A serologic analysis and the amino acid sequence of the V3 region of human immunodeficiency virus from carriers in Bangkok.
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Amino Acid Sequence↗
Biomedical subjects
K Okuda
Publications and source records attributed to K Okuda.
Rat hepatic 3 alpha-hydroxysteroid dehydrogenase: expression of cDNA and physiological function in bile acid biosynthetic pathway.
3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSD) [EC 1.1.1.213]2 plays important multifunctional roles in metabolizing steroid hormones, polycyclic aromatic hydrocarbons, and prostaglandins and also in transforming the steroid nucleus for the biosynthesis of bile acids from cholesterol in liver. To gain insight into the details and physiological functions of 3 alpha-HSD in the bile acid biosynthetic pathway, cDNA clones of 3 alpha-HSD were isolated from rat liver lambda phage cDNA libraries by using specific antibodies to 3 alpha-HSD purified from rat liver. Transfection of the 3 alpha-HSD cDNA in Simian COS7 cells resulted in the expression of an immunoreactive protein to the antibodies against the purified enzyme, and the transfected cells exhibited activities for not only 7 alpha-hydroxy-5 beta-cholestan-3-one, the intermediate of bile acid biosynthesis, but also steroid hormones and 9,10-phenanthrenequinone. Northern blot analysis on poly(A)+ RNA by selective use of different cDNA fragments of the 5'-untranslated region, the coding region, and the 3'-untranslated region as probes revealed three hybridizable bands, 3.6, 2.7, and 2.5 kb, in liver and four bands, 3.6, 2.7, 2.5, and 1.8 kb, in ovary. Of these, the 2.7- and 1.8-kb bands were predominant in liver and ovary, respectively. Northern hybridization analysis also revealed that the coding region of the various sizes of mRNA seemed to be common. Southern blot analysis of genomic DNA by the selective use of the cDNA fragments as probes indicated that the various mRNA species were derived from a single gene, probably due to an alternative splicing mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
3-Hydroxysteroid Dehydrogenases↗
Penetration of porcine oocytes during maturation in vitro by cryopreserved, ejaculated spermatozoa.
The present study was conducted to examine the penetrability in vitro of immature porcine oocytes with or without cumulus cells. Porcine oocytes were cultured for 0-36 h, at 39 degrees C in 5% CO2 in air, in modified tissue culture medium 199 (TCM-199B at pH 7.4) supplemented with 10 IU eCG/ml, 10 IU hCG/ml, and 1 microgram estradiol-17 beta/ml. At various times after the beginning of culture, some oocytes were freed from the cumulus (cumulus and corona cells), and cumulus-intact or cumulus-free oocytes were inseminated with cryopreserved ejaculated spermatozoa in TCM-199B (pH 7.8) containing 5 mM caffeine. When cumulus-free oocytes were examined 14 h after insemination, high proportions (69-84%) were penetrated and there were no significant differences among different periods of maturation culture. The incidence (47-68%) of polyspermy and the number (1.7-3.1) of spermatozoa that penetrated per oocyte were also not significantly different among oocytes cultured for 0-36 h. In cumulus-intact oocytes, however, the first evidence of penetration (15%) was observed in oocytes cultured for 6 h. The penetration rates increased significantly as the period of culture was prolonged up to 24 h. Almost all (95-100%) oocytes were penetrated when they were inseminated 24-36 h after the beginning of culture, by which time the cumulus masses showed moderate to complete expansion except for the corona radiata. A similar correlation was also observed for incidence of polyspermy and number of spermatozoa penetrated per oocyte. The presence of well-expanded cumulus around the oocyte during fertilization promoted male pronuclear formation in penetrant oocytes.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
In vitro fertilization of bovine oocytes in a chemically defined, protein-free medium varying the bicarbonate concentration.
Bovine cumulus-enclosed oocytes were matured in culture, freed from cumulus cells, and inseminated with frozen-thawed spermatozoa in a chemically defined protein-free medium containing 5 mM caffeine and 10 micrograms/ml heparin. No penetration of oocytes was observed in the medium without polyvinyl-alcohol (PVA); but when the medium was supplemented with 0.1-5 mg/ml PVA, penetration rates (9-16%) significantly increased. Sperm motility was also stimulated during incubation for 2 h in the presence of PVA. In the medium with 1 mg/ml PVA, a high penetration rate (24 of 62 = 39%) was observed at a sperm concentration of 10 x 10(6) cells/ml. When the bicarbonate concentration was changed in the fertilization medium containing 1 mg/ml PVA and 10 x 10(6) spermatozoa/ml, a high penetration rate (47 of 67 = 70%) and a high proportion (44 of 47 = 94%) of oocytes in which male and female pronuclei had developed were obtained at 46 mM NaHCO3. However, the penetration rate (58-95%), the incidence of pronuclear formation (64-96%), and the proportion of polyspermy (9-21%) varied according to the animal (five different bulls). Spermatozoa obtained from two bulls started to penetrate oocytes 5 h after insemination in the presence of 46 mM NaHCO3. This is the first report indicating that induction of capacitation of bull spermatozoa and penetration of oocytes matured in culture are possible in a chemically defined, protein-free medium.
Animals↗
Hepatitis C virus antibodies, viral RNA and genotypes in sera from patients on maintenance haemodialysis.
Patients on maintenance haemodialysis in four dialysis centres were tested for markers of hepatitis C virus (HCV) infection. Antibody to HCV (anti-HCV) was detected by the second-generation enzyme immunoassay in 142 (26%) of the 543 patients and HCV RNA in 117 (22%) of whom four were without detectable anti-HCV in serum. Seventy-seven (66%) were infected with HCV of genotype II/1b, 31 (27%) with genotype III/2a and eight (7%) with genotype IV/2b, in a distribution similar to that in blood donors who carried HCV asymptomatically. Haemodialysis patients had high HCV RNA titres comparable to those of patients with chronic hepatitis C. HCV RNA was detected in 96 (26%) of the 365 patients with a history of transfusion more frequently than in 21 (12%) of the 178 without previous transfusion (P < 0.001). In transfused patients, frequencies of anti-HCV and HCV RNA increased in parallel with the duration of haemodialysis. The frequency of anti-HCV in non-transfused patients, however, did not change appreciably with the duration of haemodialysis up to 22 years. The patients with anti-HCV had a higher frequency of HCV RNA in serum than symptom-free blood donors with anti-HCV (113/142 or 80% vs 109/166 or 66%, P < 0.01) and the patients with HCV RNA had a lower frequency of elevated aminotransferase levels than blood donors with HCV RNA (5/113 or 4% vs 27/109 or 25%, P < 0.001). These results indicate that transfusion is a significant cause of HCV infection in patients on maintenance haemodialysis, and that these patients are prone to establish the HCV carrier state after infection.
Adult↗
A longitudinal study from prepuberty to puberty of gingivitis. Correlation between the occurrence of Prevotella intermedia and sex hormones.
A longitudinal study of 24 subjects progressing normally from prepuberty to puberty was undertaken to evaluate the effects of sex hormone levels on clinical and microbiologic parameters and on serum antibodies. During elementary school, at the beginning of the longitudinal monitoring, 2 groups, 12 subjects with gingivitis and 12 gingivitis-free subjects, were selected and observed through puberty. Bone ages and self-assessment of secondary sex characteristics were used to confirm puberty. A statistically significant increase in the proportions of Prevotella intermedia including Prevotella nigrescens and serum antibody levels against P. intermedia was seen in gingivitis group throughout the longitudinal study. Serum levels of testosterone in boys and estradiol and progesterone in girls was positively correlated with levels of P. intermedia and P. nigrescens. In puberty, a slight but significant increase in GI scores over prepuberty has been shown; however, there was no significant change in PlI from prepuberty to puberty. Our study confirmed that there was a statistically significant increase in gingival inflammation and in the proportion of P. intermedia and P. nigrescens in puberty relative to the baseline value, except in the gingivitis-free male group. Our findings suggest that these increases are correlated with elevation in systemic levels of the sex hormones.
Adolescent↗
Infection by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, and antibody responses at different ages in humans.
This study examined the serum IgG and IgM responses against Porphyromonas gingivalis and 3 serotypes of Actinobacillus actinomycetemcomitans, and the correlations of these responses with age and homologous infection. A total of 90 individuals were included in this study: 40 subjects with gingivitis, 40 periodontally healthy subjects, and 10 adult periodontitis subjects. The subjects in the gingivitis and periodontally healthy groups were divided into 4 stages based on their physiological age: early childhood, school age, puberty, and adult. In the gingivitis group, there was a positive correlation between increase in age and increase in serum IgG antibody levels against P. gingivalis until puberty. However, no statistically significant difference was found between the puberty stage and the adult stage. The average level of IgG antibodies against A. actinomycetemcomitans in the school age gingivitis group was significantly higher than that in the early childhood gingivitis group for all serotypes (p < 0.01). In serotype c, IgG antibody levels in the school age gingivitis group were significantly higher than in the early childhood gingivitis group or the adult gingivitis group (p < 0.01). With both P. gingivalis and A. actinomycetemcomitans, positive correlations between elevated IgG level and infections by these microorganisms were found in the puberty gingivitis and adult periodontitis groups.
Adolescent↗
Characterization of genes in the cellulose-synthesizing operon (acs operon) of Acetobacter xylinum: implications for cellulose crystallization.
The synthesis of an extracellular ribbon of cellulose in the bacterium Acetobacter xylinum takes place from linearly arranged, membrane-localized, cellulose-synthesizing and extrusion complexes that direct the coupled steps of polymerization and crystallization. To identify the different components involved in this process, we isolated an Acetobacter cellulose-synthesizing (acs) operon from this bacterium. Analysis of DNA sequence shows the presence of three genes in the acs operon, in which the first gene (acsAB) codes for a polypeptide with a molecular mass of 168 kDa, which was identified as the cellulose synthase. A single base change in the previously reported DNA sequence of this gene, resulting in a frameshift and synthesis of a larger protein, is described in the present paper, along with the sequences of the other two genes (acsC and acsD). The requirement of the acs operon genes for cellulose production was determined using site-determined TnphoA/Kanr GenBlock insertion mutants. Mutant analysis showed that while the acsAB and acsC genes were essential for cellulose production in vivo, the acsD mutant produced reduced amounts of two cellulose allomorphs (cellulose I and cellulose II), suggesting that the acsD gene is involved in cellulose crystallization. The role of the acs operon genes in determining the linear array of intramembranous particles, which are believed to be sites of cellulose synthesis, was investigated for the different mutants; however, this arrangement was observed only in cells that actively produced cellulose microfibrils, suggesting that it may be influenced by the crystallization of the nascent glucan chains.
Amino Acid Sequence↗
Growth and survival in small hepatocellular carcinoma.
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Carcinoma, Hepatocellular↗
Regulation of messenger ribonucleic acid expression of 1 alpha,25-dihydroxyvitamin D3-24-hydroxylase in rat osteoblasts.
We have reported that PTH inhibits 25-hydroxyvitamin D3-24-hydroxylase messenger RNA (mRNA) expression induced by 1 alpha,25-dihydroxyvitamin D3 [1 alpha, 25-(OH)2D3] in rat kidney but not intestine. In the present study, we examined whether the suppression of 24-hydroxylase mRNA expression by PTH occurs commonly in tissues and cells which have PTH receptors. Administration of 1 alpha, 25-(OH)2D3 into rats fed a synthetic vitamin D-repleted diet containing adequate calcium greatly increased serum levels of calcium and 1 alpha, 25-(OH)2D3. Also, there was a 4-fold increase in bone 24-hydroxylase activity in response to 1 alpha, 25-(OH)2D3 administration. In rats fed a low calcium diet, renal 24-hydroxylase activity was suppressed probably due to secondary hyperparathyroidism. In contrast, the low calcium feeding did not suppress bone 24-hydroxylase activity. The expression of 24-hydroxylase mRNA in rat osteoblastic C-26 and C-11 cells was similar and attained maximal levels 24 h after cells were incubated with 10(-8) M 1 alpha, 25-(OH)2D3. Induction of 24-hydroxylase mRNA expression by 1 alpha, 25-(OH)2D3 was much greater and earlier in immature C-26 cells than mature C-11 cells. Simultaneous addition of PTH, prostaglandin E2, or cAMP together with 1 alpha, 25-(OH)2D3 did not down-regulate mRNA expression of 24-hydroxylase induced by the vitamin in both C-26 and C-11 cells. Of the three osteoblastic cells (C-26, C-20, and C-11) examined, C-26 cells showed the least mRNA expression of vitamin D receptors, in spite of the highest expression of 24-hydroxylase mRNA. These results suggest that unlike in the kidney, bone 24-hydroxylase mRNA expression is not down-regulated by PTH despite of the presence of PTH receptors. They also suggest that the degree of the induction of 24-hydroxylase mRNA by 1 alpha, 25-(OH)2D3 is not explained simply by the vitamin D receptors content.
Adenylyl Cyclases↗
An investigation of diterpenes from the leaves of Rabdosia trichocarpa and their antibacterial activity against oral microorganisms.
The ethanolic extract of Rabdosia trichocarpa leaves showed antibacterial activity against cariogenic mutans streptococci and periodontopathic Porphyromonas gingivalis. Chromatographic separation and purification of the extract afforded ten diterpenes, including one novel ent-kauren type compound named trichoranin. Some of these compounds possess potent antibacterial activity against these oral micro-organisms, indicating that these diterpenes may be useful natural substances for the maintenance of oral health.
Anti-Bacterial Agents↗
Detection of exercise-induced silent ischemia and the sequence of ischemic events in coronary artery disease by radionuclide ambulatory ventricular function monitoring.
Twenty-seven patients with stable coronary artery disease were continuously monitored with an ambulatory radionuclide ventricular function monitor (VEST) during exercise to determine the prevalence of silent ischemia and the temporal sequence of events during ischemic episodes. Exercise-induced ejection fraction abnormality was considered a < 6% increase in the control value lasting for more than 60 sec. Patients performed exercise for 424 +/- 111 sec during VEST recording. Seventeen exercise-induced ejection fraction abnormalities were observed in 17 patients, of which eight (47%) were silent electrocardiographically and 12 (71%) were silent symptomatically. In all of the patients, exercise-induced ischemia occurred in a temporal sequence of ejection fraction abnormalities, ST depressions and then symptoms. In nine patients with ejection fraction and ST abnormalities, ejection fraction abnormalities occurred earlier (199 +/- 87 sec) than ST depressions (321 +/- 117 sec; p < 0.01). In five patients with symptoms (399 +/- 151 sec), the sequence was ejection fraction abnormalities (205 +/- 64 sec; P < 0.05) followed by ST depressions (266 +/- 101 sec; P < 0.05) and symptoms (399 +/- 151 sec; P < 0.01). After termination of exercise, ejection fraction (49 +/- 12 sec) recovered sooner than symptoms (102 +/- 27 sec; p < 0.01) or ST abnormalities (191 +/- 96 sec; p < 0.01). Thus, hemodynamic monitoring by VEST may be helpful in detecting ischemic episodes in coronary artery disease which remain electrocardiographically or symptomatically silent. Furthermore, exercise-stress induces a cascade of temporal changes in ischemic events which may be helpful in understanding the pathophysiology of ischemia.
Aged↗
[Group cardiac rehabilitation for patients with ischemic heart disease in recovery phase].
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Adult↗
Hereditary abnormality of luteinizing hormone resulting in discrepant serum concentrations determined by different assays.
We herein report familial defects in the molecular structure of luteinizing hormone (LH). The propositus was a 29-year-old women with repeated abortion in whom the serum LH concentration was extremely low determined by an immunoradiometric assay utilizing two monoclonal antibodies for the intact LH dimer and beta-subunit of LH (SPAC-S kit). Further studies on the serum LH concentration in this propositus by five different assay systems gave various results ranging from low to normal values. Bioassay of the propositus's LH using C57 black mice showed normal in biological activity. These data suggest that LH in the propositus is abnormal in terms of its molecular structure located in the bond region between alpha and beta subunits. A family study showed that the bioactivity of LH was normal in all family members. The serum LH was not detected in either the propositus or her brother with the SPAC-S kit even after the administration of LHRH, while the serum LH concentrations in the father, mother and sister were approximately half the normal range, indicating that the defect is hereditary and the mode of inheritance is autosomal dominant: the propositus and brother were homozygously affected, and the parents and sister heterozygously affected.
Abortion, Habitual↗
Short-term starvation increases calcidiol-24-hydroxylase activity and mRNA level in rat kidney.
The renal mitochondrial calcidiol-24-hydroxylase activity and the corresponding cytochrome P-450 mRNA level were measured in rats subjected to short-term starvation alone or in combination with calcitriol treatment. Short-term starvation of 24 and 48 h increased the mRNA level by five- and six-fold, respectively. The 24-hydroxylase activity increased by five- and threefold, respectively. Treatment with calcitriol markedly increased the enzyme activity about 20-fold and the mRNA level about six-fold. In rats subjected to calcitriol treatment combined with 24 h of starvation, a significant further increase in enzyme activity was observed. The mRNA levels increased but the difference was not significant statistically. The results indicate that the mechanism by which starvation stimulates the enzymes is different, at least in part, from that behind the stimulatory effect of calcitriol.
Animals↗
Development of rat one-cell embryos in a chemically defined medium: effects of glucose, phosphate and osmolarity.
Rat one-cell embryos recovered from naturally mated females were cultured in modified hamster embryo culture medium 1 without amino acids. In the presence of 0.4 mmol phosphate l-1 (NaH2PO4), no embryos developed beyond the two-cell stage, regardless of the presence of 5.0 mmol glucose l-1. This inhibition was dose dependent at very low concentrations of phosphate in the medium supplemented with 7.5 mmol glucose l-1 and osmolarity adjusted to 244 mosmol; development to the blastocyst stage was not inhibited at 0.001-0.01 mumol phosphate l-1, but development to the morula and four-cell stages was markedly inhibited at 0.1 and 1.0 mumol phosphate l-1. In the medium without phosphate, glucose did not inhibit or promote development to the morula stage, but adequate concentrations of glucose were necessary for the development of morulae to the blastocyst stage; the percentage of one-cell embryos that developed to the blastocyst stage at 7.5 mmol glucose l-1 (67%) and 10.0 mmol glucose l-1 (60%) were not statistically different from the percentage at 5.0 mmol glucose l-1 (46%), but was significantly greater than the percentage at 2.5 mmol l-1 (33%). When osmolarity of the medium with 5.0 mmol glucose l-1 was varied by adjusting the amount of NaCl added, more (82-98%) of the one-cell embryos developed to the four-cell stage at 212-276 mosmol, but development was greatly inhibited at 304 mosmol. Development to the blastocyst stage was largely dependent on osmolarities; at 244 mosmol, 61% of embryos developed to the blastocyst stage, although this percentage was not significantly different from the percentage (43%) at 264 mosmol.
Animals↗
Flow-cytometric identification and detection of Porphyromonas gingivalis by a LPS specific monoclonal antibody.
The purpose of this study was to identify Porphyromonas gingivalis (P. gingivalis) by flow cytometry (FCM) using a monoclonal antibody (MAb) OMR-Bg1E directed to P. gingivalis-specific lipopolysaccharide (LPS). The P. gingivalis strains ATCC 33277, 381, ESO75, W50, and A7A1 were selected for the study. Fusobacterium nucleatum (F. nucleatum), Prevotella intermedia (P. intermedia), Campylobacter rectus (C. rectus), Streptococcus sanguis (S. sanguis) and Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) served as controls. A suspension of 10(7) bacteria/ml of each bacteria was prepared and then reacted with a P. gingivalis specific MAb OMR-Bg1E and fluorescein isothiocyanate (FITC) labeled second antibody. These samples were analyzed by FCM. Bacterial specific binding aggregate on data was separated out by the forward- and side-angle-scatter characteristics, while non-specific binding (NSB) was eliminated by excluding the region with mouse IgG-positive and second antibody-positive area. FCM detected a mean range of 56.2% to 97.2% P. gingivalis strains. There was a 5.1% non-specific binding using FCM to non-P. gingivalis strains. When the P. gingivalis concentration was adjusted to 10(2), 10(4), and 10(6) bacteria/ml, a detection rate of 35.7%, 48.1%, and 91.4%, was respectively observed. The lower sensitivity of the flow cytometric assay was 10(2) bacteria/ml. When P. gingivalis was added to P. intermedia suspension at 1, 20, 40, 60, and 80%, the MAb-positive fraction yielded by FCM displayed a coefficient of determination of 0.967 with the actual percentage of P. gingivalis and could be regressed to a linear function.(ABSTRACT TRUNCATED AT 250 WORDS)
Analysis of Variance↗
Locally delivered minocycline and guided tissue regeneration to treat post-juvenile periodontitis. A case report.
A case of post-juvenile periodontitis in a 28-year-old female patient is described along with new periodontal treatment modalities. Administration of minocycline-HCl with local drug delivery system was introduced as a part of initial periodontal therapy following microbiological and immunological examinations. The lesions were subsequently treated by guided tissue regeneration, which resulted in considerable gain of attachment with minimal recession of marginal gingiva. This observation suggests that the local delivery of antibiotics and regenerative therapy may prove to be effective alternative modalities in treatment of post-juvenile periodontitis.
Administration, Topical↗