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Biomedical subjects

H Inoue

Publications and source records attributed to H Inoue.

At least 1,117 records · Page 62Linked to original sources
PubMed1994

MTT colorimetric assay system for the screening of anti-orthomyxo- and anti-paramyxoviral agents.

A rapid and sensitive method was developed for screening potential antiviral agents against orthomyxo- and paramyxoviruses, using the MTT method with cell culture suspensions. The cell lines used for the assay were as follows: MDCK cells for the influenza A virus (Fluv. A), HeLa cells for the respiratory syncytial virus (RSV), and Vero cells for the measles virus (MSV). Test compounds were diluted and plated in 96-well round-bottomed microtiter plates. Trypsinized cell suspensions and viruses were added to each well, the plates were then centrifuged (700 x g, 5 min, room temperature), and incubated for several days. The MTT assay was carried out after the degeneration of virus-infected cells became evident. The optical density (OD) of formazan was determined using a computer-controlled microplate reader. With this assay system, the EC50 values of Ribavirin (used as the reference compound) were 3.7 micrograms/ml for Fluv. A, 4.5 micrograms/ml for RSV, and 12.3 micrograms/ml for MSV, respectively. These EC50 values were equivalent to those obtained using the plaque reduction assay. The confluent cell culture system was inadequate for antiviral assays against RSV and MSV when the MTT method was used, because the inhibition of formazan formation was not observed in viral-infected cells. Moreover, the suspension method is more sensitive to the cytotoxicity of antiviral agents than the confluent cell culture system.

Amides↗
PubMed1994

Enhanced induction of lymphokine-activated killer activity following a single dose of cisplatin in cancer patients.

The effects of intravenous cisplatin (CDDP) administration on the generation of lymphokine-activated killer (LAK) activity in peripheral blood mononuclear (PBM) cells were investigated in cancer patients. The ability of PBM to generate LAK activity was significantly augmented 3, 5 and 7 days after a single dose, 50 mg m-2, of CDDP injection when compared to that before injection. NK activity of PBM was not altered. The distribution of lymphocyte subsets exhibited no significant change following CDDP injection, except CD2+ cells. However, the ability of monocytes in PBM to produce TNF-alpha was significantly enhanced 5 days after the drug administration, although IL-1-alpha and IL-1-beta production was not augmented.

Cisplatin↗
PubMed1994

Cystic radio-lucency of carpal bones in haemodialysis patients. An early indicator of the onset of carpal tunnel syndrome.

Patients receiving haemodialysis for more than 10 years were selected for this study in order to clarify an apparent sequential association of cystic lesions of carpal bones and carpal tunnel syndrome. X-rays and computed tomographs of 138 hands of 69 patients revealed cystic radiolucency of carpal bones in 35% of the hands. Radiographs were classified into three groups: Group A--cyst growing, Group B--cyst not growing, and Group C--cyst absent. The prevalence of carpal tunnel syndrome was 100% (27/27) in Group A, 5.6% (1/18) in Group B, and 6.5% (6/93) in Group C. Growth of the cyst precedes the development of carpal tunnel syndrome by about 2 or 3 years. Growth of the bone cyst indicates that inflammation had already extended to the tenosynovium and median nerve. Cystic radio-lucency of the carpal bones appears to be a useful indicator of the onset of carpal tunnel syndrome.

Adult↗
PubMed1994

Cubital tunnel syndrome in a patient in long-term haemodialysis.

The onset mechanism of cubital tunnel syndrome and carpal tunnel syndrome may be similar in haemodialysis patients. Carpal tunnel syndrome is well recognized as a consequence of dialysis-associated amyloidosis. This case report documents the development of cubital tunnel syndrome in a patient on haemodialysis treatment for 10 years. Proliferating granulation tissue at the elbow had entrapped and displaced the ulnar nerve. This was corrected surgically, and the patient experienced immediate relief of the numbness and the "tingling", but the muscular atrophy had not improved after 8 months.

Humans↗
PubMed1994

Malunited Colles' fracture. Analysis of stress distribution.

To try to explain disability in the wrist after malunited Colles' fractures, an experiment was designed to evaluate pressure distribution through the wrist joint with varying degrees of angulation of the distal radius. The results of computer simulation analysis of stress concentration using finite element methods were compared with those of pressure distribution studies on the radial articular surface of cadaveric radii in various positions using pressure-sensitive film. Stress existed in volar regions of the radio-lunate joint under normal pressure in the neutral position, but shifted to dorsal regions of the joint, and was concentrated when dorsal angulation reached 30 degrees. We believe that concentrated loads result in pain and may lead to early degenerative joint disease. Early additional reduction of a displaced fragment or a post-fusion osteotomy should be considered when dorsal angulation exceeds 30 degrees in malunited distal radial fractures to decrease abnormal loading across the wrist joint.

Adolescent↗
PubMed1994

Mutagenesis and epistatic grouping of the Neurospora meiotic mutants, mei-2 and mei-3, which are sensitive to mutagens.

To understand the possible roles of the Neurospora meiotic genes mei-2 and mei-3 in DNA repair, the frequencies of spontaneous and UV-induced mutation at the ad-3 loci were investigated and double mutants between mei mutants and other DNA-repair mutants were analyzed for mutagen sensitivity. Spontaneous mutation frequency in mei-2 was similar to that of the wild type, while the frequencies were high in both of two mei-3 strains which contained different mei-3 alleles. In addition, the frequency of spontaneous mutation in mei-3 varied greatly from experiment to experiment, which clearly showed a mutator phenotype for mei-3 mutation. UV irradiation increased mutation frequencies in both mei-2 and mei-3. The mutagen sensitivity of the double mutant, mei-2 mei-3, was no greater than that of the single mutants when treated with UV and methyl methanesulfonate (MMS). Thus, both mei genes appear to be involved in the same repair group. When analyzed in combination with other mutations, both mei mutations showed an epistatic relationship to uvs-6 and a synergistic relationship to mus-18 and uvs-2. However, uvs-3 was epistatic to mei-2, but the relationship of uvs-3 to mei-3 could not be tested directly, because the double mutant was barely viable. These results indicate that mei-2 and mei-3 are involved in the same repair group as uvs-6, and uvs-3 is possibly involved in the same group. Alternatively, the mei genes, or uvs-3 itself, may be related to more than one DNA repair group.

Crosses, Genetic↗
PubMed1994

Minimal increase in serum interleukin-6 levels during laparoscopic cholecystectomy.

The chronologic changes in the serum levels of interleukin-6 (IL-6), a mediator for acute-phase inflammation, were compared between laparoscopic cholecystectomy (LC) and open cholecystectomy (OC), since these two types of operations were considered to be a unique model for examining the role of local tissue injury in postoperative inflammatory reactions. The increase in the serum IL-6 level during LC was found to be significantly smaller than that during OC and resulted in a smaller extent of postoperative elevations for C-reactive protein. These results suggest that laparoscopic surgery associated with minimal tissue injury can help limit an increase in the serum IL-6 level during surgery, thus contributing to a reduction in surgical stress.

Biomarkers↗
PubMed1994

Coexistence of MALT-type lymphoma and squamous cell carcinoma of the larynx.

A 66-year-old Japanese man was diagnosed as having a MALT-type lymphoma by histopathological examination. The lesion involved the vocal folds bilaterally, occupying the larynx and extending beyond it, as shown by computed tomography (CT). A course of radiation therapy with moderate doses was given. Six months later, a squamous cell carcinoma was found in the larynx and total laryngectomy was then performed. The patient remained well, without recurrence, 46 months after the operation. thus, MALT-type lymphoma may coexist with a squamous cell carcinoma of the larynx.

Aged↗
PubMed1994

v-src transformation of rat embryo fibroblasts. Inefficient conversion to anchorage-independent growth involves heterogeneity of primary cultures.

To clarify whether a single oncogene can transform primary cells in culture, we compared the transforming effect of a recombinant retrovirus (ZSV) containing the v-src gene in rat embryo fibroblasts (REFs) to that in the rat cell line 3Y1. In the focus assay, REFs exhibited resistance to transformation as only six foci were observed in the primary cultures as opposed to 98 in 3Y1 cells. After G418 selection, efficiency of transformation was again somewhat lower with REFs compared to that with 3Y1 cells, but the number of G418-resistant REF colonies was much greater than the number of foci in REF cultures. Furthermore, while 98% of G418-resistant colonies of ZSV-infected REFs were morphologically transformed, only 25% were converted to anchorage-independent growth, as opposed to 100% conversion seen in ZSV-infected 3Y1 cells. The poor susceptibility of REFs to anchorage-independent transformation did not involve differences in expression and subcellular distribution of p60v-src, or its kinase activity in vitro and in vivo. It rather reflected a property of the primary cultures, as cloning of REFs before ZSV infection demonstrated that only 2 out of 6 REF clones tested were permissive for anchorage-independent growth. The nonpermissive phenotype was dominant over the permissive one in somatic hybrid cells, and associated with organized actin filament bundles and a lower growth rate, both before and after ZSV infection. These results indicate that the poor susceptibility of REFs to anchorage-independent transformation by p60v-src reflects the heterogeneity of the primary cultures. REFs can be morphologically transformed by p60v-src with high efficiency but only a small fraction is convertible to anchorage-independent growth. REF resistance seems to involve the presence of a suppressor factor which may emerge from REF differentiation during embryonic development.

Actins↗
PubMed1994

Self-assembling process of cylindrical virus coat proteins as observed by synchrotron small-angle X-ray scattering.

The self-assembly process of cucumber green mottle mosaic virus (CGMMV) protein and tobacco mosaic virus (TMV) protein was examined by the thermodynamic analysis of small-angle X-ray scattering (SAXS) data. Each polymerization step of the coat proteins was assumed to be specified by a single equilibrium constant, and the equilibrium constant was evaluated by fitting the size and shape of the constituents observed by SAXS to those calculated from an assumed polymerization scheme. The logarithmic plots of the equilibrium constant against the inverse of temperature were fitted with a straight line at each buffer concentration and the thermodynamic quantities were evaluated from its intercept (yielding entropy) and slope (yielding enthalpy). The enthalpy and entropy values of TMV protein were found to be independent of buffer concentration, whereas those of CGMMV protein depended strongly on buffer concentration. In the limit, as ionic strength tends to infinity, both the enthalpy and entropy values of CGMMV protein approach those of TMV protein. The higher negative surface charge of CGMMV protein is considered to be responsible for the formation of stable single-layered disks, and for the slow polymerization process even at higher temperature and higher buffer concentrations.

Biopolymers↗
PubMed1994

Shock-induced refractory period extension and pharmacologic modulation of defibrillation threshold.

Shock-induced refractory period extension (RPE) has been suggested as a mechanism of electrical defibrillation. We measured RPE caused by localized field stimulation measured before and during infusion of disopyramide (n = 5), flecainide (n = 5), or E-4031 (n = 5) in anesthetized dogs and determined the effect of the drugs on the internal defibrillation threshold (DFT). In the baseline state (n = 15), 16 V/cm S2 field stimulation prolonged the effective RP by 36 +/- 15 ms (22 +/- 12% of RP without S2), whereas 4 and 8 V/cm S2 stimuli did not cause marked RPE. The RPE normalized by the RP without S2 was not significantly influenced by any drug (16 V/cm: disopyramide 30 +/- 11 vs. 27 +/- 11, flecainide 25 +/- 5 vs. 19 +/- 12, and E-4031 18 +/- 13 vs. 22 +/- 14%). Disopyramide did not alter the defibrillation threshold (4.2 +/- 0.6-4.4 +/- 0.6 J). In 2 dogs given flecainide, ventricular fibrillation became refractory to defibrillation. In contrast, E-4031 lowered the threshold from 4.5 +/- 2.4 to 2.2 +/- 1.2 J (p < 0.01). The results suggest that flecainide and E-4031 do not modulate defibrillation efficiency through their effects on RPE.

Animals↗
PubMed1994

Attenuation of antifibrillatory effects of lidocaine by its metabolite, glycylxylidide: application of modulated receptor hypothesis.

Lidocaine, one of the drugs effective in treating ventricular arrhythmias in acute myocardial infarction (AMI), sometimes loses its efficacy after prolonged administration, possibly owing to the counteraction of glycylxylidide, one of the metabolites of lidocaine, through modulation of binding of lidocaine to sodium channels. To determine whether glycylxylidide interferes with the antiarrhythmic action of lidocaine, we compared the antifibrillatory effects of lidocaine, glycylxylidide, and their combination in 14 anesthetized open-chest dogs. Although glycylxylidide alone prolonged intraventricular conduction time (CT) and did not affect ventricular effective refractory period (VERP), it had different effects when added to lidocaine; i.e., it had no effect on intraventricular conduction time but shortened VERP. Although glycylxylidide alone did not change ventricular fibrillation threshold (VFT), the increase in VFT induced by lidocaine was decreased by addition of glycylxylidide, possibly as a result of competition for the same cardiac sodium channels between lidocaine and glycylxylidide with similar onset but different offset kinetics, which may explain, at least in part, the drug-resistance phenomena that ensue from prolonged lidocaine administration.

Animals↗
PubMed1994

Expression of the multidrug resistance gene (MDR1) in non-small cell lung cancer.

To ezamine the clinical relevance of P-glycoprotein, encoded by the human multidrug resistance gene (MDR1), to multidrug resistance in lung cancer, we examined the expression of MDR1 in 107 non-small cell lung cancer (NSCLC) specimens and 20 corresponding specimens of normal lung tissues. We also evaluated the relationship between MDR1 expression and the histopathology and pathological staging of NSCLC. The tumors consisted of 60 adenocarcinomas, 38 squamous cell carcinomas, 8 large cell carcinomas, and 1 adenosquamous carcinoma. MDR1 expression was semi-quantified by use of the reverse transcriptase-polymerase chain reaction method. We subclassified the NSCLC into 3 grades according to the MDR1 expression level (-, +, ++). Sixty-one of the 107 tumor specimens (57%) and 18 of the normal lung tissue specimens (90%) expressed various levels of the MDR1 gene. Only one tumor specimen showed higher MDR1 expression than the corresponding normal lung tissue. The relationship between pathological stage and MDR1 expression levels was not significant. These results suggest that the level of MDR1 expression in lung cells is decreased as cells progress from the normal to the transformed state.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1994

Lymphokine-activated killer cell function of peripheral blood mononuclear cells, spleen cells and regional lymph node cells in gastric cancer patients.

Lymphokine-activated killer (LAK) cells generated by culture of peripheral blood mononuclear cells (PBMC), spleen cells (SPC) and regional lymph node cells (LNC) with IL-2 for 4 days were examined for their functional capabilities in 29 patients with gastric carcinoma. The cytotoxic activity of LAK cells induced from LNC was significantly lower than that from either PBMC or SPC, although there was no difference between PBMC or SPC. The induction of mRNA of interferon-gamma (IFN-gamma) or tumour necrosis factor-alpha (TNF-alpha) and the production of these cytokines in the non-adherent LAK cells from LNC were also significantly reduced compared with those from PBMC or SPC. Further, the LAK cells from LNC secreted significantly lower levels of these cytokines when stimulated with tumour target, Raji cells, although the production of these cytokines was markedly increased by stimulation with the targets in all three cell populations. Phenotypic analysis of each cell population revealed a decreased proportion of the cells mediating natural killer (NK) activity, including CD16+, CD56+, and CD57+ cells in LNC either before or after culture, although OKIa1+ and CD25+ cells were uniformly increased in all cell populations after culture. Changes in subpopulations of CD4+ and CD8+ cells in LNC were not apparently different from PBMC or SPC. These results indicated the differential reactivity of each lymphocyte population to IL-2 and the reduced LAK cell function of LNC compared with PBMC or SPC in patients with gastric carcinoma.

Adult↗
PubMed1994

Analysis of the uveitogenic determinant in repeat structure of retinal interphotoreceptor retinoid-binding protein (IRBP).

IRBP is a glycolipoprotein with a four-fold partially homologous repeat structure approximately 300 residues in length, and is one of the retinal antigens capable of inducing experimental autoimmune uveoretinitis (EAU) in susceptible animals by their active immunization. The most immunopathogenic peptide of bovine IRBP for EAU in Lewis rats is reported to be the sequence 1169-1191 (PTARSVGAADGSSWEGVGVVPDV) with two immunogenic motifs common to T cell epitopes (underlined). The uveitogenic site of peptide 1169-1191 was localized at the carboxyl terminus (peptide 1182-1191) and not at the amino acid terminus (peptide 1169-1182). Repeat peptides of sequence 1179-1191 containing the four homologous residues (1182W, 1186G, 1187V and 1189P), that is the peptides 271-283, 579-591 and 880-892, all elicited EAU. Peptide 579-591 could not stimulate proliferation of lymphocytes from rats immunized with IRBP, but had the capacity to adoptively transfer EAU. The role of the homologous residues was examined using analogues of the uveitogenic peptide 1182-1194, in which each homologous residue was substituted by glycine (G) or leucine (L) (1182W-->G, 1186G-->L, 1187V-->G, and 1189P-->G). One analogue (1186G-->L) strongly diminished the ability to induce EAU, while the other three analogues completely abolished the ability, indicating that these homologous residues were essential for the induction of EAU. In addition, the uveitogenic peptides tested in this study were found not to contain the major epitope for antibody production.

Amino Acid Sequence↗
PubMed1994

Involvement of apamin-sensitive K+ channels in antigen-induced spasm of guinea-pig isolated trachea.

1. In order to examine whether K+ channels play a role in antigen-induced airway responses, the effect of K+ channel blockers on antigen-induced airway smooth muscle contraction and mediator release was examined in vitro in guinea-pigs actively sensitized with ovalbumin (OA). 2. Tracheal strips from sensitized animals were suspended in organ baths under a resting tension of 1 g and isometric tension was continuously measured. Cumulative concentration-response curves to OA (0.1-1000 ng ml-1) or histamine (10 nM-1 mM) were obtained in the presence and absence of K+ channel blockers. 3. OA (10, 100 or 1000 ng ml-1) was incubated with minced lung tissues from the same animals for 15 min in the presence and absence of K+ channel blockers, and released histamine and leukotriene C4 (LTC4) in the incubating medium were measured. 4. Apamin, a small conductance Ca(2+)-activated K+ channel (PK,Ca) blocker, (0.1, 0.3 and 1 microM) significantly inhibited OA-induced smooth muscle contraction, while charybdotoxin (ChTX, 10 nM), an intermediate and large conductance PK,Ca blocker, and iberiotoxin (IbTX, 3 nM), a large conductance PK,Ca blocker, were without effect. Apamin (0.3 microM) had no effect on exogenously administered histamine-induced airway smooth muscle contraction, suggesting that the inhibition of OA-induced contraction by apamin did not occur at the smooth muscle level. 5. The inhibition of OA-induced contraction by apamin (0.3 microM) was not significantly affected by pretreatment with a leukotriene antagonist, ONO-1078 (10 microM), but was abolished by pretreatment with a histamine H1-receptor blocker, pyrilamine (1 microM). 6. Apamin by itself (up to 0.1 MicroM) had no effect on spontaneous histamine release from minced lung tissues. Histamine release induced by low and intermediate concentrations of OA (10 and 100 ng ml-1)was significantly suppressed by apamin pretreatment (P<0.05 and P<0.001), whereas LTC4 release was not affected. ChTX (0.1 MicroM) and IbTX (10 nM) had no significant effect on either spontaneous or OA (100 ng ml-1)-induced histamine release.7. These results suggest that apamin partially but substantially inhibits antigen-induced smooth muscle contraction, presumably by inhibiting antigen-induced histamine release from airway mast cells through small conductance PKca closure.

Animals↗
PubMed1994

Immunochemical study of polysaccharide antigen in Streptococcus sobrinus and Streptococcus downei with a cross-reactive monoclonal antibody.

A monoclonal antibody (mAb h-448) was prepared after cell fusion of mouse myeloma cells (SP2/0-Ag-14) to the spleen cells of mice immunised with serotype h strain (MF25) of Streptococcus downei. The antibody (IgM class) reacted in enzyme immunoassay only with whole cells as well as purified polysaccharide (PS) antigen of Streptococcus sobrinus (types d and g) and Streptococcus downei (serotype h), but not with cells or purified PS antigen from any other serotypes of the mutans group of streptococci. mAb h-448 also quantitatively precipitated in solution with the purified antigens. Competitive hapten inhibition tests demonstrated that beta-methylgalactopyranoside inhibited the reaction most strongly. Although rhamnose also showed a substantial inhibitory effect, the results of this study indicate that the antigenic determinant of the PS antigen has a structure similar to the beta-methylgalactopyranoside molecule.

Antibodies, Bacterial↗