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Biomedical subjects

Y Sekine

Publications and source records attributed to Y Sekine.

At least 109 records · Page 6Linked to original sources
PubMed1997

Molecular cloning of anti-SS-A/Ro 60-kDa peptide Fab fragments from infiltrating salivary gland lymphocytes of a patient with Sjögren's syndrome.

Anti-SS-A/Ro antibodies are commonly found in systemic autoimmune diseases such as Sjögren's syndrome and systemic lupus erythematosus, and some of these antibodies appear to be responsible for certain pathological lesions including congenital heart block in neonatal lupus. In this study, we generated three human antibody Fab fragments that specifically bind to SS-A/Ro 60-kd peptide from salivary gland lymphocytes of a patient with Sjögren's syndrome by using a phage-display technique. Sequence analysis demonstrated that two of the three Fab clones (E-42 and E-60) used homologous heavy chains derived from the germline VH gene DP73 in combination with different light chains which were derived from germline V kappa gene L6 and V lambda gene DPL23. The third Fab clone (E-56) used another heavy chain derived from the germline VH gene DP31 in combination with the identical light chain as that of E-42. All three Fab clones revealed a high number of somatic mutations that likely occurred in the context of antigen selection. These findings suggest the restricted usage of VH and VL genes of anti-SS-A/Ro antibodies in salivary gland lymphocytes of the patient.

Amino Acid Sequence↗
PubMed1997

Ocular vascular endothelial growth factor levels in diabetic rats are elevated before observable retinal proliferative changes.

Vascular endothelial growth factor (VEGF) is a potent angiogenic factor. VEGF levels in ocular tissue of 6-, 12-, 18- and 28-week-old Goto-Kakizaki (GK) rats, a well-known model of non-insulin-dependent diabetes, were evaluated by highly sensitive ELISA. VEGF concentrations in the GK rat as well as in non-diabetic Wistar rat significantly decreased from the age of 6 weeks to 18 weeks. However, although VEGF concentrations in the Wistar rat continued to fall significantly from 18 to 28 weeks of age, the levels were maintained between 18 and 28 weeks of age in GK rats. Levels were significantly different between the GK and Wistar rats at 28 weeks of age. Results of immunohistochemical studies of the eyes of Wistar and GK rats at 28 weeks of age suggest diffuse distribution of this cytokine in cells of neural origin. Weak to moderate VEGF immunoreactivity was exhibited mainly in the ganglion cell layer, inner plexiform layer and inner/outer nuclear layers in rats with and without diabetes. However, in the retinal optic nerve fiber layer, retinal pigment epithelium and choroid, strong VEGF immunoreactivity was noted only in the GK rat. In conclusion, increased VEGF production in certain ocular tissue, similar to that in humans, is observed quite early, at least before the appearance of observable retinal changes in the diabetic GK rat. This also suggests that the GK rat can be used as a model of initial or latent phase diabetic retinopathy.

Aging↗
PubMed1997

Donor-specific cytotoxic lymphocyte activity from bronchoalveolar lavage during acute canine lung allograft rejection.

OBJECTIVE: We investigated the relationship between acute lung rejection and donor-specific cytotoxic activity (DSCA) in recipient's lymphocytes obtained from bronchoalveolar lavage (BAL). METHODS: A total of 26 mongrel dogs underwent left lung allotransplantation. Dogs received either no immunosuppressive treatment (group I), cyclosporine (group II), or cyclosporine and methylprednisolone for evidence of acute rejection (group III). DSCA was measured by a 51Cr release assay, using lymphocytes from BAL samples as effector cells and 51Cr-labeled donor skin fibroblasts as target cells. The pathologic findings of the transplanted lungs were classified according to the working formulation for classification and grading of pulmonary rejection. In addition, the degree of cellular infiltration in the perivascular, peribronchial, interstitial, and intraalveolar areas was determined based on an infiltration score. RESULTS: DSCA in BAL samples was elevated during mild, moderate and severe acute rejection. The accuracy of the diagnosis of mild or moderate rejection was 92.3% at effector:target (E:T) ratios of 100:1 and 50:1. The DSCA in the BAL fluid and the total infiltration score were correlated closely with correlation coefficients of 0.859 and 0.828 at E:T ratios of 100:1 in group I and group II dogs, respectively. Lung aeration improved and DSCA decreased with methylprednisolone therapy in three of four dogs with grade 2 rejection. CONCLUSION: There is a direct relationship between the DSCA in BAL fluid and the degree of tissue damage caused by acute rejection. The DSCA can be detected by a 51Cr release assay which may hold promise for future clinical applications.

Animals↗
PubMed1997

Inhibition of transpositional recombination by OrfA and OrfB proteins encoded by insertion sequence IS3.

BACKGROUND: An insertion element IS3 is flanked by terminal inverted repeat (IR) sequences. IS3 encodes two, out-of-phase, overlapping open reading frames, orfA and orfB, from which three proteins are produced. OrfAB is a transframe protein produced by -1 translational frameshifting between orfA and orfB, and it is known to be IS3 transposase. OrfA and OrfB are the proteins produced without frameshifting, but their functions have not been elucidated. RESULTS: A plasmid carrying an IS3 mutant that produces only transposase generates miniplasmids--which are the IS3-mediated intramolecular transposition products--as well as characteristic IS3 circles and linear IS3 molecules. OrfA inhibited the generation of these small molecules to a lesser degree, but OrfB did not. OrfB, together with OrfA, however, inhibited the generation more strongly than OrfA alone. OrfA also inhibited the intermolecular transposition of mini-IS3 with the chloramphenicol-resistance gene flanked by IRs to a reduced frequency, and OrfB together with OrfA inhibited it almost completely. OrfA and/or OrfB did not, however, repress transcription from the promoter in the left-terminal region preceding orfA. CONCLUSIONS: The results obtained above show that OrfA and OrfB are not repressors but are inhibitors of transpositional recombination promoted by transposase. OrfA with an alpha helix-turn-alpha helix DNA-binding motif may compete with transposase to bind to terminal IRs. OrfA, together with OrfB that has a DDE motif conserved in retroviral integrases, may inhibit the formation of an active transpososome consisting oftransposase, two terminal IRs and target DNA for the strand transfer reaction. IS3 with a limited size, 1258 bp in length, uses strategies of translational frameshifting and coupling to produce transposase as well as negative regulators to make its copies at a low level, which minimizes a deleterious effect of transposition on bacterial hosts.

Bacterial Proteins↗
PubMed1997

[Evaluation of intrapulmonary nodules by thoracoscopic ultrasonography].

We evaluated the usefulness of thoracoscopic ultrasonography for diagnosing intrapulmonary nodules. We studied 7 patients undergoing thoracoscopic operations. Three had benign lung tumors, one had metastatic lung tumors from a renal cell cancer, and three had primary lung cancers. During thoracoscopy three benign tumors were seen as masses protruding from the visceral pleura in to the pleural space, and two primary lung cancers were seen as pleural indentations. However, the presence of one metastatic lung tumor and one primary lung cancer could not be confirmed by thoracoscopy because of the lack of pleural changes. Furthermore, intrapulmonary spread of malignant tumors in four cases could not be seen by thoracoscopy. All tumors were seen clearly and their characteristics were identified by thoracoscopic ultrasonography. Partial lung resection during thoracoscopy was successful in all patients. We found thoracoscopic ultrasonography to be useful for confirming the presence of peripheral solid nodules, and for deciding where to make incisions for partial lung resection.

Adult↗
PubMed1996

Surgical management of total anomalous pulmonary venous connection. Thirty-year trends.

BACKGROUND: Reports of surgical correction of total anomalous pulmonary venous connection (TAPVC) over the past 30 years indicate a general improvement in operative survival. However, prevention of late pulmonary venous obstruction continues to be a cornerstone of successful repair. The purpose of the study was to identify factors associated with improvement in perioperative mortality and to determine risk factors for death and reoperation due to pulmonary vein stenosis after repair of TAPVC. METHODS AND RESULTS: Using univariate and multiple regression analysis, we analyzed risk of early and late mortality and need for reoperation in 105 patients operated on between April 1966 and June 1995. Despite increased frequency of neonatal repair in the most recent time period (29% in 1966 through 1985; 55% in 1991 through 1995, P < .05), operative mortality declined (13% in 1966 through 1985; 0% in 1991 through 1995). The incidence of postoperative pulmonary hypertensive episodes and death related to pulmonary hypertension decreased significantly over the study period (P < .001). Aggressive preoperative elective medical stabilization and prophylaxis of postoperative pulmonary hypertensive episodes may have contributed to this improvement. By univariate analysis, preoperative pulmonary hypertension (P < .02) and preoperative pulmonary vein obstruction (P < .01) correlated with early mortality up to 1990 but not in the past 5 years. Multiple logistic regression analysis showed that only a small pulmonary confluence associated with diffuse pulmonary vein stenosis was an independent risk factor for early (P < .001) and late (P = .01) death as well as need for reoperation (P = .007). Type of TAPVC was not a significant risk factor throughout the three decades of our experience. At a median follow-up of 87 months, late survival was 98% (93 of 95 operative survivors), and all are NYHA class I. CONCLUSIONS: Improvements on surgical technique as well as preoperative and postoperative management account for the reduction in mortality and need for reoperation for most types of TAPVC. However, the presence of a small venous confluence and diffuse pulmonary vein stenosis remains a risk factor for adverse outcome.

Cardiac Surgical Procedures↗
PubMed1996

Identification and characterization of the linear IS3 molecules generated by staggered breaks.

Insertion sequences IS3 encodes two, out-of-phase, overlapping open reading frames, orfA and orfB. The OrfAB transframe protein that is IS3 transposase is produced by -1 translational frameshifting between orfA and orfB. Efficient production of the IS3 transposase in the cells harboring the IS3-carrying plasmid has been shown to generate miniplasmids as well as characteristic minicircles, called IS3 circles, consisting of the entire IS3 sequence and one of the 3-base pair sequences flanking IS3 in the parental plasmid. Here, we show that the IS3 transposase also generates the linear molecules of IS3 with 3-nucleotide overhangs at the 5'-ends. The nucleotide sequences of the overhangs are the same as those flanking IS3 in the parental plasmid, suggesting that the linear IS3 molecules are generated from the parental plasmid DNA by staggered double strand breaks at the end regions of IS3. The linear IS3 molecules are likely to be the early intermediates in the transposition reaction, which proceeds in a non-replicative manner.

Base Sequence↗
PubMed1996

Thymus-related liposarcoma: report of a case and review of the literature.

We report herein the case of a 77-year-old woman who was admitted to our hospital for further evaluation of an abnormal shadow in the right upper mediastinum that had been revealed by a routine chest X-ray. Computed tomography (CT) and magnetic resonance imaging (MRI) scans confirmed the presence of a thymic tumor, but did not show any evidence of invasion into the central vessels or adjacent structures. Thus, an extended thymectomy with resection of the tumor was performed. Microscopically, normal thymic tissue was found among the tumor cells, and a pathological diagnosis of well-differentiated liposarcoma was established. From the operative and microscopic findings, it is apparent that the tumor was related to the thymus. Postoperative irradiation of 60 Gy was delivered to the operative site and the patient has remained free of disease for 29 months since the operation.

Aged↗
PubMed1996

Estrogen and apoptosis in the developing sexually dimorphic preoptic area in female rats.

Effect of estrogen on apoptotic cell death was studied in the two sexually dimorphic neuronal groups in the developing rat preoptic area (POA): the anteroventral periventricular nucleus of the POA (AVPvN-POA); and the sexually dimorphic nucleus of the POA (SDN-POA). A specific labelling of nuclear fragmentation was performed by terminal deoxynucleotydyl transferase(TdT)-mediated dUTP-biotin nick end-labeling method (TUNEL method) to demonstrate apoptosis. In the AVPvN-POA whose size is larger in females than in males, the number of TUNEL-positive cells was not significantly different between day 5 control and female pups sacrificed 10 h after 25 micrograms estradiol benzoate (EB) injection. However, TUNEL-positive cells showed a significant increase in the female pups sacrificed 24 h after EB injection, compared to that shown in the control female pups. In the SDN-POA whose size is larger in males than in females, EB injection significantly decreased TUNEL-positive cells in the female pups sacrificed 24 h after EB injection, compared to that in controls. These results suggest that estrogen regulates the neuronal number by facilitating apoptotic cell death in the developing AVPvN-POA or by inhibiting it in the developing SDN-POA.

Animals↗
PubMed1996

Pulmonary hypertension after operations for congenital heart disease: analysis of risk factors and management.

BACKGROUND: Management of pulmonary hypertension, a potentially fatal complication of operations to correct congenital heart disease, has evolved through the last 15 years. Monitoring of pulmonary arterial pressure and mixed venous saturation became available, and prophylactic use of alpha-blockers and other vasodilators increased. This study examines risk factors for morbidity and mortality from pulmonary hypertension after operations to correct congenital heart disease and evaluates the impact of management changes on outcomes. METHODS: By means of multivariable logistic regression analysis, 880 high-risk patients with congenital heart disease (of 2484 patients undergoing cardiopulmonary bypass between January 1980 and December 1994) were analyzed to determine which were at risk for postoperative pulmonary hypertension and its associated morbidity and mortality. RESULTS: Patients with atrioventricular canal (n = 182), truncus arteriosus (n = 47), total anomalous pulmonary venous connection (n = 90), transposition of great arteries (n = 97), hypoplastic left heart syndrome (n = 50), and ventricular septal defect (n = 414) demonstrated a higher risk of postoperative pulmonary hypertension. By multivariable logistic regression, preoperative pulmonary hypertension (p < 0.0001), absence of mixed venous saturation monitoring (p < 0.0001), and absence of prophylactic alpha-blockade (p = 0.0004) significantly increased postoperative pulmonary hypertension. Preoperative pulmonary hypertension (p < 0.001) and absence of prophylactic alpha-blockers (p = 0.0004) were significant risk factors for in-hospital death related to pulmonary hypertension. Repair at older age (except in the case of total anomalous pulmonary venous connection) was a significant risk for postoperative pulmonary hypertension (p = 0.03). CONCLUSION: Mixed venous saturation monitoring and alpha-receptor blockade reduced the incidence of pulmonary hypertension after operations for congenital heart disease. Early definitive repair reduced morbidity and mortality from postoperative pulmonary hypertension.

Cardiac Surgical Procedures↗
PubMed1996

Structure-related inhibitory effect of antimicrobial enoxacin and derivatives on theophylline metabolism by rat liver microsomes.

Enoxacin, an antimicrobial fluoroquinolone with a 7-piperazinyl-1, 8-naphthyridine skeleton, is a potent inhibitor of cytochrome P-450-mediated theophylline metabolism. The present study was designed to clarify, using seven enoxacin derivatives, the molecular characteristics of the fluoroquinolone responsible for the inhibition. Three derivatives with methyl-substituted 7-piperazine rings inhibited rat liver microsomal theophylline metabolism to 1,3-dimethyluric acid to an extent similar to that of enoxacin (50% inhibitory concentrations [IC50s] = 0.39 to 0.48 mM). 7-Piperazinyl-quinoline derivatives, 8-hydroenoxacin (8-Hy) and 1-cyclopropyl-8-fluoroenoxacin (8-F1), which have a hydrogen and a fluorine at position 8, respectively, more weakly inhibited metabolite formation (IC50s = 0.88 and 1.29 mM, respectively). Little inhibition (IC50 > 2 mM) was observed in those with 3'-carbonyl and 4'-N-acetyl groups on the piperazine rings. The substrate-induced difference spectra demonstrated that the affinities of enoxacin, 8-Hy, and 8-F1 to cytochrome P-450 were parallel with their inhibitory activities. The substituent at position 8 was found to determine the molecular conformations of the fluoroquinolones, and the planarity in molecular shape decreased in the same order as the inhibitory activity (enoxacin > 8-Hy > 8-F1). Moreover, the 3'-carbonyl and 4'-N-acetyl groups decreased the basicity of their vicinal 4'-nitrogen atoms when judged from their electrostatic potentials, which showed a remarkably broadened negative charge around the nitrogens. As a result, the planarity of the whole molecule and the basicity of the 4'-nitrogen atom of enoxacin are likely to be dominant factors in the inhibition of theophylline metabolism by cytochrome P-450.

Animals↗
PubMed1996

Survey of risk factors for expulsive choroidal hemorrhage: case reports. Substantiation of the risk factors and their incidence.

To evaluate the interaction of risk factors for expulsive hemorrhage, we evaluated 3 cases of expulsive hemorrhage and 2 of acute intraoperative choroidal effusion that occurred at the University of Tsukuba Hospital over a 16-year-period and investigated the common findings in these 5 cases. We also selected randomly 500 cataractous cases operated at the University of Tsukuba Hospital and investigated the probabilities of risk factors for expulsive hemorrhage in these control cases. The 5 affected patients exhibited more than four risk factors of expulsive hemorrhage. The probability of more than four risk factors being involved in any cataract operation was approximately 0.33% (0.17 approximately 0.67%; 95% confidence interval). The incidence of expulsive hemorrhage at our hospital was approximately 0.2%, which is similar to the probability rate. Thus, we concluded that the development of expulsive hemorrhage was proved to be related to an accumulation of the risk factors for expulsive hemorrhage.

Aged↗
PubMed1996

Factors influencing survival following pulmonary resection for metastatic colorectal carcinoma.

The results of pulmonary resection and clinical factors relating to long-term survival were evaluated in 29 patients with histologically proven pulmonary metastasis from colorectal carcinoma. The patients were followed up from 4 months to 120 months, with an average of 43.7 months. Overall 5- and 10-year survival rates were 41% and 34% respectively. The 5- and 10-year survival rates in patients with metastatic nodules 30 cm or less in diameter were 77% and 58%, respectively, a significantly better result than in those with metastatic nodules over 30 mm in diameter. Survival in patients without nodal involvement was significantly better than that in patients with lymph node metastases. Multivariate analysis according to Cox proportional hazards model demonstrated that the size of the metastasis was an independent prognostic factor.

Adult↗
PubMed1996

[Mast cell leukemia evolved from RAEB-T (5q-syndrome) in a 12 year-old girl].

A 12-year-old, female 5q- syndrome case of refractory anemia with excess of blasts in transformation (RAEB-T) evolving to mast cell leukemia is described. This case was admitted because of general fatigue, when her peripheral blood count revealed anemia and leukocytosis with basophil-like cells. RAEB-T was diagnosed based on the laboratory findings of her peripheral blood and bone marrow aspiration, which revealed over 10% peripheral blast cells and dysmyelopoietic changes in all three lineages. Chromosomal analysis of the bone marrow cells showed 46, XX, 5q-. Six months later, the RAEB-T phase evolved to acute leukemia, despite prednisolone, vitamin D3, oxymetholone and low-dose cytosine arabinoside treatment. She had remarkable pancytopenia, hemorrhage, and hepatosplenomegaly, which were not responsive to daunomycin, enocitabine, etoposide, and 6-mercaptopurine, and eventually died. This case was unique in that her karyotype changed to normal; 46, XX, and her blast cells were mast cell lineage during the overt leukemic phase. Interestingly, some blasts were intermediate cells possessing the ultrastructural features typical of both basophils and mast cells.

Anemia, Refractory, with Excess of Blasts↗
PubMed1996

[Effect of superoxide production on nitric oxide release from the pulmonary vascular endothelium during reperfusion after warm ischemia].

It has been reported that reperfusion injury results in the diminished nitric oxide (NO) release from the pulmonary vascular endothelium. However, this mechanism is completely unknown. We examined the effect of superoxide dismutase (SOD) on NO release at reperfusion. A rabbit lung was perfused with Krebs-Henselit buffer in a recirculation system. Cyclic GMP (cGMP) content in the lung effluent from the left ventricle was assayed in a time-dependent manner. In the control group, the lung was immediately perfused without interventing ischemia. In the warm ischemia and reperfusion group, the lung was reperfused after 30 min of warm ischemia. In the SOD group, SOD (60,000 U and 120,000 U) was administered at reperfusion after warm ischemia. The cGMP release in the warm ischemia and reperfusion group significantly decreased compared with the control group. In the SOD group, cGMP release was reversed with increasing doses of SOD, and the cGMP content was significantly reversed at all time points with SOD administration of 120,000 U. The impairement of cGMP release by reperfusion injury was restored by SOD administration. This data suggests that NO release from the endothelium may be quenched by superoxide anion generated at reperfusion. In lung transplantation, efforts must be pursued to preserve the endothelial function such as NO pathway.

Animals↗
PubMed1995

Stable and general-purpose chemiluminescent detection system for horseradish peroxidase employing a thiazole compound enhancer and some additives.

A stable and highly sensitive chemiluminescent detection system for horseradish peroxidase (HRP)/luminol/hydrogen peroxide using a newly designed thiazole compound enhancer has been established. Some additives for the chemiluminescent reaction were explored to overcome some defects of the reaction such as rapid decay and high background of light emission. Recrystallization of luminol and the addition of several detergents into the reacting solution were effective to increase specific light emissions. The addition of skim milk into the reacting solution reduced the background. Consequently, skim milk combined with a detergent increased the signal to noise ratio about 20 times compared with the reactions in the absence of both additives. The optimal concentration of enhancer and the addition of egg albumin stabilized the emission. In the new method, 6x 10(-18) mol of HRP was detectable. This would be the most sensitive enhanced chemiluminescent detection system for HRP. Furthermore, we could detect picogram per milliliter (10(-17) mol) concentrations of a trace component in biological materials such as endothelin-1 by employing this reaction.

Endothelins↗
PubMed1995

IS1-encoded proteins, InsA and the InsA-B'-InsB transframe protein (transposase): functions deduced from their DNA-binding ability.

Insertion sequence IS1 encodes a transframe protein, InsA-B'-InsB, which is produced from two out-of-phase reading frames, insA and B'-insB, by translational frameshifting at a run of adenines. Unless the frameshifting event occurs, the InsA protein is produced from IS1. We found that cells harboring a plasmid carrying an IS1 mutant with a single adenine insertion in the run of adenines contained miniplasmids. Cloning and DNA sequencing analyses of the miniplasmids revealed that they had a deletion extending from an inverted repeat (IR) at the left end of IS1. This indicates that they were generated by IS1-mediated deletion due to efficient production of the InsA-B'-InsB transframe protein that is IS1 transposase. Both the InsA protein and transposase were partially purified as a fusion protein with collagen-LacZ by LacZ-specific affinity column chromatography. The InsA* and the collagenolyzed InsA* were found to bind specifically to a 24-bp region within each of the IRs at the ends of IS1. The transposase Tnp* and the collagenolyzed Tnp* were found to bind to the sequence with or without IR, but preferentially to that with IR. The nonspecific DNA-binding ability of transposase may be involved in recognition of the target DNA, an important process of transposition of IS1. Both InsA and transposase have the IR-specific DNA binding ability and a common polypeptide segment containing the alpha-helix-turn-alpha-helix motif, supporting the previous indication that InsA competes with transposase to bind to IRs and thus becomes a transposition inhibitor. Based on the observations described in this article, we speculate that transposase of IS1 consists of at least two domains, the N-terminal half, which almost entirely overlaps InsA, and the C-terminal half, which almost entirely overlaps B'-InsB. The frameshifting event adds the latter domain to the former to give the transposase activity recognizing IRs and the target sequence to initiate the transposition reaction.

Amino Acid Sequence↗