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Biomedical subjects

M Ueno

Publications and source records attributed to M Ueno.

At least 181 records · Page 10Linked to original sources

p53 gene mutation in 150 dissected lymph nodes in a patient with esophageal cancer.

For thoracic esophageal cancer, we perform extended three field lymph node dissection, and have achieved nearly 50% of overall 5-year survival. However, patients sometimes develop lymph node recurrences in spite of having no lymph node metastases found by conventional histopathologic examination. In a patient with esophageal squamous cell carcinoma, we sequenced all the p53 cDNA translated regions (exon 2-10) of primary carcinoma, and confirmed one p53 nonsense mutation in exon 10. Then we extracted genomic DNA from 150 surgically dissected lymph nodes from that patient, and performed polymerase chain reaction analysis (PCR-RFLP) to detect the same p53 mutation in the lymph nodes. PCR-RFLP analysis showed the same p53 mutation in six lymph nodes. One node was located along the right recurrent laryngeal nerve, where no positive nodes was identified by conventional histopathologic examination. The p53 mutational diagnosis of metastatic cancer may be useful in detecting minimal residual disease.

Carcinoma, Squamous Cell↗

Abnormal neurohumoral responses to exercise in patients with heart disease: inhibition of an increase in endothelin-1 production during exercise.

We have reported that the plasma endothelin-1 (ET-1) level is significantly increased by exercise in healthy athletes and that it is elevated in the circulation of the non-working leg but not the working leg, suggesting that ET-1 plays an important role in redistribution of blood during exercise. This study was designed to compare alterations of neurohumoral substances by exercise in normal subjects and patients with heart disease. Study patients comprised three groups: eight patients with congestive heart failure (CHF) due to Ebstein's anomaly or single-ventricle heart after Fontan operation; six patients with complete transposition of the great arteries (TGA) after an anatomic surgical correction who may be candidates for ischemic heart disease; and five age-matched normal subjects. All patients were in New York Heart Association functional class I. All subjects performed symptom-limited treadmill exercise. It is suggested that patients with CHF or TGA have a manifest or latent exercise intolerance, respectively. In failed to increase plasma ET-1 level, although it caused a greater increase in norepinephrine, angiotensin II, and arginine vasopressin than in the controls. Exercise also caused a delay in the increased response of plasma ET-1 levels in patients with TGA after an anatomic surgical repair. On the other hand, plasma brain natriuretic peptide (BNP) level was augmented by exercise in patients with CHF and patients with TGA but not in the controls. The present results suggest that an increase in ET-1 production during exercise is absent in patients with heart disease. The mechanisms of inhibition of ET-1 production during exercise in patients with heart disease remain to be elucidated. However, the present study suggests that ET-1 plays an important role in redistribution of blood during exercise, and proposes the possibility that failure of an increase in ET-1 production results in exercise intolerance in patients with heart disease.

Child↗

Blood-brain barrier damage in reperfusion following ischemia in the hippocampus of the Mongolian gerbil brain.

Vascular permeability to intravenously injected horseradish peroxidase (HRP) was qualitatively examined in the hippocampus of ischemic Mongolian gerbil brains by light and electron microscopy. After 30 min of right common carotid artery occlusion followed by 90 min of reperfusion, the animal was perfused with a fixative and killed. Before the perfusion of the fixative, HRP was injected into the femoral vein. HRP was visualized with tetramethyl benzidine (TMB) and diamino-benzidine (DAB) for light and electron microscopy, respectively. Staining reaction with TMB for HRP appeared in medial or dorsal portions of the operated side of the hippocampus, especially around some vessels along the hippocampal fissure. Ultrastructural examination in the vessels along hippocampal fissure revealed that the endothelial cytoplasm contained HRP-filled vesicles or vacuoles in close proximity to the basal lamina, and seemed to be slightly electron-dense. Swollen pericytes, swollen astrocytic foot processes and perivascular cells with HRP-filled cytoplasm were also observed in that area. In this study, it was clearly demonstrated that intravascular macromolecules leaked transendothelially, through vessel walls in the hippocampal fissure, from the blood stream in the medial portions of the hippocampus during reperfusion following ischemia. These findings suggest that the blood-brain barrier in some vessels along the hippocampal fissure in the medial parts of the hippocampus is more vulnerable to ischemic insults than those in other brain areas.

Animals↗

Renal haemodynamic characteristics in patients with lupus nephritis.

OBJECTIVE: To clarify the characteristics of renal haemodynamics in patients with lupus nephritis (LN). METHODS: The glomerular filtration rate (GFR) and renal plasma flow (RPF) of 37 patients with active LN were studied longitudinally over an interval of 8 to 144 weeks during treatment with corticosteroids or cytotoxic drugs, or both. All patients had clinical renal disorders and underwent renal biopsies. RESULTS: Analysis of renal biopsy specimens showed that 31 patients had class IV LN. Class II, III, and VLN were present in two patients each. The average GFR increased significantly from 65.4 (SD 33.0) in the pretreatment stage to 86.6 (31.6) ml/min in the post-treatment stage, accompanied by an improvement in urinary or immunological abnormalities, or both. On the other hand, RPF decreased significantly from 625.2 (243.0) to 519.8 (179.0) ml/min. Therefore, the filtration fraction (FF) increased significantly from 10.7 (4.3)% to 16.8 (3.7)%. Low FF was recognised predominantly in patients with class IV LN, but was also observed in patients with other classes. The FF returned towards normal irrespective of the degree of GFR recovery. No significant changes were observed in the levels of blood pressure. CONCLUSION: A reduction in GFR out of proportion to the reduction in RPF as demonstrated by the low FF values was related to the severity of LN or disease activity, or both. Therefore, relative evaluation of GFR and RPF, namely the determination of FF, may be a useful clinical parameter to determine the status of LN.

Acute Disease↗

Antiallergic action of betotastine besilate (TAU-284) in animal models: A comparison with ketotifen.

The effects of betotastine besilate (betotastine: TAU-284), a novel antiallergic drug, on homologous passive cutaneous anaphylaxis (PCA), mediator-induced cutaneous reaction, antigen-induced asthmatic responses and platelet-activating factor (PAF)-induced airway eosinophilia in several animal models, were compared to ketotifen. Betotastine (0.1 mg/kg, p.o.) and ketotifen (1 mg/kg, p.o.) inhibited both rat PCA and histamine-induced cutaneous reaction, whereas they showed little effect on serotonin-induced cutaneous reaction. Betotastine (0.3 mg/kg, p.o.) and ketotifen (1 mg/kg, p.o. ) significantly inhibited antigen-induced bronchoconstriction in guinea pigs which had been passively sensitized with guinea pig IgE antibody. In actively sensitized guinea pigs, the immediate and late phase increase in airway resistance (Rrs) were observed within 5 min and between 4 and 7 h after the aeroantigen challenge. Betotastine (1 mg/kg, p.o.) inhibited both responses. Ketotifen (1 mg/kg, p.o.) inhibited the immediate phase response, but did not affect the late phase response. Exposure of guinea pigs to aerosolized PAF increased the number of eosinophils in bronchoalveolar lavage fluid 24 h after the stimulation. Betotastine (3-10 mg/kg, p.o.) dose-dependently inhibited PAF-induced accumulation of eosinophils in the bronchoalveolar cavity. In contrast, cetirizine (10 mg/kg, p.o.) showed a tendency to inhibit eosinophil accumulation, and ketotifen (10 mg/kg, p.o.) and terfenadine (10 mg/kg, p.o.) did not have any affect. These results indicate that betotastine could be useful in the treatment of allergic disease such as bronchial asthma.

Animals↗

Significance of glomerular deposition of protein S in various glomerulopathies.

To elucidate the relationship between glomerular deposition of protein S (PS) and renal lesions or dysfunction, 30 patients with various glomerulopathies were examined. Glomerular PS deposition was found in 20 patients (group A), and other 10 patients showed no deposition (group B). PS was found mainly along the capillary loops and segmentally in the mesangium. Group A showed significantly more severe proteinuria than group B (p < 0.05). Group A patients showed significant decreases in glomerular filtration rate (p < 0.01). Patients in group A had significantly lower plasma levels of plasmin-alpha2-plasmin inhibitor complexes (p < 0.05) and thrombin-antithrombin III complexes (p < 0.01) than those in group B. Group A showed significant decreases in the mean values of plasma total PS (p < 0.01) and protein C (PC) antigens (p < 0.01) and C4b-binding protein (C4bp; p < 0.05) as compared with group B patients. There was a positive correlation between plasma PS and C4bp (p < 0.02). Histologically, group A showed a significantly higher incidence of glomerular deposition of factor XIII (subunit a), alpha2-plasmin inhibitor, PC (p < 0.05), and C4bp (p < 0.01). The present study demonstrates that glomerular PS deposition indicates the existence of PC and C4bp in the glomeruli and suggests that the glomerular PS deposition may modify the activation of fibrinolytic and coagulation systems within the glomeruli in various glomerulopathies.

Adult↗

Molecular cloning of cDNA for BRab from the brain of Bombyx mori and biochemical properties of BRab expressed in Escherichia coli.

From a brain cDNA library of Bombyx mori, we cloned cDNA for BRab, which encoded a 202-amino-acid polypeptide sharing 60-80% similarity with rab1 family members. To characterize its biochemical properties, cDNA for BRab was inserted into an expression vector (pGEX2T) and expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant protein was purified to homogeneity with glutathione S-Sepharose. The purified GST-BRab bound [35S]-GTP gamma S and [3H]-GDP with association constants of 1.5 x 10(6) M-1 and 0.58 x 10(6) M-1, respectively. The binding of [35S]-GTP gamma S was inhibited with GTP and GDP, but with no other nucleotides. The GTP-hydrolysis activity was evaluated to be 5 m mole/min/mole of BRab. In the presence of 6 mM MgCl2, bound [35S]-GTP gamma S and [3H]-GDP were exchanged with GTP gamma S most efficiently. These results suggest that BRab, having a higher affinity for GTP than GDP, converts from the GTP-bound state into the GDP-bound state by intrinsic GTP hydrolysis activity and returns to the GTP-bound state with the exchange of GDP with GTP.

Amino Acid Sequence↗

Microdialysis assessment of 5-fluorouracil release from thermosensitive magnetoliposomes induced by an electromagnetic field in tumor-bearing mice.

The current study was designed to evaluate the properties of thermosensitive magnetoliposomes (TMs), a new drug carrier proposed by the authors, in an electromagnetic field pertaining to their selective heating and drug release under an in vivo condition. TMs containing 5-fluorouracil (5-FU) were prepared by reverse-phase evaporation, injected into the tumor mass of B 16-BL6 melanoma in mice, and selectively heated by a 500-kHz electromagnetic field. The release profile of 5-FU from TMs was examined by using a microdialysis technique. The temperature of TMs in the tumor was effectively elevated to 42 degrees C and maintained at this temperature, overcoming the "cooling effect" of blood flow and surrounding tissues. The release kinetics of 5-FU from TMs was successfully analyzed by physiological modeling, which allows the prediction of intratumor drug concentrations during electromagnetic field exposure under various conditions. In conclusion, this study first demonstrated an in vivo evidence for the electromagnetic field-induced thermosensitive release of a drug from TMs in a tumor with the use of microdialysis.

Animals↗

[Anti-proliferative effects of heating on the human prostatic carcinoma cells in culture].

Prostatic cancers are well-known to be sensitive to heat stress. However, the mechanism by which the cancer cells are killed by high temperature remains poorly understood. The present study was undertaken to determine the anti-proliferative effects of heat stress on the prostatic cancer cells in culture. Heat shock at 43 degrees C inhibited the cell growth of three different prostatic cell lines. Flow cytometrical analysis using BrdU and PI showed a decrease in the proportion of cells in an S phase, accompanied by cell accumulation in G1 and G2, in both JCA-1 and PC-3 but not in LNcap. Both JCA-1 and PC-3 presented a strong expression of hsp70 at 37 degrees C. The heat shock caused apparent enhancement of the expression of hsp70 through the cell cycle. A treatment at 43 degrees C for 8 hours resulted in not only an apparent increment of positive hsp70 cells, but cells with subdiploid DNA content in LNcap. Flow cytometrical analysis by FITC-labeled Annexin V showed increment of apoptotic cells at 43 degrees C for 8 hours in LNcap cells. The results suggest that apoptosis is an important pathway of heat-induced killing of these cells. In conclusion, the cell growth of prostatic cancers may be affected by the temperature through relationship of the cell cycle and hsp70.

Apoptosis↗

[Traumatic aortic regurgitation: report of two surgical cases].

Two successful surgical cases with aortic regurgitation due to blunt trauma are reported. Case 1: A 48-year-old female was transferred to our hospital after a traffic accident. She was intubated and mechanically ventilated. Twelve days later, echocardiography demonstrated massive aortic regurgitation. The next day, aortic valve replacement was performed using a 21 mm SJM mechanical valve. The central part of the non-coronary cusp was torn and perforated 15 mm long. She is in good condition three years after surgery. Case 2: A 57-year-old male was transferred to the nearest hospital because he was involved in a snowslide accident. A month after the injury, catheterization showed massive aortic regurgitation and pulmonary hypertension. An aortotomy revealed that the intima around the right coronary ostium was completely torn. A perforation with a diameter of 3 mm was also observed on the right coronary cusp. After coronary cusps were excised, interrupted mattress sutures buttressed with pledgets were passed through the aortic wall at the distal level of the intimal laceration and then to the aortic annulus. A 25 mm SJM prosthetic valve was seated at the aortic ring. As the right coronary ostium was included in the plicated aortic wall, aortocoronary bypass was constructed using the great saphenous vein. He is doing well 14 months after surgery. Aortic regurgitation due to blunt trauma is rare and difficult to diagnose. Sometimes cardiac failure progresses rapidly. Therefore, urgent surgery is recommended as soon as the diagnosis is confirmed.

Aortic Aneurysm↗

Association between tumor necrosis factor in serum and cachexia in patients with prostate cancer.

The present study was undertaken to evaluate the relationship between serum tumor necrosis factor (TNF) and cachexia in patients with prostate cancer. TNF levels were determined in 110 serum samples from prostate cancer patients by an enzyme immunoassay. Serum TNF activity was positive in 76% of the patients with relapsed disease, whereas only 11% of the untreated patients and 0% of the patients in remission as a result of endocrine therapy were positive. The serum total protein and albumin levels, hemoglobin levels, and body mass index of the patients with elevated serum TNF levels were significantly lower (P < 0.05) than the corresponding values in patients with undetectable serum TNF levels. The serum TNF levels of patients with serum albumin levels of <3.5 g/dl, serum total protein levels of <7.0 g/dl, hemoglobin levels of <11.0 g/dl, and a body mass index of <21 kg/m2 were significantly higher (P < 0.05) than the values in their respective counterparts. There was a significant correlation between the detectability of serum TNF and performance status (P < 0.05). Patients with elevated serum TNF levels had a significantly higher mortality rate (P < 0.05) than those with undetectable serum TNF levels. These findings suggest that TNF may be one of the factors contributing to the complex syndrome of cachexia in patients with prostate cancer.

Adenocarcinoma↗

Analysis of renal pathology and drug history in 158 Japanese patients with rheumatoid arthritis.

To clarify the characteristics of renal pathology and its correlation with disease-modifying antirheumatic drugs (DMARDs) in patients with rheumatoid arthritis (RA), renal biopsy findings from 158 Japanese RA patients with urinary abnormalities and/or renal dysfunction were analyzed retrospectively in the period between 1979 and 1996. Urologic abnormality and urinary tract infection were ruled out in all patients. Light and immunofluorescence (IF) microscopy were performed in all patients. Mesangial proliferative glomerulonephritis (MesPGN) was diagnosed in 54 patients, membranous nephropathy (MN) in 49, and secondary amyloidosis (AM) in 30. Renal dysfunction was more frequent in patients with AM (22/30) than in patients without (40/128). Forty of 49 MN patients developed renal disorders during DMARDs therapy. The prevalence of DMARD-related MN increased during the period of observation. The fact that DMARDs are of very frequent use in recent Japanese RA patients may reflect the prevalence of MN in this study. Two thirds of patients with MesPGN developed renal disorders when no DMARDs were used. One half of 54 MesPGN patients demonstrated IgA glomerulonephritis (GN) by IF. The prevalence of primary renal diseases in Japan may reflect the frequency of IgA GN in Japanese RA patients. Furthermore, diffuse thinning of the glomerular basement membrane (GBM) was observed in 30 of 81 patients with electron microscopy. RA itself may underlie the pathogenesis of GBM thinning, and drugs used for RA treatment may also accelerate the development of this lesion. In conclusion, although MesPGN, MN, and AM may be relatively more common, IgA GN and GBM thinning also were other frequent entities in Japanese RA patients. No correlation was observed between DMARDs and renal disorders excepting MN.

Adolescent↗

A suppressive role of p125FAK protein tyrosine kinase in hydrogen peroxide-induced apoptosis of T98G cells.

Protein tyrosine phosphorylation was examined on a human glioblastoma cell line, T98G, after exposure to oxidative stress in vitro. Hydrogen peroxide (1 mM) markedly induced tyrosine phosphorylation of a 125 kDa protein at 30 min after stimulation. The 125-kDa molecule phosphorylated was revealed to be a focal adhesion kinase (FAK). Tyrosine phosphorylation of p125FAK continued at least up to 5 h, and decreased after 8 h concomitant with apoptosis. Tyrosine phosphorylation of p125FAK was blocked by herbimycin A, a potent inhibitor of protein tyrosine kinases, while apoptosis was accelerated. When T98G cells were incubated with FAK antisense oligonucleotide, apoptosis was also accelerated. These results suggest that tyrosine phosphorylation of p125FAK plays a suppressive role in hydrogen peroxide-induced apoptosis.

Antibiotics, Antineoplastic↗

Extensive RNA editing and possible double-stranded structures determining editing sites in the atpB transcripts of hornwort chloroplasts.

Three nonsense codons and an unusual initiation codon were located within the putative coding region of the atpB gene of chloroplast DNA of the hornwort Anthoceros formosae. Nucleotide sequencing of cDNA prepared from transcripts revealed extensive RNA editing. The unusual initiation codon ACG was changed to AUG and three nonsense codons were converted into sense codons. In total 15 C residues of the genomic DNA were replaced by U residues in the mRNA sequences, while 14 U residues were replaced by C residues. This is the highest number of editing events for a chloroplast mRNA reported so far. Partial editing was also shown in a cDNA clone where 23 sites were edited but six sites remained unedited, representing the existence of premature mRNA. The expected two-dimensional structure of the mRNA shows the existence of a sequence complementary to every editing site, which can produce continuous base pairing longer than 5 bp, suggesting that mispairing in the double strand is the site determinant for RNA editing in Anthoceros chloroplasts. Comparison of the cDNA sequence with other chloroplast genes suggests that the mechanism arose in the first land plants and has been reduced during evolution.

Amino Acid Sequence↗

Effect of dicetylphosphate or stearic acid on spontaneous transfer of protein from influenza virus-infected cells to dimyristoylphosphatidylcholine liposomes.

Membrane proteins, such as viral spike, were transferred spontaneously from influenza virus-infected cells to various liposomes. The protein transfer was enhanced by the presence of negative charged component dicetylphosphate (DCP) or stearic acid (SA) in dimyristoylphosphatidylcholine (DMPC) liposomes. The lowering of membrane fluidity did not relate to the effect of DCP or SA on protein transfer in this study. We considered that the alteration of membrane properties, such as construction of the surface or stability of transferred protein in liposomes, due to the specific structure of DCP or SA is responsible for the enhancement of spontaneous protein transfer by the presence of the amphiphilic components.

Animals↗

A novel function of transferrin as a constituent of macromolecular activators of phagocytosis from platelets and their precursors.

Macromolecular activators of phagocytosis from platelets (MAPP:s-MAPP, 1500kDa and 1-MAPP, 300kDa) are glycoproteins released from human platelets and activate leukocyte phagocytosis via the Fc gamma receptors. Their production can be shown in CPD-stored platelets which have lost MAPP releasing activity by incubation with plasma-derived precursos of MAPP (pre-MAPP : pre-s-MAPP, 150kDa and pre-l-MAPP, 300kDa) in the presence of Ca++. Partial amino acid sequence analysis of s-MAPP revealed that it had homogeneity with transferrin (TF). Affinity chromatography using anti TF immunosorbent column showed that all of pre-MAPP and MAPP had immunoreactivity with anti TF antibodies. S-MAPP and 1-MAPP rich preparations from platelet release products lost their activity after treatment with ATP at acidic pH, in which condition iron atoms could be removed from holo-transferrin molecules. In the experiment using polymerized TF and stored platelets, it was shown that platelets incubated with dimer and tetramer TF could produce MAPP function. These results suggest that dimer and tetramer transferrin are precursors of s-MAPP and 1-MAPP, respectively and that iron atoms are necessary for their phagocytosis activating function.

Amino Acid Sequence↗