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Biomedical subjects

S Hashimoto

Publications and source records attributed to S Hashimoto.

At least 811 records · Page 45Linked to original sources

Mechanism of calcium ionophore and phorbol ester-induced T-cell activation. Accessory cell requirement for T-cell activation.

We examined the role of monocytes in T-cell activation induced by phorbol myristate acetate (PMA) and calcium ionophore ionomycin. Depletion of monocytes from peripheral blood mononuclear cells (PBMC) was associated with the loss of interleukin-2 (IL-2) production, IL-2 receptor (IL-2R) expression and proliferation, in response to either PMA or ionomycin. Addition of monocytes to highly purified T cells resulted in the complete reconstitution of IL-2 production, IL-2R expression and proliferation by PMA-stimulated lymphocytes. Exogenous IL-2, but not interleukin-1 (IL-1), could reconstitute the T-cell responsiveness. Addition of monocytes to highly purified T cells stimulated with ionomycin resulted in partial reconstitution of IL-2 production, IL-2R expression and proliferation. Similarly, the addition of exogenous IL-2 to ionomycin-stimulated T cells only partially reconstituted the response compared with PBMC. These results suggest that monocyte-T-cell interactions contribute to IL-2 production and IL-2R expression and are crucial events for PMA-induced T-cell proliferation. With ionomycin, monocytes play a role, in part, in inducing IL-2 production, IL-2R expression and proliferation. However, IL-2 is not a sufficient signal to induce T-cell proliferative response to ionomycin, suggesting that an IL-2-independent mechanism may exist in ionomycin-induced T-cell proliferation.

Cell Division↗

Vacuoles and vesicles in the rat junctional epithelium: a study with serial ultrathin sections.

To investigate whether the electron-lucent structures resembling vesicles and vacuoles in the rat molar junctional epithelium (JE) are in fact intracellular or extracellular, a study using serial ultrathin sections was carried out. In one series of experiments, the animals were not treated before the tissues were conventionally fixed; in another, anesthetized animals were administered horseradish peroxidase 20 min before the tissues were fixed. A large number of electron-lucent structures resembling vesicles and vacuoles were detected in both the peripheral and central cytoplasm of the JE localized at enamel and connective tissue sites. These were 70 to 800 nm in diameter and had a lucency similar to that of the extracellular space in untreated specimens fixed with conventional fixative. Serial ultrathin sectioning revealed that the electron-lucent structures gradually became part of the extracellular space in the following sections. There were also found in the middle portion of the cytoplasm in specimens pretreated with horseradish peroxidase. Numerous vacuole-like structures containing peroxidase-positive materials were found to be contiguous with the extracellular space. A small number of vesicles, also containing peroxidase-positive materials, did not appear in the previous or following sections. These results indicate that almost all electron-lucent structures resembling vesicles and vacuoles in the JE are located at the end of a long infolding, and are still in contact with the extracellular space.

Animals↗

Cholera enterotoxin production in Vibrio cholerae O1 strains isolated from the environment and from humans in Japan.

Vibrio cholerae O1 strains isolated from various sources in Japan over the years 1977 through 1987 were examined to confirm the presence or absence of the cholera enterotoxin (CT) gene and production of CT and to determine the kappa-phage type. The CT gene was detected in none of 225 isolates from natural waters but was present in all of the 10 isolates from environmental waters implicated in domestic cholera cases, in 64 strains (26.6%) of the 241 isolates from imported seafoods, in 43 strains (95.6%) of the 45 isolates from domestic cholera cases, and in 119 strains (93.7%) of the 127 isolates from imported cholera cases. The results suggest that the CT gene-positive strains of V. cholerae O1 have been imported into Japan through seafoods and/or by travelers. Sporadic cholera cases have resulted in contamination of the surrounding environment, but the CT gene-positive strains may not have persisted in natural waters to serve as a reservoir for epidemic cholera. The commercially available VET-RPLA kit (a latex agglutination kit for immunological detection of CT) detected production of CT in all of the CT gene-positive strains, indicating that there was no silent CT gene in the test strains. There was a strong correlation between the kappa-phage type and the presence or absence of the CT gene, suggesting a significant clonal difference between CT gene-positive and -negative strains. Five CT gene-negative strains isolated from imported cholera cases (travelers with mild diarrhea) induced a considerable amount of fluid accumulation in rabbit and/or suckling mouse intestines, indicating production of an enterotoxic factor(s) other than CT.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fastidious human adenovirus type 40 can propagate efficiently and produce plaques on a human cell line, A549, derived from lung carcinoma.

Human adenovirus type 40 (Ad40) cannot propagate in conventional established human cell lines such as KB or HeLa cells. However, it has been shown that Ad40 DNA replicates in KB18 cells which express Ad2 E1B genes, suggesting that Ad40 is defective in the E1B gene function in KB or HeLa cells. We show here that Ad40 can propagate and produce plaques on A549 cells which do not contain Ad E1B genes. Our experiments show that the levels of replication of Ad40 DNA and production of infectious Ad40 virus in A549 cells are the same as or higher than those in 293 or KB18 cells. Dot blot analysis shows that the levels of Ad40 E1A and E1B mRNAs expressed in A549 cells at early to intermediate times postinfection are at least 10-fold higher than those in KB or KB18 cells. Northern (RNA) blot analysis shows that large E1B mRNA species (approximately 24S to 26S) are synthesized prior to the onset of DNA replication in A549 cells. No E1B mRNA species are synthesized in KB or KB18 cells at early times postinfection, and no differences in the expression of E1B mRNAs are seen between KB and KB18 cells. The experiment suggests that A549 cells have a cellular factor(s) which activates Ad40 E1B mRNA synthesis and that the E1B mRNA synthesis helps Ad40 propagation. In contrast, Ad40 can propagate in KB18 cells by using Ad2 E1B gene products that are constitutively expressed in this cell line. Furthermore, this result shows that Ad40 cannot propagate in KB cells because of the failure in the expression of E1B genes at early times postinfection.

Adenovirus Early Proteins↗

Vascular viscoelasticity of perfused rat hindquarters.

To determine viscoelastic features of the rat hindquarters vasculature, we measured pressure-volume curves. Male Wistar rats were transected at the lumbar level, and the perfused hindquarters were oxygenated with a hollow fiber artificial lung. The blood volume was measured by counting 51Cr-labeled red cells led to a gamma counter through an extracorporeal circuit at a constant rate. With continuous monitoring of the venous pressure and circulating blood volume, saline was infused into the circuit from a venous branch for 5 min [1.2 +/- 0.3% (SD) of tissue weight] followed by a 10-min recovery phase. In the recovery phase, the venous pressure promptly declined to the preinfusion level, whereas the circulating blood volume decreased more slowly. This implied vascular stress relaxation of the hindquarters. Maxwell's viscoelastic model, consisting of a spring component and a viscous component, was applied to analyze the venous pressure-volume diagram. With a curve-fitting method, the calculated vascular compliance and relaxation time (a time constant of stress relaxation) were 1.31 +/- 0.14 ml.mmHg-1.kg-1 and 15.7 +/- 4.0 min (means +/- SE), respectively. The value of compliance of the hindquarters was smaller than those of visceral organs reported. In addition, the value for relaxation time suggests that the viscous response of the vasculature simultaneously overlaps change in blood volume due to extravascular fluid shift during the postinfusion period.

Animals↗

Renal adaptation to dietary phosphate deprivation: role of proximal tubule brush-border membrane fluidity.

With dietary phosphate (Pi) restriction, fluidity of renal proximal tubule brush-border membranes (BBM) and Na-dependent Pi transport (Na-Pi) are increased, suggesting that changes in BBM fluidity are critical for adaptation to Pi restriction. To test this hypothesis, the temporal relationship between Na-Pi transport and changes in BBM fluidity was assessed after Pi deprivation in rats. Renal cortex was obtained from rats fed either a 0.03% (-P) or a 0.6% (+P) Pi diet for 4 h or 7 days, and BBM were prepared. Na-Pi uptake by BBM was measured by use of rapid filtration, and BBM fluidity was assessed by use of the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH). After 4 h on the diets, Na-Pi uptake was 439 +/- 142 (SD) and 984 +/- 184 pmol.mg protein-1.5 s-1 in +P and -P, respectively (P less than 0.01, n, = 8). Na-dependent proline uptake was unchanged. DPH anisotropy and total cholesterol were similar between groups: 0.204 +/- 0.025 and 0.401 +/- 0.047 nmol/mg protein, respectively, in +P and 0.205 +/- 0.015 and 0.392 +/- 0.037 in -P (P greater than 0.05, n = 8-10). After 7 days, Na-Pi uptake was 841 +/- 291 in +P and 2,168 +/- 848 pmol.mg protein-1.5 s-1 in -P, P less than 0.01, n = 8. DPH anisotropy and BBM cholesterol were 0.175 +/- 0.019 and 443 +/- 132 nmol/mg protein, respectively, in +P and 0.162 +/- 0.020 (n = 8) and 341 +/- 128 (n = 3) in -P (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

[Systems analytical investigation of the effect of countermeasures against the AIDS epidemic among homosexuals].

In Japan, the AIDS (Acquired Immunodeficiency Syndrome) epidemic among haemophiliacs has been the most urgent issue, because of the large number of haemophiliacs with AIDS. However, after governmental approval of the production of heated coagulating agents in 1985 and 1986, the prime object of prevention against the AIDS epidemic shifted from transmission through coagulation agents to that through sexual contacts. In order to investigate the most appropriate countermeasures against the AIDS epidemic among homosexuals in Japan, the numbers of HIV (Human Immunodeficiency Virus)-infected cases and AIDS cases in the future were estimated, and changes of the future numbers of HIV-infected persons and AIDS cases by behavior of homosexuals were compared using a systems analytical method. The methods of estimation and comparison are similar to those of R. M. Anderson and others, using numerical analysis of a mathematical model consisting of differential equations. They assumed a closed homosexual group whose members seldom have contact with members of other homosexual groups, but we assumed an open homosexual group whose members are being infected by other groups, because this assumption was more appropriate to the situation of homosexual society in Japan. The results showed that the prevalence number of HIV-infected cases would be about 1,800, that the prevalence number of AIDS cases would be about 100 among 100,000 homosexuals at 20 years after the beginning of the AIDS epidemic, and that the most effective countermeasure was reducing the frequency of sexual contacts among members or taking prophylactic measures during sexual contacts. These prevalence numbers of HIV-infected cases and AIDS cases would be reduced to between 1/4 and 1/2 of the above-mentioned calculated values by promotion of that countermeasure.

Acquired Immunodeficiency Syndrome↗

Low-density lipoprotein uptake and cholesterol accumulation by cultured renal cells.

The uptake of low-density lipoprotein (LDL) and the accumulation of cholesterol were assessed in opossum kidney (OK) and Madin-Darby canine kidney (MDCK) cells. OK and MDCK cells were grown to confluency on Millicell well inserts. The uptake of human LDL across the apical and basolateral surfaces of OK and MDCK cells was assessed by the degradation of internalized (125I)LDL to trichloroacetic acid-soluble products. LDL uptake via the apical surface of OK cells increased linearly with LDL concentration, indicating nonreceptor-mediated uptake. In contrast, LDL uptake via the basolateral surface of OK cells and both apical and basolateral surfaces of MDCK cells followed a saturable pattern. In addition, (125I)LDL bound to the apical membrane of MDCK cells, but not to the apical membrane of OK cells, was displaced by heparin and by excess of unlabeled LDL. Exposure to LDL (100 mg/mL) resulted in an increase in total cholesterol content of OK and MDCK cells (23 and 18%, respectively). Most of the increase in total cholesterol content with LDL exposure resulted from increased free cholesterol content in MDCK cells and esterified cholesterol in OK cells. The differences in cholesteryl ester formation were consistent with the slower rates of (14C) oleate incorporation into cholesteryl ester and lower cholesterol esterifying activity observed in MDCK cells compared with that in OK cells. These results demonstrate that LDL uptake can be receptor or nonreceptor mediated, depending upon the renal cell type and the surface exposed to LDL, and that LDL exposure leads to increased cholesterol content in OK and MDCK cells.

Animals↗

Regulation of vascular compliance and stress relaxation by the sympathetic nervous system.

We measured the changes in central venous pressure (CVP) and circulating blood volume (CBV) in dogs consequent to fluid infusion under halothane anesthesia, and compared the CVP and CBV responses to those obtained after blocking the autonomic nervous system by total spinal anesthesia (TSA) and stimulating the alpha receptor with methoxamine (MTX). Under TSA, the change in CVP consequent to fluid infusion was less than that under halothane anesthesia, while with MTX, the change in CVP was larger than that under halothane anesthesia. The recovery time of CVP response toward the baseline level after the end of fluid infusion was fastest under halothane anesthesia, while the recovery time of CVP was two times longer under TSA and MTX. Based on the relationship between changes in CVP and BV, we quantified effective vascular compliance and stress relaxation using mathematical analysis. The effective vascular compliance increased to 13.3 +/- 3.2 ml.mmHg-1.kg-1 under TSA as compared to 5.6 +/- 0.3 ml.mmHg-1.kg-1 under halothane anesthesia, and it decreased to 2.6 +/- 0.2 ml.mmHg-1.kg-1 with MTX. Stress relaxation was determined as the time constant in the unit response of CVP. The time constant for stress relaxation was 39 +/- 7 min under halothane, 74 +/- 12 min with TSA, and 92 +/- 25 min with MTX. These results suggest that the autonomic nervous system modifies cardiac preload by changing effective vascular compliance and stress relaxation.

Anesthesia, Spinal↗

Strategy for the diagnosis of small acoustic neuromas.

Twenty small (extra-meatal size less than 15 mm) acoustic neuromas have been diagnosed since high-resolution (HR) magnetic resonance imaging (MRI) became available in our clinic. Among these tumors, 18 had sensorineural hearing loss and 16 enlarged internal auditory meatus in X-ray photo, but only 8 tumors out of 18 tested showed diminished caloric response. Auditory brainstem responses (ABR) has been believed as the most reliable test for the diagnosis of acoustic neuroma despite several reports of false-negatives. In our series, 4 tumors out of 18 tested had normal ABR. The false-negative rate was 22%, which is much higher than expected. In CT, only 11 tumors were recognized. Although the total number is not large, present results clearly suggest the limits of these examinations. At present, HR-MRI is the most reliable diagnostic method for acoustic neuromas with no false-negative reported; ordinary MRI may have false-negatives. For the effective use of MRI, the results of audiometry, X-ray photo, ABR and other examinations must be evaluated properly.

Audiometry↗

Reciprocal expression of NF-IL6 and C/EBP in hepatocytes: possible involvement of NF-IL6 in acute phase protein gene expression.

The initial phase of inflammation is accompanied by dramatic changes in the concentrations of certain plasma proteins. Interleukin-6 (IL-6) is an important inducer of these acute phase proteins at the transcriptional level. The recently cloned nuclear factor NF-IL6, a potent trans-acting regulator of IL-6 gene expression, has a region that is highly homologous to the liver-specific transcriptional factor C/EBP. Both factors recognize the same nucleotide sequence. In this study the recombinant NF-IL6 was shown to interact with the IL-6-responsive elements (IL-6REs) identified in the promoter region of several acute phase protein genes whose activity increases during the acute phase reaction. Furthermore, in competition experiments, formation of all the DNA-protein complexes by the IL-6RE and IL-6-treated hepatoma cell extracts was specifically decreased by adding either the 14-bp NF-IL6 binding motif identified in the IL-6 promoter or the antibody against the recombinant NF-IL6. NF-IL6 was expressed at a minor level in mouse liver, but was dramatically induced after stimulation with IL-6. In contrast, the amount of C/EBP mRNA decreased considerably after IL-6 stimulation. These results indicate that the NF-IL6 that regulated IL-6 expression was also involved in regulation of expression of the acute phase protein genes. The ability of NF-IL6 to replace C/EBP may explain the positive and negative acute phase responses induced by IL-6.

Acute-Phase Proteins↗

[A method of interval estimation for number of patients in the nationwide epidemiological survey on intractable diseases].

A method for estimating the intervals for the number of patients with intractable diseases from nationwide epidemiological surveys was developed under the assumption that response is independent of frequency of patients. This method is based on data utilizing the number of response hospitals classified by their reported number of patients. The approximate 95% confidence intervals of numbers of several intractable diseases patients were estimated using this method.

Data Interpretation, Statistical↗

[Trends in length of stay of psychiatric inpatients based on "patient survey"].

The average length of occupancy for psychiatric beds in Japan has been pointed out to be among the longest in the world. However, neither the average length of occupancy since admission of psychiatric resident patients present at a facility at a given point of time (e.g. at the end of every October) nor the average length of stay of psychiatric patients until discharge have been reported in national health statistics. Using data from the "Patient Survey (1974-1984, 1987)" by the Ministry of Health and Welfare, estimates of trends for 1) length of hospitalization since admission of resident patients at the end of every October and 2) length of hospital stay from admission to discharge in patient cohorts admitted each year from 1974 to 1984, were determined. A remarkable difference in distribution was observed in the length of hospitalization in psychiatric residents and length of stay until discharge. For psychiatric residents surveyed at the end of October 1987, 13% had been resident for less than 3 months, 27% within one year, and 54% within 5 years. On the other hand, 33% of the patient cohort admitted in 1984 were discharged within one month, 59% within 3 months, and 85% within one year. Six percent of the cohort, however, had remained hospitalized for more than 3 years, some of whom were potential 'new' long-stay patients. The proportion of patients staying more than ten years shows an increase from 17% in 1974 to 30% in 1987, with the distribution of length of hospitalization in residents demonstrating a definite shift to increased length. On the other hand, length of stay until discharge shows a slight shortening.

Hospitals, Psychiatric↗

[Immunoscintigraphy of colorectal cancer with 111In labeled anti-CEA monoclonal antibody (ZCE-025)].

Clinical trials with 111In labeled anti-CEA monoclonal antibody (ZCE-025) was initiated. Five patients with colorectal cancer suspected were given an intravenous injection of 1 mg of 111In labeled ZCE-025. Planar and SPECT images were obtained 24 and 72 hours after injection. Surgical operation was performed on all patients between 7 and 10 days post injection. Of 4 primary sites, all were clearly visualized. Intrahepatic metastasis was visualized as higher activity than normal liver in one of two patients. In one patient whose imaging was negative, no residual cancer was found at surgery. Persistent accumulation of 111In in the lymph nodes was also observed in one patient. Surgical exploration of these lymph nodes showed no gross or microscopic evidence of metastases of colon cancer. No side effects were encountered, although HAMA were detected in all 5 patients by 4 weeks after the administration of ZCE-025. Immunoscintigraphy appears useful in distinguishing recurrent tumor from postoperative granuloma. Further investigation directed to the causes of 111In accumulation in tumor-free lymph nodes is required.

Aged↗

[Relation of baseline examination results to death from ischemic heart disease, cerebro-vascular disease and sudden death].

The relation of variables obtained from a baseline examination to death from ischemic heart disease (IHD), cerebro-vascular disease (CVD) and sudden death (SUD) was analyzed in a case-control study. From questionnaire survey of approximately 180,000 subjects who underwent baseline health examinations in 1971-1986 at Aichi prefectural center of health care, 148 deaths were selected for this study. The number of cases on IHD, CVD and SUD was 36, 60, and 52, respectively. Mean age of cases was 54.8 years old and the mean follow up interval between baseline examination and death was 3.7 years. Four controls matched according to year of baseline examination, age and sex were chosen arbitrarily for each case, and odds ratios for the three diseases were estimated. In some of the matched sets, odds ratios at a follow up examination were compared with that at the first examination. The results were as follows: 1) Variables showing positive relationships to death from each of the three diseases were hypertension, high fasting blood sugar, abnormality of cardio-thoracic ratio, ST-T abnormality in ECG, left ventricular hypertrophy in ECG. The odds ratio for ST-T abnormality in ECG was significant for all three causes of death. 2) High total cholesterol showed a significant positive relation only to death from IHD. As to death from CVD and SUD, albuminuria and sclerotic changes in fundus oculi were positively and significantly related. Risk factors differed for deaths from the three diseases. 3) In death from IHD and CVD, odds ratio at the second examination was apt to be higher than that at baseline examination. In death from SUD, however, odds ratios at the first and the second examination showed no significant difference.

Adult↗

[An autopsy case of neoplastic angioendotheliosis mainly affecting the lung and presenting with atypical cells in peripheral blood].

Neoplastic angioendotheliosis (NAE) is a rare condition characterized by proliferation of atypical cells in small vessels of various organs. In the past, atypical cells seen in this condition were considered to originate from vascular endothelial cells. However, recent immunohistochemical studies have established that the tumor cells in this condition mainly originate from B-lymphocytes. NAE is likely to affect the central nervous system and skin. Cases of NAE with a primary lesion in the pulmonary vessels are rare. One such rare case of NAE was recently encountered by us. The patient was a 77-year-old woman who was admitted to our department because of fever and interstitial shadow in both lungs. After admission, fever did not subside, and the pulmonary shadow became more marked. Laboratory tests upon admission disclosed elevated LDH. Of isozymes, LDH2 and LDH3 were high. Anemia and thrombocytopenia gradually became aggravated, and atypical cells with large basophilic cell body and irregularly shaped nucleus subsequently appeared in peripheral blood. Thereafter, the patient's general condition worsened rapidly and she died about 5 months after admission. Post-mortem examination revealed pleomorphic atypical cells filling the lumen of small vessels in various regions of the body. This finding was particularly marked in pulmonary vessels. With various immunohistochemical stains, these cells were identified as cells of B-lymphocyte origin.

Aged↗