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Biomedical subjects

K Kishi

Publications and source records attributed to K Kishi.

At least 289 records · Page 16Linked to original sources
PubMed1996

pH gradient electrophoresis of basic ribonucleases in sealed slab polyacrylamide gels: detection and inhibition of enzyme activity in the gel.

A simple method for the separation and specific detection of basic ribonucleases (RNases) was developed. The separation was achieved by polyacrylamide gel electrophoresis in a pH gradient generated by a carrier ampholyte (Pharmalyte 8-10.5) and arginine. In order to prevent interference from atmospheric carbon dioxide, the pH gradient was formed in sealed vertical gel slab. Human nonsecretory-type RNase, bovine pancreatic RNase A, and other basic proteins could be resolved without expensive equipment or complicated procedures. For activity detection after electrophoresis a zymogram technique was applied, using dry agarose film containing ethidium bromide plus RNA as substrate. This approach affords two advantages: (i) Basic RNase activities can be detected within 15 min, even in crude materials. The sensitivity is better than 0.5 ng of purified human nonsecretory-type RNase. (ii) An inhibition test of RNase activities in the gel, using human placental-type RNase inhibitor, can be performed.

Alkalies↗
PubMed1996

Alpha-2-HS-glycoprotein (AHSG) polymorphism in semen and saliva.

Polymorphism of alpha-2-HS-glycoprotein (AHSG) was demonstrated in human semen and whole saliva samples by thin-layer polyacrylamide gel isoelectric focusing (IEF) and immunoblotting. Although the seminal AHSG IEF patterns were found to differ from those of plasma AHSG from the corresponding donors, incorporation of Nonidet P-40 into the IEF gel (pH 4.2-4.9) enabled us to phenotype seminal AHSG correctly. Salivary AHSG, however, exhibited IEF patterns similar to those of the corresponding plasma AHSG. By treating the samples with neuraminidase, it was possible to determine the AHSG types using 2-5 microL semen and 50-100 microL whole saliva samples. The AHSG types determined separately in 47 sets of semen, whole saliva, urine and plasma samples from the same donors correlated perfectly with each other. AHSG typing could, therefore, provide an additional discriminant characteristic in the forensic examination of semen and saliva samples.

Blood Proteins↗
PubMed1996

Successful deoxyribonuclease I (DNase I) phenotyping from a small piece of used sock.

The forensic application of the DNase I polymorphism for individualization of used socks is described. We devised a technique that combines a simple extraction method, including a partial purification step, using a Phenyl-Sepharose CL-4B column, isoelectric focusing and activity staining, for DNase I typing from socks. As a result, DNase I types were determined from socks kept at room temperature for a month or more in all of the samples tested. The DNase I types determined from socks exactly matched those determined from corresponding sweat and urine samples. By contrast, AB0 groupings could not be determined from these samples, using the conventional serological method. Therefore, we have shown that the DNase I polymorphism is the first genetic marker found to be well suited for individualization of used socks.

ABO Blood-Group System↗
PubMed1996

Purification of mammalian ribonuclease using immobilized human ribonuclease inhibitor.

Affinity chromatography on immobilized ribonuclease (RNase) inhibitor was developed for purification of mammalian RNase. Human placental RNase inhibitor was conjugated to CNBr-activated Sepharose in the presence of dithiothreitol. About 80% of the immobilized RNase inhibitor was capable of binding bovine pancreatic RNase A. The bound RNase A was eluted with 3 M NaCl at pH 5.0. Two 25-kDa and 18-kDa RNases, which were obtained from human liver using a cellulose phosphate column, were bound to the immobilized RNase inhibitor and recovered in a pure and active form after treatment of the resin with p-hydroxymercuribenzoate. These enzymes were considered to be nonsecretory-type RNases with different sugar contents.

Amidohydrolases↗
PubMed1996

Histo-blood group Lewis genotyping from human hairs and blood.

The expression of histo-blood group Lewis antigens is determined by the Lewis-type alpha 1-->3/4fucosyltransferase (Le enzyme) encoded by Fuc-TIII gene (Le gene). The genotyping of Le genes by the PCR-RFLP methods established recently and partly modified in this study was found to be useful not only for determining the genuine Lewis blood types of samples such as human hairs and blood stains but also for distinguishing non-genuine Lewis-negative phenotypes frequently observed in pregnant women from genuine ones. The availability of the present PCR-RFLP methods for the paternity tests was also discussed.

Base Sequence↗
PubMed1996

Identification of the nucleotide substitution that generates the fourth polymorphic site in human deoxyribonuclease I (DNase I).

In addition to the three polymorphic sites responsible for protein polymorphism, a new polymorphic site has been identified in intron 7 of the human deoxyribonuclease I (DNase I) gene. Three phenotypes were observed on single-strand conformational polymorphism analysis of a 266-bp polymerase chain reaction-amplified fragment containing exon 7 and part of intron 7 of the human DNase I gene. DNA sequencing analysis demonstrated that a C-G substitution occurred at position 1978 in intron 7. This substitution was confirmed by restriction fragment length polymorphism analysis, since a new Msp1 site is created by the substitution. Population and family studies showed that the inheritance of the genotypes for DNase I C1978G polymorphism is controlled by two codominant alleles, tentatively designated DNASE1*1978C and *1978G. The gene frequencies in a Japanese population were significantly different from those in a Caucasian (German) population. The C1978G polymorphism is in linkage disequilibrium with the common DNase I protein phenotypes 1, 1-2, and 2.

Base Sequence↗
PubMed1996

Maltitol increases transepithelial diffusional transfer of calcium in rat ileum.

We explored the mechanism whereby maltitol causes an increase of calcium absorption in the lower small intestine using everted ileal segments of rats. Under the calcium concentrations tested (1-20mM), which enabled us to assess the diffusional calcium transfer, maltitol in the mucosal-side medium (100mM) caused a 155% greater transepithelial calcium transfer as compared with the segments incubated without sugars. The maltitol-induced increment of calcium transfer was significantly higher than those elicited by glucose, sorbitol and maltose, and this increase was completely inhibited by W7, a calmodulin antagonist. Thus, our results suggest that maltitol might modulate the permeability of calcium through paracellular path, which possibly involves the activation of calmodulin.

Animals↗
PubMed1996

Comparative immunophenotypic analysis of human mast cells, blood basophils and monocytes.

Mast cells (MC), blood basophils (Ba) and monocytes (Mo) are of haemopoietic origin. Lineage-relationships and transdifferentiation between MC and Mo, or MC and Ba, have been considered, based on common expression of antigens. In this study, comparative phenotypic analyses on MC, Ba and Mo and on respective cell lines were performed using monoclonal antibodies (mAb) to previously defined and novel CD antigens (CD1-130). By cluster analysis, the overall (all 130 CD) phenotypic relationships (given as similarity indices, SI), between primary cells (MC, Ba and Mo) and corresponding cell lines (HMC-1, KU-812, U937) were 0.716, 0.779 and 0.757, respectively. When primary cells were compared, lower SI values were found (MC versus Ba, 0.509; MC versus Mo, 0.625; Mo versus Ba, 0.698). More distant relationships were found between MC versus Ba and MC versus Mo, compared with Ba versus Mo, for adhesion receptor (R)-, complement R- and cytokine R profiles. Analysis of cytokine R revealed most significant dissimilarities between MC versus Ba and MC versus Mo (SI < 0.2). Moreover, in contrast to other CD subgroups and other lineages, MC and HMC-1 differed from each other in cytokine R expression (SI = 0.286). Cytokine R detectable on HMC-1 but not MC were granulocyte-macrophage colony-stimulating factor (GM-CSFR)alpha(CD116), CD40, Apo-1/FAS(CD95) and gp130(CD130). Cytokine R detectable on Ba but not MC, were interleukin-3 (IL-3)R alpha(CD123), IL-1RII(CD121b), IL-2R alpha(CD25) and CD40. In summary, MC, Ba and Mo display a unique CD profile with MC being the most distantly related cell. The most significant mismatch within a given lineage is the loss of cytokine R on mature MC as compared with normal myeloid progenitors and HMC-1 cells.

Antigens, CD↗
PubMed1996

Analysis of K-ras gene mutation in hyperplastic duct cells of the pancreas without pancreatic disease.

BACKGROUND & AIMS: We and others have previously shown that the mutation of K-ras codon 12 was found in the majority of pancreatic adenocarcinomas. The mutation has also been identified in the pancreatic duct with mucous cell hyperplasia in association with chronic pancreatitis. Ductal hyperplasia is also frequently found in the pancreas free from pancreatic carcinoma or chronic pancreatitis. The aim of this study was to assess the incidence and types of mutations in hyperplastic foci in these cases. METHODS: The nucleotide sequence of the K-ras gene at codon 12 of the DNA extracted from microdissected hyperplastic epithelium of the pancreatic duct obtained at autopsy in patients without pancreatic adenocarcinoma or chronic pancreatitis was analyzed. RESULTS: Of 38 patients with 79 hyperplastic foci, 12 patients (with 19 hyperplastic foci) had mutations. None of the 16 normal ducts in 12 specimens had this mutation. The nucleotide sequence of the codon in 53% of ductal hyperplastic foci was TGT or AGT, both of which were not found in 30 cases of adenocarcinoma. CONCLUSIONS: These results suggest that the ras gene mutation occurs frequently in multifocal hyperplastic foci of pancreatic duct and that the mutations may not have direct relevance to the carcinogenesis of pancreatic cancer.

Adult↗
PubMed1996

Geranylgeranylacetone induces heat shock proteins in cultured guinea pig gastric mucosal cells and rat gastric mucosa.

BACKGROUND & AIMS: An antiulcer drug, geranylgeranylacetone (GGA), rapidly induces resistance of gastric mucosal cells to irritants in vivo and in vitro. The aim of this study was to elucidate the mechanism of this action. METHODS: Heat shock proteins (HSPs) were detected by immunoblotting with antibody against HSP90, HSP70, heat shock cognate protein 70, or HSP60. HSP70 messenger RNA level was measured by Northern hybridization with an HSP70 complementary DNA probe. Activation of the heat shock factor was detected by gel mobility shift assay with the heat shock element oligonucleotide. RESULTS: GGA induced resistance of cultured guinea pig gastric mucosal cells against ethanol-induced exfoliation and damage within 30 minutes, proportionally to the induction of the HSPs. This protection was blocked by cycloheximide but not by indomethacin. GGA caused rapid activation of heat shock factor 1 and expression of HSP70 messenger RNA in the cells. Intragastric administration of GGA to rats induced HSPs in gastric mucosa. The administration of GGA additionally enhanced the heat shock response and reduced ulcer formation in rats subjected to restraint and water-immersion stress. CONCLUSIONS: GGA may induce transcriptional activation of HSP genes, and this novel action may increase gastric mucosal defense at conditions of stress.

Animals↗
PubMed1996

Case Report: amyopathic dermatomyositis associated with transformed malignant lymphoma.

Amyopathic dermatomyositis is a disease of unknown origin characterized by the specific skin lesions of dermatomyositis but without clinical or laboratory evidence of myopathy. During the past 15 years, a great controversy between the different reports concerning a possible association of dermatomyositis with malignancy has been noted. In this report, the authors describe a patient with amyopathic dermatomyositis who presented first with a benign hyperplasia of the lymph node, which finally transformed into frank malignant lymphoma. In addition to follow-up care, screening tests to search for occult malignancy in patients with amyopathic dermatomyositis (or dermatomyositis) are recommended.

Aged↗
PubMed1996

Differentiation induction therapy with all-trans retinoic acid (ATRA) in a patient with secondary 11q23 leukemia.

PURPOSE: Frequently recurrent leukemia is encountered in childhood and is usually resistant to multidrug therapy. We evaluated the efficacy of differentiation induction therapy in a child with leukemia. PATIENT AND METHODS: We administered all-trans retinoic acid (ATRA) alone, and then a combination of low-dose 1-beta-D-arabinofuranosylcytosine (Ara-C), to a 7-year-old boy who had leukemia secondary to etoposide with chromosomal abnormality 11q23. RESULTS: The patient exhibited a good response to ATRA and had a complete remission after combination therapy of ATRA and low-dose Ara-C. CONCLUSION: Differentiation induction therapy with ATRA was effective in treating a patient with recurrence of leukemia without t(15;17) translocation, and which is resistant to conventional cytotoxic treatment.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1996

High-dose chemotherapy with peripheral blood stem-cell transplantation for hormone-refractory advanced carcinoma of the prostate: experience of two cases.

Two patients with hormone-refractory advanced prostate cancer were treated with high-dose chemotherapy and peripheral blood stem-cell transplantation. A satisfactory number of stem cells were collected by appropriately timed leukapheresis in these patients. The stem-cell samples collected from the first patient were positive for prostate-specific antigen messenger ribonucleic acid expression; such expression was not detected in the samples from the second patient. The first patient has remained in complete remission as of December 1995. The second patient died of the disease.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1996

Presence of prostaglandin EP4 receptor gene expression in a rat gastric mucosal cell line.

RGM-1 is an epithelial cell line established from gastric mucosa of adult Wistar rats. In this study, we characterized this newly established cell line by Northern blot analysis. We also investigated deoxyribonucleic acid (DNA) synthesis and hexosamine production in RGM-1 by PGE2. Northern blot analysis did not detect any transcript of proton pump, gastrin receptor, histidine decarboxylase, somatostatin and pepsinogen 1, indicating the absence of characteristics of parietal, ECL, D and chief cells in RGM-1 cells. However, this periodic acid-Schiff (PAS)-positive cell line expressed prostaglandin EP4 receptor mRNA but not EP1 and EP3 receptor mRNAs. [3H]-thymidine incorporation into DNA of the cells was not increased by PGE2. In contrast, PGE2 increased hexosamine content in RGM-1 cells. These results suggest that RGM-1 may be a useful model of gastric mucosal cells and that PGE2 plays a role on mucin synthesis in RGM-1 cells possibly via EP4 receptors.

Animals↗
PubMed1996

Erythropoietin stimulates proliferation of rat-cultured gastric mucosal cells.

Most anemic patients with chronic renal failure have gastric mucosal lesions. However, these gastric lesions are often improved after the administration of recombinant human erythropoietin (rHuEPO). We have used the rat gastric mucosal cell line RGM-1, to examine the possibility that rHuEPO might directly stimulate the growth of gastric mucosal cells in vitro. Our results show that rHuEPO dose-dependently increased [3H]thymidine incorporation into RGM-1 cells and their expression of c-myc gene. In addition, 125I-rHuEPO specifically bound to RGM-1 cells, and moreover, erythropoietin receptor gene expression was detected by RT-PCR. We conclude that rHuEPO has a direct growth-promoting effect on RGM-1 cells, suggesting possible usefulness of rHuEPO administration for the treatment of gastric mucosal damage in patients with chronic renal failure.

Animals↗
PubMed1996

Induction therapy with all-trans retinoic acid for acute promyelocytic leukemia: a clinical study of 10 cases, including a fatal [correction of fetal] case with thromboembolism.

Ten patients with acute promyelocytic leukemia (APL) were treated with all-trans retinoic acid (ATRA). Eight of 10 patients achieved complete remission (CR), and among the 8 newly diagnosed cases, 7 achieved CR. Five of 8 CR cases remained in CR after 8 to 30 months. Except for hypotension and a large gastric ulcer resulting from hyperhistaminemia, the adverse effects of ATRA were generally mild. Severe thrombotic tendency occurred in a patient treated with ATRA combined with tranexamic acid. Intensive chemotherapy consisting of daunorubicin (DNR) and other agents was scheduled for the patients who achieved CR with ATRA.

Adult↗