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Biomedical subjects

T Yamane

Publications and source records attributed to T Yamane.

At least 253 records · Page 14Linked to original sources

Molecular analysis of the poly(3-hydroxyalkanoate) synthase gene from a methylotrophic bacterium, Paracoccus denitrificans.

A 3.6-kb EcoRI-SalI fragment of Paracoccus denitrificans DNA hybridized with a DNA probe carrying the poly(3-hydroxyalkanoate) (PHA) synthase gene (phaC) of Alcaligenes eutrophus. Nucleotide sequence analysis of this region showed the presence of a 1,872-bp open reading frame (ORF), which corresponded to a polypeptide with a molecular weight of 69,537. Upstream of the ORF, a promoter-like sequence was found. Escherichia coli carrying the fusion gene between lacZ and the ORF accumulated a level of poly(3-hydroxybutyrate) that was as much as 20 wt% of the cell dry weight in the presence of beta-ketothiolase and acetoacetylcoenzyme A reductase genes of A. eutrophus. The ORF was designated phaCPd. A plasmid vector carrying the phaCPd'-'lacZ fusion gene downstream of the promoter-like sequence expressed beta-galactosidase activity in P. denitrificans. When a multicopy and broad-host-range vector carrying the ORF along with the promoter-like sequence was introduced into P. denitrificans, the PHA content in the cells increased by twofold compared with cells carrying only a vector sequence.

Acyltransferases↗

Selective radiofrequency catheter ablation of the slow pathway for common and uncommon atrioventricular nodal reentrant tachycardia.

The utility of selective radiofrequency catheter ablation of the slow pathway for the treatment of common and uncommon atrioventricular nodal reentrant tachycardia (AVNRT) was studied in 110 consecutive patients, 94 with slow-fast form common AVNRT, and 11 and 5, respectively, with the fast-slow and slow-slow forms of uncommon AVNRT. Ablation sites were determined by mapping a late and spiky "slow pathway potential" in the posterior right atrial septum in common AVNRT, and also the earliest retrograde atrial activation over the retrograde slow pathway in uncommon AVNRT. AVNRT was successfully eliminated in all patients with a mean number of radiofrequency pulses of 2.9 +/- 3.0 and a mean total energy applied of 3536 +/- 2996 joules. There were no early or late complications, except for transient AV block for 15 sec immediately after energy application in one common AVNRT patient, and no recurrence of AVNRT in a mean follow-up period of 24 +/- 13 months. There were no significant differences between common and uncommon AVNRT in success rate, mean application number and total energy applied. However, the AVN physiology post-ablation was different. Slow pathway conduction was eliminated in only 32% of the patients post-ablation in common AVNRT, while it was elininated in 100% in uncommon AVNRT. Selective radiofrequency catheter ablation of the slow pathway can cure common and uncommon AVNRT effectively and safely. Common AVNRT can be eliminated irrespective of the persistence of slow pathway conduction, while uncommon AVNRT can be eliminated by the eradication of slow pathway conduction.

Adolescent↗

Identification and antimicrobial activity of urinary metabolites of a rifamycin derivative in dog.

1. Three metabolites of the antimicrobial agent 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin (KRM-1648) were isolated from dog urine obtained after administration of a single oral dose. These metabolites of KRM-1648 were identified by mass spectrometry and 1H and 13C-nmr spectrometry. 2. Three metabolites of KRM-1648 were identified as 25-deacetyl KRM-1648, 30-hydroxy KRM-1648 and 25-deacetyl-30-hydroxy KRM-1648. 3. The antimicrobial activities of 25-deacetyl KRM-1648 were comparable with those of the parent compound, whereas 30-hydroxy KRM-1648 was equipotent and 2-8-fold less active than the parent compound against bacteria and mycobacteria, respectively.

Administration, Oral↗

In vitro metabolism of a rifamycin derivative by animal and human liver microsomes, whole blood and expressed human CYP3A isoform.

1. In vitro metabolism of a rifamycin derivative, benzoxazinorifamycin KRM-1648, was studied using mouse, rat, guinea pig, dog, monkey and human liver microsomes. 30-Hydroxy-KRM-1648 (M2) was produced in mouse, dog, monkey and human microsomes. 25-Deacetyl-KRM-1648 (M1) was produced in dog and human microsomes, but not in mouse or monkey microsomes. Neither M1 nor M2 was detected in rat or guinea pig microsomes. 2. In dog and human liver microsomes the formation of M2 was dependent on NADPH, but the formation of M1 was not. 3. In vitro metabolism of the parent compound was studied in whole blood in some species. Only M1 was detected in mouse and rat blood, and not in dog and human blood. 4. These findings demonstrated that the metabolite pattern in dog resembled that in man, and suggested that the 30-hydroxylation of KRM-1648 was mediated by cytochrome P450, but that the 25-deacetylation was not. 5. Among the ten recombinant human P450 isoforms used, only the cell lysates including CYP3A3 and CYP3A4 catalysed the M2 formation from KRM-1648.

Animals↗

Differences in the influence of AIN-purified and non-purified diets on the development of hypertension in SHR and DOCA-salt hypertensive rats.

The influence of non-purified and AIN purified diets on the development of hypertension was examined in deoxycorticosterone acetate (DOCA)-salt hypertensive rats and SHRs. For DOCA-salt hypertensive rats, the development of hypertension was slower in rats fed the AIN 76-purified diet than in those fed the non-purified diet throughout the experimental period of about five weeks. In an experiment using spontaneously hypertensive rats (SHRs), 4-week-old rats were fed either a non-purified diet, a AIN76-purified diet or a AIN93G-purified diet for about nine weeks. In the first 1-2 weeks, the blood pressure was lower in the SHRs fed the AIN93G-purified diet than in those fed the non-purified diet. However, no significant difference in blood pressure was observed within the SHR group thereafter.

Animal Feed↗

[A case of cranial hypertrophic pachymeningitis with intracranial hemorrhage].

A case of cranial hypertrophic pachymeningitis of unknown etiology in a patient with 15-year history of headaches, cranial nerve palsies, and gait disturbance is reported. A 77-year-old woman was brought to our institute in a coma. CT revealed intracerebral hemorrhage in the right temporal lobe and thickening of the falx and tentorium. Fifteen years previously the patient had undergone CT scanning because of headaches, cranial nerve palsies, and progressive gait disturbance and a thickened tentorium, mild hydrocephalus and edematous change in the right temporal lobe had been reported. Since the etiology of her symptoms was unclear at the time, she did not receive adequate treatment. Her symptoms gradually progressed thereafter, and her visual acuity and hearing deteriorated. MR imaging in 1994 showed the thickened tentorium as a hypointense area with hyperintense edges on Gd-DTPA enhanced images. Angiography revealed narrowing of posterior portion of the superior sagittal sinus. The patient's condition rapidly deteriorated due to the intracranial hypertension and she subsequently died. Autopsy revealed a thickened tentorium with xanthochromic surface. This hypertrophic change was also seen in the dura mater of the posterior and middle cranial fossa. Microscopic examination of the thickened tentorium revealed extensive fibrous tissue with a chronic inflammatory infiltrate, predominantly of lymphocytes. No specific lesions were revealed by staining with hematoxylin-eosin, PAS, Gram's or Ziehl-Neelsen stains. The patient had no inflammatory or infectious diseases of other organs, and a diagnosis of idiopathic cranial hypertrophic pachymeningitis of unknown etiology was made. Considering the above findings, the thickened tentorium depicted as a hypointense area on the T1- and T2-weighted images and the Gd-enhanced edges of the tentorium are thought to be represent fibrous tissue and inflammatory regions, respectively.

Aged↗

[LGL lymphoma].

We reported in this article two patients with large granular lymphocytic lymphoma (abbreviated as LGL lymphoma). One was the patient with LGL leukemia/lymphoma (patient 1) and other was the patient with NK-LGL lymphoma (patient 2). Because the gene of TCR delta was rearranged in the patient 1, the clonality of the LGL leukemia/lymphoma was confirmed. However, it is not determined yet whether the lineage of tumor cells is T cells or NK cells. The cytochemical features of the lymphoma cells of the patient 2 were studied. It was found that NK cell-derived lymphoma cells of the patient were positively stained with these two monoclonal antibodies that are reactive with T cell; one is anti-CD45RO (UCHL-1) and other is anti-CD3. Judging from the result, malignant NK-LGL cells in some patients are cytoplasmic CD3+ and UCHl-1+. It is emphasized that May-Grünwald-Giemsa stain of biopsied specimen of the lymphoma is required for making the diagnosis of LGL lymphoma.

Adult↗

Constitutive expression of the thrombopoietin gene in a human hepatoma cell line.

The gene of thrombopoietin (TPO) has been cloned and identified to be identical to gene of the c-mpl ligand. It is known that the mRNA of TPO is expressed in liver and kidney. However, it is not clarified which cells in the liver produce TPO. Using a human hepatoma cell line, HepG2, we demonstrated that the TPO mRNA was expressed by liver parenchymal cells without any stimulation. To clarify the regulation of the expression of the TPO mRNA in HepG2 cells by cytokines, we assessed the effects of 5 cytokines, transforming growth factor-beta 1, activin A, platelet-derived growth factor, hepatocyte growth factor, and interleukin-6. These cytokines have no significant regulative effect on the expression of the TPO mRNA in HepG2 cells. Our results suggest that liver parenchymal cells may be the TPO producing cells and also suggest that some hepatoma cells may produce TPO constitutively.

Base Sequence↗

Analysis of beta-ketothiolase and acetoacetyl-CoA reductase genes of a methylotrophic bacterium, Paracoccus denitrificans, and their expression in Escherichia coli.

The beta-ketothiolase gene (phaA) and acetoacetyl-CoA reductase gene (phaB) were isolated from Paracoccus denitrificans. Nucleotide sequence analysis showed that they encoded proteins of 391 amino acids with a molecular mass of 40,744 Da and of 242 amino acids with a molecular mass of 25,614 Da, respectively. The predicted gene products exhibited high amino acid identities with those from other bacteria: 64.4-74.0% for the phaA gene product and 47.6-80.6% for the phaB gene product, respectively. Both genes were co-transcribed in a recombinant Escherichia coli. In addition, promoter activity was detected upstream of the phaA gene. Hence, the two genes are organized as an operon, phaA-phaB, in P. denitrificans. NADH was preferred to NADPH as a cofactor of acetoacetyl-CoA reductase.

Acetyl-CoA C-Acyltransferase↗

Inhibition of N-methyl-N'-nitro-N-nitrosoguanidine-induced carcinogenesis by (-)-epigallocatechin gallate in the rat glandular stomach.

Recently, an epidemiological study showed a lower risk of gastric cancer among people who consume a large amount of green tea. (-)-Epigallocatechin gallate (EGCG), one of the main constituents of green tea, inhibited tumor promotion by teleocidin in a two-stage carcinogenesis experiment with the use of mouse skin. The inhibitory effect of EGCG on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced carcinogenesis of the glandular stomach in rats was examined. The percentage of tumor-bearing rats in the group treated with MNNG plus EGCG was 31%, compared to 62% in the MNNG group. The difference was statistically significant (P < 0.05). To assess the effect of p.o. administration of EGCG, the gastric mucosal cellular kinetics was examined with the use of the bromodeoxyuridine labeling index, ornithine decarboxylase activity, and tissue polyamine levels. The labeling index of the EGCG treatment group decreased significantly (P < 0.05) compared to the EGCG plus MNNG treatment group. The ornithine decarboxylase activity and tissue spermidine levels were also decreased. On the other hand, the tissue putrescine and spermine levels were partly increased. These findings suggest that EGCG inhibits the cellular kinetics of the gastric mucosa during the promotion stage of MNNG-induced gastric carcinogenesis. EGCG may be useful in preventing gastric carcinogenesis. Moreover, EGCG may be applied clinically without any harmful effects and at a low cost.

Adenocarcinoma↗

Immunohistochemical staining and activity of ornithine decarboxylase in colorectal cancer.

Using a new anti-human ornithine decarboxylase (anti-hODC) monoclonal antibody, the relationship between the immunoreactivity of ODC and its activity was analyzed in 21 human colorectal cancer tissues, 42 adjacent non-tumorous mucosa specimens, and 10 normal rectal mucosa samples from frozen sections and paraffin-embedded samples. A statistical significant correlation was found between the antibody reaction and the enzymic activity (P < 0.01). The immunohistochemical staining for ODC provides a new and simplified procedure for studying the activity of ODC as compared to previous methods using radioisotopes. It offers the advantages of retrospectively determining the amount of ODC in samples previously embedded in paraffin.

Aged↗

A long-lived batch reaction system of cell-free protein synthesis.

Several reaction conditions of cell-free protein synthesis such as temperatures, buffers, tRNAs, and creatine phosphate were intensively investigated and optimized to prolong protein synthesis and make it more efficiently in a batch system. As a result of these modifications, the protein synthesis reaction continued for 10 h so that about 30 micrograms of dihydrofolate reductase (DHFR) protein derived from Escherichia coli was synthesized in 1 ml of reaction mixture. In this improved system, translational reactions of other mRNAs such as rabbit beta-globin, Xenopus beta-globin, and tobacco mosaic virus RNA also continued for about 10 h. In addition, protein synthesis directed by uncapped dhfr mRNA containing a viral cap-independent translation initiation-mediating sequence continued for 10 h, resulting in the synthesis of 18 micrograms of DHFR protein per milliliter of reaction mixture.

Adenosine Triphosphate↗

Structure of a new alkaline serine protease (M-protease) from Bacillus sp. KSM-K16.

An alkaline serine protease, M-protease, from Bacillus sp. KSM-K16 has been crystallized. Two morphologically different crystal forms were obtained. Crystal data of form 1: space group P2(1)2(1)2(1), a = 47.3, b = 62.5, c = 75.6 A, V = 2.23 x 10(5) A(3), Z = 4 and V(m) = 2.09 A(3) Da(-1). Crystal data of form 2: space group P2(1)2(1)2(1), a = 75.82 (2), b = 57.79 (2), c = 54.19 (1) A, V = 2.29 (2) x 10(5) A(3), Z = 4 and V(m) = 2.15 A(3) Da(-1). The crystal structure of M-protease in form 2 has been solved by molecular replacement using the atomic model of subtilisin Carlsberg (SBC) which is 60% homologous with M-protease, and refined to the crystallographic R-factor of 0.189 for 7004 reflections with F(o)/sigma(F) > 3 between 7 and 2.4 A resolution. The final model of M-protease contains 1882 protein atoms, two calcium ions and 44 water molecules. The three-dimensional structure of M-protease is essentially similar to other subtilisins of known structure. The 269 C(alpha) positions of M-protease have an r.m.s. difference of 1.06 A with the corresponding positions of SBC. The crystal data of form 2 are close to those of SBC, though the structure determination of form 2 made it clear that it is not isomorphous to the crystal structure of SBC. The deletions of amino acids occur at the residues 36' and 160'-163' compared with SBC (numerals with primes show the numbering for SBC). The deletion of the four residues (160'-163') may significantly affect the lack of isomorphism between M-protease and SBC.

Journal Article↗

Ossification of the posterior longitudinal ligament. Autosomal recessive trait.

STUDY DESIGN: This study analyzed the mode of inheritance of ossification of the posterior longitudinal ligament (OPLL) from the pedigree of a family. OBJECTIVES: The results were correlated to provide a new mode of inheritance of OPLL. SUMMARY OF BACKGROUND DATA: Although a nation-wide multicenter survey of OPLL has been carried out in 347 subjects and 1030 relatives in Japan since 1981, no parental consanguinity has been reported. METHODS: In the family, three siblings and one other member of a family underwent operations for OPLL in our department, and the clinical information regarding other family members was obtained from interviews with these four patients or relatives. RESULTS: The parents of three affected siblings and another unaffected sister were first cousins, and the father was suspected to be affected as well. CONCLUSION: Transmission of OPLL as an autosomal recessive trait in this family, which has not been reported, is suspected; although, the possibility that it is a dominant trait can not be excluded.

Aged↗

Differential cloning using in-gel competitive reassociation.

We describe the principle and actual processes of a differential cloning procedure designed for cloning of anonymous restriction DNA fragments whose molecular sizes differ between two genomic DNA preparations from higher organisms as a result of DNA rearrangement, polymorphism, etc. The procedure, which was extensively modified from the original one and still employs in-gel competitive reassociation (IGCR) as the basic principle, aims for cloning of DNA fragments which exist in one copy or less per mammalian genome. The modified procedure consists of dissociation and reassociation of biotinylated restriction digests of target DNA fragments (from which clones are to be isolated) in the presence of a large excess of reference (competitor) DNA in gel after electrophoresis, which is followed by absorption of the target DNA fragments to streptavidin-coated tubes and solid-phase polymerase chain reaction. After repeating these steps we attained substantial enrichment of altered DNA fragments which were originally present in one copy or less per complex mammalian genome.

Animals↗