Biomedical subjects

E Konishi

Publications and source records attributed to E Konishi.

At least 37 records · Page 2Linked to original sources

PubMed1998

Nurses' attitudes towards developing a do not resuscitate policy in Japan.

Two questionnaire surveys are reported describing the attitudes of 127 Japanese nurses towards developing a do not resuscitate (DNR) policy. The background information features the Japanese health care situations: a lack of policies for end-of-life care decisions; frequent life-prolonging treatments initiated without the patient's knowledge or consent; ethical dilemmas confronting nurses in relation to such treatments; and the public's growing concern over end-of-life care. A hypothetical DNR policy was used in which a health professional asked patients about their decision regarding DNR. The respondents supported this model DNR policy as an ideal. However, they noted that this policy was at variance with Japanese culture. Recommendations for Japanese nurses emerged from the data. In order to make an impact on end-of-life decision making, nurses should take culturally appropriate actions: (1) teach patients about their rights; and (2) work with physicians to provide patients with accurate information about their condition.

Advance Care Planning↗

PubMed1998

[Predictability of Gleason score and reduction time (tau) of prostatic volume after castration for the prognosis of prostatic cancer].

PURPOSE: The efficacy of the reduction time (tau) after castration as a prognostic factor was examined by comparing to Gleason score. MATERIALS AND METHODS: The change of prostatic volume after castration was observed from the castration to 3 months after in 24 cases of prostatic cancer. Prostatic volume was examined by transrectal ultrasonography of the prostate. Survival curves was calculated by Kaplan-Meier method. Differences among survival curves were analyzed using Cox-Mantel test. RESULTS: tau had a close relationship to the prognosis of each case (Wilcoxon test: p < 0.05, Cox-Mantel test: p < 0.05). Gleason score had a weak relationship to prognosis (Wilcoxon test: N. S., Cox-Mantel test: p < 0.05). CONCLUSIONS: tau was efficient as prognostic factor compared to Gleason score.

Aged↗

PubMed1997

Poxvirus-based Japanese encephalitis vaccine candidates induce JE virus-specific CD8+ cytotoxic T lymphocytes in mice.

Recombinant Japanese encephalitis (JE) vaccine candidates based on a highly attenuated vaccinia virus (NYVAC-JEV) and a canarypox virus (ALVAC-JEV) were evaluated for their ability to induce specific antibodies and cytotoxic T lymphocytes (CTLs) in mice. Six- to eight-week-old male Balb/c mice that received one or two intraperitoneal inoculations with these JE vaccine candidates at a dose of 1 x 10(7) PFU per mouse produced neutralizing antibody and antibodies to the envelope (E) and nonstructural 1 (NS1) proteins as determined by radioimmunoprecipitation. Immunization with either of these vaccine candidates also induced JE virus-specific T lymphocytes that proliferated in response to stimulation with infectious virus and/or noninfectious viral antigens. Mice maintained detectable levels of neutralizing antibody and JE virus-specific memory T cells for at least 6 months after immunization with NYVAC-JEV and for 4 months after immunization with ALVAC-JEV. Cells induced to proliferate after stimulation with live virus contained specific CD8+ CTLs that lysed primary Balb/c mouse kidney cells infected with JE virus and P815 mastocytoma cells infected with a recombinant vaccinia virus expressing the premembrane (prM), E, and NS1 proteins. These CTLs also lysed P815 cells infected with vaccinia recombinants expressing prM and E, and those expressing E and NS1, but did not lyse P815 cells infected with a recombinant virus expressing only NS1, indicating that the CTLs mainly recognized E, but did not recognize NS1. These results demonstrate that both recombinant JE vaccines, NYVAC-JEV and ALVAC-JEV, induce JE virus-specific antibody and CTLs in mice.

Animals↗

PubMed1997

Particulate vaccine candidate for Japanese encephalitis induces long-lasting virus-specific memory T lymphocytes in mice.

We previously reported that extracellular particles (EPs) composed of premembrane (prM) and envelope (E) proteins were released from cells infected with recombinant vaccinia viruses encoding Japanese encephalitis (JE) virus prM and E genes. In the present study, EPs were evaluated for induction of JE virus-specific antibody and specific T lymphocytes in mice. Six- to 8-week-old male Balb/c mice were inoculated intraperitoneally once or twice (at a 3-week interval) with purified EPs containing 1 microgram of E without adjuvant. Neutralizing antibody was detected and spleen cells proliferated against JE viral antigen 3 weeks after the second immunization with EPs. Neutralizing antibody and JE virus-specific T lymphocytes were also detected 10 months after immunization with EPs containing 2 micrograms of E. Spleen cells obtained from EP-immunized mice and stimulated in vitro with live JE virus, expressed JE virus-specific cytotoxic activity. The cytotoxic activity was reduced by treatment with anti-CD3 antibody and complement. These results indicate that immunization with EPs induces long-lasting specific antibody and memory T cells in mice.

Animals↗

PubMed1997

Monoclonal antibodies multireactive with parasite antigens produced by hybridomas generated from naive mice.

Hybridoma clones producing IgM naturally occurring (natural) antibodies were generated from naive Balb/c mice and characterized for reactivity against parasite antigens. Ascites of mice injected with these hybridomas reacted with Toxoplasma gondii soluble antigen at levels approximately 1000-10,000 times higher than serum pooled from mice used for generating hybridomas as determined by a conventional enzyme-linked immunosorbent assay. Western blot analysis indicated that these monoclonal antibodies reacted with multiple antigen molecules of T. gondii with patterns similar to that of pooled mouse sera. These antibodies also reacted with multiple antigens of Plasmodium yoelii, Entamoeba histolytica, Ascaris lumbricoides, Toxocara canis, Trichuris vulpis, Fasciola hepatica, Schistosoma mansoni, and Dipylidium caninum. When one monoclonal antibody was absorbed with these parasite antigens, its reactivity with T. gondii antigen molecules was consistently reduced, independent of parasite species. These natural antibodies failed to kill T. gondii tachyzoites in vitro or to protect mice from lethal challenge with T. gondii. These results indicate that natural antibodies detected in sera of naive mice are secreted from certain B cell populations and that these antibodies are multireactive with parasite antigens and have low affinity.

Animals↗

PubMed1997

Dengue hemorrhagic fever in Jakarta, Indonesia in 1988: isolation of dengue virus from patient whole blood using cell cultures.

During an outbreak of dengue hemorrhagic fever (DHF) in Jakarta, Indonesia in 1988, we attempted to isolate dengue virus using mosquito cells and a medium containing heparin. Whole blood, immediately after being drawn from patients, was inoculated into Aedes albopictus cell cultures temporarily maintained in the heparin-containing medium. The overall virus isolation rate was 25% (17 of 69) samples collected within three days after admission of the patients to hospital. No virus was obtained thereafter. The successful virus isolation was apparently not related to titers of anti-dengue virus hemagglutination-inhibiting antibodies present in patients' sera. The viruses were recovered from cases of each of the four World Health Organization grades of DHF without significant differences. The technique is simple and easily performed at bedside.

Aedes↗

PubMed1997

Prevalence of antibody to Toxoplasma gondii in Hyogo Prefecture, Japan: comparison at a 10-year interval.

We have previously surveyed antibody to Toxoplasma gondii among Japanese populations. In this report, we studied the current situation of antibody prevalence and compared it with the situation approximately 10 years before. A total of 1,800 human serum samples were collected at the Central Laboratory of Kobe University Hospital from June through September, 1995, and were examined for Toxoplasma antibody using enzyme-linked immunosorbent assay. The overall prevalence was found to be 16.4% with similar seropositivities in males (16.8%) and females (15.9%). Prevalence of antibody increased with age, reaching 41.0% in males and 59.0% in females at 80 years or more. Comparison with data obtained at the same hospital in 1981 to 1982 indicated that seropositivities at 30-39 years significantly decreased in both males and females. A positivity currently obtained at a certain age group was similar to a positivity previously obtained at the corresponding age group (10 years younger), indicating that the previous age prevalence curve shifted to the current position. A serological survey among stray cats collected in Hyogo Prefecture in 1990-1991 indicated that 44 (19.0%) of 231 cats were positive for Toxoplasma antibody as determined by the latex agglutination test. These results suggested less transmission of Toxoplasma to humans in Hyogo Prefecture at least for the last 10 years comparing with the situation in the past, although the transmission cycle among cats was still active in nature.

Adolescent↗

PubMed1996

Enzyme-linked immunosorbent assay using recombinant antigens for serodiagnosis of Japanese encephalitis.

Recombinant Japanese encephalitis (JE) virus proteins were evaluated as antigens for serodiagnosis of JE using an enzyme-linked immunosorbent assay (ELISA). The premembrane/membrane (prM/M) and envelope (E) proteins of JE virus were expressed in HeLa cells infected with a recombinant vaccinia virus that encodes the JE virus prM and E genes and were released from cells in a particulate form. The particulate antigens were partially purified from culture fluid from the infected cells by precipitation of particles with polyethylene glycol and then dissociated from the particles with 0.1% Triton X-100. This antigen preparation was used to evaluate one preimmune and two postvaccination sera from 20 volunteers given three inoculations of the commercial JE vaccine (Biken vaccine) by a conventional ELISA. The results from this assay correlated with neutralization data. The results of an IgM capture ELISA carried out with the recombinant antigen also correlated with the results of an existing IgM capture ELISA performed with JE virus-infected mouse brain, when tested with 29 serum and 13 cerebrospinal fluid samples from JE patients. These results indicated that recombinant JE virus antigens are useful for ELISA as an antigenically equivalent, highly productive, and safe alternative to authentic JE virus antigens.

Animals↗

PubMed1996

Characterization, molecular cloning and expression of megakaryocyte potentiating factor.

We examined whether the conditioned media of 64 kinds of cell lines, which have been maintained by a protein-free culture system, could produce megakaryocyte potentiating (Meg-POT) activity. In these cell lines, HPC-Y5, established from human pancreatic cancer, was shown to have the highest level of activity. The megakaryocyte potentiating factor (MPF) was purified from its conditioned medium by a combination of ion-exchange chromatography, gel filtration and reversed-phase HPLC. The purified MPF showed Meg-POT activity almost equal to human (Hu) interleukin 6 (IL-6) in the presence of murine IL-3 in a colony-forming assay with mouse bone marrow cells. The molecular weight of MPF was estimated to be 33 kDa by SDS-PAGE. Glycopeptidase F digestion and amino sugar analysis of the factor demonstrated that MPF is a glycoprotein carrying at least one N-linked sugar chain. The N-terminal amino acid sequence of MPF was determined to be Leu-Ala-Gly-Glu-Thr-Gly-Gln-Glu-Ala-Ala-Pro-Leu-Asp-Gly-Val-Leu-Ala-Asn. The same or homologous amino acid sequence has not been found in known proteins, demonstrating that MPF may be a novel cytokine which has Meg-POT activity. Then, we isolated HuMPF cDNA from an HPC-Y5 cDNA library using polymerase chain reaction and plaque hybridization methods. The HuMPF cDNA encodes a polypeptide consisting of 622 amino acids, including a signal peptide of 33 amino acids, and with a deduced molecular weight of 68 kDa, although HPC-Y5 cells secrete a 33 kDa form of HuMPF. HuMPF cDNA does not show any significant homology with other known sequences. The cDNA was expressed in COS-7 and Chinese hamster ovary (CHO) cells, and Meg-POT activity was detected in their culture supernatant. The COS-7 cells secreted only a 33 kDa recombinant (r)HuMPF, however, an additional 30 kDa form was detected in the culture medium of CHO cells. The 33 kDa rHuMPF from CHO cells showed Meg-POT activity, but not the purified 30 kDa rHuMPF. The difference in structure and activity between the 33 and 30 kDa forms of HuMPF was ascribed to the existence in the 33 kDa form of the C-terminal 25 amino acid residues. The expression of MPF mRNA was examined by Northern blot analysis using labeled MPF cDNA as a probe. MPF mRNA was detected in HPC-Y5 cells, with an approximate molecular size of 2.4 kb. We also examined the expression of the MPF gene in various human tissues, and the 2.4 kb band was detected only in lung. Then, the immunohistocytochemical analysis and in situ hybridization revealed that MPF-producing cells were identified as lung macrophages. MPF may exhibit other biological activities such as regeneration of the lung tissues.

Amidohydrolases↗

PubMed1996

Pharmacokinetics of KRM-1648, a new benzoxazinorifamycin, in rats and dogs.

The pharmacokinetics of 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl) benzoxazinorifamycin (KRM-1648) in rats and dogs given a single oral dose of 3, 30, or 100 mg/kg of body weight were studied. In the rats, the concentrations of KRM-1648 in plasma, whole blood, and tissues peaked between 2.0 and 24.0 h, with elimination half-lives ranging from 6.2 to 19.5 h. The peak concentrations and the areas under the concentration-versus-time curves (AUC) for whole blood and tissues were 2 to 277 times higher than those for plasma. The high levels of KRM-1648 in tissues were consistent with its large volume of distribution (in excess of 10 liters/kg). A nonlinear increase in peak concentrations and AUCs for plasma, whole blood, and tissues occurred as the dose was increased and was consistent with the dose-dependent decrease in bioavailability. In the dogs, KRM-1648 levels in plasma and whole blood also exhibited a late time to the peak concentration (ranging from 4.0 to 11.2 h), a long elimination half-life (ranging from 15.2 to 24.0 h), and nonlinear kinetics. KRM-1648 exhibited high levels of plasma protein binding (more than 99%) and a high degree of affinity for lipoproteins in the plasma of both animals. After administration of KRM-1648, measurable levels of its metabolites, 25-deacetyl KRM-1648 in rats and 25-deacetyl KRM-1648 and 30-hydroxy KRM-1648 in dogs, were found in the biological samples tested. Thus, KRM-1648 is characterized by a high tissue affinity, a long elimination half-life, and nonlinear pharmacokinetics.

Animals↗

PubMed1996

Detection of Plasmodium vivax and Plasmodium falciparum circumsporozoite antigen in anopheline mosquitoes collected in southern Thailand.

During a 13-month study on the ecology of malaria vectors in five villages in southern Thailand, Anopheles specimens collected on human-bait, bovid-bait, and in light traps were tested for the presence of Plasmodium vivax and P. falciparum circumsporozoite antigen by enzyme-linked immunosorbent assay. Plasmodium vivax antigen was detected in seven specimens and P. falciparum in 21 specimens, together representing 0.4% of the 7,938 specimens tested. In one village, Palao-U, circumsporozoite antigen was detected in 16 (0.7%) of the 2,196 specimens tested. In this village, combined rates of infection with P. falciparum and P. vivax were 0.6% for An. minimus, 1.1% for An. sawadwongporni, and 1.5% for An. maculatus. Circumsporozoite antigen was also detected in An. dirus, An. nivipes, An. barbirostris group, and An. hyrcanus group specimens. Combined P. falciparum and P. vivax entomologic inoculation rates in the wet season (March-October) were 0.05 for An. minimus, An. maculatus, and An. dirus, but 0 for An. sawadwongporni. Rates were higher in the dry season (November-February): 0.26 for An. minimus, 0.13 for An. maculatus, 0.13 for An. sawadwongporni, and 0 for An. dirus. The vectorial capacity, calculated based on human biting rate and rate of survival, of An. minimus during the dry season was more than two-fold higher than that of An. maculatus, the species with the second highest vectorial capacity.

Animals↗

PubMed1996

[Occurrence of numerical aberrations of chromosomes in metachronous liver metastases of colorectal cancers].

To investigate the change of numerical aberrations of chromosome 17 or 18 during progression of Colorectal carcinoma, we applied FISH, using chromosome specific probes, to 3 colorectal primary carcinomas and a total of 4 metachronous metastatic lesions in the liver. We also investigated the relationship between the DNA content of whole nucleus and the number of chromosome 17 by multiparametric analysis using autostage cytofluorometry. When compared with the results for primary lesions, in 4 metastatic lesions, the population of tetrasomy 17 cells increased in 2 lesions, and that of monosomy 18 cells increased in 1 lesion. The nuclear DNA contents histograms for disomy 17 and aneusomy 17 cells analyzed individually for 3 primary and 4 metastatic lesions, showed almost no appreciable difference of peak DNA values. These results indicate that the numerical aberrations of chromosome 17 and 18 accumulate during the progression of colorectal carcinomas, and that the subpopulation of aneusomy 17 consist mainly of those cells that involve the numerical aberrations in a few chromosomes.

Chromosome Aberrations↗

PubMed1996

[Clonal analysis of hepatocellular carcinoma].

We investigated the cell clonality of 12 cases of female solitary hepatocellular carcinoma (HCC) that were associated with hepatitis virus infection. The clonal origin of HCC could be assessed by the method based on restriction fragment length polymorphism (RFLP) of X-chromosome-linked androgen receptor gene (AR) and phosphoglycerate kinase (PGK) gene, taking advantage of random inactivation of one of two X-chromosomes by methylation in females. We extracted DNA samples from both fresh and paraffin-embedded specimens of the same lesion as a source of DNA sample for polymerase chain reaction (PCR). Consequently, it was possible to use methylation-sensitive restriction enzymes and PCR to study differential methylation patterns among alleles of these genes for both DNA samples. The RFLPs of AR gene and PGK gene were found in eight of 12 cases and five of 12 cases, respectively. There were two cases which had no RFLPs in either AR gene or PGK gene. All cases of HCC which had RFLP in either AR gene or PGK gene demonstrated monoclonal origin of the tumor regardless of their histologic patterns.

Aged↗

PubMed1996

Observations on nocturnal biting activity and host preference of anophelines collected in southern Thailand.

Over a 13-month period, 5,127 adult female mosquitoes were collected on human bait during multiple collecting periods between 1800 and 0600 h in 5 villages in southern Thailand. There were marked differences in the biting activity of different species at each of the villages studied. Anopheles maculatus and Anopheles sawadwongporni were collected most often at dusk or in the first hours of darkness between 1800 and 2100 h. Although specimens of Anopheles dirus were collected consistently between 1900 and 0400 h, peak collections were made between 2000 and 2300 h. Anopheles minimus was collected consistently throughout the night without a clearly discernible peak. Of more than 2,400 Anopheles mosquitoes collected in CDC light traps, 133 specimens (5.4%) contained blood, nearly all of which was identified by enzyme-linked immunosorbent assay (ELISA) to be of bovine origin. Ten specimens contained blood from more than one host species.

Animals↗

PubMed1996

Seasonal abundance and parity rates of Anopheles species in southern Thailand.

Mosquitoes were collected with human bait, animal bait, and CO2-baited light traps during a 13-month period in 5 villages located along the Myanmar border in southern Thailand. Altogether, 11,608 adult females consisting of 21 anopheline species were collected. Abundance of species varied according to the village where collections were made. In one village that was studied most intensely, Anopheles minimus was the most commonly collected species, representing 55.6, 10.6, and 56.5% of the human bait, cattle bait, and light trap collections, respectively. In addition, An. minimum was abundant throughout the year, whereas Anopheles maculatus was most commonly collected in July and August coincident with the rainy season. Anopheles sawadwongporni, abundant at the beginning and the end of the rainy season, was most commonly collected in June. Anopheles dirus was most abundant in April; however, it represented only 3.6% of all mosquitoes collected that month. Of 813 females collected in Palao-U Village and dissected, 38.9% were parous.

Animals↗

PubMed1995

Distribution of Dermatophagoides mite (Acari: Pyroglyphidae) antigens in homes of allergic patients in Japan.

Dust samples collected in 61 homes of patients with mite allergies and 11 homes of non-allergic people as controls were examined for antigen levels of Dermatophagoides farinae Hughes and Dermatophagoides pteronyssinus (Trouessart) to reveal the current distribution of allergenic mites in homes in Japan. Patient homes had higher antigen levels than control homes in comforters and pillows but not in futons, carpets, tatamis and wooden floors. Samples with antigen levels of > or = 10 micrograms m-2 were more frequent in and around summer (approximately April-October) than other seasons of the year in most materials and the differences between patient and control homes in comforters and pillows observed in yearly totals was also observed during this period. Wooden structures showed higher antigen levels than concrete structures in comforters, futons, pillows, carpets and tatamis in patient homes. Mite contamination in patient homes in relation to environmental factors was discussed.

Adult↗

PubMed1995

A novel proteinase, glucagon-degrading enzyme, secreted by a human pancreatic cancer cell line, HPC-YO.

Sixty-four kinds of cell lines were examined as to their ability to degrade glucagon using conditioned-media obtained from their protein-free cultures. Two human tumor cell lines were shown to produce this activity, and the cell line, HPC-YO, established from a human pancreatic carcinoma was shown to produce the highest level of activity. The glucagon-degrading enzyme (GDE) was purified from HPC-YO conditioned-medium by a combination of ion-exchange, gel filtration, and hydroxylapatite column chromatographies. The purified GDE also degraded vasoactive intestinal polypeptide (VIP) and secretin, however, it did not cleave EGF, gastrin, insulin, somatostatin, substance P, neurotensin, or growth hormone. The molecular weight of GDE is 83,000, as determined on SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of GDE was blocked, and the five partial amino acid sequences obtained on lysyl-endopeptidase digestion were determined to be N-L-T-E-E-Y-D-V-S-D-G-E-I-E-L-L-Y-E-K, V-E-T-Y-Y-D-L-L-F-E-K, L-Y-W-F-L-D-E-A-K, S-N-S-T-S-Y-V-K, and Y-Y-A-S-T-S-Y-D-D-T-Y-K. The same or homologous amino acid sequences have not been found in known proteins, demonstrating that GDE is a novel peptidase that degrades the secretin family: glucagon, VIP, and secretin.

Amino Acid Sequence↗

PubMed1995

Japanese encephalitis virus-specific proliferative responses of human peripheral blood T lymphocytes.

The T lymphocytes play an important role in prevention and recovery from viral infections. To characterize T lymphocyte responses to Japanese encephalitis (JE) virus infections, we analyzed JE virus-specific T lymphocytes in peripheral blood mononuclear cells (PBMC) obtained from seven JE patients and 10 vaccinees who had received a formalin-inactivated, purified JE virus vaccine (Biken vaccine). These PBMC were examined for proliferative responses against live JE virus, a glutaraldehyde-fixed lysate of cells infected with JE virus, and extracellular particles (EPs; subviral membrane vesicles released from cells infected with recombinant vaccinia viruses encoding the JE virus premembrane and envelope proteins). Japanese encephalitis virus-specific T cell proliferation was demonstrated with PBMC from both patients and vaccinees after stimulation with infectious JE virus or the lysate of JE virus-infected cells. Proliferating PBMC included CD4+ T lymphocytes and CD8+ T lymphocytes in responses to either form of JE viral antigens. Responses to EPs were observed only with PBMC from some American vaccinees whose PBMC also responded to the virus and lysate. These results indicate that JE virus infection and immunization with an inactivated JE vaccine induce JE virus-specific CD4+ and CD8+ T memory lymphocytes that can be induced to proliferate by infectious JE virus and noninfectious JE antigens.

Antibodies, Viral↗