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Biomedical subjects

S Fujii

Publications and source records attributed to S Fujii.

At least 541 records · Page 30Linked to original sources

Immunohistochemical localization of adult T-cell leukaemia-derived factor, a human thioredoxin homologue, in human fetal tissues.

An immunohistochemical study of the expression of adult T-cell leukaemia-derived factor (ADF), a human thioredoxin homologue, was performed using a rabbit antibody against the C-terminal peptides of ADF. Tissues were obtained from human fetuses between 9 and 23 weeks of gestation. It was revealed that ADF was widely distributed in different organs and tissues during the fetal period. The ADF antibody reacted selectively with medullary cells of the thymus, lung epithelium, the epithelium of the digestive tract, hepatocytes, bladder epithelium, peripheral nerve cells, hair follicles, sebaceous glands, osteoblasts and the proximal tubules of the kidney. It also reacted with cells destined to differentiate into ciliated cells in the fallopian tube and efferent ductules of the testis, interstitial cells in the ovary, Leydig cells of the testis, and dendritic cells in the spleen and lymph nodes. This is the first report on the thioredoxin system in human cells during the early fetal period. The selectivity of ADF staining in fetal tissues suggests that, during early fetal life, ADF expression correlates well with the cellular function of certain tissues.

Cytokines↗

Prevalence of chronic liver diseases and anti-HCV antibodies in different districts of Saga, Japan.

The authors examined the contribution of Hepatitis C Virus (HCV) to the morbidity of chronic liver diseases (CLD) in selected districts of Saga, Japan, one group with low (L) and the other with a high (H) mortality rate of CLD. Age and sex-matched epidemiological studies showed an extremely high morbidity of CLD in the H-district (5.3%) and a low one in the L-districts (2.1%). Randomized selected studies of anti-HCV antibodies showed an extremely high frequency of 10.8% in the H-district and a frequency of 4.6% in the L-district. In addition, the number of subjects with both CLD and positive anti-HCV antibodies was significantly higher in subjects older than the fifth decade, in the H-district. The high prevalence of HCV may be related to the high morbidity and mortality rate of CLD in these districts of Japan.

Adult↗

Secondary müllerian system and endometriosis.

The pathogenesis of endometriosis is controversial. The two principal histogenetic theories are (1) metastases of endometrial tissues to an ectopic location (metastatic theory) and (2) metaplastic development of endometrial tissue at the ectopic site (metaplastic theory). Studies on the development of the coelomic cavity and the müllerian duct, as well as expression of CA 125 during the fetal period and in the adult, strongly suggest that coelomic epithelium-related tissues and müllerian-derived epithelia of the adult have a shared embryologic ancestry. Many manifestations of müllerian-directed metaplasia suggest that tissues derived from the coelomic epithelium and its accompanying mesenchymal cells, referred to as the "secondary müllerian system," have the potentiality to differentiate into müllerian-directed epithelium and stroma. This metaplastic potentiality of the secondary müllerian system constitutes a basic concept in the pathogenesis of endometriosis.

Antigens, Tumor-Associated, Carbohydrate↗

Expression of stress fibers in bullfrog mesothelial cells in response to tension.

The relationship between stress fibers and tension in mesothelial cells of the bullfrog small intestine was examined by fluorescence cytochemistry using en face mesothelial cell preparations. In nontreated controls, actin revealed by rhodamine-phalloidin staining was localized only along the margins of the mesothelial cells. On the other hand, many stress fibers were formed in the mesothelial cells within 5-7 min after stretching of the intestinal wall in a given direction. The orientation of stress fibers within the cells was coincident with the direction of the tension applied. These cytoplasmic fibers disappeared almost completely from the mesothelial cells within 30 min after the release of tension. According to a difference in the intensity of tension necessary for stress fiber expression, the intestinal mesothelial cells were classified into two groups. Furthermore, cells containing stress fibers in each group showed a rapid increase in number once a given value of tension was applied. The present results indicate that the mesothelial cells of bullfrog small intestine may develop stress fibers to counteract tension exerted on the intestinal wall. Such stress fibers may serve to maintain cellular integrity by strengthening the cellular attachment to subepithelial tissue.

Animals↗

Bovine adrenal cytochrome P-450(11 beta)-mediated conversion of 11-deoxycortisol to 18- and 19-hydroxy derivatives; structural analysis by 1H-NMR.

Incubation of 11-deoxycortisol with a cytochrome P-450(11 beta)-reconstituted system yielded, in addition to cortisol, several new steroid products. In this study, structures of the three steroid products were elucidated. Retention time of the first product (Peak 2 substance) coincided with that of authentic 18-hydroxycortisol on reverse phase HPLC. To further confirm the chemical identity of this product, the purified sample was subjected to 1H-NMR analysis. The spectrum was essentially identical to that of 18-hydroxycortisol. The retention time of the second product (Peak 3 substance) did not coincide with those of commonly occurring steroids. The one- and two-dimension 1H-NMR spectra provided strong evidence for its structure of 19-hydroxy-11-deoxycortisol. The retention time of the third product (Peak 4 substance) did not coincide with those of commonly occurring steroids. The 1H-NMR spectrum showed the presence of signals of 19-CH3 and 18-CH2 protons. There was also evidence that this product is not hydroxylated at the 11-position. Further analysis of the COSY spectra identified its structure as 18-hydroxy-11-deoxycortisol. From these results, we conclude that bovine P-450(11 beta) can catalyze the hydroxylation of 11-deoxycortisol at 11 beta-, 18- and 19-positions and produce cortisol, 18-hydroxy-11-deoxycortisol, 18-hydroxycortisol and 19-hydroxy-11-deoxycortisol.

Adrenal Cortex↗

On the ligand-protein and ligand-flavin interactions in NADPH-adrenodoxin reductase as studied by 31P- and 13C-NMR. Use of 13C-enriched FAD as a probe.

The interaction between 2',5'-ADP and NADPH-adrenodoxin reductase from bovine adrenocortical mitochondria was examined by titrating the enzyme with 2',5'-ADP, while the 31P-signals of 2',5'-ADP were being monitored by 31P-NMR. From the titration profile, the dissociation constant for the complex of the enzyme with 2',5'-ADP was estimated to be 0.22 +/- 0.05 mM. Adrenodoxin reductase was reconstituted with 13C-enriched FADs. The 13C-enriched FADs used were [2-13C]-, [4,10 alpha-13C2]-, and [4 alpha-13C]FAD. The 13C-NMR spectra of these reconstituted enzyme preparations showed 13C-resonance peaks corresponding to the enriched carbon atoms at 160.6 , 165.1, 136.6, and 152.4 ppm (2-, 4-, 4 alpha-, and 10 alpha-13C atoms, respectively). When 2',5'-ADP was bound to the reconstituted enzyme, these 13C-resonance peaks did not shift appreciably from those of the unbound enzyme, whereas in the complex of the reconstituted enzyme with NADP+, the signals for 4- and 10 alpha-13C shifted to higher fields by 2.1 and 0.7 ppm, respectively and the 4 alpha-13C signal shifted to a lower field by 1.4 ppm. These results suggest that in the complex of the enzyme with NADP+ the pyridine moiety is located in the vicinity of C(4 alpha)-C(4) region and that the pi-electron density of the 4 alpha-position of flavin is decreased in the enzyme-NADP+ complex. This argues in favor of the electron transfer from the dihydropyridine moiety of NADPH to the electron-deficient N(5) = C(4 alpha) region of flavin.

Adenosine Diphosphate↗

Kinetics of the hydrolysis of monodispersed dihexanoylphosphatidylcholine catalyzed by bovine pancreatic phospholipase A2: roles of Ca2+ binding and ionizations of amino acid residues in the active site.

Phospholipases A2 (PLA2s) are classified into two groups, I and II, according to differences in the polypeptide-chain length and intramolecular disulfide bondings. The hydrolysis of monodispersed 1,2-dihexanoyl-sn-glycero-3-phosphorylcholine (diC6PC), catalyzed by bovine pancreas PLA2 (Group I) was studied at 25 degrees C and ionic strength 0.2, and the initial velocity data were analyzed by means of the Michaelis-Menten equation. The Michaelis constant, Km, was found to be practically independent of Ca2+ concentration and also of pH value. The latter result indicates no participation of the ionizable groups in the active site in the substrate binding. The pH-dependence curve of the logarithm of the catalytic center activity, kcat, obtained in the presence of a practically saturating amount of Ca2+, showed three transitions ascribable to the participation of three ionizable groups with pK values of 5.00, 8.40, and 9.50. The respective groups were tentatively assigned to the catalytic group His 48, the N-terminal alpha-amino group, and invariant Tyr 52, which is located in close proximity to the imidazole ring of His 48. Deprotonation of His 48 and protonation of Tyr 52 were shown to be essential to the catalysis. The importance of the ionization state of the alpha-amino group was also indicated.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Telecanthal approach for meningiomas in the ethmoid and sphenoid sinuses.

In three cases involving meningiomas in the ethmoid and sphenoid sinuses, transbasal spreading of the interocular distance (telecanthal approach) was used for tumor removal and reconstruction of the skull base. This telecanthal approach involves 1) bilateral en bloc removal of the superior lateral rim of the orbit, the nasal bone, and the posterior lateral wall of the orbit; 2) detachment of the medial canthal ligaments; and 3) spreading of the interocular distance. This approach provides a wide working space beneath the anterior half of the midline skull base, and needs neither a facial incision nor significant retraction of the brain. The surgical technique and its modification are described. The discussion focuses not only on comparisons with other techniques, but on the indications for this approach. Meningiomas originating in the paranasal sinuses are rare; a brief review of the literature concerning the clinicopathological features and pathogenesis is also given.

Adult↗

Serial changes of serum CA125 levels during menstrual cycles.

Serial changes of serum CA125 levels during 32 menstrual cycles were examined in 32 healthy young women (19-21 years of age) charting basal body temperature (BBT) and measuring serum estradiol and progesterone levels. Analysis of BBT charts and serum progesterone levels revealed that the 32 menstrual cycles could be classified into either an ovulatory cycle with a sustained BBT temperature increase for at least 10 days (type I: 9 cycles), an ovulatory cycle with a sustained BBT temperature increase for less than 10 days (type II: 17 cycles), or anovulation (type III: 6 cycles). All 32 cycles exhibited basal CA125 levels of less than 35 u/ml throughout the cycle, except during the period of menstruation. At menstruation, 7 of the 9 cycles of type I showed a marked, transient elevation of CA125 levels from previous basal levels (a mean net increase). On the other hand, 12 of 17 cycles of type II showed only a slight elevation of CA125 levels during the period of menstruation. None of the 6 cycles of type III showed any apparent increase in CA125 levels during the period of menstruation. The mean net increase of CA125 levels during the period of menstruation in the cycles of type I was significantly higher than that of the cycles of type II (p less than 0.05) and type III (p less than 0.05). These results imply that a transient elevation of serum CA125 levels during the period of menstruation occurs in cycles with ovulation, and the levels of elevation seem to be closely associated with the duration of the sustained BBT temperature increase.

Adult↗

Mechanisms contributing to increased synthesis of plasminogen activator inhibitor type 1 in endothelial cells by constituents of platelets and their implications for thrombolysis.

We recently hypothesized that after pharmacologically induced coronary thrombolysis, increased activity of plasminogen activator inhibitor type 1 (PAI-1) retards recanalization, contributes to early reocclusion, or both. This hypothesis was based on the increased elaboration of PAI-1 that we observed in cultured liver cells exposed to growth factors releasable from platelets activated at sites of thrombosis in vivo. PAI-1 released locally is particularly likely to attenuate lysis of thrombi that are targets of thrombolytic drugs. Accordingly, the present study was performed to determine whether synthesis of PAI-1 by endothelial cells is augmented by products of platelets. Lysates from platelets (0.5-8.0 x 10(4)/mm3 media, i.e. less than 10% of the concentration of platelets in blood) increased synthesis and release of PAI-1 into both the extracellular matrix and conditioned media (by 2.8-fold and 3.3-fold within 6 and 24 hours, respectively). Synthesis of neither tissue-type plasminogen activator nor overall protein increased. Increased synthesis of PAI-1 was confirmed by immunoprecipitation of [35S]PAI-1 after metabolic labeling of cells. The increased elaboration of PAI-1 was consistent with increased transcription as reflected by the observed increase in PAI-1 mRNA of 2.2-fold in 4 hours. Effects of platelet lysates were simulated by transforming growth factor beta (TGF-beta), known to be present in platelet alpha-granules and released with platelet activation. Antibody to TGF-beta reduced the stimulation of PAI-1 synthesis by TGF-beta, as expected, by 82%.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗

Effect of human chorionic gonadotropin on the expression of progesterone receptors and estrogen receptors in rabbit ovarian granulosa cells and the uterus.

To investigate whether LH/human CG (hCG) or progesterone acts as a regulator of estrogen receptors (ER) and progesterone receptors (PR) in granulosa cells, we studied the immunohistochemical expression of both ER and PR in the ovary and the uterus of mature rabbits, during the induction of ovulation by FSH followed by administration of hCG, progesterone, or a progesterone antagonist (RU486) and hCG. Granulosa cells pretreated with FSH for 3 days showed ER staining, but negligible PR staining. The staining pattern for ER and PR changed in animals pretreated with FSH followed by hCG injection; by 6 h after hCG injection, we observed the disappearance of ER and the appearance of PR, and by 3 days after hCG injection, we observed the reappearance of ER and the disappearance of PR. However, the expression of ER and PR in the granulosa cells of animals pretreated with FSH followed by progesterone administration instead of hCG was almost the same as that of animals pretreated with FSH alone. In addition, the expression of ER and PR in the granulosa cells of animals pretreated with FSH followed by RU486 and hCG was almost the same as that of animals pretreated with FSH followed by hCG administration. The uterine glandular epithelium, in contrast, began to show decreased appearance of ER and PR by 48 h after hCG injection, and we observed the disappearance of both receptors by 3 days after hCG administration. These results suggest that the expression of ER and PR in granulosa cells is not regulated by the action of progesterone, but by that of LH/hCG.

Animals↗

[Purification and some properties of 3 alpha-hydroxysteroid dehydrogenase from pig adrenal cytosol].

A 3 alpha-reducing activity of 5 alpha-dihydrotestosterone (5 alpha-DHT) was found in pig adrenal cytosol. The enzyme (3 alpha-hydroxysteroid dehydrogenase: 3 alpha-HSD) has been purified to homogeneity from pig adrenal cytosol by ammonium sulfate precipitation followed by DEAE-cellulose, 2', 5'-adenosine diphosphate-Sepharose and Sephadex G-100 column chromatographies. The molecular weight was estimated to be 33,000 and 39,000 by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Isoelectric point was estimated to be 8.5 by isoelectric focusing. The Km and Vmax values for 5 alpha-DHT in the reduction were 10.2 microM and 10.6 nmol/min/mg. The enzyme utilized reduced nicotinamide adenine dinucleotide phosphate (NADPH) or reduced nicotine amide adenine dinucleotide (NADH) in the reduction as a cofactor, but it preferentially required NADPH rather than NADH. Furthermore, the purified enzyme catalyzed not only 3 alpha-reduction of 5 alpha-DHT (9.65 nmol/min/mg), but also catalyzed 20 alpha-reduction of 17 alpha-hydroxyprogesterone (0.58 nmol/min/mg). The enzyme activity of 3 alpha-HSD was strongly inhibited by Hg2+, but it was not inhibited by medroxyprogesterone acetate and some anti-inflammatory agents. No remarkable differences was demonstrated between 3 alpha-HSD and 20 alpha-HSD activity under the influence of heat treatment, divalent cation, anti-inflammatory agents and some inhibitory steroids. These results strongly suggest that 3 alpha-HSD purified from pig adrenal cytosol is a bi-functional enzyme catalyzing 3 alpha- and 20 alpha-HSD activities.

20-Hydroxysteroid Dehydrogenases↗

Altered reactivity of hemolysate-treated cultured smooth-muscle cells from rabbit basilar artery determined by digital imaging microscopy.

During culture, smooth-muscle cells obtained from rabbit basilar arteries were examined for contractile activity by means of differential interference microscopy with a video analysis system (digital imaging microscopy system). This system proved useful for observing the contraction and ultrastructural changes of the living cells. Hemolysate-treated cells showed augmented responses to 5-hydroxytryptamine and leukotriene C4, but not to KCl. This augmented response diminished gradually during the culture period. Both a phospholipase C blocking agent, 2-nitro-4-carboxyphenyl-n,n-diphenylcarbamate (NCDC), and a myosin light chain kinase blocking agent, 1-(5-chloronaphthalenesulfonyl)-1H-hexahydro-1,4-diazepine (ML-9), suppressed this augmented response. Protein kinase C activity of the cells, as measured by Western blot analysis, did not increase during the period of culture with hemolysate. The results obtained suggest that hemolysate had the following effects on the cells: 1) acute but gradual contraction of the cells; 2) augmentation of cellular responses to vasoactive agents; and 3) progressive contraction and morphological alteration of the cells. Possible mechanisms by which hemolysate exerts these effects are discussed, taking into consideration the interrelationship between these effects.

Animals↗

The medical evaluation of patients with diabetes mellitus in exercise therapy.

To investigate myocardial ischemia during exercise, submaximal exercise testing with electrocardiogram and thallium 201-myocardial scintigraphy was performed in non-insulin-dependent diabetics. The relationship between plasma atrial natriuretic polypeptide (ANP) levels in response to exercise and cardiac function assessed by echocardiography was also examined. The following results were obtained: 1) Twelve of the 66 diabetics (18.2%) had ischemic ST-segment changes in exercise ECGs, compared with 4 of 75 control subjects (5.3%)(p less than 0.05). Of the 12 diabetics with positive exercise ECGs, 10 were asymptomatic. 2) The washout rate in thallium-201 myocardial scintigraphy was significantly less in diabetics with positive exercise ECGs (36.5 +/- 4.1%) than in controls (51.9 +/- 2.8%)(p less than 0.05). 3) Although no difference in heart rate were observed between diabetics and controls, diabetic patients had significantly higher systolic blood pressure than controls during all stages of exercise and at maximal exertion. Rate pressure products in stages 1 and 2 were significantly higher in diabetics than in controls. 4) The increase of plasma ANP concentration during moderate exercise (VO2 max 60%) was significantly greater in diabetics than in controls (32.3 +/- 11.9 pg/ml vs. 15.0 +/- 1.7 pg/ml)(p less than 0.02). 5) There was a significant correlation between the increase in plasma ANP during exercise and the levels of left ventricular diastolic function (A/R:r = 0.504, p less than 0.05, IRT:r = 0.587, p less than 0.02), assessed by echocardiography, in diabetics. In conclusion, this study demonstrated that the frequency of asymptomatic myocardial ischemia during exercise was higher in diabetics than in controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrial Natriuretic Factor↗

[Rupture of hepatocellular carcinoma with multiple pulmonary metastasis successfully treated by transcatheter arterial embolization (TAE) of tumor: a case report].

A 66-year-old woman was hospitalized in a state of shock with rupture of hepatocellular carcinoma and multiple pulmonary metastasis. Her bleeding was successfully controlled by emergency transcatheter arterial embolization with Lipiodol (Lp-TAE). Treatments with UFT, OK-432 and two additional Lp-TAE caused the disappearance of pulmonary metastasis with AFP levels decreased and natural killer cell activity increased. The patient died one and a half years after the emergency Lp-TAE. The disappearance of pulmonary metastatic lesions seemed to be caused by improvement of the patient's immunity, which related to the regression of primary tumor after Lp-TAE. It was suggested that Lp-TAE is worth undertaking even in rupture of hepatocellular carcinoma with remote metastatic lesions.

Aged↗

The molecular recognition mechanism of DNA by bleomycin.

The interaction between the antibiotic bleomycin (BLM) and an oligonucleotide d(CCACCTAGGTGG) was studied by 1H NMR. The DNA oligomer was titrated with BLM-VO(III), which is an 'inactive analogue' of BLM-Fe(II). A marked upfield shift was observed for the T10N3H signal, but no other imino proton signals either shifted or broadened. When BLM-VO(III) was titrated with d(CCACCTAGGTGG), a large upfield shift (0.27 ppm) was observed for the delta NH signal of butylguanidinium residue (G). Analysis of NOESY spectra of a 1:1 complex of DNA:BLM-VO(III) strongly suggests that the connectivities of the sequential NOEs of the DNA duplex are the same as those of free DNA. These results indicate that BLM does not intercalate between the base-pairs but binds to DNA in a different manner. A model of 'minor-groove binding' is more likely to explain our NMR data.

Base Sequence↗