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Biomedical subjects

M Murai

Publications and source records attributed to M Murai.

At least 181 records · Page 10Linked to original sources

Modification of alpha 2-macroglobulin into a macrophage-activating factor through the action of liposome-stimulated B-cell membranous glycosidases.

The serum glycoprotein alpha 2-macroglobulin can be converted into a potent macrophage-activating factor that promotes the Fc gamma receptor-mediated phagocytosis of macrophages, through modification of the sugar moiety with liposome-treated B-cell glycosidase(s). This paper discusses the activation mechanism of B-cell membranous glycosidase by liposomes using mouse splenic B cells. B-cell membranous beta-galactosidase and beta-N-acetylglucosaminidase were significantly activated by liposome treatment, and this process can be regulated by trypsin-sensitive protein. To clarify the contribution of trypsin-sensitive protein to enzyme activities, the B-cell surface antigen receptor was studied. With the addition of a Fab' fragment of anti-mouse IgM but not IgD antibody, the activation of both glycosidases induced by liposomes was significantly inhibited and was essentially the same as that of saline-treated glycosidase activities. Consequently, interactions of liposomes with cell-surface IgM may cause B-cell membranous glycosidase activation. A significant decrease in membrane fluidity, particularly near the membrane surface rather than deep within the membrane, was observed in liposome-treated B cells using electron spin resonance. Liposomes would thus appear to interact with B cells via cell-surface IgM, with a consequent decrease in membrane fluidity, as well as the activation of B-cell membranous glycosidases, causing alpha 2-macroglobulin to be converted into a macrophage activating factor.

Animals↗

Identification of the serum factor required for liposome-primed activation of mouse peritoneal macrophages. Modified alpha 2-macroglobulin enhances Fc gamma receptor-mediated phagocytosis of opsonized sheep red blood cells.

Culture supernatant, prepared by incubating fetal calf serum and liposome-treated B cells, augments mouse peritoneal macrophage Fc gamma receptor-mediated phagocytosis of IgG-opsonized sheep red blood cells. The activation process was hypothesized to be as follows. B-cell membranous glycosidases stimulated by liposomes, convert serum factor to a macrophage-stimulating active serum factor. As the active serum factor loses its activation potential by the addition of mannose or by digestion with alpha-mannosidase, mannose residues at the terminal present in the active serum factor are hypothesized to contribute to macrophage activation. The active serum factor was purified on a concanavalin A-Sepharose 4B column, and identified as a modified form of alpha 2-macroglobulin by immunochemical analysises. On non-denaturing polyacrylamide gel electrophoresis, modified alpha 2-macroglobulin showed a slow form, while alpha 2-macroglobulin-trypsin and alpha 2-macroglobulin-methylamine complexes, which bind specifically to alpha 2-macroglobulin receptors, showed a fast form and did not activate macropahges. These findings demonstrate that alpha 2-macroglobulin is the essential serum factor in liposome-primed macrophage activation, and that modified alpha 2-macroglobulin with mannose residues at the terminal sugar chain binds to macrophage mannose receptors, but not alpha 2-macroglobulin receptors, and increases Fc gamma receptor-mediated phagocytosis of IgG-opsonized sheep red blood cells.

Animals↗

Technical improvements in laparoscopic adrenalectomy.

Laparoscopic adrenalectomy has been believed to be the most difficult operation because of the deep location of the adrenals but also because of the lack of standard technology and specific instruments. Herein, we introduce the technique using flexible electron scope (EL2-TF2, Fujinon) and laparoscopic coagulation shears (LCS; Harmonic Scalpel). Use of these two instruments provides a better view on the monitors and faster completion of surgery because of easy dividing procedures of abundant fatty tissues around the adrenal. Our results show shorter operating time compared to the group using conventional instruments. These results reflect an improvement of quality of life of the patients with functional and/or non-functional adenomas in the adrenals.

Adenoma↗

Biochemical and genetic heterogeneity of staphylococcal protein A.

We investigated the biochemical and genetic heterogeneity of protein A from Staphylococcus aureus. SpA genes (spas) of various strains were heterogeneous when detected as DraI and EcoRV fragments of chromosomal DNA. Polymerase chain reaction using primers to detect DNA encoding the IgG-binding domains in spa revealed that they numbered between 2 and 5. Protein A from several S. aureus strains showed two types of reactivities to immunoglobulins in normal canine serum according to the number of active domains.

Base Sequence↗

Activation of Fc receptor-mediated phagocytosis by mouse peritoneal macrophages following the intraperitoneal administration of liposomes.

The effects of liposomes on the phagocytic activity of mouse peritoneal macrophages were investigated using IgG-opsonized sheep red blood cells (SRBC). The highest ingestion index of opsonized SRBC via Fc receptors of macrophages from BALB/c mice was observed for macrophages harvested on day 4 following the intraperitoneal injection of liposomes (2.27 mumol lipid/mouse). An increase in the ingestion index was observed irrespective of liposomal charge. Binding parameters of Fc receptors of macrophages from liposome- or saline-injected mice were determined using horseradish peroxidase-conjugated IgG, and an increase in the number of binding sites with the same binding constant was observed in macrophages from liposome-injected mice. The activation mechanism of mouse peritoneal macrophages by liposomes differed from that by lipopolysaccharides. Liposomes thus appear to contribute to the activation of immune response.

1,2-Dipalmitoylphosphatidylcholine↗

A new scoring system based on the histological behavior and proliferative activity of tumor cells for grading the malignant potential of bladder cancers.

Urothelial cancer can be considered to consist of a spectrum of diseases with diverse natural histories and the conventional morphological classifications provided by classical histology remain the basis for any decision-making process. The proliferative rate of the tumor cells may be significant since the rate of DNA synthesis is directly related to the rate of tumor growth or tumor involvement. We have previously reported that flow cytometric deoxyribonucleic acid (DNA)/bromodeoxyuridine (BrdU) bivariate analysis can provide important information about the malignant potential of bladder cancer. In the present study, we have formulated a new system based on the histological grade and tumor proliferative activity determined by the BrdU labeling index. An attempt has been made, moreover, to investigate whether this grading system can be used to determine the malignant potential of bladder cancers. A total of 86 patients with bladder tumors, histologically proven to be transitional cell carcinomas, were analyzed. Immediately after removal of tumor specimens, in vitro BrdU labeling was performed. Multivariate survival analysis was conducted using Cox's regression model, followed by estimation of the risk ratio for survival. Based on the risk ratio of the histological grade and the BrdU labeling index, a score ranging from 1 to 52 was assigned to each tumor. Forty-four patients with a score of 1 had a 100% survival rate at 3 years, compared with 42.9% for patients having a score greater than 1. Conversely, 26 patients with the highest score, 52 exhibited a survival rate of only 17.3% at 3 years.(ABSTRACT TRUNCATED AT 250 WORDS)

Bromodeoxyuridine↗

Scanning electron microscope observation of adherence of Candida albicans to cultured keratinocytes.

The role of antigen 6 in the adherence process of Candida albicans serotype A to cultured keratinocytes was examined with a scanning electron microscope. The number of adhered organisms was significantly lower for the antigen 6-deficient mutant strain than for the antigen 6-positive parent strain (P < 0.001). Fibril- or strand-like structures bridging the organisms and the keratinocytes were found to develop during the later stages of adherence.

Bacterial Adhesion↗

Cloning, functional expression and tissue distribution of human cDNA for the alpha 1C-adrenergic receptor.

We have cloned human alpha 1C-adrenergic receptor from human prostate cDNA library. The deduced amino acid sequence of the clone (P2C4) encodes a protein of 466 amino acids that showed strong sequence homology to the previously cloned bovine alpha 1C-adrenergic receptor. The radioligand binding properties of P2C4 expressed in COS-7 cells were very similar to those of bovine alpha 1C-adrenergic receptor. With reverse-transcription polymerase chain reaction assay, we observed alpha 1C-adrenergic receptor transcripts in heart, brain, liver and prostate, but not in kidney, lung, adrenal, aorta and pituitary. The data show that the clone P2C4 encodes a human alpha 1C-adrenergic receptor cDNA, and the receptor subtype is expressed not widely but localized in several human tissues.

Amino Acid Sequence↗

Studies on neurokinin antagonists. 3. Design and structure-activity relationships of new branched tripeptides N alpha-(substituted L-aspartyl, L-ornithyl, or L-lysyl)-N-methyl-N-(phenylmethyl)-L-phenylalaninamides as substance P antagonists.

As an extension of our study on discovering a novel substance P (SP) antagonist, we designed new branched tripeptides containing L-aspartic acid (2 and 5), L-ornithine (3 and 6), and L-lysine (4 and 7) by reconstructing the structure of the previously reported tripeptide SP antagonist [Ac-Thr-D-Trp(CHO)-Phe-NMeBzl (1), FR113680]. The strategy for this design was based on the postulate that the dipeptide half D-Trp(CHO)-Phe-NMeBzl in 1 is essential for receptor recognition. Molecular modeling studies implied that these newly designed tripeptides could mimic the spatial orientations of the essential dipeptide structure. As expected, all of these compounds potently inhibited 3H-SP (1 nM) binding to guinea pig lung membranes in the 10(-8) M range. The 1H-indol-3-ylcarbonyl derivatives (5-7) were slightly more potent than the corresponding 1H-indol-2-ylcarbonyl derivatives (2-4), as predicted by the molecular modeling studies. The structure-activity relationships studies on the selected 1H-indol-3-ylcarbonyl derivatives indicated that the threonine moiety at the side chain can be modified into a variety of structures without any significant loss of the activity. Furthermore in the L-lysine series, even dipeptide compounds having nothing or a simple acyl group at the epsilon-amino group, such as N alpha-[N alpha-(1H-indol-3-ylcarbonyl)-L-lysyl]-N-methyl-N-(phenylmethyl)- L-phenylalaninamide (18b), exhibited potent activity. These dipeptides belong to a new structural class of SP antagonist.

Animals↗

Effects of an NK1 receptor antagonist, FK888, on constriction and plasma extravasation induced in guinea pig airway by neurokinins and capsaicin.

The effects of FK888, an NK1 receptor antagonist, on airway constriction and airway plasma extravasation induced by neurokinins and capsaicin were investigated in guinea pigs. FK888 inhibited substance P (10(-8) M)- and neurokinin A (10(-9) M)-induced contraction of isolated guinea pig trachea, with IC50 values of 3.2 x 10(-8) and 4.2 x 10(-6) M, respectively. FK888 given i.v. inhibited substance P (13.5 micrograms kg-1)-induced airway constriction with an ED50 value of 0.40 mg kg-1 but did not inhibit neurokinin A (1.1 micrograms kg-1)- and capsaicin (3.1 micrograms kg-1)-induced airway constriction at a dose of 1 mg kg-1. On the other hand, FK888 given i.v. inhibited airway plasma extravasation induced by substance P (1.3 micrograms kg-1), neurokinin A (11 micrograms kg-1) and capsaicin (100 micrograms kg-1) with equal potency and ED50 values of 0.011, 0.0063 and 0.019 mg kg-1, respectively. When FK888 was given locally (into the airway directly) inhibitory activities were more potent than following i.v. administration. In this case FK888 inhibited substance P-, neurokinin A- and capsaicin-induced airway constriction with ED50 values of 3.2, 190 and 550 micrograms kg-1, respectively, suggesting that an about 100 times higher dose is required to inhibit neurokinin A- and capsaicin-induced airway constriction than substance P-induced constriction. FK888 given orally was also effective in substance P-, neurokinin A- and capsaicin-induced airway plasma extravasation with ED50 values of 4.2, 5.9 and 9.5 mg kg-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Efficient adsorption of lysostaphin on bacterial cells of lysostaphin-resistant Staphylococcus aureus mutant.

A simple and efficient method for the purification of staphylolytic endopeptidase (lysostaphin) contained in culture supernatant of Staphylococcus simulans biovar staphylolyticus strain by adsorption of the enzyme on bacterial cells of lysostaphin-resistant S. aureus mutant was successfully devised. Lysostaphin was sufficiently absorbed on the heat-killed mutant cells derived from S. aureus Cowan I and efficiently eluted by 3 M KSCN. Enzyme preparation obtained by a single procedure of the affinity purification was pure enough for practical use. The yield of the enzyme was 25 mg from 1 liter culture and recovery rate was 64%.

Adsorption↗

Effects of cytochalasins B and D on Staphylococcus aureus adherence to and ingestion by mouse renal cells from primary culture.

Cytochalasin B (CB) and cytochalasin D (CD), inhibitors of microfilament function of host cell, were examined for their effects on Staphylococcus aureus Cowan I adherence to and ingestion by several types of the hyperosmolarity-tolerant (HOT) cells obtained from primary culture of mouse kidney. Staphylococcal adherence to the HOT cells with epithelial appearance was extraordinarily enhanced by the treatment of those cells with both 5 micrograms/ml of CB and CD. In particular, staphylococci adhered to the periphery rather than the center of each cytochalasin-treated cell. Staphylococcal ingestion by all types of the HOT cells was markedly inhibited by CD in spite of the enhanced adherence. Contrary to our expectation, inhibition by CB was incomplete, and the enhanced adherence of staphylococci to CB-treated cells resulted in the enhanced ingestion.

Actin Cytoskeleton↗