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Biomedical subjects

Y Fujii

Publications and source records attributed to Y Fujii.

At least 19 recordsLinked to original sources

Thrombosis in spinal cord injury.

Some traditional coagulation assays and several new molecular markers of hemostatic activation were measured in 37 patients with spinal cord injury (SCI). Twenty one of the patients (57%) developed deep vein thrombosis (DVT). The radiofibrinogen uptake test (RFUT) was used to diagnose DVT. Thirty eight percent of quadriplegic and 88% of paraplegic patients developed DVT (p < 0.005). No significant differences were found in platelet counts, mean platelet volumes, fibrinogen levels, von Willebrand factor (Ag) levels, platelet factor 4 and beta thromboglobulin concentrations between the groups with and without DVT. Fibrinopeptide A, thrombin/antithrombin III (TAT) complexes and plasma D-dimer levels were significantly higher in the patients with thrombosis. Most patients with DVT had elevated TAT complex levels up to three days before the RFUT became positive. D-dimer levels were highest after the diagnosis had been made.

Adolescent

Action of Escherichia coli heat-stable enterotoxin II on isolated sections of mouse ileum.

When Escherichia coli STII was applied to the serosa of the ileum at a concentration of 40 micrograms/ml, an acceleration of the spontaneous motility and a weak contraction were induced 2-3 min later. The induction was not affected by the addition of atropine (10(-6) M), but was abolished by the addition of papaverine (10(-4) M). When STII was applied to the mucosa, the acceleration of the spontaneous motility appeared 7-8 min later, but a contraction was not induced. These results suggest that STII acts directly on muscle cell of the ileum and enhances the spontaneous motility of the intestine.

Animals

CD45 isoform expression during T cell development in the thymus.

Various isoforms of leukocyte common antigen, or CD45, are expressed differentially on T cells at different stages of development and activation. We report studies on CD45 isoform expression on various subsets of human T cells using two- and three-color flow cytometry and cell depletion. Bone marrow cells that were depleted of CD3+ and HLA-DR+ cells were CD45RA-RO-. The earliest CD3-CD4-CD8-CD19- thymocytes were CD45RO- with 20%-30% CD45RA+ cells. The most prominent population of CD4+CD8+ double-positive thymocytes were CD45RA-RO+. Even the CD4+CD8+ blasts were greater than 90% CD45RO+. About 80% of single-positive thymocytes (CD4+CD8- or CD4-CD8+) were also CD45RO+. Only 4.3% of CD4+ and 18% of CD8+ single-positive thymocytes were CD45RA+. In contrast, cord blood T cells which represent the stage that immediately follows single-positive thymocytes, contained 90% CD45RA+ cells. Thus, in terms of CD45 isoform expression, single-positive thymocytes are more like double-positive cells than cord blood T cells. These results suggest the following sequence of CD45 isoform switching during T cell development: CD45RA-RO- or RA+RO- (double-negative thymocytes)----RA-RO+ (double-positive and most single-positive thymocytes)----RA+RO- (cord blood T cells), the last switch from CD45RO to CD45RA occurring as a final step of maturation in the thymus.

Antigens, CD

CD45RA-R0+ subset is the major population of dividing thymocytes in the human.

CD45, or leukocyte common antigen, is expressed in different isoforms on different subsets of thymocytes, suggesting its involvement in the process of T cell development in the thymus. We report studies on CD45 isoform expression on human thymocytes at various stages of development using three-color flow cytometric analysis and cell cycle analysis. Among CD45R0+ cells 18.4% were in S+G2/M phase and represented more than 80% of the dividing cells in the thymus. Among the CD45R0- cells 10.9% were also in cell cycle. Because the CD45R0+ population is almost exclusively CD45RA-, the CD45RA-R0+ subset constitutes the major portion of dividing cells in the thymus. Both the CD1high and the CD3- populations were actively cycling. However, the former was almost 100% and the latter only 50% CD45RA-R0+. Dividing CD45RA-R0+ cells contain, therefore, many cells that have not yet expressed the CD3/T cell receptor complex and presumably have not yet undergone selective procedures. These results are hard to reconcile with the previously presented hypothesis that CD45R0 represents a marker for cells that are destined to die in the thymus. Instead, these results suggest an alternative possibility that CD45R0+ cells may contain cells that can mature and, thus, also constitute the thymic generative lineage.

Antigens, CD

Comparison of bone mineral content among Japanese, Koreans, and Taiwanese assessed by dual-photon absorptiometry.

Throughout the world the diagnosis and management of osteoporosis currently involves the measurement of bone mineral content. There are, however, no studies comparing bone mineral content among Asian people. This cross-sectional study was designed to quantify spine and femur bone mineral density (BMD) in Japanese and compare BMD among Asian people (Japanese, Koreans, and Taiwanese) using the same model dual-photon system (Norland Model 2600). Following a peak BMD in the third and fourth decades, the Japanese BMD values of the lumbar spine and femoral neck showed a clear decrease (annual loss of 0.99 and 0.74%, respectively) with age in females. On the other hand, Japanese BMD values were stable in males until the fifth decade. There was some decrease in BMD with age after the fifth decade, which was much less obvious than that in females. An age-dependent loss of BMD was clearly observed in Japanese and Korean but not in Taiwanese females. Korean males seemed to have a clearer age-dependent loss of BMD compared to Japanese males. Our findings indicate that differences may exist in the BMD of Asian people and that in addition to the quantitative determination of individual BMD, dual-photon absorptiometry may be useful for the comparison of BMD among different ethnic and cultural groups.

Absorptiometry, Photon

Thymus cells in myasthenia gravis: a two-colour flow cytometric analysis of lymphocytes in the thymus and thymoma.

The thymus in patients with myasthenia gravis has germinal centres and the thymic lymphocytes spontaneously produce anti-acetylcholine receptor antibody in vitro. This suggests indirectly that the myasthenic thymus contains mature T and B cells that are autoreactive. We report a two colour flow cytometric analysis of the surface antigen phenotype of lymphocytes in the thymus and in thymoma in 24 patients and in 14 normal thymuses. The results show a decline in the proportion of double positive immature thymocytes with age. This process seems to be partially disrupted in patients with myasthenia gravis. In addition titres of anti-acetylcholine receptor antibody were positively correlated with the proportion of mature single positive T cells and non-T DR positive cells.

Adolescent

Dobutamine increases diaphragmatic contractility in dogs.

The effects of dobutamine on diaphragmatic contractility were studied in 24 dogs anaesthetized with secobarbital and receiving mechanical lung ventilation. The phrenic nerves were stimulated supramaximally for two seconds with electrodes placed around the fifth and sixth cervical roots when the airway was closed at the level of FRC. The stimulating frequency ranged from 10 to 100 Hz. Transdiaphragmatic pressure gradient (Pdi) generated by the electrophrenic stimulation was used as an index of diaphragmatic contractility. The electrical activity of the diaphragm during the stimulation (Edi) was also measured with needle electrodes inserted in the right hemidiaphragm percutaneously. During an infusion of dobutamine (10 micrograms.kg-1.min-1 for 20 min), Pdi increased by 15 +/- 2.1% of control value at 20 Hz stimulation (P less than 0.01), and by 13 +/- 1.2% at 100 Hz stimulation (P less than 0.01). The Edi was not altered by dobutamine infusion. This enhancement of Pdi by dobutamine was abolished by simultaneous infusion of nicardipine, a Ca-channel blocker, but was not affected by prostaglandin E1. These results suggest that dobutamine has a stimulating effect on canine diaphragmatic contraction, and this action may be related to the increased inward movement of extracellular calcium.

Alprostadil

Reversal of CD45R isoform switching in CD8+ T cells.

Both CD4+ and CD8+ T cells express either CD45RA or CD45R0 isoform of CD45R in an exclusive way. Recent reports have shown that CD45RA+ T cells lose CD45RA and gain CD45R0 upon activation. This switching has been suggested to be irreversible although more recently, examples of reversal of CD45R isotype switching in CD4+ T cells have been reported. We report here that freshly isolated unprimed CD8+ T cells, when activated with PHA, temporarily lose CD45RA but reexpress an intermediate level of CD45RA 2-3 weeks after activation with PHA. This reversal seems to take place much more slowly in unprimed CD4+ T cells: the majority of CD4+ T cells that had lost CD45RA and gained CD45R0 remained CD45RA-CD45R0+ in 3 weeks after the stimulation. Also, long-term CD8+ CD45RA+ T cell lines stimulated with PHA or OKT3 showed even more rapid recovery of CD45RA while PPD-specific CD4+ T cell clones retained the original CD45R0 phenotype 3 weeks after stimulation with PPD or PHA.

Antigens, CD

Effect of p-chloromercuribenzoate on Clostridium perfringens beta toxin.

p-Chloromercuribenzoate (PCMB) was shown to bind to Clostridium perfringens beta toxin. Treatment of the toxin with N-ethylmaleimide (NEM), 5,5'-dithio-bis(2-nitro-benzoic acid) (DTNB), o-iodosobenzoate (OIBA) and metal ions such as Cu2+ and Ag+ decreased the lethal activity, but PCMB did not affect the lethal activity. On the other hand, the binding of PCMB to the toxin was inhibited by DTNB and NEM in a dose-dependent manner. Furthermore, the lethal activity of beta toxin pretreated with PCMB was not blocked by treatment with NEM, DTNB, OIBA, Cu2+ and Ag+. However, the PCMB-treated toxin treated with reduced glutathione, dithiothreitol, 2-mercaptoethanol, liver homogenate or serum from mice was inactivated by NEM.

Animals

Magnificent expression of asialo GM1 on thymus cells and spleen T cells of the musk shrew, Suncus murinus.

The expression of asialo GM1 (GA1) was observed on almost all thymocytes from young musk shrew, at the age of 4 weeks by flow cytometric analysis. In adult shrew aged 10 months, the ratio of GA1-negative thymocytes was increased. Among several anti-glycolipid antibodies used, anti-GM1 and anti-Forssman also reacted with the thymocytes. Protein fraction of the thymocytes was analyzed by SDS-PAGE followed by immunoblotting. Anti-GA1 and anti-GM1 showed two bands and one band, respectively, however, their mobilities were different from each other. Anti-Forssman did not stain any protein. The GA1-positive population in spleen T cell fraction was not detected in young shrew but most of the T cells were changed to GA1-positive cells in adult shrew. When mixed lymphocyte culture was performed, the GA1-negative spleen T cells in young shrew were changed to express GA1 marker on their cell surface by differentiation. Abbreviations used were as follows: GA1, Gal beta 1-3GalNAc beta 1-4Gal beta 1-4Glc-Cer; GM1, Gal beta 1-3GalNAc beta 1-4(NeuAc alpha 2-3)Gal beta 1-4Glc-Cer; Forssman, GalNAc alpha 1-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc-Cer.

Animals

Characterization of a murine monoclonal antibody, 5D-4, reacting with pancreatic cancers and islets of Langerhans.

A murine monoclonal antibody (5D-4) was prepared by immunizing mice ip with human pancreatic cancer cell line (AsPC-1). The 5D-4 MAb reacted immunohistochemically with pancreatic and gastrointestinal tract tumors as well as pancreatic islets, and to a less extent with normal tissues. The 5D-4 MAb reacted not only with ca 50 KDa and 30 KDa solubilized protein from AsPC-1 cells by Western blot analysis but also with human insulin in a competition RIA. Double immunoperoxidase staining showed that the 5D-4 MAb cross-reacted with insulin but did not react with glucagon, somatostatin or pancreatic polypeptide. Immunoelectron micrograph of Langerhans island double-stained with the 5D-4 MAb and anti-insulin Ab revealed that the 5D-4 Mab recognized human insulin and ca 50 KDa and 30 KDa antigens in B-cells of islets of Langerhans. Thus, the 5D-4 Mab may be useful for the diagnosis of islet cell tumors as well as pancreatic cancers.

Animals

Biotin derivatives of endothelin: utilization for affinity purification of endothelin receptor.

Three different types of biotinylated endothelin 1 (ET-1) derivatives, [Cys1]-biotinylated ET-1, [Lys9]-biotinylated ET-1, and [Cys1][Lys9]-dibiotinylated ET-1, were obtained when the biotinylation reaction was carried out with sulfosuccinimidyl-6-(biotinamido)hexanoate in an aqueous solvent. The binding of [Lys9]-biotinylated ET-1 to the ET receptor was as efficient as that of natural ET-1, whereas the binding of either [Cys1]-biotinylated ET-1 or [Cys1][Lys9]-dibiotinylated ET-1 was significantly reduced. When ET-1 was reacted with succinimidyl-6-(biotinamido)hexanoate in an organic solvent, ET-1 was exclusively modified at lysine 9. The ET receptor was then isolated from human placenta by affinity chromatography with [Lys9]-biotinylated ET-1 and avidin-agarose. The purified ET receptor was active in ET binding and was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis into two polypeptides with apparent molecular masses of 45 and 35 kDa. The NH2-terminal amino acid sequence indicated that the two polypeptides were from an identical subtype of the ET receptor (ETB, the ligand-nonselective type). A signal peptide from Met1 to Gly26 was missing from the 45-kDa ETB, whereas 64 amino acids at the NH2 terminus were missing from the 35-kDa ETB due to proteolytic cleavage which occurred between Arg64 and Ser65. Indeed, incubation of purified ETB with endopeptidase Arg-C resulted in degradation of the 45-kDa ETB, giving rise to the 35-kDa species by a specific cleavage at Arg64. The 35-kDa ETB was active in binding to ET-1, indicating that the NH2-terminal 64-amino-acid residues are not essential for ligand binding.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence

Long-term treatment in infantile choriocarcinoma.

The long-term treatment of a 5 month old boy with precocious puberty secondary to the production of chorionic gonadotropin (hCG) by a choriocarcinoma is described. Of 13 cases of infantile choriocarcinoma reported in the literature, none were successfully treated. The present study describes a partially successful outcome with embolization of a hepatic tumor, irradiation of lung and right submandibular tumors, chemotherapy with methotrexate (MTX), actinomycin D (ACD), cyclophosphamide (CPA) and etoposide (VP16), and splenectomy and hepatic lobectomy. Subsequently, the residual hepatic tumors were treated with high dose melphalan (HDM) followed by reinfusion of unpurged autologous marrow.

Antineoplastic Combined Chemotherapy Protocols

A case of mixed-type dysgerminoma with a high serum concentration of both human chorionic gonadotropin and alpha-fetoprotein in a child.

Dysgerminoma is divided into two types: pure and mixed. The mixed type is related to other various elements of germ cell tumors. We experienced a case of mixed type dysgerminoma with a high serum concentration of both human chorionic gonadotropin and alpha-fetoprotein. The patient was a 6 year old girl who was admitted to the Hamamatsu University School of Medicine with an abdominal mass. Laboratory investigations revealed elevated serum alpha-fetoprotein and high concentration of serum beta-human chorionic gonadotropin. The tumor originated from the left ovary. The histopathological diagnosis was dysgerminoma. Serum human chorionic gonadotropin and alpha-fetoprotein levels were useful markers in monitoring the response to treatment in this patient.

Angiography

Surfactant replacement therapy in premature babies with respiratory distress syndrome: factors affecting the response to surfactant and comparison of outcome from 1982-86 and 1987-91.

The impact of surfactant therapy on chronic lung disease remains uncertain. During the past decade (1982-91), over 300 babies with respiratory distress syndrome (RDS) weighing 501-2,500 g at birth were consecutively treated with surfactant-TA at our neonatal intensive care unit. Data on 95 RDS babies treated in the first 5 year period (Period 1, 1982-86) were compared with those on 158 RDS babies treated in the second 5 year period (Period 2, 1987-91). Overall respiratory improvement was better in Period 2 than in Period 1. In Period 2, surfactant therapy converted 98% of the babies with moderate/severe RDS to those with 'near normal' lung by 72 hr post-treatment. In Period 2, 95% of the surfactant-treated babies weighing 501-1,750 g at birth survived, 97% of which required no supplemental oxygen at 40 weeks corrected gestational age. Increased survival rate in the surfactant-treated babies during the past decade has not been followed by a parallel increase in chronic lung disease. The severity of the initial pulmonary disease per se was not the significant risk factor for chronic lung disease. Several other variables affecting the response to surfactant therapy and outcome have been identified by stepwise logistic regression analysis and include factors related to perinatal events such as birth asphyxia and infection, and other complications of prematurity.

Chronic Disease

Some properties of purified Escherichia coli heat-stable enterotoxin II.

We examined the biological properties of purified Escherichia coli heat-stable enterotoxin II (STII) using mouse intestinal loop assays and compared these properties with those of heat-stable enterotoxin I (STI) and cholera toxin (CT). The action of STII over time differed from those of STI and CT. STII did not alter cyclic GMP or cyclic AMP levels in intestinal mucosal cells. Our results supported the idea that the mechanism of action of STII in inducing fluid secretion is different from the mechanisms of action of STI and CT. This hypothesis was further supported by the fact that an anti-STII neutralizing serum did not neutralize the activities of STI and CT. Subsequently, we examined the involvement of prostaglandins in the action of STII. The level of prostaglandin E2 in the fluid accumulated as a result of the action of STII increased, and the prostaglandin synthesis inhibitors aspirin and indomethacin significantly reduced the response to STII. These results implicate prostaglandin E2 in the mechanism of action of STII.

Animals

Antihypertensive and metabolic effects of long-term treatment with amosulalol in non-insulin dependent diabetics.

In this open study, 41 hypertensive patients with non-insulin dependent diabetes mellitus were treated with the combined alpha- and beta-adrenoceptor blocker amosulalol hydrochloride for 24 weeks, either alone or added to existing antihypertensive therapy. The effects on blood pressure, glucose and lipid metabolism were examined. Daily administration of 20 to 60 mg amosulalol caused a significant reduction in both systolic and diastolic blood pressure within 2 weeks. This effect was stable, lasting for the entire trial period. The mean systolic and diastolic blood pressure decreased from 174 +/- 13/92 +/- 9 mmHg at the beginning to 148 +/- 16/80 +/- 11 mmHg at the end of the trial. Heart rate was not affected. Plasma glucose and haemoglobin Alc levels showed a tendency to decrease without any statistical significance. Total and HDL-cholesterol and triglyceride levels also remained unchanged. Although 3 patients had complained of dizziness, all were easily manageable. The results indicate that amosulalol is effective in the treatment of hypertension in non-insulin dependent diabetics and does not affect glucose and lipid metabolism.

Adult

Radioimmunoassay for the determination of 2-methoxyestriol concentration in plasma of pregnant women.

A specific assay method has been developed for the determination of 2-methoxyestriol in plasma of pregnant women. The quantitation was achieved by radioimmunoassay after extraction of the plasma with ethyl acetate and purification on a Sephadex LH-20 column. In order to obtain the immunogen for 2-methoxyestriol, 6-oxo-2-methoxyestriol was converted to its 6-(O-carboxymethyl)oxime derivative. The derivative was then coupled to bovine serum albumin by the mixed anhydride method, and rabbits were immunized with this conjugate. The antiserum obtained was partially purified by affinity chromatography on estrone 17-(O-carboxymethyl)oxime-aminohexyl Sepharose conjugate to eliminate the cross-reactive antibodies. Plasma 2-methoxyestriol concentrations in pregnancy were estimated to be mean values of 41.1 pg/ml (12th-14th week), 85.3 pg/ml (27th-29th week), and 97.5 pg/ml (37th-41st week).

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