Amelanotic acral lentiginous malignant melanoma.
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Aged↗
Biomedical subjects
M Ueda
Publications and source records attributed to M Ueda.
A case of serratiopeptidase-induced subepidermal bullous dermatosis.
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Aged↗
Unique distribution patterns of metastatic lymph nodes in patients with superficial carcinoma of the thoracic oesophagus.
BACKGROUND: Lymph node metastasis is commonly found in carcinoma of the thoracic oesophagus, even when the tumour invades only the submucosa. Although lymph node status greatly influences the outcome, the pattern of early lymphatic spread has not been investigated, and the role of lymph node dissection is still a matter of controversy. METHODS: A series of 110 patients with superficial carcinoma who underwent systematic extended lymph node dissection was investigated retrospectively. RESULTS: Lymph node involvement was found in 0 per cent (none of nine), 23 per cent (five of 22) and 49 per cent (38 of 78) of tumours that invaded the lamina propria, muscularis mucosa and submucosa respectively. Anatomically distant lymph nodes (recurrent nerve nodes and perigastric nodes) were involved more frequently than other intrathoracic nodes adjacent to the main tumour. Only three patients had involvement limited to the intrathoracic group, and in carcinoma that invaded only the muscularis mucosae, all metastatic nodes were located at the thoracocervical junction or in the abdomen. The 5-year survival rate was 89 per cent in the node-negative group and 54 per cent in the node-positive group (P < 0.0003). CONCLUSION: The recurrent nerve nodes and perigastric nodes are the principal proximal regional lymph nodes involved in superficial carcinoma of the thoracic oesophagus. Systematic lymph node dissection, which included these nodes, yielded an acceptable and favourable outcome in patients with node-positive superficial carcinoma.
Carcinoma, Squamous Cell↗
Clinical significance of telomerase activity in the non-cancerous epithelial region of oesophageal squamous cell carcinoma.
BACKGROUND: The aim of this study was to examine telomerase activity in affected and adjacent tissue in patients with oesophageal squamous cell carcinoma (SCC). METHODS: Telomerase activity was measured in oesophageal SCC cells, normal oesophageal culture cells, primary cancer tissues and adjacent normal tissues from patients with oesophageal SCC using a polymerase chain reaction-based assay. RESULTS: All oesophageal SCC cells had telomerase activity, whereas normal cultured cells showed no activity. All 57 cancer tissues showed telomerase activity compared with only five (10 per cent) of 50 normal tissues. Cancer cells infiltrating the vessels of mucosal or submucosal tissues in non-cancerous regions were detected in four of five telomerase-positive normal tissues, whereas such infiltration was detected in only three of 45 telomerase-negative normal tissues. CONCLUSION: In patients with oesophageal SCC, measurement of telomerase activity in normal epithelium is a highly sensitive method of detecting the microinvasion of cancer cells.
Carcinoma, Squamous Cell↗
Effects of hypoxic exercise conditioning on work capacity, lactate, hypoxanthine and hormonal factors in men.
1. Hypoxanthine is a purine degradation product and exercise plasma hypoxanthine can be an index of ATP supply-demand imbalance during exercise. The present study determined the effects of hypoxic exercise conditioning on work capacity, blood lactate, plasma hypoxanthine and various neurohormonal factors. 2. Blood lactate, plasma hypoxanthine and neurohormonal factors (catecholamines, renin-angiotensin system activity and natriuretic peptides) were measured at rest and after maximal cardiopulmonary exercise testing (at sea level) both at pre- and post-hypoxic exercise conditioning in six males (40 +/- 2 years). The training protocol consisted of ergometer exercise twice weekly for 40 min in a hypobaric chamber (61.7-47.2 kPa) for 3 weeks. 3. Pulmonary function and haematological and echocardiographic parameters were not altered after hypoxic exercise conditioning. Work rate at peak exercise (264 +/- 10 vs 321 +/- 31 W; P = 0.10) tended to be increased and peak O2 pulse (15.0 +/- 1.0 vs 18.4 +/- 1.4 mL/beat; P < 0.05) increased after exercise conditioning. The double product during submaximal exercise decreased and systolic blood pressure at peak exercise increased after exercise conditioning. Resting and exercise neurohormonal factors were unchanged, except for reduced resting plasma adrenaline levels. Blood lactate at peak exercise (7.4 +/- 0.7 vs 4.8 +/- 0.5 mmol/L; P < 0.05) became lower and peak plasma hypoxanthine (43.2 +/- 5.7 vs 26.4 +/- 5.0 mumol/L; P < 0.1) tended to be decreased after exercise conditioning. 4. Hypoxic exercise conditioning tended to increase maximal power output with a decrease in exercise blood lactate and a trend towards a decrease in exercise plasma hypoxanthine. These data suggest that exercise conditioning under simulated altitude may improve ATP supply-demand imbalance during exercise with less anaerobiosis, which could contribute to enhanced endurance performance.
Adult↗
Deletion polymorphism in the gene for angiotensin-converting enzyme is not a risk factor predisposing to abdominal aortic aneurysm.
OBJECTIVE: to establish whether deletion of the angiotensin-converting enzyme (ACE) gene is a risk factor predisposing to abdominal aortic aneurysm (AAA) or not. METHODS: the study included 125 patients with AAA and 153 controls randomly selected from 328 individuals. The control subjects were confirmed not to have an AAA, but matched with the AAA group for age, sex, hypertension, diabetes mellitus, and hyperlipidaemia. The presence of ACE polymorphism was detected by a polymerase chain reaction using DNA extracted from blood. RESULTS: the distribution of ACE genotypes and allele frequencies in the control and AAA groups was not significantly different. CONCLUSION: the deletion polymorphism in the ACE gene is not a risk factor for AAA.
Aged↗
A shear-induced in vitro platelet function test can assess clinically relevant anti-thrombotic effects.
Morphological features of haemostatic plugs formed in vitro under high shear forces were investigated. Electron microscopy confirmed the relevance of such haemostatic plug to a platelet-rich arterial thrombus, which is formed in vivo . In rat blood samples, the effects of anticoagulants and various antiplatelet agents on platelet reactivity (rate of haemostatic plug formation) and subsequent coagulation of the flowing blood were investigated. Haemostasis did not occur in citrated blood, and heparin greatly inhibited the shear-induced platelet reaction. Aspirin (1 mM), a thromboxane A(2) receptor antagonist (5 microM), a stable prostacyclin (0.55 nM), a stable prostaglandin E(1) (141 nM) and a phosphodiesterase inhibitor (100 microM) were tested. All these agents exerted significant inhibitory effect on shear-induced platelet reaction, including the inhibition of the very first phase of platelet plug formation, due to aggregation of shear-activated platelets. Except for the phosphodiesterase inhibitor, which prolonged clotting time, none of the above agents affected dynamic coagulation. These results suggest that the employed in vitro shear-induced thrombosis/haemostasis test can reveal in vivo the antithrombotic effect of various agents independently of their mechanism of action.
Journal Article↗
Unusual centrosome cycle in Dictyostelium: correlation of dynamic behavior and structural changes.
Centrosome duplication and separation are of central importance for cell division. Here we provide a detailed account of this dynamic process in Dictyostelium. Centrosome behavior was monitored in living cells using a gamma-tubulin-green fluorescent protein construct and correlated with morphological changes at the ultrastructural level. All aspects of the duplication and separation process of this centrosome are unusual when compared with, e.g., vertebrate cells. In interphase the Dictyostelium centrosome is a box-shaped structure comprised of three major layers, surrounded by an amorphous corona from which microtubules emerge. Structural duplication takes place during prophase, as opposed to G1/S in vertebrate cells. The three layers of the box-shaped core structure increase in size. The surrounding corona is lost, an event accompanied by a decrease in signal intensity of gamma-tubulin-green fluorescent protein at the centrosome and the breakdown of the interphase microtubule system. At the prophase/prometaphase transition the separation into two mitotic centrosomes takes place via an intriguing lengthwise splitting process where the two outer layers of the prophase centrosome peel away from each other and become the mitotic centrosomes. Spindle microtubules are now nucleated from surfaces that previously were buried inside the interphase centrosome. Finally, at the end of telophase, the mitotic centrosomes fold in such a way that the microtubule-nucleating surface remains on the outside of the organelle. Thus in each cell cycle the centrosome undergoes an apparent inside-out/outside-in reversal of its layered structure.
Animals↗
Role of inhaled nitric oxide in ischaemia-reperfusion injury in the perfused rabbit lung.
We have tested if inhaled nitric oxide (NO) is beneficial in ischaemia-reperfusion (IR) lung injury using an isolated perfused rabbit lung model. Ischaemia for 60 min was followed by reperfusion and ventilation with nitric oxide 40 ppm (n = 6) or without nitric oxide ventilation (n = 6) for 60 min. In the control group (n = 6), the lungs were perfused continuously for 120 min. Permeability coefficient (Kfc) and vascular resistance (PVR) were measured serially for 60 min after reperfusion. We also determined the left lung W/D ratio and measured nitric oxide metabolites (NOx) and cGMP concentrations in bronchoalveolar lavage (BAL) fluid from the right lung. IR increased Kfc, PVR and W/D followed by decreased cGMP. Ventilation with nitric oxide restored these changes by preventing the decrease in cGMP. Differences in NOx concentrations in BAL fluid between the control and IR groups were not statistically significant. Our results indicate that IR impaired pulmonary vascular function and resulted in microvascular constriction and leakage. Ventilation with nitric oxide from the beginning of the reperfusion period improved pulmonary dysfunction such as vasoconstriction and capillary leak by restoring cGMP concentrations.
Animals↗
CD9 is expressed in extravillous trophoblasts in association with integrin alpha3 and integrin alpha5.
The CD9 molecule is a 24-27 kDa cell surface glycoprotein, which may be related to Schwann cell migration and adhesion. In this study, we examined the expression of CD9 in human extravillous trophoblasts, which invade into the endometrium during implantation and placentation. CD9 was detected immunohistochemically on the extravillous trophoblasts in the cell columns of first trimester placentae, but not on villous trophoblasts. In the second and third trimester, CD9 was highly expressed on the extravillous trophoblasts in the basal plate of placentae, and in the chorion laeve in the fetal membrane of term placentae. The molecular mass of CD9 in the chorion laeve was shown to be 27 kDa by Western blotting. The mRNA of CD9 was also detected in the chorion laeve by reverse transcription-polymerase chain reaction (RT-PCR). Proteins were purified from chorion laeve by affinity chromatography with anti-integrin alpha3 and alpha5 monoclonal antibodies and Western blotting, revealed that CD9 was associated with both integrins. These findings indicate that CD9 is a differentiation-related molecule present in the extravillous trophoblasts. Since it is associated with integrin alpha5 which has been proposed to regulate trophoblast invasion, CD9 may be implicated in trophoblast invasion at the feto-maternal interface.
Antigens, CD↗
CD9 is involved in invasion of human trophoblast-like choriocarcinoma cell line, BeWo cells.
The CD9 molecule is expressed on human extravillous trophoblasts, which invade the endometrium during implantation and placentation. To elucidate the role of CD9 in trophoblastic function, we investigated the expression of CD9 protein and mRNA in BeWo cells, a human trophoblast-like choriocarcinoma cell line, using immunohistochemistry, Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). When BeWo cells were cultured with anti-CD9 monoclonal antibodies (mAb), their invasion through the extracellular matrices was significantly enhanced in a dose-dependent manner. Cell proliferation and human chorionic gonadotrophin production were unaffected. On the other hand, culture in the presence of mAb against integrins alpha3, alpha5 and beta1, which partially block the interaction with the extracellular matrices, inhibited BeWo cell invasion. Anti-CD9 monoclonal antibody had a stimulatory effect on BeWo cell invasion in the presence of anti-integrin alpha3 antibody. In contrast, it had no effect in the presence of mAb against integrins alpha5 and beta1, which were also highly expressed on BeWo cells. These findings suggest that CD9 has a function connected with the invasive properties of BeWo cells, which is partially mediated by integrin alpha5beta1. This may relate to the involvement of CD9 in trophoblastic invasion.
Antibodies, Monoclonal↗
CD9 is expressed on the cell surface of human granulosa cells and associated with integrin alpha6beta1.
The CD9 molecule is a 24-27 kDa cell surface glycoprotein which is reported to be involved in cell adhesion and migration. Recently, CD9 was shown to be associated with beta1-related integrins. We have previously found that integrin alpha6beta1 is expressed on human granulosa cells (GC) and regulates luteinization of GC in concert with its ligand laminin. In this study, we examined the expression of CD9 in human ovary and the relationship between CD9 and integrin alpha6beta1 in GC. By immunohistochemistry, CD9 was detected on GC in a small antral follicle of <1 mm in diameter. In growing follicles, CD9 was moderately expressed on both GC and theca interna cells (TI). The expression intensity of CD9 on GC increased in preovulatory follicles. In the early luteal phase, CD9 was expressed in both luteinizing GC and TI. The expression intensity on large luteal cells decreased in the mid-luteal phase. In the corpus luteum (CL) of pregnancy, CD9 continued to be expressed on large luteal cells, but not on small luteal cells. By flow cytometry, CD9 was detected on the cell surface in approximately 90% of the isolated GC from patients undergoing in vitro fertilization. The molecular weight of CD9 in the isolated GC was shown to be 26.5 kDa by Western blotting. CD9 mRNA was also detected in the isolated GC and CL by reverse transcription-polymerase chain reaction (RT-PCR). The proteins purified from GC by immunoaffinity chromatography using anti-integrin alpha6 monoclonal antibodies were shown by Western blotting to include CD9 as well as integrin beta1. These findings suggest that CD9 is a differentiation-related molecule of GC and TI and that it is associated with integrin alpha6beta1 on the cell surface of GC, suggesting that CD9 is implicated in the function of human GC in cooperation with integrin alpha6beta1.
Adult↗
Synthesis of telomere-mimic carbocyclic 5'-nor oligodeoxynucleotides.
Telomere-mimic S-ODNs have been synthesized and examined their effects on the proliferation of human tumor cell lines by XTT assay. Furthermore, the guanosine derivatives of carbocyclic 5'-nor nucleoside were synthesized.
Acids, Carbocyclic↗
Single large DNA molecule analysis using fluorescence microscopy.
A large DNA analysis method which enable to obtain spatial information of positions of specific sequences along DNA molecule has been developed. Making use of the phenomenon that large DNA molecule is elongated stably under alternative current field in a concentrated linear polymer solution, direct observation of elongated individual lambda DNA molecules with fluorescence probes was carried out using fluorescence microscopy. Then, the spatial positions of the fluorescence spot of the probe on the DNA molecule were determined by image analysis.
Bacteriophage lambda↗
Direct measurement of DNA by means of AFM.
We have demonstrated that the electrostatic stretch-and-positioning method is useful for the analysis of a long DNA molecule by means of atomic force microscopy (AFM). DNA molecules were stretched parallel to the field line, and immobilized onto the aluminum electrodes patterned on a glass plate. Through AFM observation, we confirmed the immobilization of individual DNA molecules, not aggregate.
Bacteriophage lambda↗
DNA analysis by microfabricated capillary electrophoresis device.
The LIGA (Lithographie Galvanoformung Abformung) process using synchrotron radiation lithography is applied to the microfabrication of capillary array electrophoresis (CAE) device. Laser-induced fluorescence detection system for the CAE device has been constructed by the modification of laser confocal fluorescence microscopy. DNA molecules were detected during migrating in the microchannels filled with polymer separation matrices under electric field to optimize the separation conditions for DNA analysis. Based on this observation, we demonstrated that microfabricated CAE device is realized the fast separation of DNA.
DNA↗
Preparation and evaluation of ODN conjugates with polycation comb-type copolymer.
Oligodeoxynucleotide (ODN) conjugates with the polylysine comb-type copolymer having an ability to promote and stabilize duplex and triplex DNA formation were prepared. 5'-Aminated ODN was succinylated with succinic anhydride. The resulting ODNs having carboxyl terminus were coupled with epsilon-amino groups of the comb-type copolymer using water soluble carbodiimide. The conjugate free from unconjugated ODNs was obtained by gel permeation chromatography. The resulting conjugate maintains ability to form duplex and triplex DNA as estimated by melting curve analysis. Both specificity and stability of the triplex DNA formation were increased by employing the ODN-copolymer conjugates compared to those with their mixture.
Chromatography, Gel↗
Three-dimensional analysis of protein aggregate body in Saccharomyces cerevisiae cells.
Modified genes of peroxisomal isocitrate lyase of Candida tropicalis (CT-ICL) were constructed and expressed in Saccharomyces cerevisiae cells. We observed subcellular localization of expressed products of the mutant CT-ICL genes by immunoelectron microscopy. An unknown structure termed a protein aggregate body (PAB) storing the expressed product was observed in cytoplasm in various mutants (Kamasawa et al. (1996) J. Electron Microsc. 45: 491-497). We chose two typical cells harbouring the mutant ICL genes delta 550 and delta 237-339 to analyse the ultrastructure and three-dimensional (3D) structure of PABs. The PABs had a homogeneous matrix with a wavy periphery in the cell image using a high-pressure freezing fixation method. Although PABs could not be separated from the cytoplasm or mitochondria under a confocal fluorescence microscope, 3D reconstruction of serial electron micrographs clearly showed the PAB was an independent structure of varying size and had the shape of an incomplete sphere. A cell was sometimes observed to have multiple PABs.
Candida↗