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Biomedical subjects

H Ohta

Publications and source records attributed to H Ohta.

At least 325 records · Page 18Linked to original sources
PubMed1996

Synthesis of polymer-bound 6-mercuric carboxylate DOPA precursors and solid phase labelling method of 6-radioiodinated L-DOPA.

In order to avoid separating unreacted mercury precursor and other mercury-containing compounds after the halodemercuration of a 6-mercury DOPA precursor, we developed a polymer-bound mercury precursor for the preparation of 6-halogenated DOPA. In this study, polymer-bound 6-mercuric carboxylate DOPA derivatives were synthesized from ion-exchange resin and Merrifield-type resin. Iododemercuration of polymer-bound 6-mercuric carboxylate DOPA derivatives gave higher yields (49-54%) compared with monomeric 6-mercuric trifluoroacetate protected DOPA. The radioiodination of the resin with no-carrier added iodine-125 afforded protected 6-[125I]I-L-DOPA with labelling efficiency of 92-97% with both polymer-bound 6-mercuric carboxylate DOPA derivatives.

Dihydroxyphenylalanine↗
PubMed1996

Synthesis and evaluation of radioiodinated 6-iodo-L-DOPA as a cerebral L-amino acid transport marker.

Regioselective radioiodination of N-trifluoroacetyl 3,4-dimethoxy-6-trifluoroacetoxymercurio-L-phenylalanine ethyl ester 1 under no-carrier-added condition gave 6-[125I]iodo protected L-DOPA with a labeling efficiency of more than 85%, and no-carrier-added 6-[125I]I-L-DOPA was obtained with a radio-chemical purity of over 95% after hydrolysis and chromatography. A nonradioactive standard of 6-iodo protected L-DOPA was synthesized by the iododemercuration of 6-mercuric trifluoroacetate protected L-DOPA 1 using I2 in chloroform. 6-[125I]I-L-DOPA showed high brain accumulation and rapid blood clearance in mice. The rat brain slice studies indicated high affinity of 6-[125I]I-L-DOPA for carrier-mediated and stereoselective active transport systems. The tissue homogenate analysis revealed that most of the accumulated radioactivity was intact 6-[125I]I-L-DOPA. Thus, 6-[123I]I-L-DOPA appears to be a suitable single photon emission computed tomography (SPECT) tracer for the selective measurement of cerebral L-amino acid transport, having no affinity for dopamine metabolism.

Amino Acids↗
PubMed1996

Decreased bone mineral density associated with early menopause progresses for at least ten years: cross-sectional comparisons between early and normal menopausal women.

To establish whether early onset of menopause carries an increased risk of osteoporosis, we compared the bone mineral density (BMD) of the second to fourth lumbar vertebrae (L2-4) between 18 women who had menopause before 43 years of age (early menopause group) and 19 women who had menopause after reaching 43 years of age (normal menopause group). Serum levels of calcium, phosphorus, calcitonin, intact parathyroid hormone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), and alkaline phosphatase activity were measured, and urine samples were analyzed to derive calcium/creatinine, hydroxyproline/creatinine, pyridinoline/creatinine, and deoxypyridinoline/creatinine (D-Pyr/Cr) ratios. Mean BMD was significantly lower in the early menopause group than in the normal menopause group, and individual BMD values in about half of the subjects in the former group were below the fracture threshold for Japanese women. Serum concentrations of LH, FSH, and E2 were slightly, but not significantly, lower in the early menopause group than in the normal menopause group. The D-Pyr/Cr ratio was significantly higher in the early menopause group than in the normal menopause group. There was no correlation between L2-4 BMD and age or the number of years after menopause in the normal menopause group, but both age and the number of years after menopause were negatively correlated with L2-4 BMD in the early menopause group. These results indicate that BMD in women who have early menopause continues to decline for up to 10 years, and that menopause and aging increase the risk of osteoporosis.

Adult↗
PubMed1996

Cause and effect between concentration-dependent tissue damage and temporary cell proliferation in rat stomach mucosa by NaCl, a stomach tumor promoter.

This study was designed to test whether concentration or dose of NaCl was responsible for the initial tissue damage (after 1 min) and resulting temporary cell proliferation at 17 h in stomach mucosa of male F344 rats after gastric intubation of 0.65, 1.3, 2.6 and 3.7 M NaCl. Histological damage was studied by dual staining combining horseradish peroxidase-labeled Griffonia simplicifolia agglutinin-II staining (HRP-GSA-II) and periodic acid cold thionin-Schiff reaction (PATS). Cell proliferation was studied by measuring replicative DNA synthesis with liquid scintillation counting and by BrdU staining. NaCl at the same overall dose of 0.8 g/kg body weight induced different degrees of response depending on the concentration. For 4 ml of 0.65 M NaCl, there was no tissue damage after 1 min nor any increase in replicative DNA synthesis after 17 h in the pyloric mucosa. Administration of 1.3 M NaCl (2 ml), 2.6 M NaCl (1 ml) and 3.7 M NaCl (0.7 ml) induced concentration-dependent damage of the surface mucous cell layer after 1 min and increased replicative DNA synthesis after 17 h (P<0.05). Concentration-dependent increase in replicative DNA synthesis at 17 h was also induced with the same volume (1 ml) of 1.3, 2.6, and 3.7 M NaCl, while a volume-dependent increase in replicative DNA synthesis at 17 h was induced with 0.4, 0.7 and 1 ml of 3.7 M NaCl. However, a greater increase in replicative DNA synthesis was always observed when using higher NaCl concentrations at the same dose. Liquid scintillation counting was well-correlated with BrdU staining. These results suggest that a high concentration of NaCl is responsible for the initial tissue damage and resulting temporary cell proliferation during stomach tumor promotion.

Animals↗
PubMed1996

Expression of the puf operon in an aerobic photosynthetic bacterium, Roseobacter denitrificans.

The effects of oxygen and light on the expression of the puf operon were investigated in Roseobacter denitrificans in a comparison with those in Rhodobacter sphaeroides. In darkness, the levels of the total puf mRNA in Ros. denitrificans were about 1.3 times those in Rb. sphaeroides at low concentrations of oxygen, reflecting the accumulation of bacteriochlorophyll and carotenoids. The oxygen tension, up to 94% saturation of dissolved oxygen, did not affect the levels of the total puf transcripts in Ros. denitrificans, whereas those in Rb. sphaeroides were reduced to 55% of the maximum level even at 50% saturation. Four puf-specific transcripts were detected: a 0.5-kb transcript was the most abundant; 1.2-kb and 1.9-kb transcripts accumulated at low levels; and a 3.5-kb transcript accumulated at very low levels under all conditions tested. The levels of the individual transcripts were barely affected by molecular oxygen. An S-1 nuclease protection assay revealed that the 0.5-kb transcript encoded the LHI-alpha and LHI-beta subunits (pufBA), the 1.2-kb transcript encoded puf-BA and part of pufL, and the 1.9-kb transcript encoded pufBAL and part of pufM. It was not clear whether the 3.5-kb transcript encoded the entire pufBALM and the gene for the polypeptide moiety of cytochrome c. The difference in levels between the 0.5-kb transcript and the other transcripts (1.2 kb, 1.9 kb, and 3.5 kb) was presumed to be due to the presence of several stem-loop structures at the 3' terminus of the 0.5-kb transcript which acted as terminators of transcription and, possibly, as protection against nucleolytic digestion. Light inhibited the expression of the puf operon in Ros. denitrificans more effectively than that in Rb. sphaeroides. The insensitivity to oxygen, as well as the sensitivity to light, of the expression of the puf operon in Ros. denitrificans, which was different from that in Rb. sphaeroides, seemed to represent a mode of adaptation that allowed the former cells to avoid photodynamic damage by light under highly aerobic conditions.

Bacteria↗
PubMed1996

A transcription factor with a leucine-zipper motif involved in light-dependent inhibition of expression of the puf operon in the photosynthetic bacterium Rhodobacter sphaeroides.

In the purple nonsulfur photosynthetic bacterium Rhodobacter sphaeroides the synthesis of components of the photosystem is regulated in response to oxygen tension and light intensity. We have purified and cloned a trans-acting protein (SPB) that binds to the promoter region of the puf operon, which encodes the apoproteins of light-harvesting complex I and the reaction center. The SPB was composed of a single polypeptide with an apparent molecular mass of 15.0 kDa. The nucleotide sequence of the spb gene was determined. The gene encoded 104 amino acid residues, which correspond to a molecular mass of 11.5 kDa. SPB exhibited 53% homology to HvrA in Rhodobacter capsulatus. The deduced amino acid sequence indicated that SPB contained a region with homology to the leucine-zipper motif of c-JUN, a transcription factor in eukaryotes, and SPB also had a DNA-binding domain on the amino-terminal side of the leucine-zipper motif. The leucine-zipper motif of SPB might contribute to the formation of a dimer. Northern analysis indicated that spb was constitutively and monocistronically transcribed in R. sphaeroides, irrespective of growth conditions. Structural and functional differences between SPB and HvrA are discussed.

Amino Acid Sequence↗
PubMed1996

Prediction of hemorrhagic complications after thrombolytic therapy for middle cerebral artery occlusion: value of pre- and post-therapeutic computed tomographic findings and angiographic occlusive site.

OBJECTIVE: To evaluate the usefulness of pre- and post-therapeutic computed tomographic (CT) findings in predicting hemorrhagic complications, we retrospectively examined 35 patients treated with intra-arterial thrombolytic therapy for middle cerebral artery (MCA) occlusion. METHODS: The presence or absence of early CT findings (loss of the insular ribbon, obscuration of the lentiform nucleus, and cortical effacement) and the presence and location of extravasation of contrast medium were evaluated on pre- and post-therapeutic CT scans, respectively. According to the angiographic occlusive site, the patients were classified into the following three groups: Group 1 (n = 13), MCA trunk occlusion involved lenticulostriate arteries; Group 2 (n = 11), occlusion of the MCA trunk without involvement of the lenticulostriate arteries; Group 3 (n = 11), occlusion of a branch of the MCA. Hemorrhagic complications (hemorrhagic transformation and/or massive brain swelling) were evaluated by reviewing CT scans obtained 3 to 14 days after thrombolytic therapy. RESULTS: No patient without extravasation (n = 17) showed hemorrhagic complications, and extravasation is the most useful finding in predicting hemorrhagic complications. There was significant correlation between extravasation and hemorrhagic complications (P < 0.01). In Groups 1 and 2, there was also significant correlation between early CT findings and hemorrhagic complications (P < 0.01), indicating that early CT findings are also useful in predicting hemorrhagic complications. In Group 1, 10 of 13 (76.9%) patients had both early CT findings and extravasation, and 6 of these 10 patients had hemorrhagic complications with clinical deterioration, suggesting the difficulty of thrombolytic therapy in this group. On the contrary, in Group 2, 8 of 11 (72.7%) patients had neither early CT findings nor extravasation and none of these 8 patients had hemorrhagic complications. In Group 3, however, early CT findings and extravasation had no correlation. Because the affected area was small in this group, it was difficult to evaluate cortical effacement. Although negative early CT findings did not always mean absence of extravasation and hemorrhagic complications in this group, the patients with hemorrhagic complications did not clinically deteriorate because of the small affected area. CONCLUSION: Hemorrhagic complications could be predicted by evaluation of angiographic occlusive site and pre- and post-therapeutic CT findings.

Aged↗
PubMed1996

Detection of K-ras point mutations at codon 12 in pure pancreatic juice for the diagnosis of pancreatic cancer by PCR-RFLP analysis.

The present study was undertaken to detect K-ras point mutations at codon 12 in pure pancreatic juice (PPJ) for the diagnosis of pancreatic cancer (PC) using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. PPJ was collected through a cannula under a duodenal fiberscope from 26 patients with PC and 32 patients with chronic pancreatitis (CP). DNA was extracted from PPJ and was used as the template for PCR. Analysis of PPJ by PCR-RFLP with BstNI revealed that the incidence of K-ras point mutations at codon 12 was 81% (21/26) in patients with PC and 6% (2/32) in those with CP. With reference to the location of PC, the incidence of K-ras mutations was 79% (11/14) in the head, 86% (6/7) in the body, and 80% (4/5) in the tail of the pancreas. The incidence of K-ras mutants was 50% (1/2) in tumor size 1 (TS1; < or = 2.0 cm in size), 71% (5/7) in TS2 (2.1 to < or = 4.0 cm), 89% (8/9) in TS3 (4.1 to < or = 6.0 cm), and 88% (7/8) in TS4 (> 6.1 cm). These results suggested that analysis of K-ras point mutations at codon 12 in PPJ using the PCR-RFLP method is a promising new genetic test for the diagnosis of PC.

Adult↗
PubMed1996

Detection of K-ras point mutations at codon 12 in pancreatic juice for the diagnosis of pancreatic cancer by hybridization protection assay: a simple method for the determination of the types of point mutation.

The present study was undertaken to detect K-ras oncogene point mutations at codon 12 in pure pancreatic juice (PPJ) by the hybridization protection assay (HPA) method for the diagnosis of pancreatic cancer (PC). This assay can be carried out within 30 min and can determine not only the presence of a mutation, but also the mutational type of K-ras at codon 12. The minimal ratio of mutant DNA detectable by the HPA was 5-10% of the total DNA. PPJ was collected through a cannula under duodenal fiberscope control from 20 patients with PC and 20 patients with chronic pancreatitis (CP). Analysis of PPJ by the HPA revealed that the incidence of K-ras point mutations at codon 12 was 55% (11/20) in patients with PC and 0% (0/20) in those with CP. Mutational types of K-ras at codon 12 in PC were aspartic acid (Asp) in nine cases, both Asp and cysteine in one case, and arginine in one case. Analysis of K-ras point mutations at codon 12 in PPJ using the HPA method seems promising as a new genetic test for the diagnosis of PC, because the HPA method is simple, and can easily determine the mutational type.

Base Sequence↗
PubMed1996

A new murine lymphocytotoxic monoclonal antibody recognizing HLA-A2, -A28 and -A9.

Monoclonal antibodies recognizing polymorphic as well as monomorphic epitopes on HLA antigens are important tools for understanding the immunobiology of HLA molecules. We immunized BALB/c mice with a HLA-A2 transfectant and screened for hybridomas which reacted with a HLA-A2 transfectant but not with a HLA-B75 transfectant. After subcloning by limiting dilution four times, a hybridoma secreting a monoclonal antibody (mAb) (IgG 2a, kappa) designated 1-145 was established. 1-145 reacted with Epstein-Barr virus transformed B lymphoblastoid cell lines (B cell lines) which expressed HLA-A2, -A28, -A23 and -A24. The titer of 1-145 in culture supernatant against HLA-A2 and -A28 antigens was similar and the titer against HLA-A23 was lower. 1-145 reacted with cells expressing HLA-A24 but the titer against HLA-A24 antigens was even lower than that against HLA-A23 antigens. The HLA-A24 antigens on the peripheral blood lymphocytes were not detected by 1-145 possibly due to the lower expression compared to the B cell lines. These differences of the titers were reflected to microlymphocytotoxicity assay in which 1-145 culture supernatant lysed all PBLs expressing HLA-A2,-A28 and -A23 but did not lyse PBLs expressing HLA-A24. Published deduced amino acid sequence data of HLA class 1 molecules indicate that Lys in position 127 may be critical for 1-145 binding.

Amino Acid Sequence↗
PubMed1996

Effect of estriol on bone loss in postmenopausal Japanese women: a multicenter prospective open study.

OBJECTIVES: To assess the effects of oral estriol on the bone mineral density (BMD) and bone metabolism in postmenopausal women. METHODS: Seventy-five natural postmenopausal women with a BMD of more than 10% below the peak bone density were treated for 50 weeks with 2 mg/day estriol (E3) cyclically and 0.8 g/day of calcium lactate continuously. BMDs at L2-L4 were measured by dual energy X-ray absorptiometry (DXA). RESULTS: The BMD increased 1.79% (p < 0.01 vs. pretreatment) after 50 weeks, accompanied with decrease of biochemical markers of bone turnover. With regard to climacteric symptoms, Kupperman's menopausal index improved (p < 0.01 vs. pretreatment) after 5 weeks of treatment. As to the incidence of adverse events genital bleeding was observed in only 8.0% of the subjects. Endometrial histology and cytology showed neither abnormalities nor hyperplasia during and after the treatment. CONCLUSIONS: Estriol prevented postmenopausal bone loss and improved climacteric symptoms effectively with low incidence of genital bleeding.

Bone Density↗
PubMed1996

Coupling of Metabolism and Bioconversion: Microbial Esterification of Citronellol with Acetyl Coenzyme A Produced via Metabolism of Glucose in an Interface Bioreactor.

Microbial esterification of citronellol with acetyl coenzyme A (acetyl-CoA) produced via metabolism of glucose was performed with Hansenula saturnus IFO 0809 in an interface bioreactor by using a hydrophilic carrier (an agar plate or an agar-coated filter pad) and a hydrophobic organic solvent (decane). An increase in the glucose concentration on an agar plate led to a decrease in oxidation of citronellol and an increase in the amount of citronellyl acetate. Fed-batch addition of citronellol efficiently alleviated substrate toxicity and resulted in the accumulation of high levels of citronellyl acetate. Furthermore, triple coupling of acetyl-CoA formation, microbial reduction of citronellal to citronellol, and esterification of the resulting citronellol with acetyl-CoA was also an efficient way to prepare citronellyl acetate. By using coupling systems, citronellyl acetate could be efficiently produced without any acetyl donor.

Journal Article↗
PubMed1996

Paraumbilical vein aneurysm. Case reports.

Two cases of paraumbilical vein aneurysm are reported. The patients were diagnosed as having cirrhotic liver with portal hypertension. Angiography and contrast-enhanced computed tomography demonstrated a dilated paraumbilical vein arising from the left branch of the portal vein. Furthermore, localized dilatation of a paraumbilical vein was demonstrated. The focal aneurysmal dilatation of the dilatated paraumbilical vein is rare.

Aneurysm↗
PubMed1996

S-methylmethionine sulfonium in fruits of citrus hybrids.

The S-methylmethionine sulfonium (MMS) concentrations in fruits of citrus hybrids were measured, and found to increase during ripening of the fruit. However, there of eleven hybrids of 'Seto unshiu' crossed with 'Morita ponkan' and four of 9 hybrids of 'Murcott' tangor crossed with 'Seto unshiu' had low MMS concentrations even at late harvest stage. Crossbreeding is useful in producing new citrus fruits that have juices with the desirable characteristics of their parents without formation of dimethyl sulfide which is an off-flavor.

Citrus↗
PubMed1996

Efficient lipase-catalyzed preparation of long-chain fatty acid esters of bile acids: biological activity and synthetic application of the products.

A highly regioselective (3-position) and efficient (quantitative yield) acylation of bile acids catalyzed by immobilized Candida antarctica lipase was established. Methyl cholate derivatives acylated with long-chain fatty acids (C12-C16) showed an inhibitory effect on the growth of some strains of Gram-positive and -negative bacteria (27-400 micrograms/ml). The anti-bacterial activity was slightly weaker than has been observed for methyl cholate, while the increased lipophilicity and lower melting points of the present derivatives are well suited for a potential germicide which would be safe and be topically applied. This enzyme-catalyzed transesterification is also demonstrated as an expeditious route to ursodeoxycholic acid, in respect of the regioselective introduction of acyl protecting groups on the hydroxyl groups of the intermediates. 7-Ketolithocholic acid, a known direct precursor of ursodeoxycholic acid, was obtained from cholic acid via chenodeoxycholic acid in a 46% yield and 9 steps.

Acylation↗
PubMed1996

[Role of trace elements on metallothionein gene expression].

Metallothionein (MT) is low-molecular-weight metal binding proteins with a high cysteine content, which provide the protection against metal toxicity and the regulation of essential trace elements such as zinc and copper. MT is inducible proteins by various metals, hormones, cytokines, and endogenous and exogenous agents. Human MT genes locate on chromosome 16, and mainly three isoforms of MT (MT-I, MT-II, MT-III) have been identified. MT genes expression is controlled mainly at the transcriptional level, and is regulated by the interaction between cis-acting elements (MRE, GRE, IRE) and trans-acting factors (MTF-1, ZRF, MRE-BF-1 and 2, MREBP). Regulation of MT genes by metals may be mediated by MTF-1 interacting with MREs and zinc functions to release MTF-1 from an inhibitor (MTI).

Chromosomes, Human, Pair 16↗