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Biomedical subjects

H Okabe

Publications and source records attributed to H Okabe.

At least 325 records · Page 18Linked to original sources

Activation of human plasma prekallikrein by Pseudomonas aeruginosa elastase in vitro.

Human plasma prekallikrein, precursor of the bradykinin-generating enzyme, was activated in a purified system under a near physiological condition (pH 7.8, ionic strength I = 0.14, 37 degrees C) by Pseudomonas aeruginosa elastase which is a tissue-destructive metalloproteinase. Compared with that, Pseudomonas aeruginosa alkaline proteinase poorly activated it with a rate as low as less than one-twentieth of that of elastase. The activation by elastase was blocked with a specific inhibitor of elastase, HONHCOCH(CH2C6H5)CO-Ala-Gly-NH2 (10 microM). Generation of kallikrein-like amidolytic activity was also observed in plasma deficient in Hageman factor by treatment with elastase, but was not in plasma deficient in prekallikrein. The kallikrein-like activity generated in Hageman factor deficient plasma as well as the generation process itself was indeed inhibited by anti-human prekallikrein goat antibody. These results suggest that the pathological activation of the kallikrein-kinin system might occur under certain clinical conditions in pseudomonal infections.

Amino Acid Sequence↗

Pseudomonas aeruginosa alkaline proteinase might share a biological function with plasmin.

Pseudomonas aeruginosa alkaline proteinase, which is a zinc-dependent bacterial endopeptidase, preferentially hydrolyzed Boc-Val-Leu-Lys-methylcoumarylamide (MCA) which was originally designed as a specific substrate of plasmin, a plasma serine proteinase. The hydrolytic capacity was resistant to tosyl-lysine chloromethylketone at a concentration as high as 1 mM, but was blocked by a treatment with metal chelator such as o-phenanthroline at the concentration of 5 mM. Kinetic parameters of the amidolytic reaction were Km = 21 microM, kcat = 0.067 s-1 and kcat/Km = 3190 M-1 s-1. A synthetic peptide inhibitor which bore a possible ligand for zinc atom at the carboxy terminal was designed. This inhibitor, Ac-Val-Leu-Lys-4-mercaptoanilide, blocked the amidolytic activity of the pseudomonal alkaline proteinase in a competitive manner with the dissociation constant (Ki) value of 24 microM. The results imply that P. aeruginosa alkaline proteinase must be an unusual zinc-dependent 'C (COOH)-type' endopeptidase, which hydrolyzes the peptide bond of certain amino acid residues at the carboxyl group side by specific recognition, like serine- and cysteine-proteinases. In comparison, P. aeruginosa elastase which is a typical 'N (NH2)-type' metalloproteinase did not hydrolyze all of the commercially available peptide-MCA substrates tested at the present study. P. aeruginosa alkaline proteinase also hydrolyzed natural substrates of plasmin, such as fibrin and fibrinogen, with similar specific activities to plasmin. The susceptible subunits of fibrinogen were the A-alpha and B-beta ones, in this order. P. aeruginosa alkaline proteinase also exhibited an anti-coagulant activity in human plasma attributed to the direct fibrinogenolytic function. Such potential anti-coagulant capacity of the P. aeruginosa alkaline proteinase might explain, at least partly, the most characteristic pathologic feature of the P. aeruginosa septicemia, hemorrhagic lesions with lacking thrombi (Fetzer, A.E. et al. (1967) Am. Rev. Respirat. Dis. 96, 1121-1130).

Blood Coagulation↗

Penetration enhancement across a model membrane by liposomally entrapped drugs using N,N'-diacylcystine as a bilayer lipid.

A liposomal system for percutaneous absorption and transdermal drug delivery was developed and evaluated using a model apparatus connecting two chambers by a membrane. When N,N'-long chain diacyl L-cystine was incorporated into the liposomal bilayer as well as lecithin and cholesterol, liposomally entrapped calcein as the model drug transferred well across the keratin membrane functioning as the skin model.

Cellophane↗

Role of leukocytes in the activation of intravascular coagulation in patients with septicemia.

To elucidate the role of leukocytes in intravascular coagulation in patients with septicemia, plasma levels of thrombin-antithrombin III complex (TAT), soluble fibrin monomer complex (SFMC) and fibrinogen (Fbg) were determined in 33 patients with septicemia. Twenty of 33 patients revealed marked leukopenia caused by suppression of hematopoiesis by the administration of chemotherapeutic agents for the treatment of hematological malignancies; the total leukocyte count of these patients was less than 1,000/microliters. Thirteen of 33 patients showed normal or increased leukocyte counts. Plasma levels of TAT and SFMC in septicemic patients without leukopenia were significantly higher than in patients with leukopenia. Although plasma TAT and SFMC levels correlated well with the number of leukocytes, a more significant positive correlation was found between the number of monocytes and the levels of TAT and SFMC. Plasma levels of Fbg were significantly lower in patients without leukopenia than in patients with leukopenia. No significant correlation was found between the number of leukocytes and the levels of Fbg. However, a significant negative correlation was found between the number of monocytes and the levels of Fbg. TAT levels did not correlate with the number of platelets. The fibrinolytic system was activated only in septicemic patients without leukopenia, which may be explained by secondary fibrinolysis following leukocyte-activated coagulation. These findings suggest that leukocytes, in particular monocytes, may play a critical role in the pathogenesis of intravascular coagulation in septicemia.

Antithrombin III↗

Automated immunoturbidimetric analysis of six plasma apolipoproteins: correlation with radial immunodiffusion assays.

We measured six apolipoproteins (apo AI, AII, B, CII, CIII, and E) by turbidimetric method using an automatic discrete biochemical analyzer and commercially available antisera. The turbidimetric method was compared with the single radial immunodiffusion method. Linearity for serum apolipoprotein assay by the automated turbidimetric method was better than by the single immuno-diffusion method. The linearity by the turbidimetric method was 2.5 G/L for AI, 1.0 G/L for AII, 4.5 G/L for B, 0.12 G/L for CII, 0.3 G/L for CIII, and 0.12 G/L for E. The presence of high concentrations of bilirubin (up to 0.15 G/L) and hemoglobin (up to 50 G/L) interfered with apolipoprotein measurement. Comparison of the immunoturbidimetric and the single radial immunodiffusion (SRID) methods showed excellent coefficients of correlation, r = 0.963, 0.896, 0.846, 0.936, 0.972, and 0.937 for apo AI, AII, B, CII, CIII, and E, respectively. Reference ranges for the six apolipoproteins were determined by using sera from 450 healthy subjects and were 1.4 +/- 0.3 G/L for AI and 0.3 +/- 0.01 for E. The observed levels of AII (P less than 0.001), B (P less than 0.01), and CIII (P less than 0.01) were significantly higher in males. The serum levels of apo B, CII, and E showed a gradual increase with age which was more prominent in females than in males. The levels of apo AI, AII decreased significantly over an 11 day period in 22 patients with myocardial infarction.

Adolescent↗

Measurement of human ventricular myosin light chain-1 by monoclonal solid-phase enzyme immunoassay in patients with acute myocardial infarction.

A monoclonal solid-phase enzyme immunoassay has been developed for the detection of human serum ventricular myosin light chain-1. Cross-reactivity of this with human skeletal muscle myosin was observed, but the enzyme immunoassay with the sera of patients with acute myocardial infarction gave similar results with radioimmunoassay. The human ventricular myosin light chain-1 levels in the healthy subjects were 0.2-6.6 ng/ml in males and 0.2-4.1 ng/ml in females. Within-run and between-run precision (CVs) of the assays was on the order of 2.3-4.7% and 4.3-8.7, respectively. Sensitivity of the assay was 1.0 ng/ml, and working range was 5-100 ng/ml. In all patients with define acute myocardial infarction, serum ventricular myosin light chain-1 levels increased three- to ten-fold the upper reference range within 6 hr after the onset of chest pains. Two types of subtrend were discovered: its levels remain elevated for 3-4 days and its levels increased and then decreased 1-2 days after the initial rise but became elevated again for the next 4-7 days after the onset of chest pain, which is in contrast to the case with all conventionally used biochemical cardiac markers.

Biomarkers↗

Production of monocyte chemoattractant protein-1 by malignant fibrous histiocytoma: relation to the origin of histiocyte-like cells.

Human malignant fibrous histiocytoma (MFH) comprise both fibroblast-like cells and histiocyte-like cells. We previously showed that the latter are not neoplastic cells, but are infiltrating macrophages. Since migration of blood monocytes into the tumor might be a response to a locally elaborated monocyte chemoattractant, we designed experiments to determine if the fibroblast-like tumor cells produced a chemoattractant for human monocytes. Malignant fibrous histiocytoma from three patients was put into culture. Cells of all three lines had a spindle shape, and showed no reactivity with antibodies against macrophages (MAC387), HLA-DR (LN3), or leukocyte common antigen. Immunohistochemically, they stained with antibody against human monocyte chemoattractant protein-1 (MCP-1). Culture supernatants of the three cell lines had chemotactic activity for monocytes. This activity was due to MCP-1, since it was absorbed by an anti-MCP-1 column. The production of MCP-1 by MFH tumor lines was confirmed by immunoprecipitation of metabolically labeled MCP-1. These results suggest that the histiocyte-like cells are the infiltrated macrophages that originate from blood monocytes attracted by tumor-derived MCP-1.

Chemokine CCL2↗

Successful treatment of disseminated cutaneous cytomegalic inclusion disease associated with Hodgkin's disease.

A case of disseminated cutaneous cytomegalic inclusion disease associated with Hodgkin's disease is described. The patient had a diffuse eruption of pruritic, erosive erythematous nodules. Histologically, many inclusion bodies were observed in perivascular areas of the skin lesions. Immunohistologically, the inclusion bodies positively stained with both anticytomegalovirus antibody and anti-factor VIII-related antibody. On electron microscopy many virus particles and dense bodies were found in the area where inclusion bodies were observed. Treatment with high-dose intravenous acyclovir and a large amount of immunoglobulin resulted in prompt healing of the skin lesions. Subsequently, the patient's Hodgkin's disease was well controlled on chemotherapy. The patient steadily improved without relapse of skin lesions 1 year after antiviral therapy was administered.

Aged↗

Immunohistochemical characterization of basal cell adenomas of the salivary gland.

Seven cases of basal cell adenomas of the salivary gland were analyzed by immunohistochemical methods with a broad panel of routinely used antibodies. Histologically the epithelial elements were classified as tubuloglandular, trabecular and solid patterns. The authors' results indicated the following: 1) The duct lining cells of tubuloglandular and trabecular patterns have distinct epithelial features with cytokeratins (KL 1, PKK 1, *PKK 2 and PKK 3), alpha-one-antichymotrypsin (alpha 1-ACT), carcinoembryonic antigen (CEA) and S-100 alpha subunit positivity. 2) The basaloid cells in the trabecular and solid patterns expressed two immunophenotypes: one had actin, neuron-specific enolase (NSE), S-100 protein and S-100 beta subunit patterns typical of myoepithelial cells in normal glands. The other basaloid cells had vimentin and S-100 protein patterns. The former cell type could be found in 4 of 7 cases and the latter was found in 7 cases. This represents a minor participation of the myoepithelial cells in the basal cell adenoma. 3) The basement membrane and stromal connective tissue around the neoplastic cells were positive for alpha-one-antitrypsin (alpha 1-AT). This antibody is a good marker in identifying the basement membrane-like material.

Adenoma↗

Neuronal differentiation of Ewing's sarcoma induced by cholera toxin B and bromodeoxyuridine--establishment of Ewing's sarcoma cell line and histochemical study.

An Ewing's sarcoma (ES) cell line was established from a metastatic bone marrow specimen in a patient with advanced disease, and some histochemical characteristics were investigated by neuronal differentiation induced with cholera toxin B (CTB) and bromodeoxyuridine (BrdU). Neuronal differentiation was investigated by the expression of neurofilament and Leu-7, and glial differentiation was observed by expression of S-100 protein. Neurofilament (NF) and Leu-7 were positive in ES cells and these were expressed more intensively by induction with CTB than with BrdU. There was no expression of S-100 protein in untreated or differentiated ES cells. ES cells became differentiated to neuronal cells with CTB and BrdU, but it was not observed, that ES cells had the potential to differentiate to glial cells. It appears that ES is of more primitive neural origin than neuroblastoma, primitive neuroectodermal tumors and other related neural tumors.

Antigens, Differentiation↗

Immunohistochemical localization of S-100 protein and its subunits in melanotic lesions in the oral mucosa and skin.

Immunohistochemical localization of S-100 protein, S-100 alpha and beta subunits was examined in 23 pigmented nevi, 20 malignant melanomas and 8 metastatic melanomas originating from the oral mucosa and skin. Primary oral mucosal melanomas demonstrated no beta subunit immunoreactivity while one metastatic lesion from oral melanoma that showed only a small number of melanoma cells reacted with the beta subunit. In contrast, most cutaneous melanomas and their metastatic lesions showed immunoreaction with the S-100 beta subunit. As for pigmented nevi, immunoreactions with the three antibodies were frequently detected regardless of the site. The differences in the expression of S-100 protein beta subunit between cutaneous and mucosal melanomas may be related to differences in inductive interactions between the melanoma cells and the connective tissues and skin and mucosa.

Cell Nucleus↗

Prognostic implication of silver-binding nucleolar organizer regions (AgNORs) in oral squamous cell carcinoma.

Silver-binding nucleolar organizer regions (AgNORs) were counted in sections from formalin-fixed, paraffin-embedded tissue blocks of oral squamous cell carcinoma. Thirty-nine cases, that comprised poor prognostic group (n = 19) and good prognostic group (n = 20), were examined with respect to the relation between AgNOR counts and histologic grading, and correlation between AgNOR counts and prognosis. The pooled mean AgNOR counts were: Grade 1 carcinomas, 6.39 +/- 1.67 (mean +/- SD; n = 35); Grade 2, 9.74 +/- 1.72 (n = 3). Mean AgNOR count of Grade 3 was 6.19 +/- 2.37 (n = 1). The pooled mean AgNOR count in poor prognostic group was higher than that in good prognostic group. Five-year survival rate of the cases with high AgNOR counts (greater than or equal to 6.5) was significantly lower than that with low AgNOR counts (less than 6.5). High AgNOR counts are highly suggestive of poor prognosis in oral squamous cell carcinoma.

Carcinoma, Squamous Cell↗

Lectin histochemistry of cystic jaw lesions: an aid for differential diagnosis between cystic ameloblastoma and odontogenic cysts.

The binding sites for Ulex europaeus agglutinin I (UEA-I), Bandeirea simplicifolia agglutinin I (BSA-I), and peanut agglutinin (PNA) were comparatively examined in the surgical materials from 41 cases of cystic and solid ameloblastomas and 42 cases of non-neoplastic odontogenic cysts including dentigerous cyst, odontogenic keratocyst, and radicular cyst. In non-neoplastic cysts, most of epithelial lining layers gave positive binding with UEA-I and BSA-I. However, no positive reactions were obtained for these two lectins in the epithelial components of ameloblastoma, except for limited UEA-I binding to markedly keratinized tumor cells in four cases. PNA binding was irregular and did not make any clear distinction between ameloblastomas and cysts. The results suggest that the lectin staining for UEA-I and BSA-I is a useful histologic aid for differential diagnosis between cystic ameloblastoma and non-neoplastic jaw cysts.

Ameloblastoma↗

Immunohistochemical investigation in odontogenic myxoma.

Three odontogenic myxomas are described immunohistochemically by a panel of poly- and monoclonal antibodies to characterize this tumor type. Three types of odontogenic myxoma cells were discriminated: spindle cells, stellate cells and hyaline cells. Neoplastic cells of myxomas were positively stained for transferrin, ferritin, alpha-1-antichymotrypsin (alpha 1-ACT), alpha-1-antitrypsin (alpha 1-AT), S-100 protein and vimentin; however, neuron specific enolase (NSE), S-100 alpha subunit, S-100 beta subunit, Factor VIII-related antigen (FVIII-AG) and cytokeratin (CK1) were negative. Spindle cells were positive for transferrin, ferritin, alpha 1-ACT, alpha 1-AT, S-100 protein and vimentin. Stellate cells were strongly positive for transferrin, alpha 1-AT, S-100 protein and vimentin. Hyaline cells reacted with alpha 1-ACT and alpha 1-AT. Myxomatous matrix showed negative reaction for all the antibodies used. These results have confirmed that odontogenic myxoma is a tumor of a dual fibroblastic-histiocytic origin.

Adolescent↗

Beneficial treatment with methyl 6-[3-(2-chloroethyl)-3-nitrosoureido]-6-deoxy-alpha-D-glucopyranoside in a patient with primary myelofibrosis.

We attempted treatment with methyl 6-[3-(2-chloroethyl)-3-nitrosoureido]-6-deoxy-alpha-D-glucopyranoside (MCNU), a novel nitrosourea derivative, in a 55-year-old man with advanced-stage primary myelofibrosis. MCNU was given intravenously at a dose of 50 mg once a month. Following MCNU treatment, his anemia and splenomegaly improved markedly. An increased dose of MCNU (100 mg, once a month) was even more effective for relieving the symptoms. Severe side effects resulting from this therapy, such as leukocytopenia or thrombocytopenia, were never observed. These observations indicate that MCNU treatment may be a beneficial management of advanced-stage primary myelofibrosis.

Antineoplastic Agents↗

Studies on the constituents of Luffa acutangula Roxb. I. Structures of acutosides A--G, oleanane-type triterpene saponins isolated from the herb.

From the herb of Luffa acutangula ROXB. (Cucurbitaceae), seven oleanane-type triterpene saponins, acutosides A--G, were isolated and their structures were determined. Acutoside A is oleanolic acid 3-O-beta-D-glucopyranosyl-(1----2)-beta-D-glucopyranoside. Acutosides B, D, E, F and G have a common prosapogenin structure, acutoside A, and only differ in the structures of the ester-linked sugar moieties. Acutoside B is a 28-O-[O-beta-D-xylopyranosyl-(1----4)-O-alpha-L-rhamnopyranosyl-(1----2) -alpha-L-arabinopyranosyl] ester, D is a 28-O-[O-beta-D-xylopyranosyl-(1----3)-O-beta-D-xylopyranosyl-(1----4)-O- alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, E is a 28-O-[O-alpha-L-arabinopyranosyl-(1----3)-O-beta-D-xylopyranosyl-( 1----4)-O-alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, F is a 28-O-[O-beta-D-xylopyranosyl-(1----3)-[O-beta-D-xylopyranosyl-(1----4)-O -alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, and G is a 28-O-beta-D-xylopyranosyl-(1----3)-[O-alpha-L-arabinopyranosyl-(1- ---3)-O-beta-D-xylopyranosyl-(1----4)]-O-alpha-L- rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester. Acutoside C is a machaelinic acid (=21 beta-hydroxyoleanolic acid) saponin having the same sugar moiety as that of acutoside B.

Carbohydrate Sequence↗

Iron-binding proteins in adenoid cystic carcinoma of salivary glands: an immunohistochemical study.

An immunoperoxidase staining technique was used for detecting three major iron-binding proteins (transferrin, ferritin and lactoferrin) in 54 adenoid cystic carcinomas (ACCs) of major and minor salivary glands. Twenty-three normal salivary glands were investigated for comparison. Transferrin staining was detected mainly in the intercalated duct and serous acinar cells, and was found inconsistently in the myoepithelial cells surrounding normal ductules. Ferritin was always absent in the normal epithelial component of salivary gland. The presence of lactoferrin was demonstrated in the serous acinar cells and intercalated duct cells of normal salivary tissues. Five histological patterns were found in ACC, and for the cellular components of each, it was possible to establish a special immunohistochemical profile. Transferrin positivity was detected in the small angular cells of 25 cases (48%), in the duct luminal cells of 19 cases (37%) and in the hyalinized stroma of 4 cases (8%). Ferritin staining was positive in the small angular cells of 23 cases (44%) and in the duct luminal cells of 15 cases (29%). Lactoferrin was detected in only duct luminal cells of 38 ACCs (73%). The comparative immunohistochemical analysis between transferrin and ferritin showed a similar distribution in this carcinoma. On the basis of the immunohistochemical data, lactoferrin might be used as a marker of glandular differentiation.

Biomarkers, Tumor↗

[Experimental study on intraoperative autotransfusion during urological operation].

The value of autotransfusion is widely recognized in the surgical community and may be of increasing importance in prevention of acquired immunodeficiency syndrome and hepatitis. The concern of possible contamination of the blood with urine, bacteria in urine or viable tumor cells has limited the wide use of intraoperative autotransfusion (IAT) in urological operation. There have been no experimental reports about protection of the blood from such contamination. To investigate separation of the blood from a contaminated mixture by using an autotransfusion machine, Haemonetic Cell Saver, a study composed of three experiments was performed. First, 200 ml of blood was mixed 200 ml of urine, and thereafter, the mixture was processed by the machine and the concentration erythrocytes were collected in a bag. Biochemical analysis of the collected erythrocyte solution (CES) was performed. Second, 200 ml of blood was mixed with 200 ml of urine that was adjusted to contain each 10(7)/ml of four bacterial strains. The bacteriological study of the CES was performed. Third, 200 ml of blood was mixed with 200 ml of urine that was adjusted to contain 10(7) cancer cells. Two cell lines, KK47 originated from human bladder cancer and ACHN originated from human renal cell carcinoma was used. The cytological study of the CES was performed. The results of these experiments were: Urine constituents were completely removed from the mixture. However, all strains of bacteria could not be separated, although the number of bacteria decreased. Cancer cells were found in the CES. In conclusion IAT should be done at urological operation in selected patients that have sterile urine and do not have tumor cells in the operation field.

Blood↗