has been officially banned in China since 1983, these results indicate a recent input of technical DDTs into the marine environment around Hangzhou Bay. Comparison of organochlorine concentrations in fishes collected from Lake Tai and Hangzhou Bay suggests the presence of local sources of HCHs, chlordanes and PCBs at Lake Tai. Higher proportions of penta- and hexa-PCB congeners in fishes at Lake Tai may suggest the use of highly chlorinated PCB product, such as PCB(5), around this lake. To our knowledge, this is a first comprehensive study to examine the present status of organochlorine contamination in various environmental media, such as sediments, soils and wildlife, in China.

Animals↗

PubMed2004

Sexually dimorphic expression of a teleost homologue of Müllerian inhibiting substance during gonadal sex differentiation in Japanese flounder, Paralichthys olivaceus.

Müllerian inhibiting substance (MIS), also known as anti-Müllerian hormone, is a glycoprotein belonging to transforming growth factor beta superfamily. In mammals, MIS is responsible for regression of Müllerian ducts, anlagen of the female reproductive ducts, in the male fetus. However, the role of MIS in gonadal sex differentiation of teleost fishes, which do not have the Müllerian ducts, has yet to be clarified. To address the role of MIS on gonadal sex differentiation in fishes, we isolated a MIS cDNA from the Japanese flounder testis and examined the expression pattern of MIS mRNA in gonads of both sexes during sex differentiation period. In this study, we present the first demonstration of sexually dimorphic expression of MIS mRNA during sex differentiation in teleost fishes, similarly to amniote vertebrates which possess the Müllerian ducts.

Amino Acid Sequence↗

PubMed2004

Identification of two teleost homologs of the Drosophila sex determination factor, transformer-2 in medaka (Oryzias latipes).

Transformer-2 (Tra2), an RNA-binding protein, is an important regulator in Drosophila sex determination. In vertebrates, however, the role of Tra2 homologues is not known. We identified two teleost homologues of Tra2, which we named Tra2a and Tra2b, in medaka (Oryzias latipes). Furthermore, we demonstrated that both Tra2 mRNAs were predominantly expressed in germ cells of both sexes before the onset of sex differentiation, suggesting that both Tra2 homologues might be involved in the sex differentiation in medaka.

Amino Acid Sequence↗

PubMed2004

An aromatase inhibitor or high water temperature induce oocyte apoptosis and depletion of P450 aromatase activity in the gonads of genetic female zebrafish during sex-reversal.

Dietary administration of a cytochrome P450 aromatase (P450arom) inhibitor (fadrozole) in genetic female juveniles of zebrafish (Danio rerio) was performed at 15-40 days post-hatching. The percentage of gonadal masculinization in the genetic all-females at 40 days post-hatching, treated with 0, 10, 100 and 1000 microg fadrozole g(-1) diet(-1) were 0, 62.5, 100 and 100%, respectively. Rearing at high water temperature in genetic all-females was performed at 15-25 days post-hatching. The percentage of gonadal masculinization in the genetic all-females at 40 days post-hatching, at water temperatures of 28.5, 35 and 37 degrees C were 0, 68.8 and 100%, respectively. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive oocytes of early diplotene and perinucleolar stages in fadrozole-treated genetic females (1000 microg g(-1) diet(-1)) were observed at 15-40 days post-hatching during sex-reversal. In contrast, apoptotic oocytes of early diplotene stage in high temperature-treated genetic females (at 35 and 37 degrees C) during sex-reversal and presumptive males of wild-type fish during sex differentiation were found at 15-27 days post-hatching. Our findings indicate that oocyte apoptosis, depletion of P450arom activity and differentiation of spermatogonia during gonadal sex-reversal are caused by treatments of aromatase inhibitor or high water temperature.

Animals↗

PubMed2004

Sexual plasticity in fish: a possible target of endocrine disruptor action.

Various genetic and molecular approaches have been used to investigate the mechanisms of sex determination, gonadal sex differentiation, and sex change in fish. We identified, for the first time in nonmammalian vertebrates, DMY, as the sex-determining gene of medaka. In tilapia, endogenous estrogens act as the natural inducers of ovarian differentiation, while DMRT1 may be important for testicular differentiation. In the protogynous wrasse, a rapid decline in serum estradiol-17beta levels may be an initial trigger of the female-to-male sex change. Both sex steroids and endocrine disrupters do not seem to act at the level of the sex-determining gene, but during gonadal sex differentiation. The Japanese flounder exhibits temperature-dependent sex determination. Some of the estrogenic endocrine disrupters induce feminization of the flounder larvae reared at the masculinizing temperature. The actions of these sex steroids and endocrine disrupters may be mediated by the actions of somatic cells within gonads. Thus, sexual plasticity of gonads during sex differentiation may be implicated through the somatic cells within gonads. Cloning and sequencing of a number of genes that are considered to be associated with gonadal sex differentiation have been performed and some are still in progress. These molecular probes provide useful tools for understanding not only the molecular mechanisms of sex determination and gonadal sex differentiation but also provide important basic information for studying the effects of endocrine-disrupting chemicals during these periods.

Animals↗

PubMed2003

Abnormal spermatogenesis at low temperatures in the Japanese red-bellied newt, Cynops pyrrhogaster: possible biological significance of the cessation of spermatocytogenesis.

In newt testis, spermatocytes never appear during winter, because secondary spermatogonia die by apoptosis just before meiosis. In the current study, we examined the effect of low temperatures on spermatogenesis. Incubation of newts at low temperatures (8, 12, 15 degrees C) induced defects in spermatogenesis in a temperature-dependent manner. At 8 degrees C, multinucleated giant cells (MGCs) were observed in spermatocytes and spermatogenesis never proceeded beyond meiosis. Although spermatocytes completed meiotic divisions at 12 degrees C, severe cell death was observed in the spermatids. At 15 degrees C both normal and abnormal spermiogenesis were observed. Under these conditions, impaired meiotic synapsis/recombination and down-regulation of the expression of the DMC1 protein, which play pivotal roles in meiotic pairing in eukaryotes, were also observed. Furthermore, to examine the quality of the sperm produced at low temperature for supporting development, artificial insemination was performed. The eggs inseminated with spermatozoa derived from newts kept at 15 degrees C demonstrated a restricted developmental capacity, even though these spermatozoa had an equal capacity for carrying out fertilization to those kept at 22 degrees C. These results suggest that meiosis at low temperatures cause the production of abnormal spermatozoa. Conservation and the significance of this phenomenon in poikilothermic vertebrates living in the temperate zones are also discussed.

Animals↗

PubMed2003

Low temperature promotes annexin V expression in newt testis.

We examined the effect of low temperatures on annexin V expression in newt testis. When newts were transferred to a low temperature (12 degrees C), up-regulation of annexin V protein was observed in secondary spermatogonia. In primary spermatocytes, high levels of annexin V expression were observed at both 12 degrees C and 22 degrees C, but at 12 degrees C the protein was localized in part of the cytoplasm of primary spermatocytes. These results indicate that in newt testis annexin V is a cold-sensitive protein, suggesting the possibility that annexin V might have a cold stress-related function in newt germ cells.

Animals↗

PubMed2003

Tributyltin causes masculinization in fish.

We examined the effect of tributyltin (TBT) on the sex differentiation process in genetically female Japanese flounder (Paralichthys olivaceus). The fish were fed an artificial diet containing tributyltin oxide (TBTO) at concentrations of 0.1 and 1.0 microg/g diet from 35 to 100 d after hatching, which includes the sex differentiation period. The ratio of sex-reversed males significantly increased to 25.7% of the flounder fed the 0.1 microg/g diet and to 31.1% of those fed the 1.0 microg/g diet compared with the control (2.2%). From morphological and histological examination of the fish in the TBT-treated groups, normal females had typical ovaries and sex-reversed males had typical testes. These results clearly demonstrated the masculinization of flounder exposed to TBTO. This is the first report of TBT inducing sex reversal in vertebrates.

Animals↗

PubMed2002

Promotion of cathepsin L activity in newt spermatogonial apoptosis induced by prolactin.

We previously showed that prolactin (PRL) induces apoptosis in newt secondary spermatogonia and indicated that caspase activity is involved in the apoptosis. Since it was recently reported that Z-VAD-fmk, a pan-caspase inhibitor, blocks activity of cysteine cathepsins as well, we examined whether cathepsin is involved in the newt spermatogonial apoptosis. We found cathepsin L activity in the testis that was elevated by PRL in organ culture of testis, while E-64d, a lysosomal cysteine protease inhibitor, and Z-VAD-fmk suppressed it and chromosomal condensation. These results suggest that cathepsin L activity play a pivotal role in PRL-induced spermatogonial apoptosis.

Animals↗

PubMed2002

Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish.

Large numbers of apoptotic early diplotene oocytes were observed during the transition from ovary-like undifferentiated gonadal tissue to testes during sex differentiation in presumptive males of the zebrafish (Danio rerio). The percentage of terminal-deoxynucleotidyl-transferase-mediated dUTP nick-end labelling (TUNEL)-positive apoptotic oocytes in the gonads of presumptive males was approximately eight- to 12-fold higher than in genetic all-females. By 29 days post-hatching, all oocytes had disappeared from the gonads of presumptive males. In these males, we also observed apoptotic somatic cells in the ovarian cavity between 23 and 35 days post-hatching. Therefore, the disappearance of oocytes and the decomposition of the ovarian cavity caused by apoptosis during sex differentiation were male-specific events. In genetic all-females, apoptosis in a proportion of early diplotene oocytes was found in the undifferentiated gonads at 15-19 days post-hatching, probably as a result of programmed oocyte loss during ovarian development. These findings suggest that oocyte apoptosis is the mechanism of testicular and ovarian differentiation in zebrafish.

Animals↗

PubMed2002

Purification and identification of a tributyltin-binding protein from serum of Japanese flounder, Paralichthys olivaceus.

Tributyltin (TBT) is an industrial chemical used as an antifoulant in marine environments. Previously, we reported that TBT accumulates in the serum or plasma of some fishes and is bound to a high molecular weight compound in the serum of the Japanese flounder, Paralichthys olivaceus. In this study, we succeeded in purifying the TBT-binding protein (TBT-bp) from the serum of Japanese flounder by using gel filtration chromatography, anion exchange chromatography, and polyacrylamide gel electrophoresis, with a 2.6% yield and a 77-fold purification. The molecular mass of TBT-bp was approximately 46.5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its isoelectric point was approximately 3.0 on isoelectric focusing-polyacrylamide gel electrophoresis. The TBT-bp contained 42% N-glycan. The cDNA nucleotide sequence of TBT-bp was determined by reverse transcription-polymerase chain reaction of Japanese flounder liver, and we deduced a sequence of 191 amino acids of mature TBT-bp. No sequence identical to the TBT-bp amino acid sequence was found within the SWISS-PROT (http://www.nig. ac.jp/) protein database; however, a lipocalin-like sequence pattern was observed. We concluded that the TBT-bp was a novel protein that has not yet been reported, although some DNA sequences from expressed sequence tags (ESTs) of Japanese flounder liver had a high identity. A high expression level of TBT-bp gene was found in the liver, but the gene was slightly detectable in the kidney and brain.

Amino Acid Sequence↗

PubMed2000

Electron paramagnetic resonance study utilizing stripping method on normal human stratum corneum.

BACKGROUND/AIM: Electron paramagnetic resonance (EPR) spectra of nitroxide spin probes are useful for studying biological membranes, and chemical-membrane interaction. The objective of this study was to assess the fluidity of stripped stratum corneum (SC) of normal human skin with EPR. METHODS: We quantified the fluidity of stripped SC (S-SC) by a single stripping with cyanoacrylate onto quartz glass, cover glass or transparent film. Spectra were compared with those of the SC of cadaver skin (C-SC). RESULTS: All S-SC spectra were similar to those of C-SC. There were no statistical differences in the order parameters S between C-SC and those on quartz cell and cover glass, but those of transparent film were significantly higher. Spectra after a month storage at room temperature at 5 degrees C and at -20 degrees C were unchanged. The order parameter S had decreased after a 3-month storage in all conditions. CONCLUSION: This method is useful for the evaluation of the fluidity measurement utilizing EPR spectra of S-SC.

Journal Article↗