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Biomedical subjects

K Takata

Publications and source records attributed to K Takata.

At least 163 records · Page 9Linked to original sources
PubMed1990

Novel forms of ferredoxin and ferredoxin-NADP reductase from spinach roots.

Ferredoxin and the enzyme catalyzing its reduction by NADPH, ferredoxin-NADP reductase (ferredoxin-NADP+ oxidoreductase or FNR), were found to be present in roots of spinach (Spinacia oleracea). Localization experiments with endosperm of germinating castor beans (Ricinus communis), a classical nonphotosynthetic tissue for cell fractionation studies, confirmed that ferredoxin and FNR are localized in the plastid fraction. Both proteins were purified from spinach roots and found to resemble their leaf counterparts in activity, spectral properties, and complex formation, but to differ in amino acid composition and amino terminal sequence. The results indicate that the primary structures of the FNR and ferredoxin of spinach roots differ from that of the corresponding leaf proteins. Together with earlier findings, the present results provide evidence that nonphotosynthetic plastids, including those of roots, are capable of reducing ferredoxin with heterotrophically generated NADPH.

Amino Acid Sequence↗
PubMed1990

Increased high-density lipoprotein levels caused by a common cholesteryl-ester transfer protein gene mutation.

BACKGROUND AND METHODS: The plasma cholesteryl-ester transfer protein (CETP) catalyzes the transfer of cholesteryl esters from high-density lipoprotein (HDL) to other lipoproteins. We recently described a Japanese family with increased HDL levels and CETP deficiency due to a splicing defect of the CETP gene. To assess the frequency and phenotype of this condition, we screened 11 additional families with high HDL levels by means of a radioimmunoassay for CETP and DNA analysis. RESULTS: We found the same CETP gene mutation in four families from three different regions of Japan. Analysis of restriction-fragment-length polymorphisms of the mutant CETP allele showed that all probands were homozygous for the identical haplotype. Family members homozygous for CETP deficiency (n = 10) had moderate hypercholesterolemia (mean total cholesterol level [+/- SD], 7.01 +/- 0.83 mmol per liter), markedly increased levels of HDL cholesterol (4.24 +/- 1.01 mmol per liter) and apolipoprotein A-I, and decreased levels of low-density lipoprotein cholesterol (1.99 +/- 0.80 mmol per liter) and apolipoprotein B. Members heterozygous for the deficiency (n = 20), whose CETP levels were in the lower part of the normal range, had moderately increased levels of HDL cholesterol and apolipoprotein A-I and an increased ratio of HDL subclass 2 to HDL subclass 3, as compared with unaffected family members (1.5 +/- 0.8 vs. 0.7 +/- 0.4). CETP deficiency was not found in six unrelated subjects with elevated HDL cholesterol levels who were from different parts of the United States. CONCLUSIONS: CETP deficiency appears to be a frequent cause of increased HDL levels in the population of Japan, possibly because of a founder effect. The results that we observed in heterozygotes suggest that CETP normally plays a part in the regulation of levels of HDL subclass 2. There was no evidence of premature atherosclerosis in the families with CETP deficiency. In fact, the lipoprotein profile of persons with CETP deficiency is potentially antiatherogenic and may be associated with an increased life span.

Adolescent↗
PubMed1990

[Clinical studies on 29 cases of testicular tumors].

Clinical studies were performed on 29 cases of testicular tumors over the past 16 years. There were 14 cases of seminoma (S), 4 cases of E+Teratoma (T), 1 case of embryonal carcinoma (E), 1 case of E+STGC, 1 of S+E+T, 1 of S+E+ yolk sac tumor (Y) +choriocarcinoma (C) +STGC, 1 of mature teratoma, 6 of non-germ cell tumor. The stage of the germ cell tumors were stage I in 13 cases and stage II A in one case among 14 cases of seminoma, and stage I in 3 cases, stage II A in 3 cases, stage II B in one case, stage III in one case among 8 cases of non-seminoma. All of the patients underwent high orchiectomy. The seminomas were treated by radiotherapy and non-seminoma cases mainly by chemotherapy after retroperitoneal lymph nodes dissection (RPLND). Twenty one of 23 cases have survived, excluding one death (E+STGC, stage II A) and one unclear case. High orchiectomy and radiotherapy are the established procedures for the treatment of stage I seminoma. The significance of RPLND should decrease because of the high efficacy of current chemotherapy in cases of stage I and II A non-seminoma.

Adolescent↗
PubMed1990

Identification of a novel alpha-amylase by expression of a newly cloned human amy3 cDNA in yeast.

A novel amylase gene (amy3) that differs in nucleotide sequence from salivary amylase gene (amy1) and pancreatic amylase gene (amy2) has been described [Tomita et al., Gene 76 (1989) 11-18], but whether this gene can ever code for an active enzyme has not been shown. We prepared cDNA of this gene from an mRNA obtained from lung carcinoid tissue, and expressed it in Saccharomyces cerevisiae under the control of an acid phosphatase promoter. The product was secreted into culture media, and showed enzymatic activity, demonstrating that this novel alpha-amylase gene (amy3) can code for a functional isozyme. We purified this enzyme, and compared its biological properties with those of salivary and pancreatic human amylases similarly expressed in yeast. We observed that the novel amylase isozyme is more heat-sensitive than others, and that its substrate specificity is different from the other two isozymes.

Acid Phosphatase↗
PubMed1990

Short- and long-term effects of cigarette smoking on heart rate variability.

The short- and long-term effects of cigarette smoking on autonomic cardiac regulation were investigated by power spectral analysis of heart rate variability under controlled respiration (15/min). The short-term effects were examined in 9 smokers without evidence of cardiopulmonary disorders after an overnight abstinence from smoking. The heart rate spectral component reflecting the respiratory sinus arrhythmia (0.25 Hz), a quantitative index of vagal cardiac control, decreased 3 minutes after smoking 1 cigarette (p = 0.0061) and the component reflecting Mayer wave sinus arrhythmia (0.04 to 0.15 Hz), which includes sympathetically mediated activity, increased after 10 to 17 minutes (p = 0.0124). The long-term effects were examined in 81 normal subjects comprising 25 nonsmokers, 31 moderate (1 to 24 cigarettes/day) smokers and 25 heavy (greater than 25 cigarettes/day) smokers after an overnight abstinence. Although the magnitude of the Mayer wave component was unaffected by the smoking status, the respiratory component in the supine position was smaller in the young (less than or equal to 30 years) heavy smokers than in the young nonsmokers or moderate smokers (p = 0.0078). Also, postural changes in the components, a decrease in the respiratory component and an increase in the Mayer wave component with standing, were observed in the nonsmokers but not in the heavy smokers. These results suggest that smoking causes an acute and transient decrease in vagal cardiac control, and that heavy smoking causes long-term reduction in vagal cardiac control in young people and blunted postural responses in autonomic cardiac regulation.

Adult↗
PubMed1990

Localization of Forssman glycolipid and GM1 ganglioside intracellularly and on the surface of germ cells during fetal testicular and ovarian development of mice.

Changes in the expression pattern and intracellular localization of Forssman glycolipid (FA) and GM1 ganglioside (GM1) in fetal mouse gonads were examined during germ cell differentiation by immunofluorescence microscopy and immunoelectron microscopy. In male germ cells from the 12th to 14th day p.c., anti-FA binding was localized in granular structures aggregated on one side of the cytoplasm and/or in the plasma membrane. On day 16 p.c., some germ cells still showed patch-like positive reactions in the plasma membrane, but by day 18 p.c., positive reactions for FA had completely disappeared. The female germ cells showed granular bindings of anti-FA scattered throughout their cytoplasm during the 13th to 16th day p.c., although the positive reactions in female germ cells on day 12 p.c. tended to be found in one side of cytoplasm and/or plasma membrane similar to those in male germ cells from 12th to 14th day p.c. On day 18 p.c., positive reactions remained in the plasma membrane of some germ cells, but these positive reactions disappeared before birth. Immunoelectron microscopic observation showed that the sites of anti-FA bindings were equivalent to the "small dense bodies" (SDB) and the Golgi lamellae both in male and female germ cells. On the other hand, GM1 was not detected in male germ cells at any time during fetal testicular development, whereas an anti-GM1 reaction was detected in the plasma membrane of female germ cells from the 16th to 18th day p.c. (oocytes in the first meiotic prophase).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

A highly sensitive enzyme-linked immunosorbent assay for etoposide using beta-D-galactosidase as a label.

A highly sensitive enzyme-linked immunosorbent assay (ELISA) for etoposide (EP) was developed, which is capable of accurately measuring as little as 40 pg EP/ml. Anti-EP sera were obtained by immunizing rabbits with EP conjugated with mercaptosuccinyl bovine serum albumin (MS.BSA) using N-[beta-(4-diazophenyl)ethyl]maleimide (DPEM) as a heterobifunctional coupling agent. An enzyme marker was similarly prepared by coupling EP with beta-D-galactosidase (beta-Gal; EC 3.2.23) via DPEM. This ELISA was specific for EP and showed a very slight cross-reactivity with its major metabolite, cis-hydroxy acid of EP (0.91%), but none with 4'-demethylepipodophyllotoxin and drugs commonly used with EP in combination chemotherapy for cancer treatment. The values for EP concentration detected by this assay were comparable with those detected by the high-performance liquid chromatography (HPLC) method. However, the ELISA was about 1,250 times more sensitive in detecting EP at lower concentrations. Using this assay, drug levels were easily determined in the blood and urine of rats for 7 h after i.v. administration of EP at a single dose of 3 mg/kg. Due to its sensitivity and specificity for EP, the ELISA should prove to be a valuable new tool for use in clinical pharmacological studies.

Animals↗
PubMed1990

Microquantification of cholesterol and cholesteryl esters in rat peritoneal macrophages by reverse-phase high-performance liquid chromatography.

A simple and rapid method for the microquantification of cholesterol and cholesteryl esters by reverse-phase high performance liquid chromatography has been established. Comparison of elution patterns of authentic cholesterol and cholesteryl esters revealed that a mu Bondasphere reverse-phase C8 (300-A) column was more suitable than a corresponding reverse-phase C4 or C18 column in terms of rapidity and sensitivity. Recovery of cholesterol and cholesteryl esters from a C8 column was greater than 98% when determined either by radioactive cholesterol and cholesteryl oleate or by cholesteryl heptadecanoate. The sensitivity of the quantification ranged from 5 ng to 50 micrograms for both cholesterol and cholesteryl esters. This method was applied to determination of cellular cholesterol and cholesteryl esters of rat peritoneal macrophages. Lipid extracts of these cells were found to contain 38.01 +/- 2.60 micrograms of cholesterol and 3.18 +/- 0.36 micrograms of cholesteryl esters per milligram of cell protein. When the cells were loaded with cholesteryl esters by incubation for 24 h with various concentrations of acetylated low-density lipoprotein, a cellular level of cholesteryl esters showed a dose-dependent increase and reached a maximal level of 106.60 +/- 3.05 micrograms/mg cell protein. Thus, the present method is useful for the microquantification of cholesterol and cholesteryl esters from lipid extracts of biological samples.

Animals↗
PubMed1990

Long-term results of anterior interbody fusion for treatment of degenerative spondylolisthesis.

Thirty-nine patients, 34 women and five men, underwent anterior decompression and interbody fusion for degenerative spondylolisthesis between February 1958 and August 1988. Their average age at surgery was 51 years (range, 34-74 years), and their average follow-up period was 12 years 7 months (range, 6 months to 30 years). Clinical evaluation was done by the score rating system of the Japanese Orthopaedic Association (JOA Score). Patients with JOA scores of 25 points or more were rated as "satisfactory." Survivorship was analyzed by the method of Kaplan and Meier to determine the cumulative percentage of patients with satisfactory results. The following results were obtained: Seventy-six percent of the patients had satisfactory results for 10 years after the anterior interbody fusion, 60% for 20 years, and 52% for 30 years. Irrespective of their age at surgery, the patients generally maintained satisfactory results up to 65 years of age.

Female↗
PubMed1990

Purification and Characterization of a Ferredoxin-NADP Oxidoreductase-Like Enzyme from Radish Root Tissues.

An enzyme able to reduce cytochrome c via ferredoxin in the presence of NADPH, was isolated, purified from radish (Raphanus sativus var acanthiformis cultivar miyashige) roots and characterized. The enzyme was purified by DEAE-cellulose, Blue-Cellulofine, Ferredoxin-Sepharose 4B, and Sephadex G-100 column chromatography. Molecular mass of the enzyme was estimated to be 33,000 and 35,000 daltons by Sephadex G-100 gel filtration and SDS-PAGE, respectively. Its absorption spectrum suggested that the enzyme contains flavin as a prosthetic group. The K(m) values for NADPH and ferredoxin were calculated to be 9.2 and 1.2 micromolar, respectively. The enzyme required NADPH and did not use NADH as an electron donor. The optimal pH was 8.4. The enzyme also catalyzed the photoreduction of NADP(+) in the spinach leaf thylakoid membranes depleted of ferredoxin and ferredoxin-NADP(+) oxidoreductase. The effect of NaCl and MgCl(2) concentration on the activity and amino acid composition of the enzyme were demonstrated. The results suggest that the enzyme is similar to ferredoxin-NADP(+) oxidoreductase from chloroplasts and cyanobacteria and is the key enzyme catalyzing the electron transport between NADPH, generated by the pentose phosphate pathway, and ferredoxin in plastids of plant heterotrophic tissues.

Journal Article↗
PubMed1990

Diurnal variations in vagal and sympathetic cardiac control.

To investigate the diurnal variation in autonomic cardiac control, the magnitudes of the power spectral components of supine and standing heart rate variability were measured during controlled respiration (15 breaths/min). Examination was performed hourly between 0700 and 2300 h in eight male subjects whose activities and food intake were controlled for 24 h in the laboratory. The respiratory component (0.25 Hz) was greater in the morning than in the late afternoon (P less than 0.05) and decreased 30 min after food intake (P less than 0.01) in the supine position, but it was unaffected by the time of day or food intake while in the standing position. The Mayer wave component (0.03-0.15 Hz) did not change with the time of day, but it increased 90 min after food intake in both supine and standing positions (P less than 0.01 and P less than 0.05, respectively). These data suggest that supine vagal cardiac control during the waking period increases in the morning and decreases 30 min after food intake and that sympathetic cardiac control increases 90 min after food intake.

Adult↗
PubMed1990

Decreased magnitude of heart rate spectral components in coronary artery disease. Its relation to angiographic severity.

We analyzed the spectral components of RR interval variability under controlled respiration (15 breaths/min) in 56 patients (age range, 35-73 years) referred for coronary angiography; 14 patients had multivessel disease (group M), 21 had one-vessel disease (group S), and 21 had nonsignificant disease or normal coronary artery (group N). There were 43 healthy controls (age range, 36-71 years) (group C). The patients had no clinical evidence of heart failure, hypertension, diabetes mellitus, or acute stage of infarction and had taken no medication for 3 days. The autoregressive power spectral density of RR interval variability contains two major components, respiratory sinus arrhythmia (RSA) (0.25 Hz) and Mayer wave-like sinus arrhythmia (MWSA) (0.04-0.15 Hz), which have magnitudes that are quantitative markers of cardiac vagal activity and sympathetic activity with vagal modulation, respectively. We represented the magnitudes by the coefficient of component variance (CCV), which provided the amplitude relative to the mean RR interval. The age- and sex-adjusted mean of CCVRSA significantly decreased with advancing angiographic severity (1.64 +/- 0.09%, 1.66 +/- 0.12%, 1.22 +/- 0.13%, and 0.81 +/- 0.16% for groups C, N, S, and M, respectively) (p = 0.0001). The CCVRSA was unrelated to left ventricular function, previous myocardial infarction, or stenosis of any specific artery including the sinoatrial and atrioventricular node arteries. The CCVMWSA decreased only in group M (p = 0.0462). These results indicate that coronary artery disease is associated with vagal dominant impairment in autonomic cardiac function and that reduction in the vagal cardiac function correlates with the angiographic severity.

Adult↗
PubMed1990

Cytotoxic activity of polyacetylene compounds in Panax ginseng C. A. Meyer.

The effects of the three polyacetylene compounds, panaxynol, panaxydol and panaxytriol, on in vitro-cell growth were studied. These compounds are much different in their water-solubility. In order to increase water-solubility, solid complexes of polyacetylene compounds with alpha-cyclodextrin (CD) were prepared. Accurate concentrations of the active compounds in a culture medium were determined by gas chromatography. All of these CD-complexes inhibited the growth of various kinds of cultured cell lines in a dose-dependent fashion. The cell growth inhibitory activity of these complexes was much stronger against malignant cells than against normal cells. A continuous contact between the compounds and target cells was not necessarily required for growth inhibition. And the inhibition was cytotoxic at high concentrations and cytostatic at low concentrations. These findings indicate that these polyacetylene compounds' mode of action is more dose-dependent than time-dependent. The panaxynol, panaxydol and panaxytriol contents in red ginseng powder were 250, 297 and 320 micrograms/g, respectively.

Alkynes↗
PubMed1990

A splice-junction mutation responsible for familial apolipoprotein A-II deficiency.

The first case of familial apolipoprotein A-II (apo A-II) deficiency was recently reported from Hiroshima, Japan, and designated apo A-IIHiroshima. The proband had no immunologically detectable apo A-II in her plasma. DNA sequence analysis showed that the proband was homozygous for a G----A transition at position 1 of intron 3 of the apo A-II gene. A sister of the proband, who had an intermediate level of plasma apo AII, was shown to be heterozygous for this base substitution. This splice-junction alteration is most likely responsible for apo A-II deficiency, since it would be expected to completely block splicing of intron 3 from the primary transcript and therefore prevent formation of functional mRNA. This deficiency seems to have little influence either on lipid and lipoprotein profiles or on the occurrence of coronary artery disease.

Apolipoprotein A-II↗
PubMed1990

[Cell growth inhibitory substance isolated from Panax ginseng root: panaxytriol].

A new type of cell growth inhibitory substance was isolated and purified from a powder of the root of Panax ginseng C.A. Meyer, which has been commonly used for the treatment of various disease as a commercial medical drug by the name of Korean Red Ginseng Powder. Data from the infrared spectra, proton and carbon-13 nuclear magnetic resonance, and high resolution mass spectra were identical with those of authentic panaxytriol. Cell growth inhibition by panaxytriol seemed to be tumor-specific. Its action was supposed to be more dose-dependent than time-dependent. In vivo antitumor efficacy of panaxytriol was tested against C57BL/6 mice transplanted with B 16 melanomas. Panaxytriol (40 mg/kg) administered intramuscularly produced significant tumor growth delays in mice (p less than 0.01).

Alkynes↗
PubMed1990

[Elective prosthetic valve reoperation to relieve unacceptably high gradient].

Pressure gradients of Lillehei-Kaster valve (13 patients), Omniscience valve (26 patients) and Omnicarbon valve (7 patients) in the aortic position were evaluated by Doppler echocardiography. Twenty-three out of 46 patients revealed unacceptably high gradients. Seven prostheses were removed because the valve design that they had high gradients across them. Clinical findings including left ventricular function and symptoms improved after the reoperation. Patients who underwent aortic valve replacement with this series of valve prostheses should be followed up cautiously.

Aortic Valve↗
PubMed1989

The role of the carboxyl-terminal 6 amino acid extension of human TSH beta subunit.

The nucleotide sequence of human thyroid stimulating hormone (hTSH) gene can encode a protein of 138 amino acids. However, the mature polypeptide is lacking 6 amino acids of the carboxyl-terminus (C-terminus), suggesting posttranslational cleavage of these residues. To analyze a possible function of these 6 amino acids, we expressed two hTSH beta cDNAs with or without the 6 codons for C-terminal extension, together with alpha subunit cDNA in CHO cells, and determined the amino acid sequence of C-terminus of hTSH beta. hTSH beta propeptides without C-terminal extension were glycosylated, associated with alpha subunit and secreted, as normal propeptides were, and its heterodimer with alpha subunit showed normal TSH bioactivity in FRTL-5 bioassay. These data indicate that the 6 amino acid C-terminal extension is not necessary for the hTSH maturation in the process of the biosynthesis and for its bioactivity.

Amino Acid Sequence↗