Biomedical subjects

T Kamada

Publications and source records attributed to T Kamada.

At least 271 records · Page 15Linked to original sources

PubMed1994

cDNA cloning of a cytosolic protein tyrosine phosphatase (RKPTP) from rat kidney.

A rat cDNA encoding a non-receptor type phosphotyrosine phosphatase (PTPase; EC 3.1.3.48) was identified. The 1608 bp cDNA contains a single open reading frame that predicts a 382 amino acid protein with M(r) 44,438. The predicted protein has no apparent signal or transmembrane sequences, suggesting that it is a cytosolic protein. The C-terminal region has a PTPase catalytic domain that has 40-50% nucleic acid homology to other known PTPases. The N-terminal region has little amino acid sequence homology to any other known sequences. The recombinant protein of the cloned cDNA expressed in Escherichia coli was shown to possess PTPase activity using myelin basic protein, tyrosine phosphorylated by p43v-abl tyrosine kinase, as a substrate.

Amino Acid Sequence↗

PubMed1994

Estrogen regulation of the insulin-like growth factor I gene transcription involves an AP-1 enhancer.

As a step toward elucidating the physiological role of insulin-like growth factor-I (IGF-I) in mediating estrogen action, we sought to determine the molecular basis of the phenomenon. In HepG2 cells expressing exogenous estrogen receptors (ER), a reporter gene plasmid containing 600 base pairs of the chicken IGF-I promoter enhanced expression of luciferase 8.6-fold in response to 10(-6) M 17 beta-estradiol, indicating that the IGF-I promoter is a target of estrogen regulation. Although no conventional estrogen-responsive element was identified within the promoter fragment, the AP-1 motif located therein was shown to be essential; the estrogen-responsive enhancement of the Fos-Jun binding to the AP-1 motif, which takes place by means of post-translational modification, mediates the estrogen action. A direct or indirect interaction between the estrogen-ER complex and the Fos-Jun complex seems to facilitate the Fos-Jun binding to the target DNA. Although ER binding to the target DNA was not considered to be involved in the signaling pathway, the DNA binding domain-deficient ER did not mediate the phenomenon, providing support for the existence of a unique function of the DNA binding domain of ER in facilitating some protein-protein interaction. In conclusion, our present observations demonstrate that the chicken IGF-I gene promoter is controlled by estrogen through a unique pathway involving Fos, Jun, and the DNA binding domain of ER.

Animals↗

PubMed1994

Expression of hepatitis C virus in hepatocellular carcinoma.

BACKGROUND: Epidemiologic studies have suggested a strong association between chronic hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC). To investigate the possible role of HCV in the pathobiology of HCC, the authors studied the expression of HCV in 10 cases of HCC with chronic HCV infection. METHODS: The core, envelope, and nonstructural (NS) 3 and 5 proteins were localized in liver and tumor tissues by the immunoperoxidase technique using mouse monoclonal antibodies to recombinant proteins or synthetic peptide of HCV. In addition, the positive and negative strands of HCV RNA were detected in the tissues by strand-specific reverse transcription/double polymerase chain reaction using primers for the 5'-nontranslated region. RESULTS: The HCV proteins were expressed in three of nine HCC specimens tested (the core protein in three HCC, the envelope, NS3 and NS5 proteins in one HCC) and in two of nine nontumorous liver specimens adjacent to the HCC (the core protein in two specimens, envelope, NS3 and NS5 proteins in one specimen). Positive-stranded HCV RNA was detected in all tumorous and nontumorous specimens except in one tumor. Negative-stranded HCV RNA was found in six of nine HCC tested and in all nontumorous livers. CONCLUSIONS: These findings suggest that HCV persists in hepatocytes during malignant transformation, although secondary infection of tumor cells by HCV cannot be excluded. Some HCC appear to support replication and expression of HCV.

Adult↗

PubMed1994

The point mutation Arg615-->Cys in the Ca2+ release channel of skeletal sarcoplasmic reticulum is responsible for hypersensitivity to caffeine and halothane in malignant hyperthermia.

Malignant hyperthermia (MH) is an autosomal dominant myopathy. Molecular genetic studies have shown that the alteration of Arg615 to Cys in the Ca2+ release channel of skeletal muscle sarcoplasmic reticulum (ryanodine receptor) is cosegregated with porcine MH (Fujii, J., Otsu, K., Zorzato, F., de Leon, S., Khanna, V. K., Weiler, J. E., O'Brien, P. J., and MacLennan, D. H. (1991) Science 253, 448-451; Otsu, K., Khanna, V. K., Archibald, A., and MacLennan, D. H. (1991) Genomics 11, 744-750). Here, using the fluorescence calcium indicator indo-1, we determined the concentration of ionized cytosolic calcium in myoblastic cells transfected with either the wild-type or mutated ryanodine receptor cDNA. The cells expressing the mutant ryanodine receptor showed higher sensitivity to caffeine, which induces Ca2+ release from the sarcoplasmic reticulum through the ryanodine receptor. Exposure to clinical doses of halothane resulted in a rapid increase of [Ca2+]i in cells expressing the mutated ryanodine receptor, whereas no [Ca2+] changes were observed in cells expressing the wild-type ryanodine receptor. These results provide definite evidence that a single amino acid mutation, Arg615-->Cys, in the ryanodine receptor is causative of MH.

Amino Acid Sequence↗

PubMed1994

Selective suppression of N-acetylglucosaminyltransferase III activity in a human hepatoblastoma cell line transfected with hepatitis B virus.

UDP-N-acetylglucosamine: beta-D-mannoside beta-1,4-N-acetylglucosaminyl-transferase III (GnT-III) is a key enzyme in the branching of asparagine-linked oligosaccharides, which are present in surface membrane proteins of various tissues and in secretory glycoproteins. The activity of GnT-III was assayed in 2 human hepatoblastoma cell lines, Huh6, which was the parental cell line, and HB611, which was established by transfection of 3 tandem copies of the hepatitis B virus genome into Huh6. A significant difference in GnT-III activity was found between Huh6 and HB611 (136 +/- 18.3 pmol/h/mg versus 6.7 +/- 2.4 pmol/h/mg; mean +/- SD, P < 0.001), whereas levels of the glycosyltransferases alpha-3-D-mannoside beta-1,4-N-acetylglucosaminyltransferase IV, alpha-6-D-mannoside beta-1,6-N-acetylglucosaminyltransferase-V, and beta-1,4-galactosyltransferase were almost the same in both cell lines. Northern blot analysis indicated that the decreased activity of GnT-III in HB611 was due to the decreased transcript. When HB611 was treated with interferon-alpha, expression of hepatitis B virus-related mRNA decreased, and the activity of GnT-III increased from 8.5 +/- 3.8 to 22.0 +/- 7.2 pmol/h/mg (mean +/- SD, P < 0.05). This increase was not found in Huh6. Binding capacity with erythrocyte phytohemagglutinin in these cells using fluorescence-activated cell sorter analysis was different, suggesting that the structure of sugar chain on the cell surface might be altered by suppression of GnT-III activity. This is the first report that hepatitis B virus selectively suppressed the GnT-III activity in hepatoblastoma cells.

Blotting, Northern↗

PubMed1994

Identification of amino-acid polymorphism within the leucine zipper motif of mouse transcription factor A1.

Mouse transcription factor A1 (A1) is a mouse homologue of human E47, a ubiquitously expressed DNA-binding protein which contains a basic region, helix-loop-helix (HLH) and leucine zipper (LZ) motifs [Walker et al., Nucleic Acids Res. 18 (1990) 1159-1166]. Analyses of the nucleotide (nt) sequences of A1 cDNAs isolated from various mouse strains revealed amino acid (aa) polymorphism in the highly conserved region within the LZ motif. Interestingly, the location and pattern of aa deletions are identical to those previously described for the aa polymorphism within the human counterpart, E47 (E12) [Kamps et al., Cell 60 (1990) 547-555].

Amino Acid Sequence↗

PubMed1994

Characterization of signaling pathways to Na+/H+ exchanger activation with epidermal growth factor in hepatocytes.

To investigate the signaling pathways to Na+/H+ exchanger activation with epidermal growth factor in hepatocytes, we measured changes in cytosolic free calcium and intracellular pH levels at the single-cell level using digital imaging fluorescence microscopy of fura-2- or BCECF-loaded hepatocytes in primary culture. Epidermal growth factor induced cytosolic free calcium oscillations consisting of periodic trains of spikes with a latency period of up to several minutes. These calcium responses were inhibited by tyrosine kinase inhibitor genistein (100 mumol/L) and abolished by emptying of intracellular Ca2+ pools with 3 mumol/L thapsigargin, an inhibitor of Ca(2+)-ATPase on the endoplasmic reticulum. Epidermal growth factor (1 nmol/L) induced an intracellular pH increase of 0.12 +/- 0.07 units from the basal level of 7.25 +/- 0.09 units after several minutes of latency. This effect was completely abolished by 1 mmol/L amiloride, an inhibitor of the Na+/H+ exchanger. The epidermal growth factor-induced intracellular pH increase was inhibited by pretreatment of hepatocytes with genistein (100 mumol/L), thapsigargin (3 mumol/L) or calmodulin inhibitor W-7 (25 mumol/L), but not with protein kinase C inhibitor H-7 (50 mumol/L) or with cyclic AMP-dependent kinase inhibitor H-8 (60 mumol/L). Phorbol ester PMA (phorbol 12-myristate 13-acetate), a potent activator of protein kinase C, induced a slight intracellular pH increase significantly smaller than that with epidermal growth factor, whereas this effect was completely blocked by pretreatment with H-7, indicating that PMA-induced intracellular pH increase is mediated by protein kinase C pathways, unlike epidermal growth factor.(ABSTRACT TRUNCATED AT 250 WORDS)

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1994

An ischemic opening of the blood-brain barrier may deteriorate brain stem auditory evoked potentials following transient hindbrain ischemia in gerbils.

To clarify the effect of vasogenic brain edema on the brainstem, the relationships between waveform changes in brainstem auditory evoked potentials (BAEP) and blood-brain barrier (BBB) disturbance following transient hindbrain ischemia were investigated. Hindbrain ischemia was induced in gerbils by bilateral occlusion of the vertebral arteries. The animals were divided into three groups subjected to 0, 5, and 30 min of bilateral vertebral occlusion (BVO-0',-5', and -30' groups; n = 4 in each group). Two hours after recirculation, Evans blue (EB) solution was injected into the saphenous vein. The brains were removed after 30 min of circulation, and all areas stained macroscopically by EB were noted and recorded. During hindbrain ischemia, BAEP disappeared within 3 min. In the BVO-5' group, BAEP reappeared and returned to normal within 10 min after reperfusion, whereas in the BVO-30' group, BAEP never returned to normal and finally disappeared within 30 min after reperfusion. In the BVO-5' group, no EB staining was visible. On the other hand, in the BVO-30' group, EB staining was seen in the medial part of the tegmentum in the midbrain in two animals, and around the vestibular nucleus in the lateral parts of the pons in three. These results demonstrate the close relationship between the reversibility of ischemia-induced changes in BAEP and BBB disturbance in the brainstem.

Animals↗

PubMed1994

Long-term clinical effect of nilvadipine in patients with chronic heart failure: a double-blind placebo-controlled study.

The long-term effect of calcium channel blockers on chronic heart failure is disappointing, probably because of reflex sympathetic activation through arterial vasodilation. However, nilvadipine may be beneficial for treatment of chronic heart failure since this drug has minimal effects on sympathetic activation. In this study, the effects of 12-week administration of nilvadipine or placebo on symptoms of heart failure and cardiac function were investigated in 23 patients with mild-to-moderate chronic heart failure in a double-blind trial. The patients were randomly assigned to either a nilvadipine group (16 mg daily) or a placebo group. Intergroup comparisons did not show significant differences in any parameters. Serious adverse effects were not observed during the study. Thus, this study failed to show any beneficial effect of nilvadipine in the long-term treatment of patients with chronic heart failure. We conclude that the long-term administration of nilvadipine (16 mg daily) is neither effective nor harmful in the treatment of patients with chronic heart failure.

Calcium Channel Blockers↗

PubMed1994

Predicting interferon therapy efficacy from hepatitis C virus genotype and RNA titer.

We classified 53 Japanese patients with chronic hepatitis C who were treated with natural interferon-alpha into genotypes and also tested the amounts of hepatitis C virus (HCV) RNA. The rate of the long-term complete response group, whose alanine aminotransferase levels remained within the normal range during the six months after therapy, was significantly higher (P < 0.01) in the type-III patients (4/5, 80.0%) than in type-II patients (4/43, 9.3%). For these long-term complete responders, the amounts of HCV RNA was less than 10(7) copies/ml serum in type-II patients, whereas two type-III patients with relatively high amounts of HCV RNA responded completely. These results confirm that the genotype of HCV is an important factor for predicting the response to interferon therapy. The amounts of HCV RNA can also predict its efficacy in type-II patients.

Adult↗

PubMed1994

Impaired gastric mucosal energy metabolism in congestive gastropathy in cirrhotic patients.

To clarify the characteristics of congestive gastropathy, we investigated gastric mucosal hemodynamics and energy metabolism in cirrhotic patients, using a reflectance spectrophotometry system and high performance liquid chromatography. The index of the gastric mucosal blood volume of cirrhotic patients with esophageal varices was significantly higher, and the index of gastric mucosal blood oxygenation significantly lower, than those in controls, thus indicating congestion and hypoxia in the gastric mucosa. Energy charge levels in the gastric mucosa of cirrhotic patients with esophageal varices were also significantly decreased. The energy charge level showed a strong linear correlation with the index of mucosal blood oxygenation in the antral (r = 0.996, P < 0.01) and body (r = 0.994, P < 0.01) mucosa of the stomach. These findings suggest that congestive gastropathy in a portal hypertensive state causes hypoxia in the gastric mucosa, leading to a mucosal energy deficit that may increase mucosal susceptibility to aggressive factors.

Chromatography, High Pressure Liquid↗

PubMed1994

Evaluation of thoracic aortic atherosclerosis by transesophageal echocardiography.

Transesophageal echocardiography (TEE) provides excellent images of the thoracic aorta, which cannot be visualized by transthoracic echocardiography. The purpose of this study was to assess atherosis and sclerosis of the thoracic aorta by TEE, to evaluate the risk factors for atherosis and sclerosis, and to assess the relationship between the two components of atherosclerosis. The mean value of the maximum thickness of the intima-media complex in the six segments of the thoracic aorta (MIMC) was used as an index of atherosis, and the stiffness parameter beta was used as an index of sclerosis. The study population consisted of 88 Japanese patients. Multivariate analysis showed that age, low-density lipoprotein cholesterol (or apolipoprotein B), and diabetes mellitus were significantly and independently related to MIMC, whereas age and hypertension were related to the stiffness parameter beta. Both components of atherosclerosis demonstrated a significant although weak relationship. The risk factors for atherosis appear to differ from those for sclerosis in the thoracic aorta, so we should evaluate these two components of atherosclerosis separately. TEE is a useful method of assessing thoracic aortic atherosclerosis because both atherosis and sclerosis can be examined simultaneously.

Adult↗

PubMed1994

Myocardial salvage: its assessment and prediction by the analysis of serial myocardial contrast echocardiograms in patients with acute myocardial infarction.

It has been difficult to assess myocardial salvage in patients with coronary reflow because of the lack of appropriate methods of determining the risk area and assessing effects of coronary reflow in patients, myocardial contrast echocardiography was performed in 28 patients with acute myocardial infarction before reperfusion, immediately after reperfusion, and in the chronic stage with the right and left coronary arterial injection of sonicated ioxaglate. Contrast-deficit and contrast-filled areas before reperfusion were defined as the risk area and noninfarct area, respectively. If the ratio of peak subtracted gray level in the risk area to that in the noninfarct area was < 0.4, the risk area was taken as a contrast defect. Contrast defect was observed even after reperfusion in 8 (29%) patients, and the defect was consistently observed in the chronic stage in all of them. Contrast defect disappeared after reperfusion in the other 20 patients but reappeared in 4 (20%) of them in the chronic stage despite the patent infarct-related vessel. Left ventricular function recovery of the risk area in the chronic stage as assessed with regional wall motion and wall thickness was better in the patients without contrast defect after reperfusion than in patients with persistent or reappeared contrast defect. In conclusion, (1) myocardial salvage is improbable in patients with contrast defect immediately after reperfusion, (2) contrast enhancement immediately after reperfusion does not necessarily imply myocardial salvage in the chronic stage, and (3) myocardial echocardiography in the chronic stage may provide clinically useful information about myocardial salvage in patients with myocardial infarction.

Adult↗

PubMed1994

Precise positioning of intracranial small tumors to the linear accelerator's isocenter, using a stereotactic radiotherapy computed tomography system (SRT-CT).

RT-CT was developed as a simulator using CT scan for radiotherapy. Following three-dimensional treatment planning using CT images, the treatment center and treatment fields are projected to the patients' surface by laser beam on the C arm. The prototype had an accuracy of 3 mm which was equivalent to conventional X-ray simulators but was not adequate for use in stereotactic radiotherapy. A new RT-CT system was developed to have a precise localization capability for stereotactic radiotherapy. Using this stereotactic RT-CT (SRT-CT) after three-dimensional planning, the treatment center is projected to the stereotactic frame automatically. In this study, the values of the x, y and z coordinates of the target center determined by SRT-CT are compared with those determined by the traditional method using CT localizing plates. The discrepancies were within 1.0 mm in 90% and 1.5 mm in 100% of 30 measurements in 16 patients. The disadvantages of SRT-CT may be that the accuracy of localization depends on the quality of calibration of laser beams. The traditional CT localizing method has superiority over SRT-CT because of its solid coordinates but its accuracy is vulnerable to alignment of CT fiducial marker plates. Therefore, the SRT-CT and traditional CT localizing methods would be complementary to each other for precise localization.

Brain Neoplasms↗

PubMed1994

Platelet activation in diabetic patients with asymptomatic atherosclerosis.

We studied 27 non-insulin-dependent diabetics without apparent atherosclerosis (AS) to investigate whether abnormal platelet function is related to asymptomatic atherosclerosis in diabetes mellitus. The degree of AS was quantitatively evaluated by determining the intimal plus medial thickness (IMT) of the carotid artery wall with ultrasound high-resolution B-mode imaging. Based on our previous finding that the upper threshold of the IMT was 1.1 mm in healthy subjects, the patients were divided into the AS-positive group with the IMT > 1.1 mm, (n = 17) and the AS-negative group with the IMT < 1.1 mm (n = 10). Among five variables measured as the factors concerned with thrombogenesis, only plasma levels of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4) were significantly higher in the AS-positive group than in the AS-negative group. Chronic administration of pentoxifylline (300 mg/day) significantly reduced the abnormally high plasma levels of beta-TG and PF4 in 7 patients of the AS-positive group to normal levels, without lowering the normal plasma beta-TG and PF4 levels in the remaining 10 patients. Pentoxifylline treatment did not affect the plasma levels of the 3 other variables, von Willebrand factor, 6-keto prostaglandin F1 alpha and thromboxane B2. This study suggests that the progress of atherosclerosis in diabetes mellitus is associated with in vivo platelet activation and platelet activation does not occur in diabetics without carotid atherosclerosis. Pentoxifylline may impede the vicious cycle in which atherosclerosis is accelerated by platelet activation.

Aged↗

PubMed1994

Abnormal coronary flow dynamics at rest and during tachycardia associated with impaired left ventricular relaxation in humans: implication for tachycardia-induced myocardial ischemia.

OBJECTIVES: This study attempted to clarify the effect of ventricular relaxation abnormalities on coronary flow dynamics at rest and during tachycardia in humans. BACKGROUND: Ventricular relaxation abnormality has been demonstrated in animals to have an adverse impact on early diastolic coronary flow dynamics. However, this relation has not been established in humans. Even if the adverse effect were latent at rest, it might become evident during tachycardia because tachycardia reduces coronary flow reserve and facilitates the production of myocardial ischemia. METHODS: Doppler phasic left coronary flow velocity pattern was obtained at rest and during tachycardia in 23 patients without coronary stenosis. The time constant of left ventricular isovolumic pressure (tau) was used to assess ventricular relaxation. RESULTS: The time to peak flow velocity of the diastolic coronary flow wave was longer, and the fraction of the first third of diastolic coronary flow was smaller, in patients with a longer tau (r = 0.58, p < 0.01; r = -0.44, p < 0.05), indicating a close relation between early diastolic coronary flow dynamics and ventricular relaxation. Although rapid atrial pacing yielded an increase in the coronary flow velocity integral per minute in all patients, diastolic coronary flow velocity integral per minute increased in 9 patients with a normal (< or = 40 ms) tau at rest but decreased in 14 patients with a longer (> 40 ms) tau at rest. CONCLUSIONS: Impaired left ventricular relaxation was associated with decreased coronary flow in early diastole at rest and decreased coronary flow throughout diastole during tachycardia in patients without coronary stenosis. These findings may provide more insight into the mechanism of tachycardia-induced subendocardial ischemia in patients with impaired ventricular relaxation but without concomitant coronary stenosis.

Adult↗

PubMed1994

Analysis of transmural trend of myocardial integrated ultrasound backscatter for differentiation of hypertrophic cardiomyopathy and ventricular hypertrophy due to hypertension.

OBJECTIVES: This study was undertaken to differentiate hypertrophic cardiomyopathy from hypertensive hypertrophy using a newly developed M-mode format integrated backscatter imaging system capable of calibrating myocardial integrated backscatter with the power of Doppler signals from the blood. BACKGROUND: Myocardial integrated ultrasound backscatter changes in patients with hypertrophic cardiomyopathy; however, it is unknown whether ultrasound myocardial tissue characterization may be useful in differentiating hypertrophic cardiomyopathy from hypertensive hypertrophy. METHODS: Calibrated myocardial integrated backscatter and its transmural gradient were measured in the septum and posterior wall in 31 normal subjects, 13 patients with hypertensive hypertrophy and 22 patients with hypertrophic cardiomyopathy. The gradient in integrated backscatter was determined as the ratio of calibrated integrated backscatter in the endocardial half to that in the epicardial half of the myocardium. RESULTS: Cyclic variation of integrated backscatter was smaller and calibrated myocardial integrated backscatter higher in patients with hypertrophied hearts than in normal subjects, but there were no significant differences in either integrated backscatter measure between patients with hypertensive hypertrophy and those with hypertrophic cardiomyopathy. Transmural gradient in myocardial integrated backscatter was present only in patients with hypertrophic cardiomyopathy (5.0 +/- 1.8 dB [mean +/- SD] for the septum; 1.2 +/- 1.6 dB for the posterior wall). CONCLUSIONS: Hypertrophic cardiomyopathy and ventricular hypertrophy due to hypertension can be differentiated on the basis of quantitative analysis of the transmural gradient in integrated backscatter.

Acoustics↗

PubMed1994

Polymorphonuclear leukocytes-induced injury in hypoxic cardiac myocytes.

Growing evidence suggests that free radicals derived from polymorphonuclear leukocytes (PMNs) play an important role in myocardial ischemia-reperfusion injury. To elucidate the cellular mechanism by which activated PMNs exacerbate ischemic myocardial damage, we investigated the extent of cell injury, assessed by the morphological deterioration, free radical generation, and lipid peroxidation in mouse embryo myocardial cells coincubated with activated PMNs. The generation of PMN-derived free radicals was related to the extent of myocardial cell injury. When myocardial cell sheets were subjected to hypoxia and glucose-free media, myocardial cells were injured (cristalysis in the mitochondria and disruption of the sarcolemma) after adding various PMN activators, and the injury extended to the adjacent cells. Chemiluminescent emission and production of thiobarbituric acid-reactive substances in the coincubated cells increased markedly compared with myocardial cells or PMNs alone. The augmented lipid peroxidation coincided with the progression of myocardial cell injury. Catalase inhibited the myocardial cell injury by 52%, the chemiluminescence by 46%, and lipid peroxidation by 50%, whereas superoxide dismutase exhibited less pronounced inhibition. These results indicate that a chain reaction of lipid peroxidation in myocardial cells induced by PMN-derived free radicals closely correlates with membrane damage and contributes to the propagation of irreversible myocardial cell damage.

Animals↗