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Biomedical subjects

Y Ohta

Publications and source records attributed to Y Ohta.

At least 109 records · Page 6Linked to original sources
PubMed2000

Desulfurization of alkylated forms of both dibenzothiophene and benzothiophene by a single bacterial strain.

Thirty-five bacterial strains capable of converting dibenzothiophene into 2-hydroxybiphenyl were isolated. Among them Rhodococcus erythropolis KA2-5-1 was chosen for further characterization because of its ability to retain high desulfurization activity stably. PCR cloning and DNA sequencing of a KA2-5-1 genomic DNA fragment showed that it was practically identical with dszABC genes from Rhodococcus sp. IGTS8, a representative carbon-sulfur-bond-targeted dibenzothiophene-degrading bacterium. KA2-5-1 desulfurized a variety of alkyl dibenzothiophenes through the specific cleavage of their C-S bonds. In addition, unexpectedly, KA2-5-1 also attacked alkyl benzothiophenes in a C-S-bond-targeted fashion. The purified monooxygenase, encoded by dszC of KA2-5-1, converted benzothiophene and dibenzothiophene into benzothiophene sulfone and dibenzothiophene sulfone, respectively, with the aid of an NADH-dependent oxidoreductase. This result raises the possibility that the same enzymatic step may be involved in desulfurization of alkylated forms of both dibenzothiophene and benzothiophene in KA2-5-1 cells.

Alkylation↗
PubMed2000

Copper-catalyzed autoxidations of GSH and L-ascorbic acid: mutual inhibition of the respective oxidations by their coexistence.

Glutathione (GSH) is known to inhibit copper-catalyzed autoxidation of L-ascorbic acid (AA); in this study, AA was found to conversely inhibit copper-catalyzed autoxidation of GSH. To elucidate the mechanism of the mutual inhibition of the autoxidations of these two reducing substances in their coexistence, we have kinetically investigated these phenomena. The study of the former phenomenon revealed that GSH forms a 1:1 chelate with Cu(+) and thereby prevents the autoxidation of AA. By the analysis of the latter phenomenon, it was postulated that the inhibition of GSH oxidation by AA is due to rapid reduction of thiyl radical of GSH by AA rather than competition of AA with GSH in the reduction of Cu(2+). The effect of GSH on the formation of hydroxyl radical by the copper-catalyzed autoxidation of AA was also studied and it was found that the hydroxyl radical formation was delayed dose-dependently by GSH with time lags comparable to those of the oxidation of AA. Because there are several lines of evidence that redox-active copper ions are released from tissues under pathological conditions, it is possible that such copper ions coexist with AA and GSH in vivo, and in such a situation, GSH may exert an inhibitory effect on the hydroxyl radical formation caused by the autoxidation of AA.

Ascorbic Acid↗
PubMed2000

Primitive synteny of vertebrate major histocompatibility complex class I and class II genes.

Major histocompatibility complex (MHC) class I and class II molecules bind to and display peptidic antigens acquired from pathogens that are recognized by lymphocytes coordinating and executing adaptive immune responses. The two classes of MHC proteins have nearly identical tertiary structures and were derived from a common ancestor that probably existed not long before the emergence of the cartilaginous fish. Class I and class II genes are genetically linked in tetrapods but are not syntenic in teleost fish, a phylogenetic taxon derived from the oldest vertebrate ancestor examined to date. Cartilaginous fish (sharks, skates, and rays) are in the oldest taxon of extant jawed vertebrates; we have carried out segregation analyses in two families of nurse sharks and one family of the banded houndshark that revealed a close linkage of class IIalpha and beta genes both with each other and with the classical class I (class Ia) gene. These results strongly suggest that the primordial duplication giving rise to classical class I and class II occurred in cis, and the close linkage between these two classes of genes has been maintained for at least 460 million years in representatives of most vertebrate taxa.

Amino Acid Sequence↗
PubMed2000

CYP2D6 polymorphism and the presence of anti-LKM-1 in patients with chronic hepatitis C.

Anti-LKM-1 autoantibodies are directed mostly at cytochrome P450 2D6 (CYP2D6) autoantigen, whose activity ranges from "complete deficiency" to "extensive metabolism" due to genetic polymorphism. We aimed to find any relevance of CYP2D6 alleles to the presence/absence of anti-LKM-1 in Japanese patients with chronic hepatitis C. The frequency of an extensive metabolizer-type allele (CYP2D6*1) in anti-LKM-1-positive patients was higher than that in anti-LKM-1-negative patients (0.800 vs 0.431; P = 0.0035), while the CYP2D6*10 allele with moderately reduced activity was less frequent in the former than the latter (0.050 vs 0.389; P = 0.0069). Moreover, the rate of homozygosity for CYP2D6*1 showed a striking difference between the two groups (70% vs 19%; P = 0.0021). These findings suggest that a genetic predisposition to produce the enzyme CYP2D6 of extensive metabolizer-type is associated with the induction of anti-LKM-1 in chronic hepatitis C patients.

Alleles↗
PubMed2000

Role of cholinergic neural transmission on airway resistance in the dog.

The unique contractile profiles of bronchial smooth muscle (Kondo et al., 1995) and its neural control were investigated by comparing responses of the bronchus and trachea to acute hypercapnia, stimulation of vagus efferent fibers before and after intravenous atropine, and intravenous acetylcholine in decerebrated and paralyzed dogs. During acute hypercapnia, airway resistance represented by peak airway pressure (Pedley et al., 1970) significantly increased as well as tracheal tension (Ttr). During electric stimulation of the vagal efferent fibers, Ttr increased and was sustained throughout the simulation period while the peak airway pressure was not maintained at the peak level. The peak Ttr and the airway resistance (Raw) calculated from ventilatory flow and airway pressure increased with increases in intensity of electric stimulation. Ttr reached its maximal level at an intensity 16 times of the threshold (T), while Raw became maximal at 4T. Although both the Ttr-stimulus intensity and Raw-intensity curves were shifted to the right by administration of intravenous atropine, the Raw curve shifted more to the right than the Ttr curve with the same dose of atropine. When muscular muscarinic receptors were directly stimulated by intravenous acetylcholine, Ttr once increased and then decreased promptly while peak airway pressure remained at a high level for a few minutes. These findings suggested that the bronchus is more sensitive to vagal efferent stimulation and susceptible to competitive antagonist of actylcholine than the trachea. In conclusion, the contractile profiles of the fifth-order bronchus we have reported (Kondo et al., 1995) were reflected in airway resistance, and the neuromuscular junction may be the site of adaptation of bronchoconstrictor response to motor nerve adaptation.

Acetylcholine↗
PubMed2000

The octapeptide repeat region of prion protein binds Cu(II) in the redox-inactive state.

The octapeptide repeat region of human prion protein is known to bind four Cu(II) ions per molecule. A peptide, Octa(4), representing this region was tested for inhibitory effects on copper-catalyzed oxidation of l-ascorbate or glutathione and on generation of OH(*) during the former reaction. The result indicated that the catalytic activity of the first Cu(II) ion bound to an Octa(4) molecule was completely suppressed. The valence state of the copper under reducing conditions was Cu(II), as determined by a newly developed method using bathocuproinedisulfonate under acidic conditions. Furthermore, it was shown that Escherichia coli cells expressing the octapeptide repeat region were significantly resistant to copper treatment compared with control cells. The results taken together indicate that prion protein can function to sequester copper ions in the redox-inactive state, rendering copper-induced generation of reactive oxygen species impossible.

Ascorbic Acid↗
PubMed2000

Completion pneumonectomy for patients with recurrent lung cancer: the impact of microvessel density on outcome.

The effectiveness of performing completion pneumonectomy (CP) to treat recurrent lung cancer was assessed in 11 patients, 6 of whom had intrapulmonary recurrence, and 5, locoregional recurrence. The pathological types of cancer were adenocarcinoma in 5 patients, squamous cell carcinomas in 5 patients, and adenoid cystic carcinoma in 1 patient. The pathological stages of the recurrent lung cancer were stage I in 8 patients, stage II in 2 patients, and stage IV in 1 patient. The microvessels in the tumors were also examined and the results compared with those in the microvessels of 61 nonrecurrent tumors. The 1- and 3-year overall survival rates after CP were 54.5% and 27.3%, respectively. The vessel densities within primary and correspondent recurrent tumors were able to be assessed in 9 patients and found to be significantly greater compared with the mean value for the nonrecurrent tumors (P < 0.0001). The vessel densities in the primary and recurrent tumors of 2 of 3 long-term survivors were found to be relatively low. These results indicate the high-grade malignant potential of recurrent tumors, which led us to conclude that the use of CP should be carefully evaluated in patients with recurrence. Assessing the vasculature in primary tumors may be a useful indicator for determining which patients could benefit from CP.

Adenocarcinoma↗
PubMed2000

TT virus (TTV) genotypes in native and non-native prostitutes of Irian Jaya, Indonesia: implication for non-occupational transmission.

We investigated prostitutes in Irian Jaya, an Indonesian territory of New Guinea, to know whether TT virus (TTV) is sexually transmitted and what genotypes of TTV exist there. An ORF2 region of the TTV genome was analyzed for 44 isolates from prostitutes (19 were Irian Jaya natives but 25 were immigrants from Java or other islands of Indonesia) and 26 isolates from women of child-bearing age in Surabaya as a control. The WX(7)HX(3)CX(1)CX(5)H motif sequence of the ORF2 was compared across the 70 Indonesian isolates with a reference of 47 Japanese isolates (filed in databases) and the prototype TTV isolate TA278. A total of 77 different sequences were generated from the comparison, but a phylogenetic analysis suggested that they could be divided into three categories: group A, group B and others (the third group was highly diverse). Interestingly, most of the Indonesian isolates belonged to group B (74%): this rate was considerably higher than that observed previously in Japanese isolates. Group B isolates were further compared for the N-terminal 95 amino acids of the ORF2, with a result that the natives of Irian Jaya had a different pattern of genotype distribution from other groups. In particular, 9 out of 19 isolates from the Irian Jaya-natives were co-classified under a distinct branch, to which none of the other Indonesian and Japanese isolates belonged. Our data indicate that TTV genotypes reflect the birth place of the infected prostitutes rather than their work environment, and thus suggest that an infection is more likely during the early period of life than through sexual transmission. In addition, the presence of an Irian Jaya-specific genotype is intriguing from an anthropological and viral evolutionary point of view, because Irian Jaya has been isolated from contact with other areas for a long time.

Adolescent↗
PubMed2000

Fibrillin gene (FBN1) mutations in Japanese patients with Marfan syndrome.

Marfan syndrome (MFS; MIM #154700) is a connective tissue disorder characterized by cardiovascular, skeletal, and ocular abnormalities. The fibrillin-1 gene (FBN1; MIM no. 134797) on chromosome 15 was revealed to be the cause of Marfan syndrome. To date over 137 types of FBN1 mutations have been reported. In this study, two novel mutations and a recurrent de-novo mutation were identified in patients with MFS by means of single-strand conformational polymorphism (SSCP) analysis. The two novel mutations are a 4-bp deletion at nucleotide 2820-2823 and a G-to-T transversion at nucleotide 1421 (C474F), located on exon 23 and exon 11, respectively. A previously reported mutation at the splicing donor site of intron 2 (IVS2 G + 1A), which is predicted to cause exon skipping, was identified in a sporadic patient with classical MFS.

Amino Acid Substitution↗
PubMed2000

Clinicopathological and biological assessment of lung cancers with pleural dissemination.

BACKGROUND: This study provides the surgical outcome of lung cancer patients with pleural dissemination, with the assessment of the clinicopathological and biological prognostic factors. METHODS: Forty-three patients who underwent operations were studied. Vascular endothelial growth factor (VEGF) and autocrine motility factor receptor (AMFR/gp78) expression was immunohistochemically evaluated. RESULTS: In total, the overall 3 and 5-year survival rates were 31.4% and 13.1%, respectively. The patients who underwent the pleuropneumonectomy had a worse outcome than those who underwent limited operations (pleurectomy plus parenchymal resections were less than pneumonectomy). VEGF and AMFR/gp78 were highly expressed in primary tumors. Among the patients who underwent limited operations, pathological types other than adenocarcinoma and high expression of VEGF were significantly associated with a worse outcome. The pathological type was the only characteristic to retain a significant independent prognostic impact on overall survival. CONCLUSIONS: The results imply the validation of limited operation for lung cancer with pleural dissemination for the local control. High frequency of VEGF and AMFR/gp78 expression conform to the interpretation that patients with pleural dissemination have a high-risk of systemic disease.

Adenocarcinoma↗
PubMed2000

Increased vascular endothelial growth factor and vascular endothelial growth factor-c and decreased nm23 expression associated with microdissemination in the lymph nodes in stage I non-small cell lung cancer.

OBJECTIVE: We examined a microdissemination of cancer cells in lymph nodes and assessed its clinical and biologic characteristics. METHODS: Both primary tumors and lymph nodes (2030 nodes) were obtained from 122 patients with primary stage I lung cancer who underwent curative operations with routine systematic nodal dissection of both the hilar and the mediastinal nodes. Immunohistochemical anticytokeratin staining was used to detect nodal microdissemination of cancer cells. Vascular endothelial growth factor, vascular endothelial growth factor type C, and nm23 expression at primary sites were also immunohistochemically studied. RESULTS: In total, 35 patients (29%) had cytokeratin-positive cells in lymph nodes. Increased expression of vascular endothelial growth factor (P =.0001) and vascular endothelial growth factor type C (P <. 0001) at primary sites were significantly associated with nodal microdissemination, and nm23 was inversely correlated with microdissemination (P =.008). The 3- and 5-year survivals for the patients with nodal microdissemination were 57% and 54%, respectively, which was a significantly worse prognosis as compared with those prognoses (83% and 76%) for the patients without nodal microdissemination (P =.006). The independent prognostic impact of nodal microdissemination was not clear; however, vascular endothelial growth factor retained independent significance. CONCLUSION: All of these findings lead us to conclude that the microspread of tumor cells in nodes detected by immunohistochemical anticytokeratin staining is definitely a metastasis with a high risk of systemic disease.

Aged↗
PubMed2000

Ventilatory responses to hypercapnia and hypoxia following chronic hypercapnia in the rat.

This study investigated the effects of an 18 week exposure to 10% CO(2) in air on minute ventilation (V(E)), breathing pattern and the chemoresponiveness of rats to hypoxic and hyperoxic stimuli. We found that V(E) remained elevated over the 18 weeks. Nonetheless, the breathing pattern changed significantly. Tidal volume increased and the durations of inspiration and the total cycle decreased. After the sustained hypercapnia the mean Pa(CO(2)) was 72.0+/-5.1 (S. D.) mmHg. Every 6 weeks the chemoresponiveness of the CO(2)-exposed rats was tested by an acute exposure sequentially to room air, then a 6% O(2), 10% CO(2) and 84% N(2) gas mixture, and finally a 90% O(2) in 10% CO(2) mixture. On either room air or the hyperoxic-hypercapnic mixture V(E) fell to its pre-hypercapnic level. On the hypoxic-hypercapnic mixture V(E) increased significantly. These results demonstrate that the initial stimulating effect of 10% CO(2) on V(E) persisted for the entire 18 weeks without altering hypoxic or hyperoxic ventilatory responses.

Animals↗
PubMed2000

Real-time fluorescence analysis on molecular mechanisms for regulation of cytochrome P450scc activity upon steroidogenic stimulation in adrenocortical cells.

Real-time fluorescence analysis revealed that the activity of cytochrome P450scc was related to Ca2+ signals arising from extracellular NADPH, ACTH and ATP stimulation in adrenocortical fasciculata cells. The side-chain cleavage reaction by cytochrome P450scc was measured with 3beta-hydroxy-22,23-bisnor-5-cholenyl ether (cholesterol-resorufin) by observing the distinct increase in fluorescence upon conversion of cholesterol-resorufin to resorufin and pregnenolone. Adrenocorticotropic hormone (ACTH) induced a relatively small stimulation of the P450scc activity. A significant production of resorufin was revealed after stimulation of cell cultures with 100 pM, 1 nM of ACTH for 3 h. On the other hand, extracellular NADPH was found to rapidly and greatly stimulate the resorufin production in intact cells immediately after the addition of 50-500 microM NADPH. The extracellular NADPH stimulation was prevented by the addition of thapsigargin and EGTA which abolished Ca2+ oscillations induced by NADPH. Suramin, a specific antagonist of the P2y type ATP receptor, also completely abolished the NADPH-induced cholesterol-resorufin conversion. These results imply that extracellular NADPH (membrane impermeable) produced Ca2+ oscillations through its binding to ATP receptor thereby stimulating the activity of P450scc. The application of 45-500 microM extracellular ATP to cells did not, however, significantly increase the resorufin production. These three stimulators produced very different types of Ca2+ signals. ACTH induced mainly a series of Ca2+ spikes superimposed on a long-lasting basal Ca2+ elevation. The Ca2+ signals induced by NADPH showed predominantly a series of Ca2+ spikes without elevation of the basal Ca2+ concentration. Only long-lasting Ca2+ elevation was induced by extracellular ATP. The stimulation of cytochrome P450scc may thus be correlated with the different patterns of Ca2+ signals.

Adenosine Triphosphate↗
PubMed2000

CD40 ligation inhibits trichilemmoma cell proliferation and induces IL-6 production.

CD40 is a member of the tumor necrosis factor receptor superfamily expressed by B cells, monocytes, dendritic cells, epithelial cells, and hematopoietic progenitor cells. Recently, CD40 has been reported to also be expressed on human epidermal cells. We have elucidated the function of CD40 on epidermal tumor cells and have found that trichilemmoma (KTL-1) cells constitutively express CD40 and respond to CD40 ligation by anti-CD40 mAb (EA-5) with a significant decrease in proliferation. We were also able to demonstrate that KTL-1 cells respond to CD40 ligation by EA-5 with the up-regulation of interleukin-6 (IL-6) mRNA expression. Together, the results suggest that CD40 on KTL-1 cells may function to regulate their proliferation associated with the induction of IL-6 production.

CD40 Antigens↗
PubMed2000

alpha2-Adrenergic-receptor response in reversible increase in hemoglobin concentration in intermittent hypoxia.

We have previously shown that intermittent hypoxia (IHx, 10% O(2), 60 min/day) leads to an increase in the splenic alpha2-adrenoceptor response and results in a splenic contraction-induced reversible increase in hemoglobin concentration ([Hb]). In the present study, we determined whether IHx of shorter duration (15 min/day (15-min) and 30 min/day (30-min)), produced this phenomenon in rats. A significant increase in [Hb] during hypoxia was observed in both the groups, but its magnitude was larger in the 30-min IHx rats. Even when the cumulative exposure time (time/dayxdays) was shorter, the [Hb] increase was larger in the rats with longer daily hypoxic exposure. The alpha2-adrenoceptor antagonist yohimbine abolished the [Hb] increase of 15- and 30-min IHx. The increase in [Hb] following administration of the alpha2-adrenoceptor agonist oxymetazoline was also higher in 30-min IHx; indicating that the higher [Hb] produced by longer daily hypoxic exposure times is the result of increases in alpha2-adrenergic-receptor response of greater magnitude. In conclusion, IHx for periods as short as 15 and 30 min/day increases the splenic alpha2-adrenoceptor response and its magnitude reaches the maximum value depending on the daily hypoxic exposure time. A reversible increase in [Hb] constitutes a useful mechanism that protects organ oxygen supply during hypoxic episodes of variable duration and intensity.

Journal Article↗
PubMed2000

Factors affecting the biodegradation of cyanide by Burkholderia cepacia strain C-3.

The utilization of cyanide as a nitrogen source for growth in a liquid minimal medium by Burkholderia cepacia strain C-3 isolated from soil was demonstrated. The effects of various parameters on the biodegradation of cyanide by the strain were investigated. Growth at the expense of cyanide occurred over a pH range from 8 to 10, and was optimal at pH 10. Growth and cyanide biodegradation were optimal at 30 degrees C. The presence of Cu2+ or Fe2+ in the liquid medium at a concentration of 1 mM inhibited both the growth of the bacterium and its cyanide degradation. The effects of Ni2+ and Co2+, and to a lesser extent Mn2+ and Mo2+, on cyanide degradation rate were concentration dependent. Addition of the cyanide-containing waste contaminants ethanol and methanol reduced the cyanide utilization of the isolate, but phenol was completely inhibitory. Microbial depletion of cyanide occurred even in the presence of other organic and inorganic nitrogen sources. Fructose, glucose, and mannose were the preferred carbon sources for cyanide biodegradation. The highest rate of cyanide degradation by B. cepacia C-3, 1.85 mg CN h(-1), was observed with 0.25% (w/v) fructose; the degradation rate decreased progressively thereafter with increasing fructose concentration.

Journal Article↗
PubMed2000

Mapping of cDNA clones on contig of Chlorella chromosome I.

Complementary DNA (cDNA) clones specific to the smallest chromosome (chromosome I) of Chlorella vulgaris C-169 were selected from cDNA libraries with probes of chromosome I DNA fragments amplified by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). A total of 15 clones was obtained, which included gene homologs for alpha-tubulin, inosine-5'-monophosphate dehydrogenase, beta-1,4-mannase, a TTG-binding protein, a heat shock protein, thioredoxin/protein disulfide isomerase, transcription factor NF-E2, an oxidoreductase, and UDP-n-acetylglucosamine enolpyruvyltransferase. These clones were definitely localized at specific sites on the chromosome I physical map constructed on the basis of overlapping cosmid clones (the contig). They were predominantly distributed within the left two-thirds of the chromosome. This contrasts with the distribution of repetitive elements such as short interspersed elements (SINEs), which are rather abundant in the right two-thirds of chromosome I. The comparative simplicity of the gene arrangement of Chlorella chromosome I suggests that it may be able to serve as a prototypic system for deciphering the complexity of huge plant chromosomes.

Journal Article↗