[Drug therapy in infectious disease of the lungs].
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Anti-Bacterial Agents↗
Biomedical subjects
Y Ichikawa
Publications and source records attributed to Y Ichikawa.
[Examination of glucocorticoid receptor].
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Humans↗
Conformational change of the heme moiety of the ferrous cytochrome P-450scc-phenyl isocyanide complex upon binding of reduced adrenodoxin.
Reduction of cytochrome P-450scc(SF) (SF, substrate free) purified from bovine adrenocortical mitochondria with sodium dithionite (Na2S2O4) or with beta-NADPH mediated by catalytic amounts of adrenodoxin and adrenodoxin reductase in the presence of phenyl isocyanide produced a ferrous cytochrome P-450scc(SF)-phenyl isocyanide complex with Soret absorbance maximum at 455 nm having a shoulder at 425 nm. On the other hand, when a preformed cytochrome P-450scc(SF)-adrenodoxin complex was reduced chemically or enzymatically under the same conditions, the absorbance spectrum showed drastic changes, i.e., an increase in intensity at 425 nm and a concomitant decrease in intensity at 455 nm. Similar spectral changes could be produced by addition of the same amount of reduced adrenodoxin afterward to the ferrous cytochrome P-450scc(SF)-phenyl isocyanide complex. Titration experiments with adrenodoxin showed that (1) a 1:1 stoichiometric saturation of the spectral change was obtained for both the absorbance increase at 425 nm and the absorbance decrease at 455 nm, (2) there was no spectral change in the presence of 0.35 M NaCl, and (3) there was no spectral change for cytochrome P-450scc(SF) whose Lys residue(s) essential to the interaction with adrenodoxin had been covalently modified with PLP. These results suggest that ternary complex formation of ferrous cytochrome P-450scc(SF)-phenyl isocyanide with reduced adrenodoxin caused a conformational change around the ferrous heme moiety. By analysis of temperature and pH dependencies of the spectral change of the ternary complex, it was suggested that this conformational change may reflect the essential step for electron transfer from reduced adrenodoxin to the ferrous-dioxygen complex of cytochrome P-450scc.
Adrenodoxin↗
Binding characteristics of N-acetylglucosamine-specific lectin of the isolated chicken hepatocytes: similarities to mammalian hepatic galactose/N-acetylgalactosamine-specific lectin.
Binding characteristics of N-acetylglucosamine- (GlcNAc) specific lectin on the chicken hepatocyte surface were probed by an inhibition assay using various sugars and glycosides as inhibitors. Results indicated that the binding area of the lectin is small, interacting only with GlcNAc residues whose 3- and 4-OH's are open. The combining site is probably of trough-type, since substitution with as large a group as monosaccharide is permitted on the C-6 side of GlcNAc, and on the C-1 side, the aglycon of GlcNAc can be very large (e.g., a glycoprotein). These binding characteristics are shared with the homologous mammalian lectin specific for galactose/N-acetylgalactosamine, suggesting that tertiary structure of the combining area of these two lectins is similar. This is understandable, since there is approximately 40% amino acid sequence identity in the carbohydrate recognition domain of these two lectins [Drickamer, K., Mannon, J. F., Binns, G., & Leung, J. O. (1984) J. Biol. Chem. 259, 770-778]. A series of glycosides, each containing two GlcNAc residues separated by different distances (from 0.8 to 4.7 nm), were synthesized. Inhibition assay with these and other cluster glycosides indicated that clustering of two or more GlcNAc residues increased the affinity toward the chicken lectin tremendously. Among the ligands containing two GlcNAc residues, the structure which allows a maximal inter-GlcNAc distance of 3.3 nm had the strongest affinity, its affinity increase over GlcNAc (monosaccharide) amounting to 100-fold. Longer distances slightly diminished the affinity, while shortening the distance caused substantial decrease in the affinity.(ABSTRACT TRUNCATED AT 250 WORDS)
Acetylgalactosamine↗
Inhibition of electron transfer from adrenodoxin to cytochrome P-450scc by chemical modification with pyridoxal 5'-phosphate: identification of adrenodoxin-binding site of cytochrome P-450scc.
Covalent modification of cytochrome P-450scc (purified from bovine adrenocortical mitochondria) with pyridoxal 5'-phosphate (PLP) was found to cause inhibition of the electron-accepting ability of this enzyme from its physiological electron donor, adrenodoxin, without conversion to the "P-420" form. Reaction conditions leading to the modification level of 0.82 and 2.85 PLP-Lys residues per cytochrome P-450scc molecule resulted in 60% and 98% inhibition, respectively, of electron-transfer rate from adrenodoxin to cytochrome P-450scc (with beta-NADPH as an electron donor via NADPH-adrenodoxin reductase and with phenyl isocyanide as the exogenous heme ligand of the cytochrome). It was found that covalent PLP modification caused a drastic decrease of cholesterol side-chain cleavage activity when the cholesterol side-chain cleavage enzyme system was reconstituted with native (or PLP-modified) cytochrome P-450scc, adrenodoxin, and NADPH-adrenodoxin reductase. Approximately 60% of the original enzymatic activity of cytochrome P-450scc was protected against inactivation by covalent PLP modification when 20% mole excess adrenodoxin was included during incubation with PLP. Binding affinity of substrate (cholesterol) to cytochrome P-450scc was found to be increased slightly upon covalent modification with PLP by analyzing a substrate-induced spectral change. The interaction of adrenodoxin with cytochrome P-450scc in the absence of substrate (cholesterol) was analyzed by difference absorption spectroscopy with a four-cuvette assembly, and the apparent dissociation constant (Ks) for adrenodoxin binding was found to be increased from 0.38 microM (native) to 33 microM (covalently PLP modified).(ABSTRACT TRUNCATED AT 250 WORDS)
Adrenal Cortex↗
Protein-losing enteropathy in systemic lupus erythematosus. Observations by magnifying endoscopy.
We report a 35-year-old man with systemic lupus erythematosus and an associated protein-losing enteropathy that was most likely due to mesenteric venulitis or thrombosis. Evaluation of the patient's intestinal abnormality was aided by the use of magnifying endoscopy; the duodenal villi were lustrous and swollen and of various size, a pattern different from that previously described for intestinal lymphangiectasia. The patient was treated with corticosteroids, resulting in a good clinical response and return of the villi to normal shape and size.
Adult↗
Remarkable effect of endocrine therapy on brain metastasis from prostatic carcinoma.
The brain metastasis of prostatic carcinoma is rare and is distinguished by its poor prognosis in cases which are not surgically resectable. Herein we report on a case of brain metastasis of prostatic carcinoma which showed a dramatic regression through bilateral orchiectomy and doses of diethylstilbestrol diphosphate and estramustine phosphate. Neurologic and psychiatric symptoms diminished within three months, and the patient is alive and well without any subjective symptoms after twelve months.
Adenocarcinoma↗
Constriction of a large coronary artery contributes to serotonin-induced myocardial ischemia in the dog with pliable coronary stenosis.
To clarify the role of serotonin in eliciting myocardial ischemia, the effects of serotonin on coronary vasculature and cardiac performance were examined in anesthetized open chest dogs. Without coronary stenosis, intracoronary infusion of serotonin (0.001 to 1.0 micrograms/kg per min) for 40 s caused dose-dependent increases in coronary blood flow up to 175 +/- 8.3%. During fixed coronary stenosis produced by a metal constrictor that restricted active vasomotion of the stenosed segment, only the highest dose of serotonin increased coronary blood flow by 23 +/- 5.7% and decreased distal coronary pressure without any changes in stenosis resistance and systemic hemodynamic variables. During dynamic coronary stenosis created by inflation of an intraluminal microballoon that preserved active vasomotion of the stenosed segment, intracoronary serotonin (0.1 and 1.0 micrograms/kg per min) evoked marked decreases in coronary blood flow and distal coronary pressure and an increase in stenosis resistance, resulting in an elevation of left ventricular end-diastolic pressure and worsening of left ventricular dP/dt. These deleterious effects were reversed by additive intracoronary infusion of nitroglycerin (10 micrograms/min). The detrimental effects of serotonin were not attenuated by pretreatment with aspirin (10 mg/kg, intravenously), which suppressed adenosine disphosphate- and arachidonic acid-induced platelet aggregation in vitro. Furthermore, intracoronary infusion of serotonin (0.1 and 1.0 micrograms/kg per min) from the site distal to coronary stenosis failed to decrease coronary blood flow during dynamic coronary stenosis. Therefore, these detrimental effects of serotonin during dynamic coronary stenosis could be caused by constriction of a large coronary artery but not by platelet aggregation. Vascular effects of serotonin without coronary stenosis and with dynamic coronary stenosis were inhibited by pretreatment with methysergide (0.3 mg/kg, intravenously), but not with ketanserin (0.5 mg/kg, intravenously) which inhibited serotonin-induced platelet aggregation in vitro. Thus, serotonin produced opposing effects on coronary vasculature, that is, small coronary artery dilation and large coronary artery constriction. Both vascular effects were mediated by non-S2-serotonergic receptor activation of the coronary artery. The data obtained in this study suggest that serotonin-elicited constriction of a large coronary artery may play a crucial role in the pathogenesis of myocardial ischemia during pliable coronary stenosis.
Animals↗
Acid phosphatase in lymphoid tissues of developing chick embryos.
Activity of acid phosphatase (ACPase) in bursa and thymus has been determined in developing chick embryos of either normal or treated with bovine serum albumin (BSA). From the study of light microscopical histochemistry, ACPase activity could be detected in cytoplasm. These ACPase activities were detected in both the follicular medulla and cortex in bursa. In thymus, moderate ACPase activities were also obtained in both the cortex and medulla. ACPase activity in the tissue homogenate was increased in the bursa but the thymic ones showed constant level during the incubation days examined in normal embryos. ACPase distribution observed at the distinct developing stage indicates that ACPase is a useful parameter for growth and development of chicken lymphoid tissues.
Acid Phosphatase↗
Microsomal cytochrome P-450-linked monooxygenase systems and lipid composition of human hepatocellular carcinoma.
The tissues of hepatocellular carcinoma were operatively resected from six patients. All four components of the systems of microsomal cytochrome P-450-linked monooxygenase of the tissues were investigated and compared to those of normal liver tissue. The concentrations of cytochromes P-450, P-420 and b5 were measured optically and the concentrations of all components except cytochrome P-450 were measured by the Western blotting method followed by immunochemical staining. In microsomes of hepatocellular carcinoma tissues, there was as much cytochrome P-450 and other redox components as in the normal liver tissues, but cytochrome P-450 in liver cancer tissues was unstable and easily converted to cytochrome P-420. The specific activities of NADPH- and NADH-ferricyanide and cytochrome c reductase of each sample were also measured. In the microsomes of the cancer tissues, the specific activities were remarkably reduced compared with those of normal liver tissues. The lipid compositions of the microsomes and the phospholipid/cholesterol ratios (w/w) were 13.1 +/- 3.13 in the cancer tissues and 43.0 +/- 6.74 in normal liver tissues. This difference of the lipid composition elucidates the instability of cytochrome P-450 molecules and the inefficiency of the electron transport of cytochrome P-450-linked monooxygenase systems.
Carcinoma, Hepatocellular↗
Purification of human alpha 2-plasmin inhibitor using monoclonal antibody column chromatography.
alpha 2-Plasmin inhibitor (alpha 2PI) was purified directly from human plasma by using a monoclonal antibody affinity column, which recognizes the reactive site of alpha 2PI. alpha 2PI was eluted from the column under mild conditions with 50% v/v ethyleneglycol containing 0.05% Tween 80 in phosphate-buffered saline, pH 7.4. The yield was around 50% and the specific activity of the purified protein was the same as that of the best product of preparation conventionally purified alpha 2PI.
Antibodies, Monoclonal↗
Expression and characterization of pro alpha 2-plasmin inhibitor.
alpha s-Plasmin inhibitor (alpha 2PI), one of the serine protease inhibitors in plasma, was expressed in baby hamster kidney (BHK) cell line. The expression vector was constructed with its genomic DNA and cDNA, and was transfected into BHK cells by the calcium phosphate method. The recombinant alpha 2PI which was secreted from the cells was estimated by SDS-PAGE to have a molecular mass of 67 kDa, which is indistinguishable from that of normal plasma alpha 2PI. The leader peptide of 12 amino acids was retained at the amino terminus of the recombinant alpha 2PI. This finding suggests that alpha 2PI has pre-pro type processing and the propeptide of 12 amino acids is not removed in BHK cells. This pro-alpha 2PI shows essentially the same inhibitory activity on plasmin and the same affinity for plasmin(ogen) as those of normal alpha 2PI. However, the cross-linking ability to fibrin is reduced to less than one-third of that of normal alpha 2PI. The cross-linking site is the glutamine residue located at the second position from the amino terminus of normal alpha 2PI. The conformational change of this region caused by the addition of the propeptide may have affected the cross-linking capacity of the inhibitor.
Amino Acids↗
A protein associated with prodigiosin formation in Serratia marcescens.
A protein associated to prodigiosin formation was found in Serratia marcescens. The protein was not found in nonpigmented strains and was correlated with the pigment level. The protein was about 100 kilodaltons (kDa) and was also found in nonpigmented bacteria of the pigmented strain grown in glucose medium, at high temperature, or under anaerobic condition. The 100 kDa protein was found not in the outer membrane and the periplasm, but in the inner membrane and/or the cytoplasm. The protein was also found singly or dominantly in pigment-protein complexes and pigment-localizing vesicles described in previous reports. These results suggest that the 100 kDa protein is associated with prodigiosin formation.
Bacterial Proteins↗
Kg, a new low-frequency red cell antigen responsible for hemolytic disease of the newborn.
Hemolytic disease of the newborn in a Japanese infant led to studies which indicate that an antibody detected in the maternal serum is recognizing a hitherto unknown red cell antigen. The antigen, which we have named Kg, was found in two generations of the family and it is inherited as a Mendelian dominant character. The maternal serum failed to react with the red cells of more than 600 random Japanese blood donors, 64 red cell samples known to possess low-frequency antigens and all but one of 75 red cell samples known to lack high-frequency antigens without recognized low-frequency antigens. The father's red cells were tested extensively for established low-frequency antigens; only Dia was demonstrated and the maternal antibody was shown not to contain anti-Dia.
ABO Blood-Group System↗
[Oral immunization against tetanus, using liposome-entrapped tetanus toxoid].
A study was made on oral immunization using a tetanus toxoid. Liposome entrapped tetanus toxoid (LTT) was p.o. administered to cats and serum antibody responses were examined. As a result it was known that the LTT antigen induced antibody production in the serum and that an antitoxin antibody titer higher than 0.1 IU/ml, a prophylactic level against tetanus, was produced. On the contrary to this, no antibody was detected in the group administered a tetanus toxoid alone throughout the test period. Also, when an absorbed tetanus toxoid (ATT) was booster-injected to cats p.o. primed with LTT, high responses of the secondary immunization were obtained. Moreover, in case LTT was p.o. administered as a booster antigen, the antitoxin titer in the serum showed a rapid, steep elevation. From these results it has been clarified that LTT on p.o. administration produces an antibody equivalent to the subcutaneous inoculation of ATT.
Administration, Oral↗
[Bronchoalveolar lavage in diffuse panbronchiolitis].
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Adult↗
In vivo responsiveness to glucocorticoid correlated with glucocorticoid receptor content in peripheral blood leukocytes in normal humans.
Dexamethasone loading tests (0.1 mg dexamethasone/kg, iv) were performed in 18 normal males to evaluate the individual responsiveness to glucocorticoid. There were inter-individual differences in increase in peripheral blood polymorphonuclear leukocyte count, decrease in peripheral blood lymphocyte count, and increase in plasma free fatty acids levels after dexamethasone injection. In addition, there was a significant correlation between the maximum increase in polymorphonuclear leukocytes and the maximum decrease in lymphocytes (r = 0.7514, p less than 0.0003). Simultaneous measurements of glucocorticoid receptor content by whole-cell assay revealed that glucocorticoid receptor content in polymorphonuclear leukocytes linearly correlated with that in the corresponding lymphocytes (r = 0.9482, p less than 0.0001). There were also significant correlations between the maximum increase in polymorphonuclear leukocytes and glucocorticoid receptor content in polymorphonuclear leukocytes (r = 0.7239, p less than 0.0007), and between the maximum decrease in lymphocytes and glucocorticoid receptor content in lymphocytes (r = 0.7703, p less than 0.0002). These results suggest that individual differences are preserved both in glucocorticoid responsiveness and in glucocorticoid receptor content in peripheral blood leukocytes in normal humans.
Adolescent↗
Composition of culture media for steroid hormone secretion by murine adrenal tumor cells, Y-1 clone.
Murine adrenal tumor cells (Y-1 clone) were stimulated by adrenocorticotropic hormone (ACTH) and cyclic adenosine 3',5'-monophosphate (cyclic AMP) to produce steroid hormone (delta 4, 3-keto steroids). The steroids were secreted into the medium immediately after synthesis. The optimum concentrations of ACTH and cyclic AMP for stimulation of steroid production were 10(-2) U/ml and 1.0 mM, respectively. In serum-free medium, ACTH and cyclic AMP stimulated steroidogenesis in Y-1 cells, but the amount of steroid hormone in the culture medium was low. However, a high level of steroid production was maintained with medium containing 10 mg/ml bovine serum albumin (BSA). In culture medium containing a higher concentration of BSA, Y-1 cells did not become spherical as is usually the case when steroid production is stimulated by ACTH or cyclic AMP. The morphological changes did not always correlate with steroid secretion by Y-1 cells.
17-Ketosteroids↗