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Biomedical subjects

H Tamura

Publications and source records attributed to H Tamura.

661 records · Page 37Linked to original sources
PubMedDate not supplied

Nuclear location of hormone-free estrogen receptors by monoclonal antibodies could be a tissue-fixation dependent artifact.

The 'two-step' model proposed by Jensen and his collaborators for explaining estrogen action conceptualized hormone-free estrogen receptors (ER) to be cytoplasmic, and hormone-filled, transformed ER to be nuclear. Applying monoclonal antibodies which recognized epitopes in ER and formaldehyde-fixed tissues, King et al demonstrated exclusively nuclear staining in target tissues utilizing immunoperoxidase technique. Recently these antibodies have become commercially available enabling other investigators to conduct studies. In this report, using these monoclonal antibodies we have demonstrated that a change in the concentration of formaldehyde alters the staining pattern yielding cytoplasmic instead of nuclear staining in calf uterus, MCF-7 cells, and ER(+) human breast cancer. In addition, neutralization of the antibody activity was not achieved with freshly prepared ER(+) cytosols. Formaldehyde-treated cytosols were essential. These results ought to caution investigators in determining in vivo location of antigens based on the staining pattern obtained in fixed tissues. Furthermore, this effect of formaldehyde on estrogen receptors may be applicable to other steroid hormone receptors.

Animals↗
PubMedDate not supplied

Processing of contrast polarity of visual images in inferotemporal cortex of the macaque monkey.

Cells in the anterior part of the inferotemporal cortex (anterior IT) respond to moderately complex stimulus features of object images. To study dependency of their responses on contrast polarity of stimulus images, we selected cells with optimal stimuli that were defined only by shape and not related to texture or color, and examined effects of reversing the contrast of the image or removing it except for edges between dark and bright parts of the image ("outlining"). The contrast reversal produced a reduction of the response to the optimal stimulus by > 50% in 60% of tested cells; the outlining, in 70%. When the two transformations were considered together, 94% of the cells showed a reduction by > 50%. Effects of the transformations on shape selectivity were also studied by comparing responses to several different shapes each of whose contours were expressed in different ways. Statistically significant changes in relative effectiveness of the different shapes as a function of contour expression were observed in more than half of the cells. These results suggest that responses of individual cells in anterior IT carry information about contrast polarity as well as about shape.

Animals↗
PubMedDate not supplied

Development of a stopped flow reactor for the rapid, high sensitivity measurement of endotoxin concentration in dialysate.

Continuous measurement of endotoxin concentration in dialysate, using a separated endotoxin-specific limulus reagent, promises rapid measurement without the complex operating procedures of the limulus reagent. To achieve high sensitivity measurements in a short period of time, an improved system featuring stopped-flow operation was developed. To prevent dispersion of the limulus reagent and residence of reacting solution containing the limulus reagent in the system reactor, the circuit in the reactor was changed from a coil configuration to a straight line, and its length was reduced. An endotoxin test solution was supplied at 760 microl/min, into which 40 microl of limulus reagent was pulse-injected. Flow was stopped at the point where the test solution entered the reactor. After the completion of the reaction, the solution was passed through a spectrophotometer and the relationship between reaction time and absorbance was determined. Peak tailing was less than that obtained by the conventional technique, good correlation was obtained from the peak height, and a decrease in sensitivity caused by broadening of the peak was suppressed. The lower detection limit of dialysate was 100 endotoxin units (EU)/L at a reaction time of 20 minutes, and 60 EU/L at 30 minutes. Change from the monitoring system to stop-flow operation made high sensitivity monitoring of endotoxin concentration with a short reaction time possible.

Animals↗
PubMedDate not supplied

Differential effects of neuropeptides on cytokine production by mouse helper T cell subsets.

Though immune outcome is known to be determined by which helper T cell response predominates, no local mechanism has yet been established which can explain how the neuronal system may control this. It is possible that the nervous system releases neuropeptides at specific local sites of infection or challenge, which triggers lymphocytes at those points to release specific cytokine profiles. These may then influence the direction of the Th1/Th2 response and therefore immune outcome. The aim of this study was to evaluate whether and if so how neuropeptides influence cytokine production by lymphocytes, especially T cells. We investigated the effects of neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), substance P (SP) and vasoactive intestinal peptide (VIP) on the production of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) by stimulating nonadherent splenocytes and helper T cell clones with antigens in vitro in the presence or absence of these peptides. NPY greatly enhanced IL-4 production and inhibited IFN-gamma. CGRP inhibited IFN-gamma production markedly in a dose-dependent manner, but had no effects on IL-4 production. SP and VIP had no effects on IFN-gamma production, but SP enhanced and VIP suppressed IL-4 production slightly but consistently. Therefore neuropeptides can influence cytokine production. This opens the door to speculations that these specific cytokine profiles might play a part in influencing the direction of the consequent Th1/Th2 cascade and immune outcome and possibly the pathogenesis of immune-related diseases.

Animals↗
PubMedDate not supplied

Cadmium effect on the Na,K-ATPase system in cultured vascular smooth muscle cells.

The present study focuses on the interaction between cadmium (Cd) and the Na, K-ATPase system in in vitro grown vascular smooth muscle cells (VSMCs) derived from the rat carotid artery. In disrupted VSMCs rendered permeable by osmotic shock, Cd inhibited Na, K-ATPase; I50 was reached at 10(-5) M Cd. Mg-ATPase was also inhibited by Cd; I50 was attained at concentrations of 10(-4) M Cd. Cd inhibition of Na,K-ATPase in the VSMCs was noncompetitive with respect to Na, K, and ATP. Rubidium transport experiments performed with intact VSMCs demonstrated that within an incubation period of 150 minutes, a concentration of 10(-4) M Cd in the extracellular fluid exerted no acute effect on the Na-K pump. Within this time interval, intracellular Cd attained a concentration eightfold higher than the extracellular Cd concentration. Thus, it appears that under acute conditions Cd exerts its inhibitory effect on Na, K-ATPase only in disrupted VSMCs. The data further suggest that, in the VSMC, conditions under which Cd inhibits Na, K-ATPase are consistent with inhibition from the cytoplasmic side of the cell membrane.

Animals↗
PubMedDate not supplied

Increased membrane permeability of skin fibroblasts from the spontaneously hypertensive rat.

Recently, we have demonstrated several abnormalities in Na+ and K+ homeostasis in cultured vascular smooth muscle cells derived from spontaneously hypertensive rats (SHR). To study whether similar defects can be identified in other cells of this rat strain, 86Rb and 22Na flux experiments as well as measurements of intracellular Na+ and K+ levels were performed in cultured skin fibroblasts of SHR and normotensive Wistar-Kyoto rats (WKY). The efflux rate constant (ke) for Rb+ (K+ analogue) was higher (p less than 0.001) in fibroblasts of SHR than in those of WKY (2.11 +/- 0.03 and 1.66 +/- 0.02 X 10-2/min; mean +/- SEM). The ouabain-insensitive influx rate constant (ki) for Rb+ was also higher (p less than 0.001) in fibroblasts of SHR than in those of WKY (13.26 +/- 0.41 and 10.71 +/- 0.27 X 10-2/min. On the other hand, the activity of the Na+-K+ pump of cells of SHR (44.81 +/- 0.81 X 10-2/min) was not different from that of cells of WKY (44.72 +/- 0.47 X 10-2/min). This parameter was obtained by calculating the ouabain-sensitive Rb+ influx rate constant. There was also no difference in the Na+ uptake (in the presence of ouabain) between cells of the two rat strains. Although there was no statistically significant difference in the measured intracellular total K+ levels between the two groups, on the basis of equilibrium distribution of 86Rb+, we calculated a significantly lower (p less than 0.001) level of exchangeable intracellular K+ in fibroblasts of SHR (98.2 +/- 1.2 mEq/L) as compared with cells of WKY (115.3 +/- 1.5 mEq/L). These findings indicate increased membrane permeability to K+ in fibroblasts of SHR and that this defect is likely to be innate to their membrane structure.

Animals↗
PubMedDate not supplied

Anemia and neutropenia due to copper deficiency in enteral nutrition.

Copper deficiency has been regarded a rare complication of total parenteral nutrition. This report describes the first known case of anemia and neutropenia caused by copper deficiency in a patient receiving long-term enteral nutrition. A 34-year-old man presented with bulbar palsy and tetraplegia after an operation for cerebellar hemorrhage on June 7, 1989. Nasopharyngeal tube feeding with a defined-formula diet (Besvion) was instituted on June 19, 1989. He developed normocytic anemia and neutropenia approximately 19 months after the initiation of the tube feeding. Serum copper and ceruloplasmin levels were markedly below normal. There were anisocytosis and erythrocyte deformities in the peripheral blood smear. Bone marrow aspirates revealed a hypocellular marrow with numerous myeloid and erythroid cells with cytoplasmic vacuoles. Because the amount of copper administered was estimated to be 2.6 to 5.1 mumol/d during the tube feeding, copper deficiency was suspected to be the cause of the hematologic disorders. The patient's daily oral copper intake was increased to 34 mumol/d. His serum copper and ceruloplasmin concentrations reached a normal level after 16 days and 23 days of copper supplementation, respectively. A marked reticulocytosis occurred after 10 days of copper supplementation, and his anemia gradually improved over the next 3 months. His blood neutrophil count also returned to normal within 2 weeks.

Adult↗
PubMedDate not supplied

The micronucleus assay with peripheral blood reticulocytes by acridine orange supravital staining with 1-beta-D-arabinofuranosylcytosine.

Dose-dependent induction of micronuclei with 1-beta-D-arabinofuranosylcytosine (ara-C) was clearly shown in CD-1 mouse peripheral blood reticulocytes (RETs) using an acridine orange (AO) supravital staining method, as well as in the conventional bone marrow assay. The maximum frequencies of micronucleated RETs (MNRETs) in peripheral blood and of micronucleated polychromatic erythrocytes (MNPCEs) in bone marrow were comparable, as shown in two laboratories independently. The maximum frequencies of MNRETs in peripheral blood lagged about 24 and 12 h behind those of MNPCEs in bone marrow in experiments with 24- and 12-h sampling intervals, respectively. The proportion of each type of RET was examined periodically after treatment with ara-C at doses ranging from 6.25 to 50.0 mg/kg. The proportion of type I RETs among total RETs decreased 24 or 48 h after treatment according to the dose level. This suggest that this ratio could be a good indicator of the bone marrow cell toxicity of test chemicals.

Acridine Orange↗
PubMedDate not supplied

Tear volume in relation to contact lens wear and age.

We examined the tear volume of the inferior conjunctival sac in relation to patient age and complications of contact lens wear using a special phenol red thread test. Records of patients who had undergone this test between 1984 and 1987 were reviewed. Based on the wet length of the thread, three groups were defined: 9 mm or less, 10-14 mm, and 15 mm or more. Those patients who showed consistent results over three or more separate testings were selected for this study. A total of 11,336 eyes met this criterion. A confidence interval of 95% was established as a basis for comparison of the groups. We found that the percentage of eyes with 9 mm or less of wet length increased with the age of the subject group. Likewise, the percentage of each age group with 15 mm or more of wet length steadily decreased among progressively older groups. Eyes wearing PMMA lenses with 9 mm or less of wet length showed the highest incidence of superficial punctate keratitis (SPK) and erosion. The group of eyes wearing high-DK lenses with 15 mm or more of wet length showed the lowest incidence of SPK and erosion.

Adolescent↗
PubMedDate not supplied

False-positive endotoxemia derives from gauze glucan after hepatectomy for hepatocellular carcinoma with cirrhosis.

BACKGROUND/AIMS: The purpose of this study was to evaluate the occurrence of false-positive endotoxemia after hepatectomy for hepatocellular carcinoma in patients with cirrhosis. The chromogenic conventional limulus test (Toxicolor test) revealed transiently increased blood endotoxin levels after hepatic resection for hepatocellular carcinoma in patients with liver cirrhosis. However, clinical signs of endotoxemia were not observed in all of the cases. METHODOLOGY: Pre- and postoperative changes of blood endotoxin levels and beta-glucan were measured in 20 patients with liver cirrhosis who underwent hepatic resection for hepatocellular carcinoma, using the Toxicolor test, the Endospecy test (which specifically reacts with endotoxin) and the Gluspecy test (which specifically reacts with beta-glucan). The changes in endotoxin levels and beta-glucan in ascitic fluid during surgery were also studied. RESULTS: The Toxicolor test revealed transiently increased blood endotoxin levels, but Endospecy test results were not elevated. The changes in the Gluspecy test were almost the same as in the Toxicolor test. During surgery, beta-glucan levels in the ascitic fluid increased remarkably due to their release from the surgical gauze. Digestion studies using 1-3-beta-D-glucanase on specimens which showed high Toxicolor and Gluspecy test values resulted in a lack of response for both tests. CONCLUSION: The cause of false-positive endotoxemia was determined to be caused by beta-glucan, which was released from the surgical gauze.

Ascitic Fluid↗