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Biomedical subjects

Y Asai

Publications and source records attributed to Y Asai.

At least 91 records · Page 5Linked to original sources
PubMed1999

Interaction of alpha-Tocopherol and Soybean Oil with Phosphatidylcholine and Their Formation of Small Dispersed Particles.

In this study, alpha-tocopherol (alpha-T) was dispersed by cosonication with soybean phosphatidylcholine (PC). The particle size in the dispersion was increased to 250 nm up to the alpha-T mole fraction (XT) 0.4. At XT = 0.5, the alpha-T/PC mixture was difficult to disperse and the macroscopic oil/water phase separation was observed. By the addition of soybean oil (SO) to alpha-T (molar ratio alpha-T:SO = 1:1), stable aqueous dispersions (diameter 50-70 nm) were obtained in the mole fraction range for the alpha-T and SO mixture (XM) 0.1-0.8. In order to clarify the dispersal mechanism, the dispersed particles were characterized and the interaction among alpha-T, SO, and PC was investigated using several physicochemical techniques. The trapped aqueous volume inside the alpha-T/PC particles was determined using the aqueous space marker calcein, and this volume was increased with the addition of alpha-T into small unilamellar vesicles of PC. The trapped aqueous volume of alpha-T/SO/PC particles was decreased remarkably with an increase in XM, and the decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the PC membrane by the addition of the quencher CuSO4. These results indicate that the interaction of alpha-T with PC bilayers and the structure of the alpha-T/PC mixture will be changed by the addition of SO. Copyright 1999 Academic Press.

Journal Article↗
PubMed1999

The interaction of ubiquinone-3 with phospholipid membranes.

The effects of ubiquinone-3 (UQ) on dipalmitoylphosphatidylcholine (DPPC) membrane were studied by surface monolayer, differential scanning calorimetry (DSC) and fluorescence techniques. DPPC and UQ are proved to be freely miscible in the mixed monolayer at an air/water interface, and to be partially miscible in bulk phase, i.e. bilayer and solid phase. There is a condensing interaction between UQ and DPPC in the UQ/DPPC mixed monolayers. The solubility of UQ in the DPPC is about 20 mole% and the solubility of DPPC in UQ is about 10 mole%. The membrane fluidity of DPPC was increased by the addition of UQ and the phase transition temperature was decreased.

Membrane Fluidity↗
PubMed1999

Na+-driven flagellar motor resistant to phenamil, an amiloride analog, caused by mutations in putative channel components.

The rotation of the Na+-driven flagellar motor is specifically and strongly inhibited by phenamil, an amiloride analog. Here, we provide the first evidence that phenamil interacts directly with the Na+-channel components (PomA and PomB) of the motor. The alterations in Mpar (motility resistant to phenamil) strains were mapped to the pomA and/or pomB genes. We cloned and sequenced pomA and pomB from two Mpar strains, NMB205 and NMB201, and found a substitution in pomA (Asp148 to Tyr; NMB205) and in pomB (Pro16 to Ser; NMB201). Both residues are predicted to be near the cytoplasmic ends of the putative transmembrane segments. Mutational analyses at PomA-Asp148 and PomB-Pro16 suggest that a certain structural change around these residues affects the sensitivity of the motor to phenamil. Co-expression of the PomA D148Y and PomB P16S proteins resulted in an Mpar phenotype which seemed to be less sensitive to phenamil than either of the single mutants, although motility was more severely impaired in the absence of inhibitors. These results support the idea that PomA and PomB interact with each other and suggest that multiple residues, including Asp148 of PomA and Pro16 of PomB, constitute a high-affinity phenamil-binding site at the inner face of the PomA/PomB channel complex.

Amiloride↗
PubMed1999

Treatment of life-threatening huge atrial myxoma: report of two cases.

We herein report two patients with left atrial myxoma who needed an emergency operation. Case 1 was a 48-year-old woman who was injured in a traffic accident and underwent an operation for a right leg fracture. Just after the operation she developed cardiac and respiratory arrest with complaints of chest pain. She was successfully resuscitated and diagnosed to have a left atrial myxoma by echocardiography. Emergency surgery was performed and a giant left atrial myxoma was thus removed from the atrial septum. Case 2 was a 54-year-old housewife who was transferred to our department under the diagnosis of a left atrial myxoma by echocardiography. She complained of dyspnea and chest discomfort. By angiography, the tumor was seen to be partially incarcerated at the diastolic phase. A huge myxoma was removed from the atrial septum which was secured by a patch closure. Cardiac echocardiography can help rule out left atrial myxoma if it is highly suspected. As early surgical mortality is low and the long-term results are good, we strongly believe that patients with cardiac myxoma should be operated on as early as possible, once a diagnosis is made.

Echocardiography↗
PubMed1999

Interaction of retinol with dipalmitoylphosphatidylcholine and their formation of small dispersed particles.

Stable aqueous dispersions of all-trans-retinol (vitamin A, VA) were obtained by sonication with dipalmitoylphos-phatidylcholine (DPPC) in the VA mole fraction range 0.1-0.7. In order to clarify the dispersal mechanism, the dispersed particles were characterized and the interaction between VA and DPPC was investigated using several physicochemical techniques. Dynamic light scattering measurements showed that the diameter of the dispersed particles was 50-70 nm. A limited amount of VA was incorporated into DPPC bilayer membranes (approximately 5 mol%). The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein and the volume in the VA/DPPC particles was decreased markedly with the addition of VA into small unilamellar vesicles of DPPC. The decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO4. These results indicate that the excess VA separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Endothelium-dependent relaxation of rabbit atherosclerotic aorta was not restored by control of hyperlipidemia: the possible role of peroxynitrite (ONOO(-)).

We determined the role of ONOO(-) in nitric oxide (NO) mediated vascular response in atherosclerosis and regression following removal of dietary cholesterol. The effect of ONOO(-) on NO-mediated vascular responses was examined in vitro. Basal and stimulated NO release was estimated by an NO-selective electrode as well as vascular response and the plasma NO metabolites. An immunohistochemical study was also carried out. Responses were compared in normal controls, atherosclerotic rabbits fed 1% cholesterol diet for 6 or 9 weeks (atherosclerotic group) and animals fed a normal diet for 6-36 weeks after the high cholesterol diet for 6 or 9 weeks (regression group). ONOO(-) impaired the basal and acetylcholine-stimulated NO release, but did not affect endothelium-independent relaxation. After 15 weeks on a normal diet, the acetylcholine-stimulated and basal NO-mediated relaxation, which was diminished in the aorta induced by 6 weeks high cholesterol diet, became restored. However, the vascular response in the 9 weeks high cholesterol diet group did not return to normal after 36 weeks on a normal diet. iNOS was observed in atherosclerotic plaques in atherosclerotic and regression groups along with ONOO(-) in the 9 weeks high cholesterol diet group, but not in the 6 weeks group. Conclusively, ONOO(-) can play a role in impairment of NO-mediated vascular response during the regression of dietary cholesterol-induced atherosclerosis, not in the initiation of atherosclerosis.

Acetylcholine↗
PubMed1999

Formation and structure of stably dispersed particles composed of retinal with dipalmitoylphosphatidylcholine: coexistence of emulsion particles with bilayer vesicles.

In order to develop an intravenous formulation of all-trans-retinal (vitamin A aldehyde, VAA) for the treatment of night blindness, VAA and dipalmitoylphosphatidylcholine (DPPC) were sonicated and the dispersions in the VAA mole fraction range of 0.1-0.7 were stable at room temperature for 3 days. In order to clarify the dispersal mechanism, the dispersed particles were characterized and the interaction between VAA and DPPC was investigated using several physicochemical techniques. Dynamic light scattering measurements showed that the diameter of the dispersed particles was 50-70 nm. A limited amount of VAA is incorporated into DPPC bilayer membranes (approximately 5 mole%). The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein and the volume in the VAA/DPPC particles was decreased remarkably with the addition of VAA into small unilamellar vesicles of DPPC. The decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO(4). These results indicate that the excess VAA separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer. The monolayer-bilayer equilibrium of VAA/DPPC mixtures was estimated by measurement of spreading and collapse pressures. The results showed that the coexistence of emulsion particles (surface monolayer of DPPC+core of VAA) with vesicular particles (bilayer) was critically important for the formation of the stably dispersed particles of the lipid mixture.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Cloning and nucleotide sequencing of the secA gene from coryneform bacteria.

Taking advantage of highly conserved domains present in the secA gene from Escherichia coli and Bacillus subtilis, we designed degenerate oligonucleotides (oligos) corresponding to these regions. These oligos were used as primers in PCR in order to amplify DNA sequences from Brevibacterium flavum MJ233 chromosomal DNA. The PCR product was used as a probe to recover genomic fragments from a lambda library of Br. flavum MJ233. The complete nucleotide sequence (nt) of the cloned 5.3-kb EcoR1 fragment containing the secA homolog from Br. flavum MJ233 indicated that the deduced gene product of the Br. flavum secA homolog is composed of 845 amino acids (aa) with a deduced molecular weight (MW) of 95429. Comparison of this aa sequence to the corresponding sequences from E. coli and B. subtilis revealed a high degree of conservation and suggested that the Br. flavum secA homolog has putative ATP binding regions.

Adenosine Triphosphatases↗
PubMed1999

p38 MAP kinase regulates RANTES production by TNF-alpha-stimulated human pulmonary vascular endothelial cells.

BACKGROUND: RANTES plays an important role in the production of allergic inflammation of the airway through its chemotactic activity for eosinophils. However, the intracellular signal regulating RANTES expression in human pulmonary vascular endothelial cells has not been determined. In the present study, therefore, we examined the role of p38 mitogen-activated protein (MAP) kinase in RANTES production by tumor necrosis factor (TNF)-alpha-stimulated pulmonary vascular endothelial cells in order to clarify the signal transduction pathway regulating RANTES production by pulmonary vascular endothelial cells. METHODS: We examined p38 MAP kinase activation, and the effect of SB 203580, as the specific inhibitor for p38 MAP kinase, on p38 MAP kinase activity and RANTES production by TNF-alpha-stimulated human pulmonary vascular endothelial cells. RESULTS: The results showed that TNF-alpha induced RANTES production and p38 MAP kinase activity in human pulmonary vascular endothelial cells. Abrogation of p38 MAP kinase activity by SB 203580 repressed TNF-alpha-induced p38 MAP kinase activity and RANTES production. CONCLUSIONS: These results indicate that p38 MAP kinase plays an important role in the TNF-alpha-activated signaling pathway which regulates RANTES production by human pulmonary vascular endothelial cells.

Cells, Cultured↗
PubMed1999

A case of anaplastic large cell (Ki-1) lymphoma of B-cell phenotype, occurring in Waldenström's macroglobulinemia.

A case of anaplastic large cell (Ki-1) lymphoma of B-cell lineage occurred in a 59-year-old male with Waldenström's macroglobulinemia. Immunostaining of the lymphoma cells showed sporadic positivity for IgM and occasional positivity for kappa chain. This immunoglobulin specificity is the same as that of plasmacytoid lymphocytes in the bone marrow; therefore anaplastic transformation of Waldenström's macroglobulinemia was strongly suggested. This seems to be the first reported case of anaplastic large cell lymphoma, confirmed by CD30 expression, arising in Waldenström's macroglobulinemia.

Antigens, CD20↗
PubMed1999

Interaction of soybean oil with phosphatidylcholine and their formation of small dispersed particles.

Stable aqueous dispersions of soybean oil (SO) were obtained by cosonication with dipalmitoylphosphatidylcholine (DPPC) in the SO mole fraction range 0.1-0.8. To clarify the dispersal mechanism, the dispersed particles were characterized, and the interaction between SO and DPPC was investigated using several physicochemical techniques. Dynamic light scattering (DLS) measurements showed that the diameter of the dispersed particles was 40-60 nm. The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein. The trapped volume in the SO/DPPC particles decreased remarkably with the addition of SO into small unilamellar vesicles of DPPC. The decline in fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO4. These results indicate that the excess SO separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer. Monolayer-bilayer equilibrium of SO/DPPC mixtures was estimated by measurement of spreading and collapse pressures. The results showed that the coexistence of emulsion particles (surface monolayer of DPPC + core of SO) with vesicular particles (bilayer) was critically important for the formation of stably dispersed particles of the lipid mixture.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Preparation of transparent injectable formulation for lipid A analog E5531.

We developed a "pH-jump method" to obtain transparent solutions of E5531, a synthetic lipid A analog, at neutral pH for pharmaceutical injection. E5531 has two pKa: pKa1 = 6.0 and pKa2 = 9.3. At pH 11.0, E5531 was dispersed as a dissociated form, and the phase transition temperature Tc of E5531 was determined to be 30 degrees C using differential scanning calorimetry (DSC). Based on these results, the pH-jump method procedure involves dispersing E5531 at pH 11.0 (above pKa2) at 50 degrees C (above Tc) and mixing with a phosphate buffer to neutralize the pH. No degradated products of E5531 were observed at 50 degrees C for 3 hr during dispersing in the alkaline solution. The turbidity of samples prepared with the pH-jump method was similar to that of water and superior to the samples dispersed directly in neutral pH. This method does not need mechanical power and is suitable for large-scale production.

Calorimetry, Differential Scanning↗
PubMed1999

Effect of divalent cations on the membrane properties of the lipid A analog E5531.

To obtain information on the effects of Mg2+ on the membrane properties of the lipid A analog E5531, we determined the size, structure, zeta potential, membrane fluidity, and micropolarity of the aggregates and the permeability of the E5531 membrane after the addition of Mg2+. E5531 forms a vesicle structure and within the molar ratio of [E5531]:[Mg2+] = 1:3, Mg2+ increased the zeta potential of the E5531 membrane, but did not change the size of the aggregates (approximately 20 nm). Within that molar ratio, Mg2+ decreased the membrane fluidity and micropolarity of E5531 and increased the phase transition temperature. Above the molar ratio of [E5531]:[Mg2+] = 1:5, the size of the aggregates was increased, but at [E5531]:[Mg2+] = 1:3, the size of the aggregates was similar to that in the absence of Mg2+ (approximately 20 nm), and we could stabilize the aggregates in rat plasma.

Animals↗
PubMed1999

Growth of starved Escherichia coli O157 cells in selective and non-selective media.

Escherichia coli O157 strains starved in sterile deionized water (SDW) and filter-sterilized natural river water (SRW) were investigated with specific reference to their culturability in selective and non-selective media. Growth of the strains starved in both SDW and SRW were markedly suppressed with time in selective liquid media such as modified trypticase soy broth supplemented with novobiocin (mTSB+n) and modified E. coli broth supplemented with novobiocin (mEC+n). This suppression was more pronounced when incubated at 42 C than at 37 C, especially with mEC+n. By contrast, such growth suppression was seldom observed when cultured at 37 C in non-selective liquid media such as trypticase soy broth (TSB) and buffered peptone water. In mEC+n at 42 C, the non-starved cells from overnight cultures with an initial density of less than 10(3) colony-forming units (CFU)/ml grew to the density of over 10(7) CFU/ml after 24 hr incubation, whereas those starved for 6 weeks in SRW were only to maintain their initial density or died off after 24 hr incubation under the same culturing conditions. These results indicated that the isolation of starved cells of E. coli O157 from water samples would be most difficult with selective enrichment or direct plating on the selective plate media. It is thus highly recommended that a "resuscitation" of the cells with non-selective enrichment should be performed as a routine practice for maximum recovery of E. coli O157 from water systems.

Animals↗
PubMed1999

Digestible energy expenditure in grazing activity of growing horses.

Ten Thoroughbred yearlings (5 females and 5 males) were used to examine the effect of time of grazing on pasture forage and digestible energy (DE) intake, bodyweight gain and DE expenditure in grazing activity. Five females were grazed for 17 h/day (LTG), 5 males were grazed for 7 h/day (STG) and they were fed differently. As a result, DE intake from pasture forage of LTG horses and STG horses was 27.3 and 12.7-13.9 Mcal/day, respectively. The average daily gain (ADG) of LTG and STG horses was 0.37 and 0.39-0.61 kg/day, respectively. The regression lines between DE intake and ADG of both groups were parallel and the difference in DE intake between them was 10 Mcal of DE. It was suggested that DE expenditure in grazing activity (entirely at night) of growing horses was 1 Mcal/h under the conditions of this study.

Animal Feed↗
PubMed1999

Phytate and phosphorus movements in the digestive tract of horses.

This experiment was conducted to study phytate degradation and the effect of dietary phytate level on phosphorus absorption in the gastrointestinal tract of horses. Six Thoroughbred horses were fed diets containing low-phytate diet or high-phytate diet for 5 days. The diets were supplemented with Cr2O3 as an unabsorbable marker. The horses were killed 3 h after the last feeding and digesta in some segments of the intestine were collected. In both dietary groups, the daily passage of phytate phosphorus was decreased in the upper small intestine and in the lower large intestine. The daily passage of phytate phosphorus in the lower large intestine did not differ between these groups. Although the passage of total phosphorus increased in the upper small intestine and in the upper large intestine of both groups, its passage decreased in the lower large intestine. These results suggest that phytate phosphorus is degraded in the upper small intestine and in the lower large intestine of both groups. Phosphorus is suggested to be mainly absorbed in the lower large intestine. Therefore, phytate phosphorus is considered to be easily absorbed in horses because the major site of phosphorus is the lower large intestine where most of phytate is already degraded.

Animal Feed↗
PubMed1999

[Isolation of Shiga toxin-producing Escherichia coli O157:H7 from processed salmon roe associated with the outbreaks in Japan, 1998, and a molecular typing of the isolates by pulsed-field gel electrophoresis].

Shiga toxin-producing Escherichia coil (STEC) O157 were isolated from processed salmon roe which had been a suspected food item in sporadic infections which occurred in Japan in 1998. A total of 45 samples of the processed salmon roe were pre-enriched in trypticase soy broth (TSB) at 36 degrees C for 6 h and novobiocin-supplemented modified EC broth (mEC-NB) at 42 degrees C for 18 h. After the pre-enrichments, the cultures were examined for possible occurrence of STEC O157, using an immunomagnetic separation (IMS) method. From the examination, a total of 84 strains of STEC O157:H7 that were positive for both stx 1 and stx 2 genes were isolated. By applying the most-probable-number technique, it was estimated that the number of STEC O157 was in the range of 0.73-1.5 per 10 g of the processed salmon roe. Subsequent analysis of the isolates by a pulsed-field gel electrophoresis (PFGE) revealed a pattern commonly seen in 82 isolates and another pattern in two isolates. Clinical isolates from 7 patients also showed an identical pattern to those of the 82 isolates and one isolate from a patient showed the other pattern identical to those of the two isolates. The isolates were found to belong to the phage type 14.

DNA, Bacterial↗