PubMed Health⌕ Search

Biomedical subjects

M Kouno

Publications and source records attributed to M Kouno.

At least 19 recordsLinked to original sources
PubMed1997

Characterization of porcine reproductive and respiratory syndrome virus hemagglutinin.

Porcine reproductive and respiratory syndrome virus hemagglutinin (HAin) was readily adsorbed on mouse erythrocytes at 4, 22, or 37 degrees C, but not on goose erythrocytes. The adsorbed HAin could not be eluted from the cells by resuspending in phosphate buffered saline, by incubating at 37 or 50 degrees C, or by incubating in the presence of neuraminidase. The hemagglutinating activity was not dependent on the pH and NaCl molarity tested. The receptor of mouse erythrocytes for the HAin was relatively stable to trypsin, neuraminidase, sodium deoxycholate (DOC), potassium periodate (KIO4), dithiothreitol (DTT), 2-mercaptoethanol (2-ME) and formalin treatments. The HAin was inactivated by 2-ME and was gradually inactivated by pepsin, formalin and DTT, but not by beta-glucosidase, trypsin, alpha-amylase, papain, phospholipase C, neuraminidase, KIO4, and ethylendiamine tetraacetic acid (EDTA) treatments. The HAin was stable at 37 degrees C or lower temperatures, but not at 56 degrees C or higher. The HAin was relatively resistant to ultraviolet irradiation and sonication. In the equilibrium centrifugation of the HAin preparation on a CsCl density gradient, the HAin activity showed a sharp peak at 1.17 g/cm2. In the SDS-PAGE analysis, the structural polypeptide of HAin in the peak fraction seems to be the nucleocapsid (N) polypeptide with molecular weight of 15 kDa.

Adsorption↗
PubMed1997

Effect of heparin on infection of cells by equine arteritis virus.

The number of plaques formed by equine arteritis virus (EAV) and Aujesky's disease virus (ADV) was reduced to 14% and 5% of the untreated control (100%), respectively, by 10 U/ml of heparin, but could not be reduced below to 13 and 4%, respectively, by use of concentration up to 100 U/ml. An inhibitory effect of heparin, at concentration up to 100 U/ml, was not observed on parainfluenza virus 3 (PIV-3). Heparinase treatment of RK13 cells reduced the number of EAV-, as well as ADV-induced plaques. On the other hand, the number of PIV-3 induced plaques did not decrease after treatment of RK13 cells with heparinase.

Animals↗
PubMed1997

Effect of heparin on infection of cells by porcine reproductive and respiratory syndrome virus.

OBJECTIVE: To investigate whether heparin has any effect on the growth of porcine reproductive and respiratory syndrome virus (PRRSV). SAMPLE POPULATION: 2 isolates of PRRSV, and as control viruses, 1 isolate of pseudorabies virus (PRV) and 1 isolate of parainfluenza 3 virus (PIV-3). PROCEDURES: Plaque assays, using a continuous cell line (MARC-145) derived from African green monkey kidney cell line (MA104), were performed for determination of inhibitory effect of heparin on PRRSV, PRV, and PIV-3. The effect of various doses of heparin and heparinase on the growth of PRRSV, PRV, and PIV-3 was evaluated and compared. In each experiment, value were expressed as the mean value for duplicate samples. RESULTS: The number of plaques formed by PRRSV and PRV was reduced to 24 to 25 and 15% of the untreated control (100%), respectively, by 1 U of heparin/ml, but could not be reduced below 6 to 7 and 3%, respectively, by use of concentrations up to 50 U/ml. An inhibitory effect of heparin, at a concentration up to 50 U/ml, was not observed on PIV-3. Delaying addition of heparin for 30 minutes after the addition of PRRSV and PRV reduced plaque formation by 48 to 51 and 68%, respectively, compared with 91 to 92 and 95%, respectively, if heparin was added at the time of infection. In addition, most PRRSV added was retained by heparin beads, as was PRV. Heparinase treatment of MARC-145 cells reduced the number of PRRSV-, as well as PRV-induced plaques. On the other hand, the number of PIV-3-induced plaques did not decrease after treatment of MARC-145 cells with heparinase. CONCLUSIONS: Addition of heparin to PRRSV or to the MARC-145 cells before virus inoculation and treatment of the cells with heparinase prevented the virus from infecting the cells.

Animals↗
PubMed1996

Slow-reacting and complement-requiring neutralizing antibody in swine infected with porcine reproductive and respiratory syndrome (PRRS) virus.

Various conditions were evaluated and modified to enhance the sensitivity of the neutralization (NT) test for detecting antibody in swine infected with porcine reproductive and respiratory syndrome (PRRS) virus. Higher NT antibody titers were consistently obtained by the addition of 10% (v/v) complement, fresh guinea pig serum, to the virus diluent and by the incubation of serum-virus mixture at 4 degrees C for 24 hr. The appearance and persistence of antibodies detected by the modified NT test showed a similar pattern to those of antibodies detected by the indirect fluorescent antibody (IFA) assay, although the antibody titers obtained by the former method were consistently lower than those obtained by the latter method. Slow-reacting complement-requiring NT antibody was detected in sera from pig 2 weeks after infection with PRRS virus. The slow-reacting complement-requiring NT antibody in the early serum samples was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting complement-requiring NT antibody in the late serum samples was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which serum-virus mixtures were incubated at 4 degrees C for 24 hr with complement; this gave an improved sensitivity over the previous incubation at 37 degrees C for 60 min.

Animals↗
PubMed1995

Purification of recombinant alpha-amylase by immunoaffinity chromatography with anti-peptide antibody.

Adsorption characteristics of an anti-peptide antibody, obtained by immunization of eight amino acids in the C-terminal region of chimeric alpha-amylase of rice alpha-amylase isozymes, were studied by use of the chimeric enzyme and the peptide used for immunization. This anti-peptide antibody adsorbed the enzyme, as well as the peptide antigen, with sufficient affinity for immunoaffinity purification and was used for purification of the enzyme secreted from yeast cells. Chimeric alpha-amylase was purified by immunoaffinity chromatography to high purity in one step from the fermentation broth. One-third of the secreted enzyme was not adsorbed by the column of anti-peptide antibody because of processing in the C-terminal region.

Amino Acid Sequence↗
PubMed1995

Smoldering adult T-cell leukemia with B-cell lymphoma and early gastric cancer.

We report a case of smoldering adult T-cell leukemia (ATL) with B-cell lymphoma and early gastric cancer. A 64-year-old man was admitted to our hospital because of proteinuria and hypergammaglobulinemia. Systemic lymphadenopathy, "flower cells" in peripheral white blood cells, and hypergammaglobulinemia with monoclonal gammopathy (IgA, lambda type) were found. As Southern blot analysis revealed monoclonal integration of human T-lymphotrophic virus type I proviral DNA in peripheral blood mononuclear cells, he was diagnosed as having smoldering ATL. The tissue specimen of an inguinal lymph node showed proliferation of abnormal lymphocytes which were stained with anti-lambda antibody, indicating B-cell lymphoma. A polypoid lesion in the stomach was histologically diagnosed as early gastric cancer.

HTLV-I Infections↗
PubMed1994

[The influence of the alcohol and the low protein diet on rat pancreas].

We investigated the influence of alcohol and the low protein diet upon rat pancreas. Rats were separated in four groups, 1) Control diet group (Cont), 2) Alcohol diet group (Al), 3) Low protein diet group (Lp), 4) Low protein and alcohol diet group (Lp+Al). They were fed on isocaloric liquid diet compulsorily through the gastric tube. They were sacrificed 3, 6, 12 weeks after. By the light microscopic observation on the rat pancreas which were fed for 12 weeks, lipid droplets in the acinar cells were observed in all groups other than Cont. Apoptosis was founded in Lp and Lp+Al groups. Protein plugs were observed in all groups, and no relation was found between the plugs and the location of the injured acinar cells. By the electromicroscopic observation, in Lp+Al group, acinar cells were typically injured (ER dilation, atrophic nuclei, mitochondria degeneration, etc.) and mesenchymal cells appeared among acinar cells. These results suggest that alcohol causes pancreatic acinar cell injury directly, and relative low protein diet helps to turn it worse.

Animals↗
PubMed1993

Clinical significance of measuring plasma concentrations of glutamine and glutamate in alcoholic liver diseases.

We investigated the significance of assaying plasma glutamine and glutamate concentrations in various liver diseases. Comparison was made with an aminogram in non-alcoholic liver disease. In alcoholic liver diseases, the plasma glutamate concentration showed a statistically significant elevation compared to the value in non-alcoholic liver diseases. In contrast, the plasma glutamine concentration was elevated in non-alcoholic liver diseases but showed no elevation in alcoholic liver diseases. The glutamate (Glu)/glutamine (Gln) ratio and plasma gamma-GT level correlated well in patients of alcoholic liver diseases. The Glu/Gln ratio became normal after alcohol abstinence. The results suggest that measuring the plasma concentrations of glutamine and glutamate may be a useful method to estimate the pathophysiological state of patients with alcoholic liver diseases.

Acute Disease↗
PubMed1993

Ethanol and hydrazine sulfate induced chronic hepatic injury in rats: the curative effect of administration of glucogenic amino acids.

There is a widespread belief that when ethanol is fed to rats for a long time, it produces only fatty degeneration without necrosis or fibrosis. In this study, hydrazine sulfate, an inhibitor of low Km ALDH and gluconeogenetic enzymes, was fed with ethanol to rats, and produced more serious pathological changes compared with those found in Lieber's model. Male Sprague-Dawley rats were fed with a low fat liquid diet as a basal diet with ethanol (4%, w/v) and hydrazine sulfate for 4 weeks. At the end of the experiment, plasma aminotransferase levels were found to be elevated. Histological examination showed not only fatty degeneration but also pericellular fibrosis. Therefore, we have evaluated the curative effect of glucogenic amino acids, alanine and glutamine, on this hepatic injury model and found them to be partially protective.

Alanine Transaminase↗
PubMed1993

[Minitracheotomy].

Twenty-four consecutive patients undergoing minitracheotomy were reviewed. Postoperative sputum retention was the major indication. In one case the procedure was not possible. Nineteen patients made an uneventful recovery, and decanulation was done in 17. In four patients minitracheotomy treatment was discontinued because formal tracheotomy was performed subsequently. This method is much simpler, less invasive, and more advantageous than formal tracheotomy. It can also be used for the patients after median sternotomy. On the other hand, in the patients with misswallowing because of recurrent nerve palsy etc., conventional tracheotomy should be performed.

Aged↗
PubMed1992

[Two cases of hepatobiliary scintigraphy of liver transplantation in rejection].

We reported two patients with liver transplantation who are performed hepatobiliary scintigraphy in rejection. In one patient hepatobiliary scintigraphy shows dilatation of bile duct but it shows good clearance from liver. In another patient hepatobiliary scintigraphy shows delayed clearance from liver. Both patients recovered from rejection. There are four main complications--vascular and biliary abnormality, infection, rejection--after liver transplantation. Hepatobiliary scintigraphy shows almost same findings except biliary complication, so it is difficult to distinguish between infection and rejection. But hepatobiliary scintigraphy may be useful to evaluate transplanted liver function in rejection.

Biliary Tract↗
PubMed1991

[A case of renal pelvic squamous cell carcinoma accompanied with humoral hypercalcemia of malignancy].

A 74-year-old male was urgently admitted to our hospital because of consciousness disturbance. Laboratory data showed remarkable hypercalcemia (7.8 mEq/L), hypophosphatemia, low % TRP, low intact PTH level, normal nephrogenic cyclic AMP and normal 1,25 (OH)2D level. Serum bone Gla protein, which was thought to express osteoblastic activity, was low. Serum tartarate resistant acid phosphatase and urinary excretion of hydroxyproline, which were thought to express osteoclastic activity, were high. CT scan showed an enlarged mass in the left renal pelvis, which was found to be a squamous cell carcinoma (SCC) by biopsy through percutaneous nephroscopy. Bone scintigram appeared normal. Therefore, we diagnosed it as renal pelvic SCC with humoral hypercalcemia of malignancy (HHM) and performed left nephrectomy. After nephrectomy, serum calcium returned to normal. But after a few weeks, lung metastasis appeared and serum calcium was reelevated. As to PTH related protein (PTHrP) which was thought to induce HHM, PTHrP content of the resected tumor measured by RIA assay was 13 pmol/g wet weight of tissue, which suggested that this tumor might have been producing PTHrP.

Aged↗
PubMed1991

[A case of adult T-cell leukemia with a defective HTLV-I proviral DNA, in which the single T-cell clone appeared to have progressed from chronic phase to crisis].

A 39-year-old woman was first admitted to our hospital with increased white cell count on May, 1983. Physical examination showed only mild splenomegaly. Hematological examination revealed leukocytosis (14,600/microliters) with ATL cells (59%). Serum anti-HTLV-I antibody was positive. Examination of HTLV-I provirus in the abnormal T cells revealed the defective type. She was diagnosed as chronic type of ATL based on the clinical features. Cytogenetic study of the ATL cells revealed 47, xx, +4. For 12 months, she was followed without any therapy. WBC reduced to almost normal range and ATL cells decreased to 3 to 6% for 8 months. On May, 1985, she was readmitted to our hospital because of leukocytosis (32,200/microliters), and increased ATL cells (57%). She was diagnosed as crisis of ATL. Investigation of the proviral DNA and chromosome showed the same results as those of the chronic phase, indicating that ATL cells in both the chronic phase and the crisis phase originated from the same clone. She died after 3 months from massive diarrhea. Postmortem examination showed the extensive infiltration of leukemic cell in the small intestine.

Adult↗
PubMed1989

Tumor-infiltrating lymphocytes and histocompatibility antigens in primary intracranial germinomas.

Subpopulations of tumor-infiltrating lymphocytes (TIL's) and the major histocompatibility complex (MHC) antigens of neoplastic cells were examined in three intracranial germinomas by an immunohistochemical method using monoclonal antibodies. About 70% to 80% of TIL's were T lymphocytes which were either infiltrating diffusely or in clusters, whereas 20% to 30% of TIL's were B lymphocytes which tend to cluster in tumor tissues. Examination of T lymphocyte phenotypes revealed both the cytotoxic/suppressor and helper/inducer T lymphocytes, as in other tumors. However, the existence of a considerable number of B lymphocytes in the TIL population was uncommon and seemed to be a characteristic feature of the intracranial germinoma, which might suggest a difference of host immune response to this neoplasm as compared to other tumors. On examination of the MHC antigens, no MHC class I or II antigens in the neoplastic cells were stained, while positive staining for both antigens was seen in the TIL and stroma tissues. From these findings, it was suggested that the degree of TIL infiltration might not be correlated with the expression of MHC antigens in neoplastic cells in cases of primary intracranial germinoma.

Adolescent↗
PubMed1988

An immunohistochemical study of cytokeratin and glial fibrillary acidic protein in choroid plexus papilloma.

Cellular localization of cytokeratin and glial fibrillary acidic protein (GFAP) was examined in two normal choroid plexuses and five choroid plexus papillomas by the peroxidase-antiperoxidase (PAP) method and double immunofluorescence (IFL) microscopy. Cytokeratin was observed in the majority of epithelial cells in all samples of normal and neoplastic choroid plexuses. On the other hand, GFAP was observed in some of the constituent epithelial cells in two cases of papilloma. Most of these GFAP-positive papilloma cells were simultaneously positive for cytokeratin, as could be seen by the PAP stainings of serial sections and by the double IFL stainings of the same sections. From these findings, it was suggested that normal and neoplastic choroid plexus epithelial cells usually express cytokeratin and that some of the neoplastic cells can simultaneously express both cytokeratin and GFAP.

Adult↗
PubMed1987

[Immunohistological analysis of infiltrating lymphocyte subpopulations in gliomas and metastatic brain tumors].

Subpopulations of infiltrating lymphocytes were studied by immunohistological method using monoclonal antibodies in gliomas and metastatic brain tumors. Thirteen specimens from 8 glioma patients, and 7 specimens from 3 metastatic brain tumor patients were used. No special therapy for brain tumor had been performed in these cases, but 3 glioma patients and all metastatic brain tumor patients had received steroid hormone. Frontal lobe obtained from the autopsy case of chest trauma was served as a normal control. Frozen sections were stained with avidin-biotin peroxidase complex method using Leu-series monoclonal antibodies for pan T-cells (Leu-1), cytotoxic/suppressor T-cells (Leu-2 a), helper/inducer T-cells (Leu-3 a) and B-cells (Leu-12). Lymphocyte infiltrates were quantitated by counting positively stained cells in 13 glioma and 7 metastatic brain tumor specimens. In normal frontal lobe, only a few T-cells infiltrated around several blood vessels in the parenchyma and subarachnoid space. But in the cases of glioma, many perivascular lymphocytic infiltrates were found and in the cases of metastatic brain tumor, many lymphocytes were found diffusely in the interstitial area between nests of tumor cells. Most of these lymphocytes were T-cells and B-cells were scarce, and Leu-2 a and Leu-3 a positive cells intermingled with each other. Len-3 a/2 a ratio ranged from 0.2 to 0.9 in the half of gliomas and 1.5 to 3.6 in another half of gliomas, three of which were treated with steroid hormone.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗