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Biomedical subjects

S Ohki

Publications and source records attributed to S Ohki.

At least 19 recordsLinked to original sources
PubMed1992

Association of lysozyme to phospholipid surfaces and vesicle fusion.

Lysozyme-induced fusion of phosphatidylserine (PS) vesicles was studied as a function of pH. Fusion, monitored by lipid-mixing, was measured by following the dilution of pyrene-labelled phosphatidylcholine, incorporated in PS vesicles, into unlabelled bilayers. It is demonstrated that lysozyme-induced fusion is pH-dependent and significant fusion is triggered at pH 5 or below. The interaction of lysozyme with the vesicle bilayer was characterized by measuring resonance energy transfer from tryptophane, present in the protein, to pyrene. It is shown that concomitant with fusion, a strong resonance energy transfer signal appears at pH 5 or below. Furthermore, in monolayer experiments it was found that addition of lysozyme to the subphase caused an increase in surface pressure, when the pH was kept below 5.5. Very low concentrations of lysozyme sufficed to bring about the observed effects. The results are taken to indicate that lysozyme-induced fusion results from penetration of protein into the hydrophobic core of the bilayer, occurring at acidic pH.

Energy Transfer↗
PubMed1992

Dextran sulfate-dependent fusion of liposomes containing cationic stearylamine.

The incorporation of the positively charged stearylamine into phosphatidylcholine liposomes was studied by measuring electrophoretic mobilities. Up to a molar ratio SA/PC = 0.5 an increase of the positive zeta potential can be observed. Addition of the negatively charged macromolecule dextran sulfate leads to a change of the sign of the surface potential of the PC/SA liposomes indicating binding of the macromolecule to the surface. This process is accompanied by an increase in turbidity, which is dependent on the molecular weight of the dextran sulfate and the SA concentration (measured by turbidimetry). Using the NBD/Rh and Pyr-PC fluorescence assays the fusion of SA containing liposomes was investigated. A strong influence of the SA content and molecular weight of dextran sulfate on the fusion extent was observed. The fusion extent is proportional to the SA content in the PC membrane and the molecular weight of dextran sulfate. PC/SA/PE liposomes exhibit a higher fusion extent after addition of dextran sulfate compared to PC/SA liposomes indicating that PE additionally destabilizes the bilayer. Freeze-fracture electron microscopy reveals that the reaction products are large complexes composed of multilamellar stacks of tightly packed, straight membranes and aggregated vesicles. The tight packing of the membranes in the stacks (and the narrow contact of the aggregated vesicles) indicates a strong adherence of opposite membrane surfaces induced by dextran sulfate.

Amines↗
PubMed1992

Effect of anionic polymers on fusion of Sendai virus with human erythrocyte ghosts.

The effect of anionic polymers (dextran sulfate, heparin and chondroitin sulfate) on fusion of Sendai virus with erythrocyte ghosts was studied. The effect of pH on the activity of these anionic polymers was also investigated. In order to examine the interaction of such polymers with the Sendai virion and erythrocyte ghost surfaces, the binding of virions to erythrocyte ghosts and the aggregation of virions and/or erythrocyte ghosts were also measured with respect to the same parameters. It was found that the anionic polymers suppressed the fusion of Sendai virus with erythrocyte ghosts. The order of effectiveness of the polymers in suppression was dextran sulfate greater than heparin greater than chondroitin sulfate, for the application of a same quantity (weight/ml) of the polymers. The lower the pH of the suspending medium, the more effective were the polymers in suppressing virion-erythrocyte ghost aggregation and fusion. The suppression of fusion was dependent on the concentration of the polymers applied: the higher the concentration of the polymer applied, the more the suppression was observed. Evidence from binding studies, turbidity measurements and electrophoretic mobility measurements indicates that the anionic polymers interact preferentially with the virion surface.

Adsorption↗
PubMed1992

[Expression of c-erbB-2 protein and vessel invasion in colorectal cancer].

Using sections of formalin-fixed, paraffin-embedded tissues from 64 colorectal cancer patients, the expression of c-erbB-2 oncoprotein was studied immunohistochemically. Twenty-seven percent of the cases with liver metastasis showed positive staining. On the other hand, only 3% of cases without liver metastasis were positive. Expression rates of c-erbB-2 protein in liver metastasis cases showed no significant difference between primary operation (26%) and recurrence (27%). Of all c-erbB-2 positive patients, 90% (9/10) had liver metastasis. Secondly, vessel invasions of 45 rectal cancer patients were studied using Victoria Blue (VB) elastic staining and endothelial staining by factor VIII-related antigen and Ulex europaeus agglutinin I (UEA-I) lectin. VB-HE double stain was efficacious to detect vascular invasion, but endothelial staining was not. There were statistically more vascular invasions in 30 patients with liver or lymph node metastases than in those without metastasis. And in cases with metastasis, many vascular invasions into the extra-muscular layer were seen. Both vascular invasions and c-erbB-2 protein were valuable indicators of possible liver metastasis.

Biomarkers, Tumor↗
PubMed1991

Effect of dextran sulfate on fusion of Sendai virus with human erythrocyte ghosts.

The effect of dextran sulfate on the fusion of Sendai virus and erythrocyte ghosts was studied as a function of dextran sulfate concentration and pH of cell suspension solutions. In order to examine the interaction of dextran sulfate with Sendai virion and erythrocyte ghost surfaces, the turbidity of cell suspensions was also measured with respect to the same parameters as above. It was found that dextran sulfate inhibited the fusion of Sendai virus with erythrocyte ghosts. The lower the pH of the suspension solution was, the more effective was dextran sulfate in suppressing virion erythrocyte ghost fusion. Dextran sulfate of a higher molecular weight was slightly more effective in suppressing fusion than that of a lower molecular weight. From turbidity measurements of Sendai virus and erythrocyte ghost suspensions, it is likely that dextran sulfate interacts preferentially with Sendai virion surfaces and inhibits interaction between Sendai virions and erythrocyte ghosts.

Depression, Chemical↗
PubMed1991

1H NMR study on amide proton exchange of calmodulin-mastoparan complex.

Amide proton exchange rates of Ca2(+)-saturated calmodulin and Ca2(+)-saturated calmodulin-mastoparan complex were studied by 1H NMR spectroscopy. Exchange rates of Gly25, Gly61, Gly98, Gly134, Ile27, Ile100, and Asn137 were determined for Ca2(+)-saturated calmodulin and for Ca2(+)-saturated calmodulin-mastoparan complex, and were found to be less than 10(-4)s-1. All these residues of which the amide proton resonances appear at lower fields were considered to form hydrogen bonds, based on the results of X-ray analysis. Exchange rates of Ile27 and Asn137 became an order of magnitude smaller when mastoparan bound to Ca2(+)-saturated calmodulin, while those of the four glycines and Ile100 did not change appreciably. The reduction in accessibility of Asn137 to water cased by mastoparan binding suggests that a part of the mastoparan binding site is probably located in or near the hydrophobic cluster of the C-terminal-half domain. The reduction in accessibility of Ile27 also suggests that another part of the mastoparan binding site is located in or near the hydrophobic cleft of the N-terminal-half domain.

Amino Acids↗
PubMed1991

[Effects and complications of continuous hepatic arterial infusion chemotherapy using implantable reservoir for liver metastases from colorectal cancer].

Continuous arterial infusion chemotherapy using implantable reservoir was performed for unresectable liver metastases from colorectal cancer and the therapeutic effects, side effects and complications were evaluated. Eleven patients were treated with four kinds of arterial infusion courses that mainly consisted of 5-FU. The arterial infusion courses were discontinued in 2 patients because of nausea and vomiting, and in one patient because of diarrhea. The catheters were dislocated in 2 patients and another 2 developed fistulous between the hepatic artery and bile duct. Three patients developed duodenal ulcer. Serum CEA was reduced in 4 patients (36%). After all, response rate was 9% (1/11). The one-year survival rates of all cases and cases treated with more than 4 courses were 36.3% and 80.0%, respectively.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1990

Surface dielectric constant, surface hydrophobicity and membrane fusion.

Membrane fusion induced by ions and its associated membrane property, surface dielectric constant, were studied with the use of acidic and neutral phospholipid vesicles. The fusion of vesicles was monitored by utilizing two fluorescence fusion assays: fluorescence content mixing method and fluorescence labelled membrane component dilution method. For the surface dielectric constant measurements, a fluorescence method was used which detected the environmental effect on the membrane surface upon the addition of various fusogenic cations. Also, the effects of poly-(ethylene glycol) on both fusion and surface dielectric properties were examined. It was found that the extent of fusion correlated well with the degree of lowering in the dielectric constant of the surface membrane, which corresponds to the increase in hydrophobicity of the membrane surface. This agrees with the previously obtained experimental results that the increase in interfacial tension of the membrane, which also corresponds to the increase in surface hydrophobicity, correlates with the extent of membrane fusion.

Calcium↗
PubMed1990

Interaction of dextran sulfate with phospholipid surfaces and liposome aggregation and fusion.

The binding of dextran sulfate to phospholipid liposomes was investigated by microelectrophoresis experiments. The polyanion binds to neutral phospholipid liposomes (DMPC and PE) only in the presence of Ca2+. If positively charged stearylamine is incorporated in the vesicles dextran sulfate is bound without Ca2+. Negatively charged phospholipids as PS do not bind dextran sulfate, even in the presence of millimolar concentrations of Ca2+. The adsorption of dextran sulfate results in an aggregation of vesicles due to a bridging mechanism. In all cases the aggregation is followed by a disaggregation toward higher dextran sulfate concentrations. The disaggregation process starts at polymer concentrations smaller than the concentration of the onset of saturation of the adsorption. By use of the probe dilution method a fusion of small DMPC and DMPC/PE vesicles in the presence of Ca2+ and dextran sulfate was found.

Calcium↗
PubMed1990

[Continuous hepatic arterial infusion chemotherapy using implantable reservoir in liver metastases from colorectal cancer].

Continuous hepatic arterial infusion chemotherapy using implantable reservoir was performed for liver metastases from colorectal cancer, and the therapeutic effects, side effects, and complications were evaluated. 9 patients with unresectable liver metastases were as follows, 1. Group A; 3 patients, MMC 2 mg.one shot + 5-FU 250 mg/day.continuous infusion x 14 days, and then 5-FU tablets 150 mg/day.p.o. x 14 days, 2. Group B; 4 patients, MMC 2 mg.one shot + 5-FU 500 mg/day.continuous infusion x 7 days, and then 5-FU tablets 150 mg/day.p.o. x 14 days, 3. Group C; 2 patients, 5-FU 500 mg/day.continuous infusion x 14 days, and then free from agents for 14 days. In 2 of 3 group A patients, the catheters became dislocated and one died of perforation of duodenum. In group A and group B, no severe side effects were noted. But both of group C patients showed nausea, vomiting and diarrhea. In 8 of 9 patients (89%), serum CEA level fell below the preoperative level. In 4 of 7 patients who underwent CT scan, the size of the tumor regressed. Total infused dose of 5-FU was 8.17 +/- 7.56 g in group A, 16.9 +/- 2.88 g in group B, and 21.0 +/- 9.90 g in group C on average. In 2 patients of group B, therapy was repeated seven times.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1990

Influence of dextran sulfate on the fusion of Sendai virus with liposomes.

Liposomes made from phosphatidylserine and labeled with the fluorescence probe pyrene-phosphatidylcholine were used for studies of the influence of dextran sulfate on the fusion process. The fusion is monitored by the increase of the fluorescence signal due to the dilution of the fluorescence probes as a result of the fusion of liposomes with the virus. Addition of dextran sulfate inhibits the fusion process. Binding of the sulfate groups of the polymer to aminogroups on the virus proteins is considered as the reason for the inhibiting effect of this polymer on fusion.

Dextran Sulfate↗
PubMed1988

Electron microscopic study of the calcium phosphate-induced aggregation and membrane destabilization of cytoskeleton-free erythrocyte vesicles.

Cytoskeleton-free vesicles derived from human erythrocytes were treated with trypsin, chymotrypsin, or neuraminidase followed by calcium, phosphate, or combined calcium/phosphate treatments in order to study the roles of cell surface proteins and glycoproteins in calcium/phosphate-induced cell aggregation and fusion. Vesicle aggregation (a necessary pre-cursor to membrane fusion) and subsequent membrane destabilization (an essential component of fusion) were examined by freeze-fracture electron microscopy. Enzymatic treatment alone had no effect on the morphology of the cytoskeleton-free vesicles. Neither did separate calcium nor phosphate treatments, although the treatment of the cytoskeleton-free vesicles with calcium did reduce their size slightly. Enzymatic pretreatment had no effect on the calcium-induced size changes. In contrast, the combination of calcium and phosphate drastically disrupted the membrane integrity of aggregated cytoskeleton-free vesicles at pH 7.8, although the effect was reduced at lower pH values. The extent of this membrane destabilization was independent of enzyme treatment. Our results indicate: (1) that the cell surface proteins and glycoproteins have only secondary effects on calcium/phosphate-induced cell aggregation and membrane destabilization, (2) that these processes primarily depend on the reaction between calcium and phosphate ions at the membrane surface, and (3) that cytoskeletal elements probably play no active (positive) role in the Ca2+/PO4(3-) induced erythrocyte membrane fusion process, apart from maintaining cell shape.

Calcium↗
PubMed1988

Relaxivity and binding of Mn2+ ions in solutions of phosphatidylserine vesicles.

We report the magnetic field dependence (NMRD profiles) of 1/T1 of solvent protons in solutions of unilamellar phosphatidylserine vesicles with added Mn2+ ions, including studies of the variation of the profiles with temperature, extent of coverage of available binding sites by Mn2+ ions, ionic strength, and competition with (nonparamagnetic) Ca2+ ions. ions. In addition, we sketch the theory of screening of the negative surface charges of the vesicles due to both specific binding of Mn2+ ions and nonspecific effects of other mobile solute ions. The major result is that the NMRD profiles, although qualitatively similar, vary systematically as the parameters of the solutions are altered, in a manner consistent with the theory of screening. The profiles of the Mn2+-vesicle complexes are much like those of Mn2+-protein complexes that have the ions in an octahedral ligand environment. In addition, we find that the profiles are similar to those reported for Mn2+ ions in packed liver cells and liver tissue, supporting a previous conjecture that available Mn2+ in liver binds to the polar head groups of cell membranes, saturating these sites before binding elsewhere. Again, it is evident that results for in vitro model systems can be extrapolated reliably to tissue behavior.

Contrast Media↗
PubMed1988

Effect of monovalent cations on polyvalent cation-induced fusion of phosphatidylserine small unilamellar vesicles.

Fluorescence internal contents mixing assay was used to monitor the fusion of phosphatidylserine (PS) small unilamellar vesicles, initiated by metal ions (Ca2+, La3+ and Tb3+), at various concentrations of monovalent cations (Li+, Na+ and K+). The influence of ionic strength (0.02-1.0 M) on the threshold concentration of "fusogenic" cations required to induce fusion was measured. The threshold concentrations increased monotonically (1 mM at 0.1 M to 3.1 mM at 1 M) with the increasing ionic strength of the solution for Ca2+, but remained unchanged for both La3+ and Tb3+. Changes in the ionic strength of the encapsulated solution did not alter the threshold concentrations for all the ions studied, in the range 0.02-0.3 M. The results are analyzed in terms of competitive binding between the monovalent ions and the "fusogenic" ions (Ca2+, Tb3+ and La3+). It is shown that there is a critical value for calcium bound-PS, below which no massive fusion occurs. Bound and free fractions of PS are calculated based on the Gouy-Chapman model, taking activities rather than concentrations of metal ions into account. Our experiments also show that monovalent ions alone do not induce fusion even at high concentrations.

Animals↗
PubMed1988

[Surgical treatment of metastatic lung cancer from colorectal cancers].

Based on conclusions obtained after the observation of 61 colorectal cancer patients with a lung metastasis, the resection of lung metastasis as a therapy was evaluated. Among these 61 patients, only 5 patients had been identified as having a lung metastasis at the time of resection of the primary lesion, whereas the other 56 patients developed the lung metastasis after the curative resection of the colorectal cancer. Only one patient with a synchronous lung metastasis and twelve patients (eleven with a solitary metastatic lesion and one with multiple metastatic lesions) with metachronous lung metastasis underwent removal of the lung metastasis. The three-year survival rate was 65.2% in the metachronous group.

Adult↗
PubMed1987

Percutaneous diagnosis and drainage of pylephlebitis: a case report.

Suppurative pylephlebitis is a rare complication of intra-abdominal inflammatory processes, but it carries a high mortality rate. Even in this modern era, diagnosis and treatment are difficult because of the nonspecificity of clinical signs and symptoms, as well as laboratory tests. We present a case in which the diagnosis of pylephlebitis was made in the radiology department by percutaneous needle aspiration of the portal venous system. Computerized tomography is very helpful in the diagnosis of pylephlebitis but requires that the radiologist be familiar with this rare entity. The computerized tomographic findings in this case are described and discussed. This patient was treated with percutaneous transhepatic drainage of the portal venous system and antibiotics only since we thought he would not survive a surgical procedure. To the best of our knowledge, there have been no previous reports of percutaneous therapy of pylephlebitis. The patient had an uneventful recovery.

Biopsy, Needle↗
PubMed1986

Effects of cations and polyamines on the aggregation and fusion of phosphatidylserine membranes.

Effects of various metal cations and polyamines on aggregation and fusion of phosphatidylserine vesicles and their associated physicochemical properties (such as surface tension and vesicle electrophoretic mobility) have been studied. It was found that metal polycations and hydrogen ion caused an increase in the surface tension of a phosphatidylserine monolayer, whereas the polyamines and other monovalent cations did not increase the surface tension of the membrane appreciably. All cations used affected the vesicle mobility roughly in the order of the number of their valencies and linearly with respect to the logarithm of their concentrations of ions; vesicle surface charge densities are reduced by adsorption and screening of the counter ions depending on their valencies and concentrations. The degree of aggregation of lipid vesicles parallels somewhat that of the reduction of vesicle electrophoretic mobilities. However, the degree of membrane fusion induced by these cations parallels that of the increase in surface tension of the membranes induced by these cations.

Animals↗