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T Kondo

Publications and source records attributed to T Kondo.

At least 253 records · Page 14Linked to original sources

Effects of EP4 solution and LPD solution vs Euro-Collins solution on Na(+)/K(+)-ATPase activity in rat alveolar type II cells and human alveolar epithelial cell line A549 cells.

BACKGROUND: Intact alveolar epithelial Na(+)/K(+)- adenosinetriphosphatase (ATPase) function is important in preventing alveolar fluid accumulation after lung transplantation. We examined whether the type of preservation solution used influences Na(+)/K(+)-ATPase activity in alveolar epithelial cells. METHODS: Rat alveolar type II cells were preserved with EP4, low-potassium dextran (LPD), or Euro-Collins solution at 7 degrees C for 5 and 20 hours. To assess cell toxicity, we measured cell viability and lactate dehydrogenase release. Na(+)/K(+)-ATPase activity was measured as ouabain-sensitive ATPase hydrolysis. We also examined the effect of terbutaline (10(-3) mol/liter) and dibutyryl cyclic adenosine monophosphate (dbcAMP) (10(-3) mol/liter) on Na(+)/K(+)-ATPase activity in A549 cells preserved for 5 hours. RESULTS: All solutions caused significant damage of rat alveolar type II cells at 20 hours. However, Na(+)/K(+)-ATPase activity was preserved at normal levels with EP4 and LPD over 20 hours. Terbutaline and dbcAMP significantly increased Na(+)/K(+)-ATPase activity in A549 cells preserved with EP4 and LPD solutions for 5 hours. However, we observed no activation in the cells preserved with Euro-Collins solution. We found no significant difference in intracellular cAMP levels after terbutaline challenge among the types of preservation solution. CONCLUSIONS: We conclude that extracellular-type solutions such as EP4 and LPD may be preferable for maintaining not only the basal activity but also the ability to activate Na(+)/K(+)-ATPase in response to beta-adrenergic agonists, in alveolar epithelial cells.

Animals↗

Determination of triazolam by GC-MS in two autopsy cases: distribution in body fluids and organs.

A detailed procedure for analysis of triazolam by GC-MS was constructed in our laboratory. At the concentration of 100 ng/ml, recoveries of triazolam in plasma and urine were 84.9 and 91.0%, respectively. The coefficients of variation in terms of its recovery were 11.5 (plasma) and 10.2% (urine). The detection limit for quantitation by the method was approximately 5 ng/g. This method was applied to two autopsy cases, giving triazolam distribution in body fluids and organs. In one case (33-year-old woman), concentrations of triazolam in the heart blood, urine, brain, lung, liver, kidney, skeletal muscles and stomach contents were 83.9, 741, 106, 165, 507, 293, 125 and 343 ng/g, respectively. From these toxicological data together with autopsy findings, her cause of death was diagnosed as triazolam poisoning. In the other case (45-year-old man), triazolam concentrations in the urine and stomach contents were 7.81 and 41.1 ng/g, respectively, but it could not be detected in the pleural blood; his cause of death was judged to be a traumatic shock, based on autopsy findings.

Journal Article↗

Rapid detection of substrate-oxidizing activity of hiochi bacteria using benzoquinone-mediated amperometric method.

The substrate-oxidizing activity of sake spoilage lactic acid bacteria (hiochi bacteria) was measured using a benzoquinone-mixed carbon paste electrode on which the bacteria had been immobilized. The electrode immersed in a buffer began to produce an anodic current following the addition of D-glucose or gluconate, reflecting the substrate-oxidizing activity of the immobilized bacteria. The time course of the first derivative of the current (dI/dt) exhibited a maximum, the magnitude of which increased with an increase in the number of hiochi bacterial cells immobilized on the electrode. When heat-treated cells were immobilized, the value at the maximum (dI/dt(max)) was lower than that obtained with the electrode on which cells which had not been heat-treated had been immobilized. The decrease in dI/dt(max) increased at higher heating temperatures and longer heating time. The relative decrease in dI/dt(max) due to heat treatment was smaller than the relative decrease in the number of colony formation due to the same treatment, suggesting that cells injured by heat treatment still retain substrate-oxidizing activity.

Journal Article↗

Periodate oxidation of crystalline cellulose.

Conversion of 1,2-dihydroxyl groups to dialdehyde by periodate oxidation is a useful method of derivatizing polysaccharides but has not been extensively utilized in derivatization of cellulose because of complicacy due to the crystalline nature of cellulose. To understand the influence of cellulose crystallinity on this reaction, we investigated how the periodate oxidation proceeds with a highly crystalline cellulose of the marine alga Cladophora sp. The crystallinity of the oxidized cellulose, determined by X-ray diffraction, decreased according to the oxidation level. The half-height widths of equatorial diffraction peaks were nearly unchanged. The solid-state 13C NMR spectra did not show peaks corresponding to aldehyde groups, but solution 13C NMR spectra showed the presence of dicarboxylic groups after subsequent oxidation by sodium chlorite. Transmission electron microscopy showed that microfibrils of Cladophora tended to be bent and more flexible than the original sample. Gold labeling of the aldehyde groups, mediated by thiosemicarbazide derivatization, revealed a highly uneven distribution of dialdehyde groups. When treated by 50% (w/v) sulfuric acid, partially oxidized Cladophora cellulose gave many short fragments of microfibril. These features indicate that the periodate oxidation proceeds by forming dialdehyde groups in longitudinally spaced, bandlike domains.

Aldehydes↗

Identification of flavone C-glycosides including a new flavonoid chromophore from barley leaves (Hordeum vulgare L.) by improved NMR techniques.

Six flavone C-glycosides were isolated from young leaves of barley. One of the C-glucosides has a new type of nucleus, a 2',4',5,5', 7-penta-OH-substituted flavone bearing a 6-C-beta-D-glucoside, which has apparently never been isolated before. One mono- and two di-C-glycosyl flavones were isolated for the first time from barley and identified as isoscoparin 7-O-beta-D-glucoside, carlinoside, and shaftoside, respectively. Other flavones were 7-O-beta-D-glucosides of isoorientin and isovitexin. The known problematic NMR structure elucidation of C-glycosyl flavonoids has been solved by using both a temperature close to the freezing point of the solvent (22.5 degrees C in DMSO-d(6)) and a high temperature (70, 90 degrees C) for comparison during NMR measurements. Structural determination of all the compounds was achieved by employing 1D and 2D NMR techniques.

Flavonoids↗

Immunohistochemical study on Fas and Fas ligand in skin wound healing.

An immunohistochemical study on the expression of Fas and Fas ligand (Fas L) was performed in order to examine the role of apoptosis through Fas-Fas L in mouse skin wound healing. After a 1-cm-long incision in the central dorsum skin, mice were sacrificed at intervals ranging from 0.5 to 240 h, followed by the sampling of wound margin. The expression of Fas and Fas L in the wound margins and in uninjured skin controls was studied using frozen sections. In uninjured skin controls, a very weak expression of Fas and Fas L was detected immunohistochemically in hair follicles, sebaceous glands and epidermal cells. In wounded specimens, polymorphonuclear cells and inflammatory mononuclear ones (round-shaped and spindle-shaped types) were evident. A single immunostaining showed that Fas or Fas L was detectable in inflammatory mononuclear cells involved in the skin wound healing process. Double immunostaining for Fas and Fas L revealed that inflammatory mononuclear cells co-expressed both antigens. In situ TUNEL combined with immunostaining showed that the inflammatory mononuclear cells expressing Fas or Fas L and the polymorphonuclear cells were TUNEL-stained, although neither Fas nor Fas L was detected in the polymorphonuclear cells. The number of TUNEL-positive, inflammatory mononuclear cells expressing Fas or Fas L per 0.01 x 0.01 cm2 was counted. The average number of 10 randomly selected microscope fields reached a peak at the fibro-proliferative phase of wound healing. These results indicate that apoptosis through Fas and Fas L may play an important role for reducing the cellularity during skin wound healing in mice.

Animals↗

Immunohistochemical study on the expression of c-Fos and c-Jun in human skin wounds.

An immunohistochemical study on the temporal expression of c-Fos and c-Jun, both of which designate proto-oncogene products, was performed on 60 human skin wounds with different post-infliction intervals. In unwounded skin, c-Fos or c-Jun was immunolocalized at the nuclei of the epidermal cells in the basal layer, hair follicle cells and sweat gland cells. During the early inflammatory phase of wound healing, the nuclei of polymorphonuclear cells (probably neutrophils), mainly infiltrating at the wound site, were labeled with anti-c-Fos or -c-Jun antibody. As the wound age increased, the neutrophils had disappeared at the wound site, and both mononuclear cells (probably macrophages) and spindle-shaped fibroblastic cells, which expressed a c-Fos or c-Jun positive reaction in the nuclei, were mainly observed. Morphometrically, the distribution of the c-Fos-positive ratio was very similar to that of the c-Jun-positive ratio; the positive ratio was considerably increased in wound specimens with a post-infliction interval of > or = 1 day, thus indicating the late inflammatory or proliferative phase. This study showed that c-Fos and c-Jun were closely involved in the inflammatory phase as well as the proliferative phase of the wound healing process.

Humans↗

Estimation of nitrate metabolism in intestinal tract by measuring breath nitrous oxide concentration in Chinese and Japanese.

We have previously reported that ingestion of vegetables containing high nitrate (NO3-) increases breath nitrous oxide (N2O) concentration, probably due to denitrification. In the present study, we estimated NO3- metabolism in the intestine by determining exhaled breath N2O concentration after the ingestion of vegetables by 16 healthy Chinese and Japanese. Breath samples were collected at 15-min intervals for 4 hr after subjects ingested 180 g of vegetable juice or 50 g of lettuce. Breath N2O concentration was measured by an IR-PAS analyzer. Lettuce but not vegetable juice increased N2O concentrations similarly in Japanese and Chinese. In control subjects who ingested nothing, there were significant differences between Chinese and Japanese in peak N2O concentrations (334 +/- 91 vs 140 +/- 24 ppb, P = 0.027) and total excretions (535 +/- 143 vs 214 +/- 36 microg, P = 0.036). Although the reason for this difference is unclear, Chinese subjects could produce breath N2O from other metabolic pathways than denitrification of dietary NO3.

Adult↗

Association of fasting breath nitrous oxide concentration with gastric juice nitrate and nitrite concentrations and Helicobacter pylori infection.

Fasting breath nitrous oxide (N2O) concentration was determined in relation to pH, nitrate (NO3-) and nitrite (NO2-) concentrations in gastric juice, and Helicobacter pylori infection in 86 successive patients. N2O was measured with an infrared-photoacoustic analyzer. NO3- and NO2- were measured using the Griess reaction. Fasting breath N2O concentration in controls (87 +/- 21 ppb) was not significantly different from that in patients with gastric ulcer or other gastric lesions. Breath N2O was significantly correlated with gastric NO3- (P < 0.01) and was higher in patients with elevated gastric NO2- (246 +/- 87 ppb) than in patients without NO2- (75 +/- 13 ppb). Breath N2O did not differ significantly between subjects who were positive or negative for H. pylori. In conclusion, fasting breath N2O concentration is in some manner related to intragastric NO3- and NO2- concentrations. The possible use of measuring breath N2O as predictors of cancer needs further research.

Adult↗

Encephalitogenic potential of the myelin basic protein peptide (amino acids 83-99) in multiple sclerosis: results of a phase II clinical trial with an altered peptide ligand.

Myelin-specific T lymphocytes are considered essential in the pathogenesis of multiple sclerosis. The myelin basic protein peptide (a.a. 83-99) represents one candidate antigen; therefore, it was chosen to design an altered peptide ligand, CGP77116, for specific immunotherapy of multiple sclerosis. A magnetic resonance imaging-controlled phase II clinical trial with this altered peptide ligand documented that it was poorly tolerated at the dose tested, and the trial had therefore to be halted. Improvement or worsening of clinical or magnetic resonance imaging parameters could not be demonstrated in this small group of individuals because of the short treatment duration. Three patients developed exacerbations of multiple sclerosis, and in two this could be linked to altered peptide ligand treatment by immunological studies demonstrating the encephalitogenic potential of the myelin basic protein peptide (a.a. 83-99) in a subgroup of patients. These data raise important considerations for the use of specific immunotherapies in general.

Adolescent↗

Targeted inversion of a polar silencer within the HoxD complex re-allocates domains of enhancer sharing.

Mammalian Hox genes are clustered at four genomic loci. During development, neighbouring genes are coordinately regulated by global enhancer sequences, which control multiple genes at once, as exemplified by the expression of series of contiguous Hoxd genes in either limbs or gut. The link between vertebrate Hox gene transcription and their clustered distribution is poorly understood. Experimental and comparative approaches have revealed that various mechanisms, such as gene clustering or global enhancer sequences, might have constrained this genomic organization and stabilized it throughout evolution. To understand what restricts the effect of a particular enhancer to a precise set of genes, we generated a loxP/Cre-mediated targeted inversion within the HoxD cluster. Mice carrying the inversion showed a reciprocal re-assignment of the limb versus gut regulatory specificities, suggesting the presence of a silencer element with a unidirectional property. This polar silencer appears to limit the number of genes that respond to one type of regulation and thus indicates how separate regulatory domains may be implemented within intricate gene clusters.

Animals↗

Moderate-intensity regular exercise decreases serum tumor necrosis factor-alpha and HbA1c levels in healthy women.

OBJECTIVE: To evaluate the effects of moderate-intensity regular exercise on serum levels of tumor necrosis factor-alpha (TNF-alpha) and glucose and lipid metabolism parameters. DESIGN: Longitudinal intervention study of a 5 month exercise training program (30-45 min/day, 4-5 days/week). SUBJECTS: Forty-one healthy Japanese women aged 41-69 y at baseline; 27 participants in the exercise program. MEASUREMENTS: Body mass index (BMI), waist-to-hip ratio (WHR), percentage body fat, and fasting levels for serum TNF-alpha, serum soluble TNF receptor p55 (TNF-RI) and TNF receptor p75 (TNF-RII), serum lipids, HbA1c, and serum insulin before and after exercise. RESULTS: In overweight to obese subjects, serum levels of TNF-alpha, TNF-RI and TNF-RII were significantly higher than those in lean subjects. There were significant correlations between log serum TNF-alpha and BMI, percentage body fat, WHR, HbA1c and log insulin. TNF-RI was significantly correlated with BMI, percentage body fat, WHR and HbA1c. TNF-RII was also associated with BMI, percentage body fat and WHR. However, the correlation between TNF-RII and HbA1c did not reach statistical significance. Neither TNF-RI nor TNF-RII was correlated with log insulin. In contrast, TNF-alpha, TNF-RI and TNF-RII were negatively correlated with HDL cholesterol. Regular exercise decreased BMI, percentage body fat, HbA1c, serum TNF-alpha, TNF-RI and TNF-RII and increased HDL cholesterol levels. In addition, exercise-induced change in serum TNF-alpha was independently correlated with changes in HbA1c and serum insulin, after being adjusted for the change in fat-free mass. CONCLUSION: Changes in serum TNF-alpha that occur with exercise may play an important role in improving glucose metabolism parameters.

Adult↗

Incidentally detected liver metastasis of well-differentiated follicular carcinoma of the thyroid, mimicking ectopic thyroid.

A case of incidentally detected liver metastasis of follicular carcinoma of the thyroid, histologically mimicking ectopic thyroid, is described. The patient was a 48-year-old woman. A 2-cm mass was incidentally detected in the left lobe of the liver by abdominal computed tomography (CT) scan. Partial liver resection was performed for diagnosis and treatment. Histologically, the liver nodule was composed of small-to-large follicles containing colloid material. The lining epithelium was flat or cuboidal and showed no cellular or nuclear atypia. Immunohistochemical studies for thyroid-specific proteins, thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4), suggested that the nodule was of thyroid origin. Therefore, a differential diagnosis of metastasis of well-differentiated thyroid cancer, ectopic thyroid tissue and teratoma was made. The patient had a history of subtotal thyroidectomy performed 8 years ago due to a thyroid tumor. The original surgical specimens of the thyroid tumor were diagnosed as follicular adenoma. Additional sections of the specimen were reviewed and an area of convincing vascular invasion was found that was suggestive of follicular carcinoma. Subsequent whole-body examination failed to find other metastases. It was determined that the liver tumor was metastasized from well-differentiated follicular carcinoma of the thyroid.

Adenocarcinoma, Follicular↗

A dynamic analysis of chest wall motions with MRI in healthy young subjects.

OBJECTIVE: The objective of this study was to analyse respiratory-related motion of the chest wall with non-invasive method. METHODOLOGY: Using magnetic resonance image (MRI), 30 sequential images (scanning time, 0.4 s per image) on sagittal, axial and coronal planes were obtained in nine healthy young subjects during quiet breathing (QB) and maximal deep breathing (MDB). The coronal planes were obtained in five of nine subjects during MDB. Ventilation was simultaneously measured with pneumotachometer. RESULTS: There was a linear correlation between instantaneous lung volume and lung cross-sectional area. Motion of the diaphragm and rib cage was also linearly related to instantaneous lung volume. The exception was lower anteroposterior (AP) diameter of the rib cage. The contribution of individual part of the chest wall motion to a unit lung volume change was assessed by slope (S) of the linear regression line. The S at the anterior diaphragm was significantly smaller than those at middle and posterior parts during MDB. The S of middle and posterior diaphragmatic motion was approximately five times that of AP motion of upper rib cage. The S of AP motion of upper rib cage was twice that of transverse motion during either QB or MDB. CONCLUSION: We concluded that dynamic MRI study with concurrent ventilation measurement is a simple and reliable method for evaluation of local chest wall motion, and that neither diaphragm nor rib cage works as a single functional unit during active ventilation.

Adult↗

Mitogen-activated protein kinase, ERK1/2, is essential for the induction of vascular endothelial growth factor by ionizing radiation mediated by activator protein-1 in human glioblastoma cells.

Vascular Endothelial Growth Factor (VEGF)/Vascular Permeability Factor plays an important role in angiogenesis and cell proliferation of cancer cells. Glioblastoma cells are most malignant and show resistance to radiation therapy inducing VEGF to cause angiogenesis and brain edema. In the present study, the regulatory mechanism of the expression of VEGF by ionizing radiation was studied in three human glioblastoma cells. Induction of VEGF mRNA by ionizing radiation was dependent on dose and incubation time. Activator protein-1 (AP-1) was activated by 10 Gy of ionizing radiation in 1 h in T98G glioblastoma cells on an electrophoretic mobility shift assay. We constructed chimeric genes containing various regions of the VEGF promoter gene and the coding region for chloramphenicol acetyltransferase (CAT) and transiently transfected them to T98G cells. CAT assay with the VEGF promoter gene containing an AP-1 site demonstrated that the promoter activity of the VEGF gene was enhanced by ionizing radiation. Immunological analysis of the activity of mitogen-activated protein kinase, ERK1/2, showed that this activity is up-regulated by ionizing radiation. These results suggest that ERK1/2 pathway is involved in the up-regulation of VEGF expression ionizing radiation mediated by AP-1, which may lead to further neovascularization and proliferation of glioblastoma cells resistant to radiation therapy.

Blotting, Northern↗

Minimal peptide length requirements for CD4(+) T cell clones--implications for molecular mimicry and T cell survival.

CD4(+) T lymphocytes usually recognize peptides of 12-16 amino acids in the context of HLA class II molecules. We have recently used synthetic peptide combinatorial libraries to dissect in detail antigen recognition by autoreactive CD4(+) T cell clones (TCC). The results of these studies demonstrated that antigen recognition by T cells is highly degenerate and that many cross-reactive ligands can be defined, some of which much more potent than the selecting autoantigen. Based on these observations, we examined the response of a myelin basic protein-specific HLA class II-restricted CD4(+) TCC to truncation variants of optimal ligands. Surprisingly, pentapeptides, tetrapeptides and even tripeptides derived from different segments of the optimal ligands were recognized by the TCC, and some were even more potent than the selecting autoantigen. In addition, these peptides enhanced the survival of the TCC at low concentration. The relevance of this finding was supported by the generation of pentapeptide-specific CD4(+) TCC from peripheral blood lymphocytes. These observations not only change existing views on the length requirements for activation of CD4(+) HLA class II-restricted T cells, but also extend our knowledge about the flexibility of TCR recognition and the potential for cross-reactivity in the immune system.

Antigen Presentation↗

The current state and problems of circadian clock studies in cyanobacteria.

Circadian rhythms have been observed in innumerable physiological processes in most of organisms. Recent molecular and genetic studies on circadian clocks in many organisms have identified and characterized several molecular regulatory factors that contribute to generation of such rhythms. The cyanobacterium is the simplest organism known to harbor circadian clocks, and it has become one of most successful model organisms for circadian biology. In this review, we will briefly summarize physiological observations and consideration of circadian rhythms in cyanobacteria, molecular genetics of the clock using Synechococcus, and current knowledge of the input and output pathways that support the cellular circadian system. Finally, we will document some current problems in the studies on the cyanobacterial circadian clock.

Biological Clocks↗