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Biomedical subjects

Y Takarada

Publications and source records attributed to Y Takarada.

At least 19 recordsLinked to original sources
PubMed2000

Applications of vascular occlusion diminish disuse atrophy of knee extensor muscles.

PURPOSE: We have previously shown that the combination of low-intensity resistive exercise and moderate vascular occlusion induces in humans a marked increase in growth hormone secretion and muscular hypertrophy. The present study investigated the effects of vascular occlusion on the size of thigh muscles in patients who underwent an operation for the reconstruction of the anterior cruciate ligament to see whether it attenuates the disuse muscular atrophy without any exercise combined. METHODS: Two sessions of occlusive stimulus, each consisting of five repetitions of vascular occlusion (mean maximal pressure, 238 mm Hg) for 5 min and the release of occlusion for 3 min, were applied daily to the proximal end of the thigh from 3rd to 14th days after the operation. Changes in the cross-sectional area (CSA) of thigh muscles were analyzed with magnetic resonance images taken on the 3rd and 14th day after the operation. RESULTS: Without occlusive stimulus (control), the CSAs of knee extensors and flexors decreased by 20.7 +/- 2.2% and 11.3 +/- 2.6% (mean +/- SEM, N = 8), whereas with the occlusive stimulus, they decreased by 9.4 +/- 1.6% and 9.2 +/- 2.6% (N = 8), respectively. The relative decrease in CSA of knee extensors was significantly (P < 0.05) larger in the control group than in the experimental group. CONCLUSION: The results indicate that the occlusive stimulus effectively diminishes the postoperation disuse atrophy of knee extensors.

Adult↗
PubMed2000

Rapid increase in plasma growth hormone after low-intensity resistance exercise with vascular occlusion.

Hormonal and inflammatory responses to low-intensity resistance exercise with vascular occlusion were studied. Subjects (n = 6) performed bilateral leg extension exercise in the seated position, with the proximal end of their thigh compressed at 214 +/- 7.7 (SE) mmHg throughout the session of exercise by means of a pressure tourniquet. Mean intensity and quantity of the exercise were 20% of 1 repetition maximum and 14 repetitions x 5 sets, respectively. In each set, the subjects repeated the movement until exhaustion. Plasma concentrations of growth hormone (GH), norepinephrine (NE), lacate (La), lipid peroxide (LP), interleukin-6 (IL-6), and activity of creatine phosphokinase (CPK) were measured before and after the exercise was finished and the tourniquet was released. Concentrations of GH, NE, and La consistently showed marked, transient increases after the exercise with occlusion, whereas they did not change a great deal after the exercise without occlusion (control) done at the same intensity and quantity. Notably, concentration of GH reached a level approximately 290 times as high as that of the resting level 15 min after the exercise. IL-6 concentration showed a much more gradual increase and was maintained at a slightly higher level than in the control even 24 h after exercise. Concentrations of LP and CPK showed no significant change. The results suggest that extremely light resistance exercise combined with occlusion greatly stimulates the secretion of GH through regional accumulation of metabolites without considerable tissue damage.

Adult↗
PubMed2000

Effects of resistance exercise combined with moderate vascular occlusion on muscular function in humans.

Acute and long-term effects of resistance exercise combined with vascular occlusion on muscular function were investigated. Changes in integrated electromyogram with respect to time (iEMG), vascular resistive index, and plasma lactate concentration were measured in five men either during or after elbow flexion exercises with the proximal end of the arm occluded at 0-100 mmHg. The mean iEMG, postexercise hyperemia, and plasma lactate concentration were all elevated with the increase in occlusion pressure at a low-intensity exercise, whereas they were unchanged with the increase in occlusion pressure at high-intensity exercise. To investigate the long-term effects of low-intensity exercise with occlusion, older women (n = 24) were subjected to a 16-wk exercise training for elbow flexor muscles, in which low-intensity [ approximately 50-30% one repetition maximum (1 RM)] exercise with occlusion at approximately 110 mmHg (LIO), low-intensity exercise without occlusion (LI), and high- to medium-intensity ( approximately 80-50% 1 RM) exercise without occlusion (HI) were performed. Percent increases in both cross-sectional area and isokinetic strength of elbow flexor muscles after LIO were larger than those after LI (P < 0.05) and similar to those after HI. The results suggest that resistance exercise at an intensity even lower than 50% 1 RM is effective in inducing muscular hypertrophy and concomitant increase in strength when combined with vascular occlusion.

Adult↗
PubMed1999

Development of an enzyme-labeled oligonucleotide probe for detecting the Escherichia coli attaching and effacing A gene.

Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC) can produce attaching and effacing (AE) lesions on intestinal epithelium in vitro and in vivo. A gene necessary to cause the AE lesion has been identified and designated Escherichia coli attaching and effacing A (eaeA) gene. In this study, an alkaline phosphatase (ALP)-conjugated oligonucleotide probe for the eaeA gene was developed and used to detect the eaeA gene among 163 strains of classical EPEC and 25 strains of EHEC O157. The prevalence rates of eaeA gene in the strains of classical EPEC and EHEC O157 were 51.5 and 100%, respectively. The eaeA-positive rate (60.0%) in strains of class I EPEC serogroups (O26, O55, O86, O111, O119, O125, O126, O127, O128ab, and O142) was significantly higher than that (22.9%) in strains of the class II EPEC serogroups (O18, O44, O114) (P<0.01). A total of 109 eaeA-positive classical EPEC and EHEC O157 were positive for fluorescent actin staining (FAS) assay, whereas 79 eaeA-negative classical EPEC were negative. Both the sensitivity and specificity of the eaeA probe versus the FAS assay positivity were 100%. Thus, use of the ALP-conjugated oligonucleotide probe for the eaeA gene would be specific and reliable in identifying the adherence capability of EPEC and EHEC.

Adhesins, Bacterial↗
PubMed1998

Serum levels of tumor necrosis factor-alpha are increased in obese patients with noninsulin-dependent diabetes mellitus.

To clarify the significance of the serum levels of tumor necrosis factor-alpha (TNF-alpha) in the mechanism of insulin resistance, we studied 12 obese patients with noninsulin-dependent diabetes mellitus (NIDDM). We evaluated the relationship of TNF-alpha levels with the visceral, subcutaneous, and total fat areas measured by computed tomography (CT), and with insulin resistance evaluated by the glucose infusion rate (GIR) observed during an euglycemic hyperinsulinemic clamp study. Controls consisted of 12 normal subjects and 12 nonobese patients with NIDDM. TNF-alpha levels were measured using a high sensitivity enzyme-linked immunosorbent assay. Following admission, all patients with NIDDM participated in a 4-week program of diet and exercise. After this treatment, we evaluated the relationship of the serum levels of TNF-alpha with the area of body fat, the GIR, and the resultant change in the TNF-alpha level. Serum levels of TNF-alpha in the obese patients with NIDDM significantly exceeded those observed in normal subjects (P < 0.01) or in the nonobese patients with NIDDM (P < 0.01). Serum levels of TNF-alpha in obese NIDDM patients showed a significant positive correlation with the area of visceral fat before (r = 0.662, P < 0.03) and after (r = 0.508, P < 0.05) the treatment; similar correlation was observed in all patients with NIDDM before (r = 0.537, P < 0.02) and after (r = 0.430, P < 0.05) the treatment. Serum levels of TNF-alpha in obese NIDDM patients showed a significant negative correlation with GIR after the treatment (r = -0.595, P < 0.05). Serum levels of TNF-alpha were significantly reduced in the obese patients with NIDDM after the treatment (P < 0.01), while those in the nonobese NIDDM patients were unchanged. These results suggest that serum TNF-alpha levels may play an important role in mechanism of insulin resistance associated with obesity.

Adipose Tissue↗
PubMed1997

Evaluation of enzyme-labeled oligonucleotide probes to identify enterohaemorrhagic Escherichia coli.

Alkaline phosphatase-conjugated oligonucleotide probes were developed to detect the gene coding for Vero toxin 1 (VT1) and Vero toxin 2 (VT2). Using these probes, 3 hr was enough to detect VT genes when suspicious colonies of enterohaemorrhagic Escherichia coli (EHEC) were obtained on an agar plate. The results of a hybridization test with 144 isolates of EHEC O157 and one isolate of Shigella dysenteriae Type 1 agreed exactly with the immunological detection, reversed passive latex agglutination (RPLA) test, of VTs in their culture supernatants. The sensitivity levels of these probes for the detection of VT genes were 100%. The specificity of these probes were also tested with a total of 1,002 strains of Escherichia coli other than EHEC and 8 strains of Shigella sp. other than Shigella dysenteriae Type 1; the results showed 100% specificity.

Alkaline Phosphatase↗
PubMed1997

Stretch-induced enhancement of mechanical work production in frog single fibers and human muscle.

The relations between the velocity of prestretch and the mechanical energy liberated during the subsequent isovelocity release were studied in contractions of frog single fibers and human muscles. During isometric contractions of frog single fibers, a ramp stretch of varied velocity (amplitude, 0.02 fiber length; velocity, 0.08-1.0 fiber length/s) followed by a release (amplitude, 0.02 fiber length; velocity, 1.0 fiber length/s) was given, and the amount of work liberated during the release was measured. For human muscles, elbow flexions were performed with a prestretch of varied velocity (range, 40 degrees; velocity, 30-180 degrees/s) followed by an isokinetic shortening (velocity, 90 degrees/s). In both frog single fibers and human muscles, the work production increased with both the velocity of stretch and the peak of force attained before the release up to a certain level; thereafter it declined with the further increases of these variables. In human muscles, the enhancement of work production was not associated with a significant increase in integrated electromyogram. This suggests that changes in intrinsic mechanical properties of muscle fibers play an important role in the stretch-induced enhancement of work production.

Adult↗
PubMed1997

Stretch-induced enhancement of mechanical power output in human multijoint exercise with countermovement.

The relation between the eccentric force developed during a countermovement and the mechanical power output was studied in squatting exercises under nominally isotonic load (50% of 1-repetition maximum). The subjects (n = 5) performed squatting exercises with a countermovement at varied deceleration rates before lifting the load. The ground reaction force and video images were recorded to obtain the power output of the body. Net muscle moments acting at hip, knee, and ankle joints were calculated from video recordings by using inverse dynamics. When an intense deceleration was taken at the end of downward movement, large eccentric force was developed, and the mechanical power subsequently produced during the lifting movement was consistently larger than that produced without the countermovement. Both maximal and mean power outputs during concentric actions increased initially with the eccentric force, whereas they began to decline when the eccentric force exceeded approximately 1.4 times the sum of load and body weight. Video-image analysis showed that this characteristic relation was predominantly determined by the torque around the knee joint. Electromyographic analyses showed no consistent increase in time-averaged integrated electromyograph from vastus lateralis with the power output, suggesting that the enhancement of power output is primarily caused by the prestretch-induced improvement of an intrinsic force-generating capability of the agonist muscle.

Adult↗
PubMed1996

Detection of the gene for toxic shock syndrome toxin 1 in Staphylococcus aureus by enzyme-labelled oligonucleotide probes.

A colony hybridisation method with enzyme-labelled oligonucleotide probes was developed to detect the gene for toxic shock syndrome toxin 1 (tst). For rapid identification, bacterial colonies were transferred from agar plates directly on to nylon membranes. These procedures took only 3 h. Results obtained by this test correlated well with those obtained by the reverse passive latex agglutination test. Thus, this method is convenient and reliable for the detection of tst in staphylococci, which could be useful for both research and clinical purposes. This method demonstrated that tst was more prevalent in methicillin-resistant Staphylococcus aureus (56%) than in methicillin-susceptible S. aureus (4%).

Bacterial Toxins↗
PubMed1996

[Detection of hepatitis B virus by using polymerase chain reaction and nonradioactive DNA probes. II. Identification of mutations in the core gene by PCR-direct sequencing and ASO probe method].

Mutants of hepatitis B virus (HBV) in the pre-core/core gene exist in patients chronically infected with HBV. The core gene of HBV DNA is preceded by the pre-core region which takes a role of synthesis and secretion of HBe antigen (HBeAg). Previous studies have suggested that products of the core gene could be immunological targets of cytotoxic T lymphocytes as well as those of variant pre-core genes and subsequently, that the clinical courses of HBV carriers might be influenced due to infected HBV variants. In this study, we therefore examined mutations in the core gene of HBV DNA on strains with or without a translational stop codon at the 28th codon of the pre-core region, using allele-specific oligonucleotide (ASO) probes and PCR-direct sequencing. The analysis of nucleotide sequences (codon 27-100) of HBV DNA in anti-HBe positive sera showed that there were two hypervariable segments of codon 31-49 and codon 87-97, where amino acid substitutions of L31I, S49T, S87G/N, K96N and I87F/1 frequently occurred regardless of the presence or absence of the mutation in the pre-core region. Meanwhile, less mutations were detected in the core gene of HBV DNA in HBeAg positive sera. Furthermore, we could identify the mutations at codon 87, 96 and 97 by using nonradioactive probes in a good coincidence with results by PCR-direct sequencing. The ASO probe method is useful for detection of mutations in the core gene in many specimens.

Amino Acid Sequence↗
PubMed1996

Use of an alkaline phosphatase-conjugated oligonucleotide probe for the gene encoding the bundle-forming pilus of enteropathogenic Escherichia coli.

An alkaline phosphatase-conjugated 29-base oligonucleotide probe was developed to detect the gene encoding the bundle-forming pilus of enteropathogenic Escherichia coli. The sensitivity and specificity of the probe versus the results of localized adherence in the HEp-2 cell assay and fluorescent actin staining assay positivity were 95.7 and 100%, respectively.

Alkaline Phosphatase↗
PubMed1995

Use of a new oligonucleotide probe for detection of colonization factor antigen III gene in enterotoxigenic Escherichia coli.

An alkaline phosphatase-labeled 30-mer oligonucleotide probe was designed to detect the gene for pilus colonization factor antigen III (CFA/III) of the human type of enterotoxigenic Escherichia coli (ETEC). The CFA/III probe was used to identify CFA/III-producing ETEC among 303 Escherichia coli obtained from subjects with traveler's diarrhea. Six isolates positive for the CFA/III gene were found. This result was confirmed immunologically by using a specific monoclonal antibody developed against CFA/III. These six isolates, isolated from travelers returning from India, Pakistan and China, were all positive for the gene of heat-labile enterotoxin and possessed an identical serotype (025:H-).

Antigens, Bacterial↗
PubMed1995

[Detection of hepatitis B virus by using polymerase chain reaction and nonradioactive DNA probes. I. Identification of mutations in the precore region by PCR-direct sequencing and ASO probe method].

Seroconversion from hepatitis B e antigen (HBeAg) to anti-HBe antibody (anti-HBe) frequently occurs in hosts chronically infected with hepatitis B virus (HBV). Further, this phenomenon is related to a point mutation from guanine to adenine at nucleotide 83 in the precore region of HBV, which, converts codon 28 for tryptophan (TGG:W) to a translational, stop codon (TAG:X). Therefore, we decided to examine HBV in sera from patients for mutations in the precore region by a simple allele-specific oligonucleotide (ASO) probe method. Direct sequencing was first performed on DNA fragments amplified by the polymerase chain reaction in order to establish whether there were mutations in the precore region. Subsequently, specific DNA probes were applied to detection of mutations in the precore region. Subsequently, specific DNA probes were applied to detection of mutations in the precore gene. Five unknown mutations (I10N, C12W, C14S, V17F and A19D), three known mutations (I9V, W28X and G29D) and for novel nucleotide insertions were identified in anti-HBe positive sera. By using seven nonradioactive probes, we could determine the mutations at codons 9, 28 and 29 in anti-HBe positive sera. The W28X mutation was found in anti-HBe positive but not in any of HBeAg positive sera. Meanwhile, wild-type strains of HBV were detectable in sera from patients who were positive to HBeAg or anti-HBe. This ASO probe assay could determine in a few days the mutations in the precore region of HBV, especially including the defect to prohibit the synthesis and secretion of HBeAg.

Amino Acid Sequence↗
PubMed1994

Gypsy phenylketonuria: a point mutation of the phenylalanine hydroxylase gene in Gypsy families from Slovakia.

A direct mutational analysis of the phenylalanine hydroxylase gene (PAH) in Gypsy families with phenylketonuria (PKU) has not yet been presented. However, they obviously represent a group at high risk for this inherited disease. We analyzed the PAH loci of 65 Gypsies originating from Eastern Slovakia by a combination of PCR amplification, direct sequencing and ASO hybridization. These studies uncovered 10 "classical PKU" patients to be homozygous for a R252W (CGG-TGG) transition, and 29 heterozygous carriers of this mutation. Fifteen control Caucasoid PKU patients from the Czech and Slovak Republics were selected. In this group we detected R252W mutation in two subjects (6.67% of all mutant alleles). Both were compound heterozygous for two different mutations. Previous haplotype studies of Welsh Gypsies with PKU were uninformative in the determination of heterozygosity. ASO hybridization served us effectively for the consequent analyses in Gypsy PKU-related families and to identify the carriers among the unrelated subjects.

Adolescent↗
PubMed1994

Demonstration and characterization of simultaneous production of a thermostable direct hemolysin (TDH/I) and a TDH-related hemolysin (TRHx) by a clinically isolated Vibrio parahaemolyticus strain, TH3766.

Simultaneous production of a thermostable direct hemolysin (TDH)-like toxin (TDHx) and a TDH-related hemolysin (TRH)-like toxin (TRHx) by a clinical isolate (strain TH3766) of Kanagawa phenomenon-positive Vibrio parahaemolyticus was demonstrated and characterized. The two hemolysins were differentially purified by column chromatography on hydroxyapatite and immunoaffinity columns. The molecular weight of the two hemolysins were estimated to be 23,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE). The purified TDHx was indistinguishable from the previously reported TDH/I (from strain TH012) but was different from the authentic TDH of a Kanagawa phenomenon-positive strain (T4750) physicochemically. The mobility of TRHx in nondenaturing PAGE differed from all the known TDHs and TRHs. The genes (tdhX and trhX) coding for TDHx and TRHx were cloned and sequenced. Homologies of nucleotide sequences of the coding regions between tdhX and tdhA (a gene for the authentic TDH) and between trhX and trh (a gene for the authentic TRH) were 98.1 and 99.1%, respectively, and homology between tdhX and trhX was 68.1%. At the amino acid level, TdhX was completely identical to TDH/I, although two base differences were found in the nucleotide sequences between tdhX and tdh/I. Two amino acid differences were observed between TrhX and Trh. Thus, these findings suggest that the TH3766 strain produces two types of hemolysins simultaneously. This is the first evidence that a strain of V. parahaemolyticus produces two types of toxins of the TDH-TRH family at the same time.

Amino Acid Sequence↗
PubMed1994

Development of enzyme-labeled oligonucleotide probe for detection of mecA gene in methicillin-resistant Staphylococcus aureus.

A DNA hybridization method with an enzyme-labeled oligonucleotide probe (mecA-ELONP) was developed to detect the methicillin-resistant gene (mecA) in methicillin-resistant Staphylococcus aureus. For rapid identification, bacterial colonies were transferred from agar plates directly onto nylon membranes. Lysis of cells, denaturation of DNA, and hybridization were performed on the membranes. These procedures required only 3 h for completion. The results obtained by this test closely corresponded with those obtained by determining the MICs of oxacillin against S. aureus. The results of the mecA-ELONP also correlated well with those of a commercially available PCR test. Thus, mecA-ELONP proved to be a reliable and convenient method for the rapid identification of methicillin-resistant S. aureus, which could be useful in clinical microbiology laboratories.

Bacterial Proteins↗