Isolation of transporter associated with antigen processing genes, TAP1 and TAP2, from the horned shark Heterodontus francisci.
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ATP Binding Cassette Transporter, Subfamily B, Mem↗
Biomedical subjects
Y Ohta
Publications and source records attributed to Y Ohta.
Clinical implications of mutations C-to-T1653 and T-to-C/A/G1753 of hepatitis B virus genotype C genome in chronic liver disease.
Among many mutational "hot spots" on hepatitis B virus (HBV) genome, A-to-T1762 and G-to-A1764 within the core promoter have been underscored in view of disease association as well as viral expression/replication. Although to a lesser extent, C-to-T1653 and T-to-V(C/A/G)1753 were also noteworthy in our previous study. To assess the clinical significance of these mutations, we determined the nucleotide sequence of an HBV DNA fragment covering these sites in HBsAg-positive blood donors (n = 160) and patients with chronic hepatitis (n = 66), liver cirrhosis (n = 45), and hepatocellular carcinoma (n = 58), most of whom were infected with genotype C HBV (subtype adr). In cases where HBe antigen was positive, the frequency of T1653 and/or V1753 showed a striking increment from chronic hepatitis patients (18%) to liver cirrhosis and/or hepatoma patients (82%), whereas that of T1762/A1764 was already high in chronic hepatitis patients (76%). In HBe antigen-negative cases, by contrast, significant difference in the frequency of T1653/V1753 mutants was found between blood donors (22%) and chronic hepatitis patients (67%). Our results suggest that T1653/V(particularly C)1753 mutants are more closely associated than T1762/A1764 with the progression of liver disease from chronic hepatitis to cirrhosis in HBe antigen-positive patients. A system of site-directed mutagenesis PCR RFLP was constructed to diagnose T1653 and C/A1753 more conveniently. Detecting T1653 and C/A1753 by this method would contribute to the differential diagnosis of HBV-associated liver disease.
Base Sequence↗
Genotypes of TT virus (TTV) compared between liver disease patients and healthy individuals using a new PCR system capable of differentiating 1a and 1b types from others*.
TT virus (TTV) lacks obvious pathogenicity; almost all of the infected hosts are symptom-free. A possibility remains, however, that certain strains can cause liver disease while most others are non-pathogenic. Genotypes 1 a and 1 b have been proposed to contain such pathogenic strains. To test this possibility, we constructed a PCR system capable of detecting TTV of the 1 a and 1 b genotypes differentially from the other genotypes, and compared the frequencies of these genotypes between patients with liver disease of unknown etiology (n=42) and healthy individuals (n=50). The assay comprised 3 steps: i) PCR to amplify a 3.2-kb fragment using universal primers; ii) 2nd-round PCR, starting from the 3.2-kb amplicon, for a approximately 280-nt fragment by use of genotype 1-specific primers; and iii) digestion of the approximately 280-nt amplicon with MboI and BanI to discriminate between 1 a and 1 b. Eventually, 40 (95%) of the patients and 47 (94%) of the controls were positive for the 3.2-kb amplicon, and the 1 a, 1 b, 1 a+1 b, and non-1 genotypes of TTV were found in 2 (5%) vs 4 (9 percent), 5 (13%) vs 4 (9%), 1 (3%) vs 1 (2%) and 32 (80%) vs 38 (81%) of the 40 patients and 47 controls, respectively: the distribution was almost identical between the two groups. The hypothesis that the genotype 1 of TTV may be more closely associated with liver disease than other genotypes was not supported by this study.
Base Sequence↗
Tumor angiogenesis and recurrence in stage I non-small cell lung cancer.
BACKGROUND: Tumor angiogenesis appears to relate to recurrence after an operation as a route for distant metastasis. We assessed the association of vascular endothelial growth factor (VEGF) expression and intratumoral microvessel density (MD) with recurrence in primary lung cancer. METHODS: Samples were randomly obtained from 104 stage I lung cancer patients who underwent curative operations (43 recurrent, 61 nonrecurrent patients). Microvessels were highlighted by staining endothelial cells for factor VIII and VEGF antigen was detected using a polyclonal antibody. RESULTS: VEGF antigen was detected in large amounts in both recurrent (100%) and nonrecurrent tumors (73.8%). The percentages of patients with the strongest VEGF stain (more than 50% of staining area in tumor cells) were 46.5% in tumors with recurrence and 11.5% in tumors without recurrence. The mean MD in recurrent and nonrecurrent tumors were 18.2+/-10.5 and 8.5+/-5.0, respectively, resulting in a significantly greater value in tumors with recurrence (p<0.0001). Although there were no significant differences in mean MD according to pathological types, in adenocarcinoma and adenosquamous carcinoma, the mean value in the recurrent group was significantly greater than that in the nonrecurrent one. CONCLUSIONS: An evaluation of VEGF expression and MD in tumors may contribute to the estimation of the risk of recurrence of non-small cell lung cancer in early stages.
Adult↗
Isolation of bacteria from cervical lymph nodes in patients with oral cancer.
Thirty patients with oral mucosal cancer were studied in relation to oral mucosal damage and bacterial translocation to the regional lymph nodes in the neck. All 30 patients (gingiva 11, tongue 13, cheek mucosa four, oral floor two) underwent extensive, clean-contaminated, head-and-neck surgery (including neck dissection) with free flap reconstruction. A total of 153 lymph nodes was harvested for microbial and histological examination. Viable bacteria were isolated from 70 lymph nodes (45.8%) from 25 patients (83.3%). Bacterial cells in the nodes were detected by scanning electron microscopy. Bacterial translocation was found more often in metastatic nodes (75.0%) than in uninvolved nodes (40.3%) (p = 0.015; chi2 test). Gingival carcinoma yielded 56.4% of bacterial growth in the regional lymph nodes compared to tongue (40.3%), oral floor (40.0%) and cheek mucosa (37.5%). As the gingival carcinoma group includes more T4 cases (11/11; 100%) than the other three groups (7/19; 36.8%), bacterial translocation in uninvolved nodes could be caused by the size and invasion of the primary oral tumor. Oral streptococci (Streptococcus intermedius, Strep. constellatus, Strep. oralis, Strep. mitis, Strep. sanguis, Strep. salivarius) were the most common isolates. Aerobic enteric bacteria (Enterococcus, Escherichia, Klebsiella etc.) were also found in the lymph nodes. Among the anaerobic bacteria, Peptostreptococcus spp. were isolated from 12 patients. Damaged oral mucosa in patients with oral cancer might allow the new bacterial colonization on the surface and subsequently drain the bacteria into the regional lymph nodes as well as the general circulation.
Bacteria↗
Modulation of stress-induced gastric mucosal lesions by exogenous L-arginine.
L-arginine, a nitric oxide (NO) precursor, can exert both ameriolative and deteriorative effects on gastric mucosal lesions. This study was designed to determine whether exogenous L-arginine modulates stress-induced gastric mucosal lesions through NO production by either constitutive NO synthase (cNOS) or inducible NO synthase (iNOS) in gastric mucosal tissues. In rats subjected to water immersion restraint stress over a 6-hour period, the concentration of gastric mucosal nitrite/nitrate, breakdown products of NO, increased with the development of gastric mucosal lesions and a decrease in cNOS activity and a drastic increase in iNOS activity in the gastric mucosal tissue. Preadministration of L-arginine (150 to 600 mg/kg intraperitoneally) attenuated the lesion development with prevention of increases in gastric mucosal nitrite/nitrate concentration and iNOS activity. In contrast, postadministration of L-arginine (150 to 600 mg/kg intraperitoneally) enhanced the lesion development with further increase in gastric mucosal nitrite/nitrate concentration. This deteriorative action of postadministration of L-arginine (300 mg/kg intraperitoneally) was prevented by pretreatment with aminoguanidine (100 mg/kg subcutaneously), a selective iNOS inhibitor, with inhibition of increases in gastric mucosal iNOS activity and nitrite/nitrate concentration. These results indicate that preadministered L-arginine protects against water immersion restraint stress-induced gastric mucosal lesions, possibly through restricted NO production by cNOS in gastric mucosal tissues, whereas postadministered L-arginine aggravates the stress-induced gastric mucosal lesions, possibly through excessive NO production by iNOS increasing in gastric mucosal tissues.
Animals↗
Synthesis and anti-influenza virus activity of novel pyrimidine derivatives.
Efficient synthetic routes of 2-amino-4-(omega-hydroxyalkylamino)pyrimidine derivatives were investigated in relation to the anti-influenza virus activity of these compounds. The derivatives in which cyclobutyl and cyclopentyl groups were introduced to the beta-position of the aminoalkyl group (especially the cyclobutyl group substituted by a phenylalkyl group at the 3'-position) resulted in improved antiviral potency: i.e. an average 50% effective concentration for inhibition of plaque formation (EC50, microM) of 0.1-0.01 microM for both types A and B influenza virus. The antiviral efficacies were in the order of amino group > hydroxyiminomethyl group > halogen substitution at the 5-position, and chlorine or methoxy group > hydrogen at the 6-position of the pyrimidine ring. The antiviral indices of these compounds were 2-6 with respect to the 50% inhibitory concentration for cell proliferation (IC50, microM) for growing cells, but > 500 to > 10(4) with respect to the IC50 for stationary cells, indicating that these compounds may be efficacious for the topical treatment of influenza virus infection.
Animals↗
Production of canthaxanthin by extremely halophilic bacteria.
Soil samples from a salt farm were used as a source for the isolation of carotenoid-producing bacteria. The conditions for optimum growth and carotenoid production were established for the isolated bacteria. Carotenoids were analysed by spectrophotometry and High Performance Liquid Chromatography (HPLC). Thirty-one red extremely halophilic bacteria were isolated from saline soil samples collected from a salt farm in Alexandria, Egypt. Among the isolated strains, strain TM exhibited the highest carotenoid-producing ability. Maximum growth of strain TM occurred in the presence of high concentrations of sodium chloride and magnesium sulfate. Growth did not occur when NaCl concentration was lower than 10% and the cells lysed at this concentration. Optimum growth of and carotenoid production by strain TM were realized at 37 degrees C in the presence of 1% yeast extract, 0.75% casamino acids, 25% NaCl, 4% MgSO4, 0.2% KCl and at pH 7.2 with shaking for 6 d. Strain TM produced 2.06 mg total carotenoids g(-1) dry cells, including 0.06 mg of beta-carotene and 0.70 mg of canthaxanthin. This is the first report of an extremely halophilic bacterium that produces canthaxanthin.
Journal Article↗
Purification and properties of hydrogen sulfide oxidase from Bacillus sp. BN53-1.
A hydrogen sulfide oxidase was purified to homogeneity from the heterotroph Bacillus sp. BN53-1 isolated from pig feces compost. The enzyme was found to be a monomer with a M(r) value of approximately 37 kDa. It required FAD for its activity, which was not replaced by FMN. The optimum reaction pH and temperature were 7.5 and 40 degrees C, respectively. The enzyme was stable between pH 6.0 and 7.0 and up to 30 degrees C. Its activity was stimulated by Ca2+ and Mn2+ and inhibited by Al3+, dithiothreitol, and 2-mercaptoethanol. The main product was elemental sulfur, and H2O2 was not detected. The N-terminal sequence of the enzyme showed similarity to other FAD-requiring enzymes.
Journal Article↗
Rapid measurement and control of the moisture content of compost using near-infrared spectroscopy.
In a compost fermentation of soybean-curd (tofu) refuse, the effects of the moisture content of the compost on the compost reaction were studied. The moisture content of the compost was a very important factor for good fermentation. Near-infrared spectroscopy (NIRS) was applied to the determination of the moisture content of the compost. The reflected rays in the wavelength range between 400 and 2500 nm were measured at 2 nm intervals. The absorption of water was observed at three wavelengths, 960, 1406 and 1888 nm. To formulate a calibration equation, a multiple linear regression analysis was carried out between the near-infrared spectral data at 960 nm (sample number, n = 50) and on the moisture content obtained using a drying method. The values of the simple correlation coefficient and the standard error of calibration were 0.987 and 1.33%, respectively. To validate the calibration equation obtained, the moisture content in the prediction sample set (n = 35) not used for formulating the calibration equation was calculated using the calibration equation, and compared with the values obtained using the drying method. Good agreement was observed between the results of the drying method and those of the NIRS method. The simple correlation coefficient and standard error of prediction were 0.979 and 1.85%, respectively. Then, the NIRS method was applied to a practical situation in which the moisture content was measured and controlled during the compost fermentation, and good results were obtained. The study indicates that NIRS is a useful method for measurement and control of the moisture content in the compost of soybean-curd refuse.
Journal Article↗
Banding of the pulmonary trunk in preparation for a Fontan operation.
Banding of the pulmonary trunk is an important surgical procedure for patients who have congenital cardiac malformations with unrestricted pulmonary flow. We propose a new concept for determining in such circumstances the most appropriate length of the band used to constrict the pulmonary trunk in preparation for a Fontan operation. We studied 14 patients undergoing banding of the pulmonary trunk and measured the following parameters: diameter of aorta, diameter of pulmonary trunk, length of pulmonary arterial band and maximum flow velocity across the banded segment. We calculated an index from our original parameter based on the formula; length of band/(diameter of aorta diameter of pulmonary trunk). The diameter of aorta was 9.5 +/- 1.4 mm, and that of the pulmonary trunk was 9.6 +/- 2.3 mm. The length of the band was 16.5 +/- 3.4 mm, giving a calculated index of 0.188 +/- 0.038. The maximum flow velocity was 4.02 +/- 0.46 m/s. No correlation was found between the length of the band and body weight, and also no correlation was found between the length of the band and maximum flow velocity. The calculated index had a negative correlation with the maximum velocity of flow across the band (y = -8.13x +/- 5.56, R = 0.74, p < 0.01). We believe that the proposed index is a useful guide in determining the length of a pulmonary band when preparing patients for a Fontan operation.
Cardiac Catheterization↗
BMIPP in angina pectoris.
Radionuclide imaging with BMIPP for assessment of myocardial metabolism can assist in clinical decision making in certain subsets of patients with coronary artery disease. In stable angina, abnormal BMIPP imaging and less BMIPP uptake than Tl-201 are more likely to be seen in the distribution of vessels with severe stenoses and related to regional wall abnormalities. BMIPP SPECT imaging is a promising approach in the evaluation of patients with suspected unstable angina during the pain-free period even after stabilization. Fatty acid metabolism is easily suppressed due to the abrupt onset of ischaemia. In addition, combined imaging with BMIPP and Tl-201 can provide insights into the understanding of abnormal metabolism in myocardium in stable and unstable angina. This review addressed the clinical applications of BMIPP imaging for evaluation of stable and unstable angina.
Angina Pectoris↗
Sulfated sialic acid-polymers inhibit the cytotoxic action of bee and snake venom.
Colominic acid is an alpha2,8-linked sialic acid polymer produced by Escherichia coli. We found that synthetic sulfated-colominic acids (SC) remarkably inhibited the cytotoxicity of bee and snake venom toward mouse fibroblast cells, but colominic acids showed no inhibition themselves, indicating the important role of sulfate groups in the inhibitory activity of SC. Other sulfated carbohydrates such as chondroitin sulfates, heparin and heparan sulfate showed no inhibition. SC also exhibited potent inhibition of melittin, a highly basic peptide, which is a major cytotoxic component of bee venom. SC did not inhibit phospholipase A2 activity in bee venom. This suggests that the inhibition of bee and snake venom by SC is due to inhibition of melittin and cardiotoxin, which is a cytolytic peptide in snake venom, respectively. SC with a higher sulfur content and a larger molecular mass showed more potent activity. The interaction between SC and melittin basically seems an ionic one, however, the conformation of SC is also likely important. For the binding of SC to melittin leading loss of its cytotoxic activity, the sulfate groups of SC must be properly arranged to interact with lysine and arginine residues of melittin molecules, which play an important role in the cytolytic activity. A higher molecular mass of SC substituted with more sulfate groups is required for more obvious inhibition of the cytotoxic activity.
Animals↗
Participation of xanthine-xanthine oxidase system and neutrophils in development of acute gastric mucosal lesions in rats with a single treatment of compound 48/80, a mast cell degranulator.
The participation of xanthine-xanthine oxidase and neutrophils in the development of acute gastric mucosal lesions was examined in rats injected once with compound 48/80, a mast cell degranulator. Gastric mucosal lesions appeared 0.5 hr after compound 48/80 injection and developed at 3 hr. The formation of gastric mucosal lesions at 0.5 hr after compound 48/80 injection was prevented by pretreatment with anti-neutrophil antiserum and NPC 14686, an antiinflammatory agent, but not with allopurinol, a xanthine oxidase inhibitor. The development of gastric mucosal lesions at 3 hr after compound 48/80 injection was prevented by pretreatment with anti-neutrophil antiserum, NPC 14686, or allopurinol. Increases in the activities of gastric mucosal xanthine oxidase and myeloperoxidase, an index of neutrophil infiltration, and the content of lipid peroxide occurred 0.5 hr after compound 48/80 injection, and these increases were enhanced at 3 hr. The increases in gastric mucosal myeloperoxidase activity and lipid peroxide content at 0.5 hr after compound 48/80 injection were attenuated by pretreatment with anti-neutrophil antiserum and NPC 14686, while only the increase in gastric mucosal xanthine oxidase activity at the same time point was arrested by allopurinol pretreatment. The increases in gastric mucosal xanthine oxidase and myeloperoxidase activities and lipid peroxide content at 3 hr after compound 48/80 treatment were attenuated by pretreatment with anti-neutrophil antiserum, NPC 14686, or allopurinol. When compound 48/80-injected rats were treated with allopurinol at 0.5 hr after compound 48/80 injection, the progression of gastric mucosal lesions at 3 hr after the injection was almost completely prevented with inhibition of the increases in gastric mucosal xanthine oxidase and myeloperoxidase activities and lipid peroxide content. These results indicate that in rats with a single compound 48/80 treatment neutrophils infiltrated into the gastric mucosa participated in the development of acute gastric mucosal lesions and that the xanthine-xanthine oxidase system in the gastric mucosa participated in the progression rather than the formation of the gastric mucosal lesions.
Acute Disease↗
VEGF and VEGF type C play an important role in angiogenesis and lymphangiogenesis in human malignant mesothelioma tumours.
The vascular endothelial growth factor (VEGF) family is a novel regulator of endothelial cell proliferation. We assessed the mRNA expression of VEGF, VEGF type C (VEGF-C) and their receptors together with the microvessel density (VD) and microlymphatic vessel density (LVD) in pursuit of their connection and prognostic value in malignant pleural mesothelioma (MPM). We used four human MPM cell lines, 54 MPM tumours and five normal pleural tissues. Expression levels for receptors and ligands were assessed by semiquantitative reverse transcriptase polymerase chain reaction analysis. Microvessels were highlighted by immunohistochemical staining for factor VIII. The discrimination of lymphatics was performed by enzyme-histochemistry for 5'-nucleotidase after adequate inhibition of non-specific activity. The expression levels of VEGF, VEGF-C and VEGFRs were high in all MPM cell lines. The percentages of tumours with higher expression compared to the mean values of normal pleural tissues were 31.5% (17/54) for VEGF, 66.7% (36/54) for VEGF-C, 20.4% (11/54) for fms-like tyrosine kinase (flt)-1, 42.6% (23/54) for kinase insert domain-containing recepter (KDR) and 59.3% (32/54) for flt-4. Significant positive correlations were found between VEGF-C and flt-4, VEGF and KDR, VEGF and flt-1 in tumour tissues. The association between LVD and VEGF-C expression level was especially strong (P< 0.0001, r= 0.63). There were also significant correlations between LVD and flt-4, and VD and VEGF. No correlation, however, was found between LVD and nodal metastasis. VD was a negative prognostic indicator in this study. The associations between VEGFNEGF-C and vessel density suggest that these factors play an important role in angiogenesis and lymphangiogenesis in this tumour, and assessment of vascularity may be a useful prognostic indicator for MPM patients.
5'-Nucleotidase↗
The effect of papaverine on morphologic differentiation, proliferation and invasive potential of human prostatic cancer LNCaP cells.
BACKGROUND: Intracellular cyclic adenosine monophosphate (AMP) level changes are thought to play an important role in inhibiting cell proliferation and inducing differentiation in several types of cells. It has been reported that cyclic AMP analogs induce terminal differentiation in human prostate cancer cells. Consequently, phosphodiesterase inhibitors may be useful in delineating the role of cyclic AMP in the differentiation of these cells. Therefore, the effect of phosphodiesterase inhibitors on morphologic differentiation, proliferation and invasive potential of human prostate cancer cells was investigated. METHODS: Three human prostate cancer cell lines PC-3, DU145 and LNCaP were treated with one of the phosphodiesterase inhibitors, papaverine, 3-isobutyl-1-methylxanthine (IBMX) or theophylline, for 6 days. Morphologic changes of these cells induced by phosphodiesterase inhibitors were observed by microscopy. Intracellular cyclic AMP levels in LNCaP cells were measured by radioimmunoassay using a cyclic AMP assay kit. The effect of papaverine on the proliferation and invasive potential of LNCaP cells were measured by cell counting and the Matrigel invasion chamber assay. RESULTS: Of the three agents, examined papaverine (10(-5) mol/L) is the most effective inducer of morphologic change and also raised intracellular cyclic AMP levels in LNCaP cells. However, unlike LNCaP cells, PC-3 and DU145 cells treated with phosphodiesterase inhibitors, including papaverine, showed little change in morphology. Additionally, proliferation and invasive potential of LNCaP cells were significantly inhibited by papaverine. CONCLUSION: The results suggest that papaverine induces terminal differentiation in LNCaP cells, which is correlated with an intracellular cyclic AMP-mediated pathway.
1-Methyl-3-isobutylxanthine↗
Ganglioside GM(1a) on the cell surface is involved in the infection by human rotavirus KUN and MO strains.
Rotavirus is the most common cause of severe gastroenteritis in infants and children worldwide. The cell attachment of most animal rotaviruses, which belong to the neuraminidase-sensitive strains, requires sialic acid residues on the host cell membranes. On the other hand, most human rotaviruses are classified as neuraminidase-insensitive strains. The involvement of gangliosides on the host cell surface in human rotavirus infection was investigated by immunostaining analysis of target cells, and by assaying the neutralization of infection by rotavirus and the blocking of target cellular receptors. In host cells (MA104 cells) pretreated with Arthrobacter ureafaciens neuraminidase, which were still infected by human rotaviruses (KUN and MO strains), GM(3) was hydrolyzed markedly by the neuraminidase, while GM(1a) was not hydrolyzed at all. Infection by the rotaviruses was strongly inhibited by exogenous ganglioside GM(1a), but not GA(1). Infection was also inhibited by pretreatment of the MA104 cells with cholera toxin B-subunit, which specifically blocked ganglioside GM(1a) on the plasma membrane. The treatment of MA104 cells with the endoglycoceramidase attenuated human rotavirus infection. From these findings, we concluded that GM(1a) on the plasma membrane of the host cells was involved in the infection by human rotavirus KUN and MO strains.
Animals↗
Effect of amino acid injection in broiler breeder eggs on embryonic growth and hatchability of chicks.
Two experiments were conducted to evaluate the effect of amino acid (AA) injections in ovo in Cobb broiler breeder eggs on hatchability and subsequent chick BW. In Experiment 1, moisture, crude fat (CF), and CP were analyzed over time during incubation (Day 0, 7, 14, and 19 of incubation). Moisture, CP, and CF of the embryo increased, and moisture, CP, and CF of eggs decreased, as incubation time increased (P < 0.05). Combined egg and embryo AA contents, except Gly and Pro, decreased (P < 0.05) as incubation time increased. However, the pattern of AA in the egg did not change as the embryo developed. In Experiment 2, AA were injected into the yolk or air cell at Day 0 and 7 of incubation. Hatchability was reduced (P < 0.05) when AA were injected at Day 0 of incubation. However, when the AA solution was injected into the yolk sac at Day 7 of incubation, hatchability was not affected, and BW of chicks increased relative to egg weight prior to incubation. These results suggest that in ovo administration of AA may be an effective method of increasing chick BW at hatch.
Amino Acids↗