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Biomedical subjects

K Ohki

Publications and source records attributed to K Ohki.

At least 91 records · Page 5Linked to original sources
PubMed1992

Metabolic fate of [14C]triglyceride-entrapped lactosylceramide-bearing liposomes after intravenous injection into mouse.

We investigated the distribution and fate of liposomes after their intravenous injection into a mouse. Liposomes were composed of dimyristoylphosphatidylcholine, cholesterol and dicetylphosphate (7:2:1, molar ratio) with or without lactosylceramide. They were characterized as small unilamellar vesicles, approximately 100 nm in diameter, using gel-exclusion chromatography on a Sephacryl S-1000, freeze-fracture electron microscope and dynamic light scattering method. Liposomes were very stable in serum as seen by the results of leakage of the entrapped marker and electrophoresis experiments. We demonstrated that liposomes were internalized by way of an endocytotic process via coated vesicles detected in the electron microscope. The increase in liver uptake of lactosylceramide-bearing liposomes was mostly accounted for by enhanced uptake in the parenchymal cells, while uptake by non-parenchymal cells was only slightly increased. This observation supported the notion that a galactose-specific receptor was involved in liver uptake of lactosylceramide liposomes. The lactosylceramide-bearing liposomes were preferentially recovered in the liver and were found to first be predominantly localized in the mitochondria-lysosomal fraction. They were then decomposed by lysosomal enzymes, and the hydrolyzed components were reincorporated into membrane phospholipids in the microsomal fraction. At the same time, a rapid and reversible exchange of phosphatidylcholine between microsomes and mitochondria was demonstrated.

Absorption↗
PubMed1992

Wear of denture teeth by use of metal plates. Part 2: Abrasive wear of posterior teeth.

An in vitro study was conducted to evaluate the abrasive wear resistance of high-strength denture teeth (HS teeth). Eight types of specimen were used in the experiments; 3 types of HS teeth, 3 types of conventional plastic denture teeth (PL teeth), porcelain teeth and metal teeth. Sliding-induced wear tests were conducted by sliding the samples on a metal plate. The abrasive wear resistance of the samples was evaluated in terms of wear depth, weight loss and SEM observation. Comparison of wear depth showed that abrasive wear resistance of HS teeth was 4.7 times that of PL teeth, 0.7 times that of porcelain teeth and 8.3 times that of metal teeth. In terms of weight loss, the corresponding values were 3.3-fold, 0.2-fold and 11.4-fold, respectively.

Dental Porcelain↗
PubMed1992

Wear of denture teeth by use of metal plates. Part 3: Abrasive wear of posterior teeth and wear of opposing metal plates.

An in vitro evaluation of abrasive wear resistance of high-strength denture (HS) teeth and wear of metal plates (Pd alloy) on the opposing side was conducted. A total of 8 types of teeth were used in the experiments including 3 types of HS teeth, 3 types of conventional plastic denture (PL) teeth, porcelain teeth and metal teeth (Pd alloy). Sliding-induced wear tests were conducted by sliding these teeth over the metal plates. Abrasive wear resistance of the teeth was evaluated in terms of wear depth and weight loss. A comparison of wear depth showed that the abrasive wear resistance of HS teeth was 4.7 times that of PL teeth, 0.7 times that of porcelain teeth and 8.3 times that of metal teeth. Weight loss showed that the abrasive wear resistance of HS teeth was 3.3 times that of PL teeth, 0.2 times that of porcelain teeth and 11.4 times that of metal teeth. The weight loss of the metal plates was minimal when they slid over HS teeth, but increased in the order PL teeth, porcelain teeth and metal teeth.

Dental Alloys↗
PubMed1992

Effects and distribution of intravitreally or subretinally injected silicone oil identified in rabbit retina using osmium tetroxide method.

Silicone oil was injected into the eyes of 28 rabbits. Utilizing the specific affinity of osmium tetroxide for lipids, the pathological features and histochemical localization of silicone oil within the retina were studied. This osmium tetroxide technique makes it possible to identify the presence of silicone oil particles in the ocular tissues. It was demonstrated that in eyes with silicone oil injected into the vitreous cavity or subretinal space, silicone oil was detected in the inner retinal layer and in phagocytes in the vitreous cavity but not in the outer retinal layer, retinal pigment epithelium and choroid. In addition, prominent subretinal and epiretinal proliferation with abundant phagocytes containing silicone oil particles were noted in the eyes with subretinal injection of silicone oil through an artificial retinal break. This study suggests that when injected subretinally silicone oil induces excessive subretinal and epiretinal proliferation.

Animals↗
PubMed1992

[An experimental study of the surgical trauma on the rabbit's lens suspensory apparatus after cataract surgery].

Extracapsular cataract extraction (ECCE) and posterior chamber intraocular lens (PC-IOL) implantation requires that the posterior capsule and the zonules remain sufficiently intact. To assess the surgical trauma on the lens suspensory apparatus of the enucleated rabbit eye, we developed a new fine pressure gauge that could quantitate its strength. The cornea and the iris of enucleated eyes were removed to allow clear visualization of the anterior surface of the lens to the equator. Afterwards several types of anterior capsulotomy were performed for ECCE and PC-IOL implantation. And then the lens of the pseudophakic eye was vertically pressed with the plastic rod of the fine pressure transducer, and the strength of the lens suspensory apparatus was simultaneously measured. The strength of the lens suspensory apparatus of the pseudophakic eye decreased approximately 35% compared with that of the phakic eye. Moreover the integrity of the lens capsule of animals with circular capsulorhexis with tears or linear capsulotomy decreased approximately 43% compared with that of continuous circular capsulorhexis (CCC) and also zonular rupture and vitreous loss occurred as a result of the expansion of radial tears in the anterior capsule. The present study indicates that the strength of the lens suspensory apparatus decreases with cataract surgery, suggesting that much more attention to cataract surgery is needed if the anterior capsule had radial tears. It is also suggested that CCC is the best type of anterior capsulectomy procedure.

Animals↗
PubMed1991

Electrostatic interaction of poly(L-lysine) with dipalmitoylphosphatidic acid studied by X-ray diffraction.

Structure of dipalmitoylphosphatidic acid (DPPA) bilayers in the presence of poly(L-lysine) is proposed from the results of X-ray diffraction obtained by a storage phosphor detector with a high resolution called an imaging plate. The small-angle X-ray diffraction pattern exhibits that DPPA/poly(L-lysine) complex forms a highly ordered multilamellar structure. The electron density profile of the DPPA/poly(L-lysine) complex draws that only one poly(L-lysine) layer is intercalated between the neighboring DPPA bilayers. The wide-angle X-ray diffraction pattern suggests that the presence of poly(L-lysine) hardly affects the nature of hydrocarbon chain packing in the DPPA bilayers. The X-ray reflection from the DPPA/poly(L-lysine) complex indicates that the poly(L-lysine) molecules adopt a beta-sheet conformation on the surface of the DPPA bilayers. The both surface areas occupied by a headgroup of the DPPA and by a lysine residue in poly(L-lysine) are estimated from the observed spacings. The number ratio of lysine residues to DPPA headgroups per unit area is greater than unity. Therefore, one DPPA headgroup interacts with more than one lysine residue electrostatically, i.e., the electric charge distributions in both the surface of a DPPA bilayer and the poly(L-lysine) beta-sheet are incommensurate.

Electrons↗
PubMed1991

Effect of substitution of hydrogen oxide by deuterium oxide on thermotropic transition between the interdigitated gel phase and the ripple phase of dihexadecylphosphatidylcholine.

Thermotropic transitions of dihexadecylphosphatidylcholine (DHPC) dispersions in hydrogen oxide (1H2O) and deuterium oxide (2H2O) were investigated by differential scanning calorimetry (DSC). In DHPC dispersions, transition temperature between interdigitated gel phase (L beta I) and ripple phase (P beta') is lower in 2H2O than in 1H2O, and transition between the ripple phase (P beta') and fluid phase (L alpha) in 2H2O occurs at a temperature slightly higher than in 1H2O. In dipalmitoylphosphatidylcholine (DPPC) dispersions, on the other hand, transition temperature between lamellar gel phase (L beta') and ripple phase is higher in 2H2O than in 1H2O. These results suggest that the interdigitated gel phase is more stable in 1H2O than in 2H2O. To account for the shift of transition temperature by the water substitution, difference of interfacial energies between these aqueous environments is discussed.

Calorimetry, Differential Scanning↗
PubMed1991

Arrangement of nitrogenase structural genes in an aerobic filamentous nonheterocystous cyanobacterium.

Members of the marine filamentous, nonheterocystous cyanobacterial genus Trichodesmium not only are capable of fixing nitrogen aerobically in the light but when grown under a light-dark cycle will fix nitrogen only during the light phase. In this study, we constructed a restriction map of the structural nitrogen fixation genes (nifHDK) in Trichodesmium sp. strain NIBB 1067. We found that the organization of the nif genes in Trichodesmium sp. strain NIBB 1067 is contiguous, as found in other nonheterocystous cyanobacteria and in heterocysts. Furthermore, the nif gene arrangement was identical when the cultures were grown with combined nitrogen or under nitrogen-fixing conditions. Therefore, no gene rearrangements occur, such as those that occur during the development of heterocysts in heterocystous species.

Blotting, Southern↗
PubMed1991

Unique modification of adenine in genomic DNA of the marine cyanobacterium Trichodesmium sp. strain NIBB 1067.

The genomic DNA of the marine nonheterocystous nitrogen-fixing cyanobacterium Trichodesmium sp. strain NIBB 1067 was found to be highly resistant to DNA restriction endonucleases. The DNA was digested extensively by the restriction enzyme DpnI, which requires adenine methylation for activity. The DNA composition, determined by high-performance liquid chromatography (HPLC), was found to be 69% AT. Surprisingly, it was found that a modified adenine which was not methylated at the usual N6 position was present and made up 4.7 mol% of the nucleosides in Trichodesmium DNA (15 mol% of deoxyadenosine). In order for adenine residues to be modified at this many positions, there must be many modifying enzymes or at least one of the modifying enzymes must have a degenerate recognition site. The reason(s) for this extensive methylation has not yet been determined but may have implications for the ecological success of this microorganism in nature.

Adenine↗
PubMed1991

Opposite effects of bacterial lipopolysaccharide on Fc-receptor-mediated phagocytosis of two bone marrow-derived macrophage cell lines, BDM-1 and BDM-1W3.

We have reported the isolation and characterization of three factor-dependent macrophage cell lines from bone marrow cells of C3H/HeN mice. We have since isolated a subclone, BDM-1W3, from one of these cell lines. We found previously that BDM-1W3 has a different sensitivity to bacterial lipopolysaccharide (LPS) for growth than its parental cell line, BDM-1. In this report, we show that LPS inhibits BDM-1W3 phagocytosis of antibody-coated sheep erythrocytes (Fc-mediated phagocytosis), whereas it enhanced Fc-mediated phagocytosis by BDM-1. It was observed that a loss of Fc-receptor capacity parallels a loss of phagocytic activity in LPS-treated BDM-1W3 cells. LPS stimulated phagocytosis of latex beads by BDM-1 and BDM-1W3, suggesting that Fc-mediated phagocytosis and phagocytosis of latex beads differ in their regulatory mechanisms. When BDM-1 cells were cultured with LPS, they underwent drastic morphological changes, whereas LPS-treated BDM-1W3 cells did not change significantly. Gamma interferon enhanced FC-mediated phagocytosis by BDM-1, while it has no significant effect on that by BDM-1W3. These cell lines should be useful for studying signal transduction mechanisms in LPS-mediated macrophage activation.

Antigens, Surface↗
PubMed1991

Constitutive production of granulocyte colony-stimulating factor by hybrids of a SV40-transformed mouse macrophage and a renal adenocarcinoma cell line.

Mouse macrophage BAM3 cells produced colony-stimulating factors (CSFs) after stimulation with bacterial lipopolysaccharide (LPS). By assaying the CSF using various interleukin 3-dependent cell lines, it was shown that most of the CSFs produced by BAM3 cells were granulocyte CSF (G-CSF). The granulocyte-macrophage CSF (GM-CSF) gene was also expressed in BAM3 cells after stimulation with LPS. When BAM3 cells were fused with the mouse renal adenocarcinoma cell line RAG which does not produce G-CSF, two of four hybrid cell lines constitutively produced large quantities of G-CSF. About 300 bp of the promoter region of mouse G-CSF chromosomal gene was inserted upstream of the Escherichia coli chloramphenicol acetyltransferase gene, and introduced into BAM3, RAG and hybrid cells. The G-CSF promoter was activated by stimulation with LPS, in BAM3 cells, but was inert in RAG cells. On the other hand, there was significant constitutive CAT activity in the hybrid cells.

Adenocarcinoma↗
PubMed1991

Noninfectious doughnut-shaped human immunodeficiency virus type 1 can induce syncytia mediated by fusion of the particles with CD4-positive cells.

A defective doughnut-shaped human immunodeficiency virus type 1 (HIV-1)-producing cell clone (designated as L-2) was isolated from persistently HIV-1-infected MT-4 cells. The syncytium-forming capacity of the cell and virus particle fractions was examined in human CD4-positive T cells. Several cell lines producing infectious HIV-1 particles, such as persistently HIV-1-infected MOLT-4 (MOLT-4/HIV-1) cells, were used as controls. Syncytia were formed within 20 h by the cell fraction of both L-2 and MOLT-4/HIV-1 and the virus particle fraction of L-2, but not MOLT-4/HIV-1. These formations were not affected by 3'-azido-3'-deoxythymidine (ZDV). In contrast, similar syncytium formation was first observed 2 days after the incubation of the virus particle fraction of MOLT-4/LAV-1 and this syncytium formation mediated by the cell fractions of MOLT-4/HIV-1 and L-2 or the virus particle fraction of L-2 differently.

Antibodies, Monoclonal↗
PubMed1991

[Attempt to the measurement of the tension of zonular fibers--influence to the lens suspensory apparatus when the lens was vertically displaced by pushing posteriorly].

A new fine pressure gauge was developed to quantitate zonular tension of the albino rabbit eye. This apparatus consists of a plastic rod to press the crystalline lens, transducer, amplifier, and recorder. The diameter of this plastic rod is 8 mm. The cornea and the iris of enucleated eyes were removed by excision in the limbus. Afterwards the lens were vertically pressed by the plastic rod of the fine pressure transducer. It was necessary to disrupt a part of zonules by approximately 106 g (2.11 g/mm2). This result was equivalent to 155 mmHg. The morphological changes of the anterior part of eye globe which consists of lens, zonules, and ciliary process were investigated using scanning electron microscopy. In the control eyes, the zonular fibers mostly originated from the superior or lateral ciliary processes and inserted straight into the equatorial region of the lens. In comparison with control eyes, the zonular fibers which were pressed by the plastic rod were almost absent within 1/6 of a circle. This disruption occurred at the region adjacent to the ciliary e process, while the region of the insertion to the lens demonstrated a normal appearance. In the unaffected area zonules were well-preserved morphologically, apart from splits of the bundles of zonules. The present study indicated that zonular fibers could resist a stretching press up to approximately 8 times normal intraocular pressure.

Animals↗
PubMed1990

Ethanol induces interdigitated gel phase (L beta I) between lamellar gel phase (L beta') and ripple phase (P beta') in phosphatidylcholine membranes: a scanning density meter study.

Effects of ethanol on dipalmitoylphosphatidylcholine (DPPC) and distearoylphosphatidylcholine (DSPC) dispersions were investigated with an automated scanning density meter and a differential scanning calorimeter (DSC). The temperature-dependent profile of specific volume measured by the density meter clearly exhibited phase transitions of the DPPC and the DSPC dispersions as drastic changes in the thermal expansion coefficients. On increasing the ethanol concentration in the DPPC dispersions, the pretransition temperature was reduced faster than the main transition temperature was. An interdigitated gel phase (L beta I) appeared as a region of lower specific volume at the pretransition temperature when the ethanol concentration reached 40 mg/ml. The L beta I phase spread both its ends in an ethanol-dependent fashion, and the high-temperature end merged to the main transition at 50 mg/ml of ethanol. The temperature-ethanol phase diagram has been determined for DPPC. The transitions L beta' to L beta I and from L beta I to P beta' were also observed on the thermograms of DSC measurements. In the DSPC dispersions, the L beta I phase was induced between the L beta' and the P beta' phases by a lower ethanol concentration (about 20 mg/ml).

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1990

Antiserum to Nitrogenase Generated from an Amplified DNA Fragment from Natural Populations of Trichodesmium spp.

A fragment of the nifH gene was amplified from natural populations of Trichodesmium spp. and cloned into a maltose-binding protein (MBP) expression vector. The peptide product of the amplified 359-bp fragment of nifH was cleaved from the fusion protein, purified, and used to generate a specific antibody to the Fe protein of nitrogenase. The antiserum recognized the MBP-nitrogenase fusion protein and the cleaved nif peptide product but not MBP. The antibody cross-reacted with nitrogenase from natural populations of Trichodesmium spp. from the Caribbean Sea and with a cultured isolate from the Kuroshio waters (Trichodesmium sp. strain NIBB1067). The same nifH fragment was amplified, cloned, and sequenced from Trichodesmium sp. strain NIBB1067 and was found to be 98% identical at both the protein and DNA levels to nifH from the Caribbean populations. Three of the six nucleotide differences between the Trichodesmium sp. strain NIBB1067 and the Trichodesmium spp. nifH sequence had also been found in a second sequence from the natural populations, indicating either that there is more than one strain of Trichodesmium sp. in natural assemblages or that there are multiple copies of nifH in the genome. This DNA fragment, which is easily amplified with the polymerase chain reaction, may provide a good indicator of species relatedness without requiring extensive cloning or sequencing. Furthermore, the use of the polymerase chain reaction in combination with a MBP protein fusion vector provides a rapid method for production of highly specific sera, starting with a small amount of DNA.

Journal Article↗