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Biomedical subjects

S Omata

Publications and source records attributed to S Omata.

At least 19 recordsLinked to original sources
PubMed2003

Development of a vibratory microinjection method.

To reduce cellular damage by pronuclear microinjection and nuclear transfer, we have recently developed a vibratory microinjection method. A micropipette was fixed to a piezoelectric ceramic with a resonance frequency of 70 kHz. When this micropipette was vibrated, it easily entered a mouse-fertilized egg without any sharp depression of the cell body, whereas a sharp, deep depression at the insertion site was observed when the micropipette was not vibrated. A depression rate defined as a rate of a depth of depression over an original cell diameter was utilized as an index of cellular deformation. The depression rates with and without vibration were 11.1 +/- 5.2% (N = 24) and 40.4 +/- 8.8% (N = 16), respectively (P < 0.0001, Student's t-test). In conclusion, the vibratory microinjection method is a new, useful option for gene transfer because it resulted in much less cellular deformation, therefore implicating less cellular damage.

Animals↗
PubMed2001

Muscle contraction and relaxation described by tactile stiffness.

We developed a tactile sensor system that measures the stiffness of objects (tactile stiffness) and used it to describe the time course of muscle contraction and relaxation. We examined fatigue resistance of the latissimus dorsi muscle (LDM), which is preconditioned for cardiomyoplasty. Time to peak, ripple of LDM, and time constant were calculated from the time course of LDM contraction and relaxation as described by tactile stiffness. We compared conditioned and unconditioned LDMs using these 3 parameters. The time course can be described by tactile stiffness. Tactile stiffness fell exponentially during LDM relaxation. In mean values, time to peak increased 230%, ripple decreased 20%, and time constants increased 424%. Significant differences were shown in 3 parameters between conditioned and unconditioned LDMs (p < 0.05). Our tactile sensor system can describe the time course of LDM contraction and relaxation. Examining the difference in time courses, we might detect the level of LDM preconditioning for cardiomyoplasty.

Animals↗
PubMed2001

Lupeol esters from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko.

Five new lupeol esters, lup-20(29)-ene-3beta-yl eicosanoate, docosanoate, tetracosanoate, hexacosanoate and octacosanoate, were isolated as a mixture from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko, together with lupeol and epifriedelinol. Their structures were determined by spectral analyses including 2D-NMR experiments.

Esters↗
PubMed2000

Myocardial tactile stiffness during acute reduction of coronary blood flow.

BACKGROUND: Evaluation of regional myocardial contractile function is of clinical importance. We have developed a new tactile sensor system for accurate measurement of myocardial stiffness in situ. We found that the myocardial stiffness measured by this sensor, which we call tactile stiffness, can be a very useful index for accurate quantification of regional myocardial function. In this study, we used a coronary stenosis model to investigate regional myocardial tactile stiffness under conditions of reduced coronary blood flow. METHODS: The myocardial tactile stiffness, coronary blood flow, and ventricular pressure and volume, of five open chest mongrel dogs weighing 15 to 17 kg, were measured. After measuring the baseline myocardial stiffness, coronary arterial stenosis was induced with a balloon occluder. RESULTS: Reducing the coronary flow to 50% and 25% of the baseline level reduced the end-systolic tactile stiffness significantly from 2.20+/-0.16 g/mm2 to 2.05+/-0.20 g/mm2 (p<0.05) and from 2.21+/-0.16 g/mm2 to 1.96+/-0.18 g/mm2 (p<0.01), respectively. Reducing the flow, to 50% and 25%, increased the end-diastolic stiffness significantly from 1.29+/-0.15 g/mm2 to 1.39+/-0.14 g/mm2 (p<0.01) and from 1.30+/-0.16 g/mm2 to 1.46+/-0.14 g/mm2 (p<0.05), respectively. CONCLUSIONS: We consider that the regional myocardial tactile stiffness will be a useful index sensitive enough to detect changes in regional contractile function under conditions of reduced coronary blood flow.

Acoustics↗
PubMed2000

Objective evaluation of liver consistency to estimate hepatic fibrosis and functional reserve for hepatectomy.

BACKGROUND: The empiric evaluation of liver consistency is currently used to plan the surgical strategy. The aim of this study was to verify the feasibility of the objective measurement of liver consistency and to check its correlation with liver fibrosis and liver functional reserve. STUDY DESIGN: Fifty-two consecutive patients who underwent hepatic resections in our department were enrolled. The indications for liver resection were hepatocellular carcinoma in 36 patients, metastatic liver tumors in 12 patients, and other conditions in 4 patients. Liver consistency was measured with a new tactile sensor. A fibrosis index was calculated as an expression of the percentage of fibrotic tissue. Liver consistency was compared with the degree of liver fibrosis observed in histologic specimens (fibrosis index) and with liver function parameters. RESULTS: Liver stiffness showed a significant positive correlation with fibrosis index (r = 0.887, p < 0.0001). Liver stiffness also showed significant positive correlation with the indocyanine green test (r = 0.631, p < 0.0001) by a univariate analysis. The indocyanine green test and platelet count were independently and significantly associated with liver stiffness by a multiple regression analysis. In five patients, the liver stiffness values measured intraoperatively differed markedly from those expected from the indocyanine green test values. In these patients, the operative procedures were finally selected based on the liver stiffness measured with the tactile sensor and good clinical outcomes were obtained. CONCLUSIONS: These results show for the first time that liver stiffness can be clinically assessed quantitatively by means of the tactile sensor. The tactile sensor adequately estimates liver stiffness and this estimation is well correlated with liver fibrosis and functional reserve. Liver consistency determined objectively in this manner may be useful for optimizing surgical decision making.

Adult↗
PubMed2000

Use of tactile stiffness to detect fatigue in the latissimus dorsi muscle.

In clinical settings, no method has been established to examine the fatigue of a latissimus dorsi muscle (LDM) preconditioned for cardiomyoplasty. We examined the feasibility of measuring muscle stiffness (tactile stiffness) to evaluate muscle fatigue in situ using our tactile sensor. We stimulated canine LDM with burst pacing and monitored both stiffness and tension to determine their relationship. In both dissected LDM and LDM in situ, the decrements of these parameters during burst pacing were compared between preconditioned and unconditioned LDM. In measurement in situ, the sensor probe was placed on the LDM through a small incision. Strong statistical correlation was shown between stiffness and tension (r = 0.935). In decrements of stiffness in situ, there were statistically significant differences between preconditioned and unconditioned LDM. Our tactile sensor system can provide an efficient method for evaluating fatigue of muscles in situ without measuring muscle tension.

Analysis of Variance↗
PubMed1999

Interaction between A beta(1-42) and A beta(1-40) in Alzheimer's beta-amyloid fibril formation in vitro.

We analyzed the interaction of two kinds of amyloid beta-peptides (A beta), i.e., A beta(1-42) and A beta(1-40), in the kinetics of beta-amyloid fibril (fA beta) formation in vitro, based on a nucleation-dependent polymerization model using fluorescence spectroscopy with thioflavin T. When 25 microM A beta(1-42) was incubated with increasing concentrations of amyloidogenic A beta(1-40), the time to proceed to equilibrium was extended dose-dependently. A similar inhibitory effect was observed when 45 microM A beta(1-40) was incubated with increasing concentrations of A beta(1-42). On the other hand, when 50 microM of nonamyloidogenic A beta(1-40) was incubated with A beta(1-42) at a molar ratio of 10:1 or 5:1, A beta(1-42) initiated fA beta formation from A beta(1-40). The lag time of the reaction shortened in a concentration-dependent manner, with A beta(1-42). We next examined the seeding effect of fA beta formed from A beta(1-42) (fA beta(1-42)) on nonamyloidogenic A beta(1-40). When 50 microM of nonamyloidogenic A beta(1-40) was incubated with 10 or 20 microg/mL (2.2 or 4.4 microM) of fA beta(1-42), the fluorescence showed a sigmoidal increase. The lag time of the reaction was shortened by fA beta(1-42) in a concentration-dependent manner. However, the time to proceed to equilibrium was much longer than when an equal concentration of fA beta formed from A beta(1-40) (fA beta(1-40)) was added to A beta(1-40). The fluorescence increased hyperbolically without a lag phase when 25 microM A beta(1-42) was incubated with 10 or 20 microg/mL (2.3 or 4.6 microM) of fA beta(1-40), and proceeded to equilibrium more rapidly than without fA beta(1-40). An electron microscopic study indicated that the morphology of fA beta formed is governed by the major component of fresh A beta peptides in the reaction mixture, not by the morphology of preexisting fibrils. These results may indicate the central role of A beta(1-42) for fA beta deposition in vivo, among the different coexisting A beta species.

Alzheimer Disease↗
PubMed1999

Stimulus-coupled interaction of tyrosine hydroxylase with 14-3-3 proteins.

Tyrosine hydroxylase (TH) is phosphorylated by CaM kinase II and is activated in situ in response to a variety of stimuli that increase intracellular Ca(2+). We report here, using baculovirus-expressed TH, that the 14-3-3 protein binds and activates the expressed TH when the enzyme is phosphorylated at Ser-19, a site of CaM kinase II-dependent phosphorylation located in the regulatory domain of TH. Site-directed mutagenesis showed that a TH mutant in which Ser-19 was substituted by Ala retained enzymatic activity at the same level as the non-mutated enzyme, but was a poor substrate for CaM kinase II and did not bind the 14-3-3 protein. Likewise, a synthetic phosphopeptide (FRRAVpSELDA) corresponding to the part of the TH sequence, including phosphoSer-19, inhibited the interaction between the expressed TH and 14-3-3, while the phosphopeptide (GRRQpSLIED) corresponding to the site of cAMP-dependent phosphorylation (Ser-40) had little effect on complex formation. The complex was very stable with a dissociation constant of 3 nM. Furthermore, analysis of PC12nnr5 cells transfected with myc-tagged 14-3-3 showed that 14-3-3 formed a complex with endogenous TH when the cultured cells were exposed to a high K(+) concentration that increases intracellular Ca(2+) and phosphorylation of Ser-19 in TH. These findings suggest that the 14-3-3 protein participates in the stimulus-coupled regulation of catecholamine synthesis that occurs in response to depolarization-evoked, Ca(2+)-dependent phosphorylation of TH.

14-3-3 Proteins↗
PubMed1999

In vitro nuclear assembly with affinity-purified nuclear envelope precursor vesicle fractions, PV1 and PV2.

Nuclear envelope precursor vesicles were affinity purified from a Xenopus egg extract by a chromatin binding method. Vesicles bound to chromatin at 4 degrees C were dissociated with a high salt buffer and further fractionated into nuclear envelope precursor vesicle fractions 1 (PV1) and 2 (PV2) by differential centrifugation. PV1 contained larger vesicles. When chromatin was incubated in a Xenopus egg cytosol fraction supplemented with PV1, vesicles bound to chromatin, fused with each other, formed a bilayered nuclear envelope, and assembled into spherical small nuclei. However, the thus assembled nuclei did not grow to the normal size. Nuclear pore complexes were not found on the thus assembled nuclei. On the other hand, PV2 contained smaller vesicles. PV2 vesicles bound to chromatin, fused little with each other in the Xenopus egg cytosol fraction, and no nuclei were assembled. When PV1 supplemented with PV2 was used for the nuclear assembly reaction, the assembled nuclei grew to the normal size. Nuclear pore complexes existed in the thus assembled nuclear envelopes. These results suggested that 1) two vesicle populations, PV1 and PV2, are necessary for the assembly of normal sized nuclei, 2) PV1 contains a chromatin targeting molecule(s) and membrane fusion machinery, 3) PV2 contains a chromatin targeting molecule(s) and a molecule(s) necessary for nuclear pore complex assembly, and 4) PV1 has the ability to assemble a nuclear membrane, and PV2 is necessary for the assembly of nuclear pore complexes and for nuclei to grow to the normal size. An in vitro nuclear assembly system constituted with affinity-purified vesicle fractions, PV1 and PV2, was established.

Animals↗
PubMed1999

Molecular shape and ATP binding activity of rat p50, a putative mammalian homologue of RuvB DNA helicase.

Based on partial amino acid sequences of p50 purified from a high-salt buffer extract of a rat liver nuclear matrix fraction, p50 cDNA was cloned and sequenced, and its amino acid sequence was predicted. The sequence contained helicase motifs, and showed homology with RuvB DNA helicase of Thermus thermophilus and an open reading frame for an unknown 50.5 k protein of Saccharomyces cerevisiae. p50 was expressed as a GST-fusion protein and antiserum against the protein was generated. p50 was localized to the nuclear matrix by cell fractionation and immunoblotting. p50 bound to ATP-Sepharose beads. Ultracentrifugation and gel filtration analyses showed that p50 in rat liver and Xenopus egg mitotic extracts exists as large complexes corresponding to 697 k and 447 k, respectively. A 50 k protein reactive with p50 antibodies was detected not only in rat liver nuclei, but also in a Xenopus egg cytoplasm fraction and a S. cerevisiae extract. This suggests that this putative DNA helicase is present in a wide variety of species ranging from yeast to mammals.

ATPases Associated with Diverse Cellular Activitie↗
PubMed1999

Molecular cloning of mouse p47, a second group mammalian RuvB DNA helicase-like protein: homology with those from human and Saccharomyces cerevisiae.

A 47k protein (p47) in a high-salt buffer extract of a rat liver nuclear matrix fraction was purified by means of a wheat germ agglutinin affinity column, reversed phase HPLC, and SDS-PAGE, and partial amino acid sequences were analyzed. Based on these sequences, the mouse cDNA of the protein was cloned and sequenced, and its amino acid sequence was deduced. Mouse p47 consists of 463 amino acid residues with a molecular weight of 51,112. The amino acid sequences of human and Saccharomyces cerevisiae p47s were also deduced from the nucleotide sequences of "expressed sequence tag" fragments and genomic DNA, respectively. These sequences contain helicase motifs and show homology to bacterial RuvB DNA helicases acting in homologous recombination. They also show homology with the putative mammalian helicases p50/TIP49 and RUVBL1. Comparison of the amino acid sequences of p47 group proteins and those of p50/TIP49 group proteins revealed the p47 group proteins to comprise a group distinct from the p50/TIP49 proteins. Ultracentrifugation and gel filtration analyses showed that p47 in the rat liver cytosol fraction exists as large complexes of 697k.

ATPases Associated with Diverse Cellular Activitie↗
PubMed1999

Molecular cloning and splicing isoforms of mouse p144, a homologue of CA150.

We previously characterized p144 bearing N-acetylglucosamine residues in a rat liver nuclear matrix fraction. Based on partial amino acid sequences of rat p144, mouse p144 cDNA was cloned and sequenced, and its amino acid sequence was predicted. The sequence revealed that p144 is a rat homologue of CA150, which is a transcription factor involved in Tat-activated human immunodeficiency virus type 1 transcription. The reported human CA150 consists of 1098 amino acids and has a leucine zipper-like motif in its carboxyl-region. However, a clone of mouse p144 cDNA encoded a CA150 consisting of 1,034 amino acids. The mouse CA150 was shorter by 64 amino acids than hitherto known human CA150 and lacked the leucine zipper-like motif. We designated the longer and shorter CA150 species as CA150a and CA150b, respectively. The partial nucleotide sequences of other mouse p144 cDNA clones were examined and it was found that some clones encode CA150a having a leucine zipper-like motif. It was suggested that CA150a and CA150b are splicing isoforms. All rat and mouse tissues examined contained transcripts for both CA150a and CA150b. Both transcripts were detected in human blood and Jurkat cells as well as mouse CD4(+) T-cells, which are the HIV-1-sensitive counterpart in humans.

Amino Acid Sequence↗
PubMed1998

Chronology and urodynamic characterization of micturition in neurohormonally induced experimental prostate growth in the rat.

The purpose of this study was to evaluate the impact of chronic urinary tract obstruction which was produced in the rat using neurohormonally induced experimental prostate growth. In this model, we considered the chronology of changes in the micturition characteristics of awake rats relative to prostate weight and stiffness. The corresponding urodynamic characteristics of both the upper and lower tracts were evaluated in anesthetized animals relative to the development and extent of the obstruction produced. Prostate growth was produced by capitalizing on the synergistic properties afforded by the combined administration of dihydrotestosterone propionate (DHT) and the alpha1 adrenoreceptor antagonist prazosin (PRZ). DHT (1.25 mg/kg/day) was dissolved in 0.1 ml sesame oil (SO) and coadministered with PRZ 30 microg/kg/day subcutaneously for 14 days to 12 experimental rats. SO alone was given to 8 control rats. Micturition studies were first performed using all 20 awake rats, which were placed unrestrained in metabolic cages. Urodynamics of the upper and lower urinary tracts were repeated following anesthesia at the 5th, 10th, and 15th weeks after initiation of hormonal or SO treatment. Following the urodynamic studies, the rats were killed and prostates were removed and weighed, and stiffness was measured. Studies with awake rats show that hormonal treatment produces a significant and progressive increase in mean frequency of micturition, ranging from 0.63+/-0.16 in controls and reaching the maximum of 2.15+/-0.40/hr by the 10th wk. Results from urodynamic studies with anesthetized rats also show typical and progressive obstructive characteristics: maximum detrusor voiding pressure (Pdetmax) increased from 52.7+/-2.03 in controls to a maximum of 77.5+/-2.2 cm H2O by the 10th week; urethral opening pressure Puo likewise increased from 52.6+/-2.7 in controls to 73.3+/-2.1 cm H2O in experimental rats. The duration of time during which the detrusor sustains contraction during voiding also rose, from 16.8+/-1.8 sec in controls to 32.0+/-3.2 sec by the 10th week. There were no significant changes in bladder capacity, baseline filling pressures, or arterial pressures. Prostate weight increased significantly from 0.76+/-0.05 g in controls to 1.17+/-0.1 g by the 15th week. Similarly, stiffness increased from control values of 1.33+/-0.18 g/cm to a maximum of 3.59+/-0.14 g/cm by the 10th week. It is concluded that neurohormonally stimulated prostate growth in the rat is a suitable animal model for the study of the development of urinary tract obstruction. Obstructive characteristics were validated in both awake rats by the increase in the frequency of micturition and urodynamically under anesthesia in terms of elevations in maximum detrusor pressures, urethral opening pressure, detrusor contraction time, and prostatic stiffness. The effect of obstruction was further shown to be associated with vesicoureteral reflux during micturition and elevated upper tract pressures.

Adrenergic alpha-Antagonists↗
PubMed1998

Myocardial tactile stiffness: a variable of regional myocardial function.

OBJECTIVES: We developed a new sensor system for in situ measurement of myocardial tactile stiffness-stiffness in a direction perpendicular to the wall-and validated its use for providing a reasonable estimation of regional myocardial function. BACKGROUND: Numerous attempts have been made to directly assess regional myocardial function. The complexity and highly invasive nature of the measuring devices have hampered their in situ application. METHODS: In open chest mongrel dogs, myocardial tactile stiffness, ventricular pressure and ventricular volume were monitored. Under the preload reduction, these variables were measured to determine the relation between the end-systolic pressure-volume relation (ESPVR) and the end-systolic tactile stiffness-volume relation (ESSVR). The changes in myocardial tactile stiffness were monitored in the regional ischemic myocardial model and infarcted model to evaluate their usefulness as indexes of regional myocardial function. RESULTS: Myocardial tactile stiffness changed cyclically and followed a time course similar to left ventricular pressure. When preload was altered, the ESSVR was as linear as the ESPVR. The slope of the ESSVR and that of the ESPVR showed a strong correlation over a wide range of contractility. These results suggest that myocardial tactile stiffness can be a good index of regional wall stress or fiber stress. End-systolic myocardial tactile stiffness of ischemic and infarcted regions decreased significantly, with a concomitant increase in end-diastolic stiffness compared with that of intact myocardium. CONCLUSIONS: Using our tactile sensor system, regional myocardial tactile stiffness of a beating heart was measured with reasonable temporal resolution. We consider myocardial tactile stiffness to be a useful index of regional myocardial function.

Animals↗
PubMed1997

The stiffness of lymph nodes containing lung carcinoma metastases: a new diagnostic parameter measured by a tactile sensor.

BACKGROUND: It is believed that the stiffness or hardness of a lymph node containing a metastasis differs from that of lymph node without a metastasis because of the difference in tissue density, which is derived from the lymph node's histopathologic features. Prior to this study, however, there had been no attempts to quantify the hardness or stiffness of lymph nodes. The authors developed a new tactile sensor and system for measuring the stiffness (g/cm) of lymph nodes accurately, and they studied its utility as a tool for diagnosing lymph node metastases. METHODS: Clinical specimens were obtained from 14 patients who underwent lobectomy or pneumonectomy with hilar and mediastinal lymph node dissection for nonsmall cell lung carcinoma at the University of Tokyo between January and July 1996. With the tactile sensor developed by the authors, 212 resected lymph nodes were measured for their stiffness. RESULTS: Among these 212 resected lymph nodes, 57 were diagnosed as containing metastases (38 from adenocarcinomas and 19 from squamous cell carcinomas). The mean stiffness of the lymph nodes that contained metastases was 3.35 +/- 1.57 g/cm, and that of lymph nodes without metastases was 1.23 +/- 0.50 g/cm (P < 0.001). Receiver operating characteristic analysis revealed that the area under the curve was 0.93, indicating excellent accuracy of the method. When the cutoff was 1.5 g/cm, the sensitivity was 91.2% and the specificity was 78.1% for detection of lymph node metastases. CONCLUSIONS: Measurement of the stiffness of resected lymph nodes was confirmed as an accurate approach to diagnosing lymph node metastases without knowledge of other factors, such as lymph node size or color.

Aged↗
PubMed1997

Methylmercury alters the tyrosination status of tubulin in the brains of acutely intoxicated rats.

Tyrosination/detyrosination, a post-translational modification at the carboxyl terminus of alpha-tubulin, was investigated in the brain cytosol fraction of rats treated with methylmercury (MeHg) chloride (10 mg/kg per day, for 7 days). The amount of detyrosinated tubulin species, determined as the incorporation of 14C-tyrosine at the carboxyl-terminal end of alpha-tubulin, was significantly decreased throughout the experimental period of MeHg intoxication. Furthermore, the activity of tubulin-tyrosine ligase, as well as the amounts of tyrosinatable tubulin determined and calculated by a method involving pancreatic carboxypeptidase A, also decreased in the latent and symptomatic periods. Tubulin-tyrosine carboxypeptidase activity did not change during the MeHg intoxication. The total amounts of alpha- and beta-tubulins, as determined by densitometry and immunoblotting, did not show significant changes during the intoxication. These results suggest that MeHg treatment may produce perturbation of cellular activities associated with the tubulin/microtubule system by altering the tyrosination status of tubulin in the rat brain.

Animals↗