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Biomedical subjects

H Doi

Publications and source records attributed to H Doi.

At least 37 records · Page 2Linked to original sources
PubMed1996

Hepatitis C virus (HCV) subtype prevalence in Chiang Mai, Thailand, and identification of novel subtypes of HCV major type 6.

Subtype analysis of hepatitis C viruses (HCVs) obtained from patients with chronic liver disease in Chiang Mai, Thailand, was performed. Of 46 HCV isolates, 13 (28%) were shown to belong to HCV subtype 3a (HCV-3a), 10 (22%) to belong to HCV-1a, 7 (15%) to belong to HCV-1b, 1 (2%) to belong to HCV-3b, and 1 (2%) to belong to a variant group, as determined from partial nucleotide sequences of the NS5B region of the viral genome. Analysis of 5' untranslated region sequences identified five other isolates (11%) of HCV type 1 and two other isolates (4%) of type 3. Detailed phylogenetic positions for the variant described above and those previously obtained from blood donors and drug addicts in Chiang Mai were determined by a six-parameter neighbor-joining method on the basis of core, E1, and NS5B region sequences. The results revealed that those sequence variants represent novel subtypes of HCV type 6. The HCV type 6 isolates appear to be antigenically different from isolates of HCV types 1 and 2, as determined by a serotyping method that utilizes recombinant peptides corresponding to a portion of the NS4 protein. The significance of subtype analysis around this area is discussed.

Adult↗
PubMed1996

Suppression of atherosclerotic changes in cholesterol-fed rabbits treated with an oral inhibitor of neutral endopeptidase 24.11 (EC 3.4.24.11).

Neutral endopeptidase 24.11 (NEP), widely distributed in the body, hydrolyzes and inactivates a number of endogenous vasoactive peptides, some of which could alter various functions of cells present in the arterial wall. Recently NEP has been found to exist in the vascular endothelium. The aim of this study was to assess the influence of chronic NEP inhibition by daily administration of UK79300 (candoxatril), an orally active NEP inhibitor (NEPI), on the development of atherosclerotic changes in high-cholesterol-fed rabbits. Male New Zealand White rabbits were fed for 8 weeks as follows: normal rabbit diet (Normal, n = 15), 1.5% cholesterol diet (Cholesterol, n = 15), or 1.5% cholesterol diet containing NEPI (20 mg.kg-1.d-1) (Cholesterol+NEPI, n = 15). At the end of the dietary period, NEPI treatment was found to suppress the surface area of the aorta covered by plaques (% surface area: Cholesterol, 59 +/- 6 versus Cholesterol+NEPI, 36 +/- 7, P < .01) and decreased contents of cholesterol and cholesterol esters in the aortas. NEPI also reduced plasma total cholesterol by 27% of Cholesterol rabbits (1781 +/- 130 mg/dL). The endothelial function, estimated by the endothelium-dependent relaxation of the isolated aortas in response to acetylcholine, was preserved in Cholesterol+NEPI rabbits compared with that in Cholesterol rabbits. NEP enzymatic activities in plasma and the particulate fraction of the homogenates from the aortas in Cholesterol rabbits were both increased, 3.1- and 3.9-fold, respectively, above those in Normal rabbits, but the activities in Cholesterol+NEPI rabbits were significantly lower than those in Cholesterol rabbits. UK73967, an active form of UK79300, or phosphoramidon partly reversed the atherosclerotic impairment of relaxation of the isolated thoracic aortic rings from Cholesterol rabbits in response to exogenous additions of C-type natriuretic peptide (CNP) and substance P, which are NEP substrates known to exist endogenously in the vascular endothelium. The results suggest that the increased NEP activity plays a significant role in atherogenesis, and NEPIs might be therapeutically useful in the prevention of atherosclerosis. Reduction of plasma cholesterol and suppression of degradations in the arteries of endogenously released CNP, substance P, or possibly other kinins known to have anti-atherosclerotic actions may at least partially contribute to the inhibitory effects of NEPIs on atherosclerotic changes.

Administration, Oral↗
PubMed1996

Prolongation of the life span of cardiomyopathic hamster by the adrenergic beta 1-selective partial agonist denopamine.

Influence of cardiotonic agents on the prognosis of heart failure depends on the individual therapeutic agents, and favorable and unfavorable effects of these agents have been reported in clinical trials. We studied the effect of the cardiotonic agent denopamine on the life span of cardiomyopathic hamsters (BIO 14.6 strain) in the heart failure period. Non-treated hamsters started to die at 40 weeks of age, and their survival rate decreased to 23.8% at the age of 65 weeks. Hamsters treated with denopamine (400 ppm in diet) from 36 weeks of age did not die until the age of 52 weeks, except in cases of accidental death. The survival rate of this group at 65 weeks of age was about 40%. Survival rates of these 2 groups were significantly different (P < 0.05) when animals with accidental death were excluded. To elucidate the mechanism of the effect of denopamine, we performed several experiments after dietary treatment with denopamine for 4 to 6 weeks from 37 weeks of age. Denopamine treatment lowered plasma levels of noradrenaline and dopamine (P < 0.05), but affected neither the cardiac contractility nor the beta-adrenoceptor density. In summary, denopamine significantly decreases the mortality of cardiomyopathic hamsters. Its effect to lower the plasma catecholamine levels may be responsible for the beneficial effect of denopamine.

Adrenergic beta-1 Receptor Agonists↗
PubMed1996

[Flow cytometric measurement of DCFH-oxidation of neutrophils in peripheral whole blood, isolated leukocytes and cerebrospinal fluid cells].

Using 2',7'-dichlorofluorescein diacetate (DCFH-DA) as an indicator for the intracellular formation of H2O2 and free radicals, we measured DCFH-oxidation (DCF fluorescence) of neutrophils by flow cytometry. The DCF fluorescence intensity of unstimulated neutrophils in whole blood was the same as the autofluorescence of neutrophils. However, that of unstimulated isolated neutrophils was higher than autofluorescence. The DCF fluorescence of isolated neutrophils was higher than that of neutrophils in whole blood with or without stimuli, and was decreased to the intensity of neutrophils in whole blood by the addition of plasma or erythrocytes. The DCF fluorescence of neutrophils in whole blood indicates the oxidative state of neutrophils in vivo. The determination of DCFH-oxidation of neutrophils in isolated leukocytes is also necessary to evaluate precisely the oxidative state of neutrophils. In cerebrospinal fluid (CSF) cells from patients with meningitis, the DCF fluorescence intensity of unstimulated neutrophils was higher than the autofluorescence of CSF neutrophils. The DCF fluorescence of CSF neutrophils stimulated with PMA showed increased intensity over that of unstimulated CSF neutrophils. The DCF fluorescence of CSF neutrophils, both unstimulated and stimulated with PMA, were decreased with the addition of plasma but not with the addition of CSF.

Cerebrospinal Fluid↗
PubMed1996

[Comparison of biochemical properties of human airway tryptase isolated from mucoid sputum with those of lung mast cell tryptase].

We found a novel trypsin-like enzyme (tryptase) in sputum from patients with chronic airway diseases, and named this enzyme human airway tryptase (HAT). To clarify its physiological significance in the airway, we compared biochemical properties of purified HAT with those of purified lung mast cell tryptase (MCT). Studies with model peptide substrates showed that both the HAT and MCT preferentially cleaved the COOH-terminal side of arginine residues of certain peptides, but substrate specificities to nine synthetic model substrates of HAT differed from those of MCT. Effects of protease inhibitors on the two enzymes were examined at a concentration of 10 microM. Both the HAT and MCT were strongly inhibited by the trypsin inhibitors leupeptin, antipain, and aprotinin. An alpha-1-protease inhibitor inhibited HAT by 50%, but it did not inhibit MCT. In contrast, a secretory leukocyte protease inhibitor strongly inhibited MCT, but not HAT. Mucoid sputum from patients with chronic bronchitis contained much more HAT than MCT. These differences in biochemical properties between HAT and MCT indicate that they play different physiological roles in the airways.

Asthma↗
PubMed1996

[Simultaneous aortic root and total arch graft replacement in a patient with aortitis syndrome].

The patient was a 45-year-old female diagnosed with aortitis syndrome and aortic regurgitation (AR). She had been taking steroid therapy since 1975. She had recently developed congestive heart failure due to AR while both the ascending aorta and aortic arch were enlarged. She had no inflammatory reaction on admission. An aortogram showed heavy dilation of both the ascending aorta and aortic arch and maximum diameters was 11 cm in the ascending aorta and 4.5 cm in the descending aorta. There was an obstruction of the left subclavian artery. Moderate AR was seen on an echocardiogram. She had a simultaneous graft replacement of aortic root and total arch. The aortic root was replaced with composite graft and coronary arteries were implanted using Carrel's patch technique, and the aortic arch was also replaced with a graft with two side branches. The postoperative course was uneventful without complication of cerebral infarction or paraplegia. The postoperative aortogram showed stenosis of the left carotid artery, but no abnormality of the coronary orifices and graft anastomosis. She returned home with disappearance of symptoms of congestive heart failure.

Aorta↗
PubMed1995

Induction of TCR-gamma delta+ cells from thymocytes stimulated by a fetal liver-derived hepatocyte clone.

We have previously reported that a fetal liver-derived hepatocyte clone, FHC-4D2, can support hematopoiesis in vitro. Here, we show that fetal thymocytes (FT) or adult thymocytes (AT) proliferate on the monolayer of FHC-4D2 cells in the presence of rIL-2. Fresh thymocytes contained few TCR-gamma delta+ cells (< 4% for FT and < 1% for AT); significant numbers of TCR-gamma delta+ cells were detected (2-11% for FT and 15-33% for AT) after the coculture with FHC-4D2 and rIL-2. Although FT-derived TCR-gamma delta+ cells predominantly used the V gamma 5 chain, the major population in AT-derived TCR-gamma delta+ cells used V gamma 1, V gamma 4, or V gamma 7 chains. Both FT- and AT-derived TCR-gamma delta+ cells killed FcR-bearing target cells when incubated with anti-TCR-gamma delta Ab. Half of FT-derived TCR-gamma delta+ cells were CD4-CD8 alpha+8 beta-; the rest were CD4-CD8 alpha-8 beta-. AT-derived TCR-gamma delta+ cells expressed neither CD4 nor CD8 molecules. Separation of thymocytes from FHC-4D2 cells with a membrane filter reduced the proliferative response by two- to threefold. Taken together, these results demonstrate that a fetal hepatocyte clone supports thymocytes to develop preferentially into TCR-gamma delta+ cells in cooperation with rIL-2 through cell-cell contact, that the repertoire and the phenotype of induced TCR-gamma delta+ cells are determined by the age of the mice, and that hepatocytes might thus play an active role in T lymphopoiesis in the fetal liver.

Age Factors↗
PubMed1995

Diverse incidences of individual oligopeptides (dipeptidic to hexapeptidic) in proteins of human, bakers' yeast, and Escherichia coli origin registered in the Swiss-Prot data base.

Oligopeptidic permutations of the 20 amino acid residues give rise to proteins of diverse functions. Our long-term goal is to produce a lexicon of oligopeptides, classifying them into at least five categories: (i) ubiquitous, (ii) function specific, (iii) group specific, (iv) species specific, and (v) nonexistent. To begin with, we report on the varying frequencies of individual oligopeptides (dipeptidic to hexapeptidic in length) found among 2862 human proteins, 1942 Saccharomyces cerevisiae proteins, and 2672 Escherichia coli proteins registered in the Swiss-Prot data base (version 29.0, released in June 1994). At all lengths (dipeptides to hexapeptides), homooligopeptides were very prominent among the most frequently occurring varieties in proteins of human and bakers' yeast origins. However, this was not the case with E. coli. While all of the expected 20(3) varieties of tripeptides were found among human proteins, three tripeptides (Cys-Cys-Trp, Trp-Trp-Cys, and Trp-Trp-His) were missing from the bakers' yeast proteins. Three tripeptides (Cys-Ile-Trp, Cys-Met-Tyr, and Cys-Trp-Trp) were also absent from E. coli proteins. Inasmuch as the Swiss-Prot data base already contained 67% of the expected total of 4000 E. coli proteins, it is virtually certain that 96,000 varieties of hexapeptides containing at least one or another of the three missing tripeptides noted above shall be nonexistent in E. coli. Furthermore, the observation of missing tripeptides in the bakers' yeast proteins suggests that nonexistent hexapeptides shall be highly phylum specific. Because of the sample size, only a small fraction of the 20(6) varieties of hexapeptides were expected to be encountered in the present survey. Indeed, only 1.2-1.5% of the possible hexapeptides were found, and the average copy number of observed hexapeptides varied between 1.06 and 1.25. Nevertheless, 33 varieties of hexapeptides occurred in 102-169 copies among human proteins. Furthermore, 15 of the 33 varieties contained such rarely used residues as Tyr, His, Cys, and Trp.

Amino Acid Sequence↗
PubMed1995

Activation of thymic B cells by signals of CD40 molecules plus interleukin-10.

We have previously found that thymic B cells, particularly thymic CD5+ B cells, show low responsiveness to the usual B cell stimulants such as lipopolysaccharide or anti-IgM plus interleukin (IL)-4, although they proliferate and produce antibodies after direct interaction with major histocompatibility complex class II-restricted T blasts. These findings raise the possibility that a CD40-CD40 ligand (L) interaction is involved in the activation of thymic B cells. In the present study, we therefore examine this possibility using CD40L-transfected Chinese hamster ovary (CHO) cells or anti-CD40 monoclonal antibody (mAb). When B cells in the spleen and peritoneal cavity were stimulated, they proliferated and produced immunoglobulin (Ig) in the presence of CD40L-CHO cells or anti-CD40 mAb alone. However, another signal delivered by IL-10 in addition to CD40L-CHO cells or anti-CD40 mAb was found to be necessary for thymic B cells to proliferate and secrete Ig. Other interleukins acting on B cells, such as IL-4, IL-5, and IL-6, had no effect on the activation of thymic B cells, which thus have unique characteristics not found in peripheral B cells. This report discusses the physiological significance of IL-10- and CD40-driven signals in the activation of thymic B cells.

Animals↗
PubMed1995

Abnormalities of B cells and dendritic cells in SAMP1 mice.

The age-related changes in the function of antigen-presenting cells (APC) were examined using a substrain of senescence-accelerated mouse (SAMP1). In the primary mixed lymphocyte reaction (MLR), dendritic cells (DC) from aged SAMP1 mice showed less stimulatory activity than those of age-matched BALB/c or young SAMP1 mice. In the secondary MLR, the stimulatory activity of B cells was found to be lower in aged SAMP1 mice but not in age-matched BALB/c or young SAMP1 mice. In addition, these age-related decreases in the stimulatory activity of APC were found to be related to changes in the surface density of major histocompatibility complex class II and intercellular adhesion molecule-1 (ICAM-1) (but not B7-1 or B7-2 molecule) on APC (DC and B cells).

Aging↗
PubMed1995

Mechanical properties of the binary titanium-zirconium alloys and their potential for biomedical materials.

Mechanical properties of titanium-zirconium binary alloys were investigated in order to reveal their possible use for new biomedical materials and to collect useful data for alloy design through a hardness test, a tensile test, and optical microscopy. The hardness of the alloy containing 50% zirconium was approximately 2.5 times as large as the hardness of pure titanium and pure zirconium. Tensile tests showed a similar tendency. No changes between hardness of as cast specimens and as homogenized specimens were observed, nor were changes in microstructures noted. Comparisons between the Ti-6Al-4V alloy and the Ti-Zr-6Al-4V alloy indicated that a titanium-zirconium alloy could provide a base material for a new biomedical alloy. From these results, it was concluded that new alloys for biomedical materials should be designed as titanium-zirconium base alloys.

Alloys↗
PubMed1995

The amino acid sequence required for 5' --> 3' exonuclease activity of Bacillus caldotenax DNA polymerase.

We studied the 5' --> 3' exonuclease activity of Bacillus caldotenax DNA polymerase by site-directed mutagenesis. Among seven mutants constructed, two mutant DNA polymerases with an amino acid substitution of Gly184 --> Asp or Gly192 --> Asp were confirmed to be deficient in this exonuclease. The two positions corresponded to those of the Escherichia coli DNA polymerase I mutants defective in 5' --> 3' exonuclease, polA480ex and polA214. These results provide experimental support for the proposed amino acid sequence essential for the 5' --> 3' exonuclease activity associated with eubacterial polymerase I-like DNA polymerases (family A), including E.coli and Thermus aquaticus.

Bacillus↗
PubMed1995

Utility of Tc-99m GSA SPECT imaging in estimation of functional volume of liver segments in health and liver diseases.

The authors examined whether there was a difference in liver function among hepatic segments in liver cirrhosis cases, and in cases of hepatocellular carcinoma (HCC) associated with liver cirrhosis. If the average counts in the lateral segment of the left lobe were set at 1, the average counts in the right upper and lower segment of the liver were 0.75 approximately 1.02 (0.89 +/- 0.09, mean +/- SD) in normal cases, 0.38 approximately 2.24 (1.01 +/- 0.39) in liver cirrhosis cases, and 0.61 approximately 2.85 (1.15 +/- 0.58) in HCC cases. There is a significant difference between normal cases and liver cirrhosis cases or HCC cases (P < 0.001). Also, in HCC cases, if the average counts in the cancer-bearing segment of the liver were set at 1, the average counts in the noncancerous segment of the liver were 0.55 approximately 2.85 (1.23 +/- 0.58), and many average counts in the cancer-bearing segment were equal to, or lower than those in the noncancerous segment. It has been found that there were significant differences in function among hepatic segments in liver cirrhosis cases, and in HCC cases. Furthermore, the liver function in the cancer-bearing segment tended to be worse due to the existence of carcinoma compared with that in the noncancerous segment.

Adult↗
PubMed1995

Prediction of pulmonary function after resection of primary lung cancer. Utility of inhalation-perfusion SPECT imaging.

To help determine whether preoperative perfusion and inhalation SPECT imagings are useful in predicting postoperative lung function, Tc-99m MAA perfusion SPECT imaging, CT scans, and pulmonary function tests were prospectively performed in 33 patients with primary lung cancer before and after lobectomy or pneumonectomy. Tc-99m Technegas inhalation SPECT imaging was performed in 6 of 33 patients as well. The authors also studied changes in radioactivity on the operated and nonoperated sides before and after surgery, examined the lowest limit value for adaptability to the operation, and made a comparison of both perfusion and inhalation SPECT imaging. The predicted postoperative values obtained from the preoperative Tc-99m MAA SPECT images correlated more closely with the measured 6-month postoperative values than with the measured 3-month postoperative values. The highest correlation coefficient (r = 0.86) was observed between the predicted forced vital capacity (FVC) value and the measured 6-month postoperative FVC value. In many cases, there was not a great difference between the 6-month and 3-month radioactivity on the operated side obtained from Tc-99m MAA SPECT images. This appears to indicate that pulmonary blood flow on the operated side has completely recovered by 3 months after surgery. However, radioactivity in both the upper and lower lobes of the nonoperated side increased soon after surgery compared with that before the operation, and had not returned to preoperative levels 6 months after surgery. The radioactivity in the right middle lobe did not change before and after surgery.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗
PubMed1995

Frequent detection of hepatitis C virus subtype 3a (HCV-3a) isolates in Thailand by PCR using subtype-specific primers.

By means of a polymerase chain reaction (PCR) method using subtype-specific primers for hepatitis C virus (HCV) subtypes 1a, 1b, 2a, 2b and 3a, the prevalence of each subtype among HCV isolates in Chiang Mai, Thailand, was determined. HCV-3a appeared to be the most common subtype in blood donors, and was also frequently found in patients with liver disease. HCV-1b, but not HCV-2a or -2b, was also commonly found in this area, while a considerable percentage of the total HCV isolates still remained unclassifiable by the above methods. Serotype analysis of the HCV isolates using C14-1 and C14-2 recombinant peptides revealed that HCV-3a was likely to carry an antigenic determinant(s) different from those of the major types 1 (HCV-1a and -1b) and 2 (HCV-2a and -2b).

Adult↗
PubMed1995

The hyperthermophilic archaeon Pyrodictium occultum has two alpha-like DNA polymerases.

We cloned two genes encoding DNA polymerases from the hyperthermophilic archaeon Pyrodictium occultum. The deduced primary structures of the two gene products have several amino acid sequences which are conserved in the alpha-like (family B) DNA polymerases. Both genes were expressed in Escherichia coli, and highly purified gene products, DNA polymerases I and II (pol I and pol II), were biochemically characterized. Both DNA polymerase activities were heat stable, but only pol II was sensitive to aphidicolin. Both pol I and pol II have associated 5'-->3' and 3'-->5' exonuclease activities. In addition, these DNA polymerases have higher affinity to single-primed single-stranded DNA than to activated DNA; even their primer extension abilities by themselves were very weak. A comparison of the complete amino acid sequences of pol I and pol II with two alpha-like DNA polymerases from yeast cells showed that both pol I and pol II were more similar to yeast DNA polymerase III (ypol III) than to yeast DNA polymerase II (ypol II), in particular in the regions from exo II to exo III and from motif A to motif C. However, comparisons region by region of each polymerase showed that pol I was similar to ypol II and pol II was similar to ypol III from motif C to the C terminus. In contrast, pol I and pol II were similar to ypol III and ypol II, respectively, in the region from exo III to motif A. These findings suggest that both enzymes from P. occultum play a role in the replication of the genomic DNA of this organism and, furthermore, that the study of DNA replication in this thermophilic archaeon may lead to an understanding of the prototypical mechanism of eukaryotic DNA replication.

Amino Acid Sequence↗
PubMed1995

Association between maternal antibodies to the external envelope glycoprotein and vertical transmission of human T-lymphotropic virus type I. Maternal anti-env antibodies correlate with protection in non-breast-fed children.

Vertical transmission of human T-lymphotropic virus type I (HTLV-I) depends primarily on breast-feeding; substitution of bottle-feeding has reduced the transmission rate from 20% in breast-fed children to 3% among bottle-fed. To determine the correlates of transmission for long breast-feeding (> or = 6 mo), short breast-feeding (< 6 mo), and bottle-feeding mothers, the antibody titers of transmitter (T) mothers and non-transmitter (nT) mothers were analyzed by using synthetic and recombinant epitopes representing the immunodominant epitopes of gag (Gag1a, r24), env (Env1/5, MTA1, RE3), and tax (Tax8/22-24) proteins. Seroreactivity to gag and tax epitopes was not significantly different except for anti-r24 antibody titer, which was significantly higher among T-mothers (geometric mean 134) when compared with nT-mothers (62) in the long-feeding group (P < 0.001). Profiles of antibody titers against env epitopes were different. Within the long-feeding group, Env1/5, MTA1, and RE3 titers were significantly higher among T-mothers (258, 1,476, and 738, respectively) when compared with nT-mothers (106, 279, and 320, respectively) (P < 0.01 for all three epitopes). In contrast, within the bottle-feeding group, antibody titers to Env1/5 (269) and RE3 (418) among nT-mothers were significantly higher than those among T-mothers (80 and 113, respectively) (P < 0.01). These data confirm that high-titered anti-HTLV-I antibodies in the long-feeding group correlate with milk-borne transmission of HTLV-I and, more importantly, imply that maternal anti-env antibodies may reduce the risk of non-milkborne infection.

Amino Acid Sequence↗