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Biomedical subjects

M Akashi

Publications and source records attributed to M Akashi.

At least 55 records · Page 3Linked to original sources
PubMed1999

Apatite formation on/in hydrogel matrices using an alternate soaking process: II. Effect of swelling ratios of poly(vinyl alcohol) hydrogel matrices on apatite formation.

In our previous study, we reported a novel method of apatite formation on/in a three-dimensional hydrogel matrix. Using this method, bone-like apatite could be formed on/in the hydrogel matrix under normal conditions in vitro. A poly(vinyl alcohol) (PVA) gel was used as a model matrix. The method consists of two steps: first, water is transformed in a PVA gel with a CaCl2/Tris-HCl aqueous solution (pH 7.4) and second, the gel is soaked in a Na2HPO4 aqueous solution. In the present study, we report a detailed study of the effects of the swelling ratios of PVA gels on apatite formation. Cross-sectional observations and gravimetric measurements of PVA gels with various swelling ratios were done. The amount of apatite formed on/in PVA gels increased almost linearly with an increase in the reaction cycles. The rates of apatite formation on/in PVA gels largely depended on the swelling ratios, which were approximately 0.48, 0.61, 1.28, and 1.55 mg per cycle for swelling ratios of 4.1, 10.4, 16.8, and 30.1, respectively. The apatite content in PVA-apatite composites that was obtained by this method also increased with an increase of the reaction cycles. After six reaction cycles, a PVA gel with a high swelling ratio contains approximately 70 wt% of formed apatite in the composite. On the other hand, a gel with a low swelling ratio contains about 15 wt% of formed apatite in the composite. Cross-sectional views of the PVA gels after each cycle showed that apatite crystals were formed, not only on the surface of the gel but also within it after fifteen reaction cycles. The hydrogel-apatite composites that were obtained using an alternative soaking process will be useful as not only bone substitute materials but also as soft tissue adhesive materials.

Apatites↗
PubMed1999

Synthesis of polystyrene nanospheres having lactose-conjugated hydrophilic polymers on their surfaces and carbohydrate recognition by proteins.

We prepared a poly(vinylamine)-grafted polystyrene nanosphere by the free radical polymerization of poly(N-vinylacetamide) macromonomer and styrene with AIBN in ethanol and subsequent acid hydrolysis. We conjugated lactose on the nanosphere by an amide linkage and found that the lactose was recognized by a galactose-specific lectin RCA120, which was analyzed by enzyme-linked lectin assay (ELLA). The binding ability of the lectin to lactose on the nanosphere was at least 10(2) times larger than that to a monomeric lactose. Binding ability was clearly affected by the lactose density on the nanosphere and showed a maximum value at the suitable surface density of lactose. We found that the carbohydrate-conjugated nanosphere would be a useful biomaterial for a site specific drug carrier.

Carbohydrate Metabolism↗
PubMed1999

Ca-adsorption and apatite deposition on silk fabrics modified with phosphate polymer chains.

Silk fabric was modified with polymethacryloyloxyethylphosphate (pMOEP) by graft copolymerization. Ca-adsorption onto pMOEP-grafted silk fabric was significantly enhanced compared to that onto original silk fabric. SEM observation indicated that some crystallites were deposited on the pMOEP-grafted silk fabric after 1 week of immersion in simulated body fluid, whereas no change occurred on the surface of the original silk fabric. X-ray diffraction showed that this crystallite contained hydroxyapatite. These results indicate that pMOEP-grafted silk fabric induce hydroxyapatite formation more effectively than the original silk fabric.

Adsorption↗
PubMed1999

Apatite formation on/in hydrogel matrices using an alternate soaking process (III): effect of physico-chemical factors on apatite formation on/in poly(vinyl alcohol) hydrogel matrices.

The aim of this study is to clarify the physico-chemical factors which influence apatite formation on/in a hydrogel during a novel alternate soaking process. A poly(vinyl alcohol) (PVA) gel was used as a model matrix. The amount of apatite formed on/in PVA gels decreased with an increase in the reaction temperature during the same reaction cycles. This suggested that the equilibrium swelling ratios decreased with increasing reaction temperatures; that is, the diffusion of calcium and phosphate ions reduced at high reaction temperature. However, the crystallinity of apatite formed on/in PVA gels was greater at higher reaction temperatures. The amount of apatite formed on/in PVA gels increased with an increase in the calcium and phosphate solution concentrations, and increased by shaking at the first three reaction cycles. A few influences could be observed when the solution volume was changed, however, the soaking order was not effective in this study. These results indicate that the amount of apatite formation on/in PVA gels can be controlled by changing the reaction temperature and the Ca- and P-solution concentrations, and that the crystallinity of apatite can be also changed by controlling the reaction temperatures.

Apatites↗
PubMed1999

Chromosome aberrations in bone marrow cells from Japanese patients with thorotrastosis.

Exposure of bone marrow cells to alpha-particle radiation causes various types of chromosome abnormalities and hematological malignancies. We performed chromosome analysis of hematopoietic stem cells from the bone marrow of 52 Japanese patients with thorotrastosis and 21 age-matched controls. The frequency of cells with stable chromosome abnormalities was significantly higher in the patients with thorotrastosis. Further studies found 14 clonal chromosome aberrations in cells from 11 patients (21.2%); clones observed in the cells from 2 of these patients had high frequencies of chromosome abnormalities. In one case, 68 to 100% of the cells analyzed had a large partial loss in the short arm of chromosome 1 and a translocation between the short arms of chromosomes 2 and 3 [46,XY,1p-,t(2p+;3p-)]. The cells from the other patient contained a clone with partial loss of both the short and long arms of chromosome 5 (46,XX,5p-,5q-). The frequency of this clone has been constant for the last 15 years (6-24%). We also analyzed bone marrow mononuclear cells from 17 of the patients for mutations of the TP53 tumor suppressor gene (formerly known as p53). However, no mutation was found in any of the cells, including those from the 2 patients with abnormal clones. Moreover, repeated medical examinations showed no evidence of leukemia or myelodysplasia in these patients. Our study suggests that exposure of bone marrow cells to alpha-particle radiation may induce clonal chromosomal aberrations at a high frequency.

Aged↗
PubMed1998

Control of endotoxin shock by the dried preparation of low virulent Streptococcus pyogenes OK-432.

Bacterial endotoxin, lipopolysaccharide (LPS), is a causative agent of Gram-negative septic shock. However, if preadministered at a low dose, LPS makes mice resistant to subsequent endotoxin challenge, the phenomenon known as LPS tolerance. Here we demonstrated that the pharmaceutical preparation of Gram-positive Streptococcus pyogenes, OK-432, also induced a state analogous to LPS tolerance if administered 6-48 h prior to LPS challenge. The preadministration of OK-432 increased the lethal dose of LPS threefold in BDF1 mice, and this was accompanied by reduced gene expression of IL-6, IFN-gamma, inducible nitric oxide synthase, and IL-10 in spleen and peritoneal cells. Serum concentrations of IL-6 and IFN-gamma were also suppressed by the preadministration of OK-432. In contrast to the LPS tolerance, the levels of TNF-alpha mRNA were not suppressed in OK-432-administered mice, and their peritoneal cells produced high levels of TNF-alpha and soluble TNF receptor p75 in response to LPS in vitro. Peritoneal cells from OK-432 but not LPS-administered mice were hyporesponsive to IFN-gamma in terms of nitric oxide synthesis, and this hyporesponsiveness to IFN-gamma was abrogated by anti-IL-10 antibodies. Likewise, peritoneal cells from both OK-432- and LPS-administered mice were hyporesponsive to LPS, serum, TNF-alpha, IFN-gamma, and PMA in terms of IL-6 production. Anti-IL-10 antibodies increased IL-6 production eightfold in cells from OK-432-administered mice, but marginally in cells from LPS-administered mice. Even in peritoneal cells from OK-432-administered mice, anti-IL-10 antibodies failed to fully restore IL-6 production. Thus, the hyporesponsive state of peritoneal cells was mediated by both IL-10-dependent and -independent mechanisms. These results demonstrated that OK-432 controlled endotoxin shock by blocking the cytokine cascade from TNF-alpha.

Animals↗
PubMed1998

Preparation of a novel functional hydrogel consisting of sulfated glucoside-bearing polymer: activation of basic fibroblast growth factor.

Sulfated poly(glucosyloxyethyl methacrylate) [poly(GEMA)-sulfate] and its hydrogel were synthesized and evaluated to develop novel tissue regeneration assist devices. The poly(GEMA)-sulfate hydrogel was obtained by gamma-ray irradiation. Basic fibroblast growth factor (bFGF) was expected to be subsequently incorporated with hydrogel matrices and activated or released from the matrices. A sustained release of bFGF from the poly(GEMA)-sulfate hydrogel was not realized; however, bFGF from poly(GEMA)-sulfate hydrogel was effectively activated. Poly(GEMA)-sulfate hydrogels are thought to be potential component materials for a tissue engineering matrix.

Cell Adhesion↗
PubMed1998

Concanavalin A-immobilized polystyrene nanospheres capture HIV-1 virions and gp120: potential approach towards prevention of viral transmission.

To establish an effective tool for the prevention of HIV-1 transmission, lectin-immobilized polystyrene nanospheres were synthesized and examined for their HIV-1 capture activity. When concanavalin A (Con A) was immobilized on the surface of polystyrene nanospheres (400 nm in diameter) with poly(methacrylic acid) branches and incubated with HIV-1 suspension at room temperature for 60 min, the nanospheres (Con A-NSs) achieved a >3.3 log and a 2.2 log reduction of viral infectivity in HIV-1 (IIIB strain) suspension at a concentration of 2 and 0.5 mg/ml, respectively. Meanwhile, Con A-free nanospheres, which were not immobilized with Con A, achieved only a 0.29 log reduction at 0.5 mg/ ml. Con A-NSs (2 mg/ml) could also reduce the gp120 level of III(B) and HE strains to <7.1% and 5.5% of each control, respectively. The combination of Con A-NS treatment followed by filtration with a microporous membrane efficiently removed virion-free gp120 as well as infectious viral particles from HIV-1 suspension. Electron microscopic examination demonstrated that HIV-1 virions were trapped on the surface of Con A-NSs. Thus, Con A-NSs can capture HIV-1 virions and gp120 with high affinity, and may have potential as an effective tool for the prevention of HIV-1 transmission.

Concanavalin A↗
PubMed1998

Base-specific separation of oligodeoxynucleotides by capillary affinity gel electrophoresis.

Capillary affinity gel electrophoresis was applied to sequence-specific and base composition-specific recognition of oligodeoxynucleotides, utilizing the formation of heteroduplexes between a nucleic acid analogue immobilized into the capillary gel and soluble oligodeoxynucleotides with different sequences. Capillary affinity gel electrophoresis using capillaries filled with a conjugated gel of polyacrylamide and a synthetic nucleic acid analogue [poly(9-vinyladenine)] was effective for the selective separation of hexathymidylic acid from a mixture of four homopolymers of A6, C6, G6, and T6 and for the complete resolution of five heteropolymers of hexadeoxynucleotides (TAAAAA, TTAAAA, TTTAAA, TTTTAA, TTTTTA). We also demonstrated that capillary affinity gel electrophoresis was useful for the selective and the sensitive sequence-specific recognition of sequence isomers of DNA (TTTTAA, TTTTAAT, TTTATA, and TTTTAA).

Affinity Labels↗
PubMed1998

Li-Fraumeni syndrome and the role of the p53 tumor suppressor gene in cancer susceptibility.

Mutation of the tumor suppressor gene p53 is a molecular genetic event frequently observed in human cancer, and inactivating missense mutations usually are accompanied by the resultant overexpression of mutant p53 protein. In gynecologic cancers, p53 is also often altered; the frequency varies depending on types of cancers and where they develop. Further, human papillomavirus oncoproteins that inactivate p53 and Rb proteins play important roles in the development of several gynecologic cancers. Individuals who are heterozygous for germline mutations of the p53 gene are strongly predisposed to a variety of cancers. The identification of these individuals may have profound value in the future when therapies or chemopreventive agents specific for the p53 alteration are available. The role of p53 tumor suppressor gene in gynecologic cancers and heritable cancer susceptibility syndromes including Li-Fraumeni and Lynch II syndromes is an active and important area of study.

Female↗
PubMed1998

Anticoagulant and antiprotease activities of a heparinoid sulfated glucoside-bearing polymer.

We studied anticoagulant and antiprotease activities of the poly(glucosyloxyethyl methacrylate) sulfate [poly(GEMA)-sulfate] in plasma and purified enzyme systems in order to evaluate the anticoagulant behavior of a heparin-like sulfated glucoside-bearing polymer. As a result, we found that poly(GEMA)-sulfate can inhibit some coagulation proteases, although its antiprotease behavior differed from those of heparin and dextran sulfate. Poly(GEMA)-sulfate could not enhance antithrombin activity; therefore, we did not observe any significant inhibition of Factor Xa via antithrombin. However, we found that poly(GEMA)-sulfate was able to inhibit thrombin through the activation of heparin cofactor II. In addition, poly(GEMA)-sulfate was able to inhibit Tenase. In our previous research. we found that the anticoagulant activity of poly(GEMA)-sulfate is due primarily to the formation of an insoluble complex with fibrinogen. This paper showed that the antiprotease activities of poly(GEMA)-sulfate contribute to some extent to its anticoagulant activity.

Anticoagulants↗
PubMed1998

Inhibition in a microgravity environment of the recovery of Escherichia coli cells damaged by heavy ion beams during the NASDA ISS phase I program of NASA Shuttle/Mir mission no. 6.

We participated in a space experiment, part of the National Space Development Agency of Japan (NASDA) Phase I Space Radiation Environment Measurement Program, conducted during the National Aeronautics and Space Administration (NASA) Shuttle/Mir Mission No. 6 (S/MM-6) project. The aim of our study was to investigate the effects of microgravity on the DNA repair processes of living organisms in the in orbit. Heavy ion beam radiation- or ç-irradiation-damaged biological samples of Escherichia coli and the radioresistant bacterium Deinococcus radiodurans were prepared and placed in a biospecimen box, which was loaded into the RRMD III sensor unit of the Space Shuttle. Two identical sets of samples were left in the Spacehab's Payload Processing Facility (SPPF) in Florida, USA, as a control. (flight No. STS-84) was launched from NASA John F. Kennedy Space Center (KSC) in Florida, USA, on May 15, 1997. The mission duration was 9.22 days. An astronaut activated the biological samples in the biospecimen box in the Spacehab during orbit in order to start repair of the DNA damaged by heavy ion beams or ç-irradiation and the samples were incubated for 19 h 35 min at about 22ûC, the cabin temperature. The control specimens in the SPPF were subjected to the same treatment under terrestrial gravity. After returned to earth, we investigated cell recovery by comparing the repair of the radiation-damaged DNA of E. coli and D. radiodurans in the microgravity environment in space with that on Earth. The results indicated that the DNA repair process of E. coli, but not of D. radiodurans, cells was inhibited in a microgravity environment.

Cell Survival↗
PubMed1997

Endogenous production of tumour necrosis factor is required for manganese superoxide dismutase expression by irradiation in the human monocytic cell line THP-1.

Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that scavenges superoxide (O2-) ions. We studied the regulation of MnSOD gene expression by irradiation and the mechanisms in human monocytic cell line THP-1. We found that irradiation induced expression of the MnSOD gene through the autocrine mechanism, involving the production of tumour necrosis factor (TNF). Irradiation increased TNF production in THP-1 cells, and TNF increased the levels of MnSOD transcripts. Supernatant from irradiated THP-1 cells induced the expression of MnSOD mRNA, and anti-TNF antibody blocked the induction of MnSOD mRNA. Irradiation also increased the levels of MnSOD mRNA in other myelocytic cell lines, HL60 and KG-1, and the ovarian cancer cell line SK-OV-3. Moreover, increased levels of MnSOD mRNA were observed in mature myeloid cells, including macrophages and granulocytes, as well as in immature cells. However, irradiation did not increase the level of MnSOD mRNA in THP-1 cells with prolonged exposure to PMA. We also found that irradiation increased the rate of MnSOD transcription, and irradiation stabilized MnSOD mRNA in THP-1 cells. Our results indicate that the endogenous production of TNF is required, at least in part, for the induction of MnSOD mRNA expression by irradiation in THP-1 cells, and the increased levels of MnSOD transcripts on irradiation occur through a pathway involving protein kinase C activation. Our results also indicate that the increase in MnSOD mRNA caused by irradiation is regulated by both transcriptional and post-transcriptional mechanisms.

Autocrine Communication↗
PubMed1997

Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis. II: Study of HSP 70 mRNA expression.

In order to investigate how cells recognize biomaterials, mRNA that was expressed in attached HeLa S3 cells on various substrates was evaluated. As culture substrates, cellulose, ethylene-vinyl alcohol copolymer (EVAL), nylon, tissue culture polystyrene (TCPS), high-density polyethylene (PE), silicone rubber, and tetrafluoroethylene-hexafluoropropylene copolymer (6F) were used. HeLa S3 cells were cultured on these substrates for 24 h. The expressed HSP 70s mRNA was then isolated and detected using the RT-PCR method. As a result, the expression of HSP 70B mRNA was largely induced in cells that adhered to hydrophilic surfaces. On the other hand, on hydrophobic surfaces, the HSP 70B mRNA expression was low. It is concluded that HSP 70B mRNA expression is sensitive to differences in the hydrophilicity-hydrophobicity of the substrates.

Biocompatible Materials↗
PubMed1997

The mechanism of anticoagulant activity of a novel heparinoid sulfated glucoside-bearing polymer.

In our previous study on glycoside polymer, it was discovered that sulfated poly(glucosyloxyethyl methacrylate) [poly(GEMA)-sulfate], which bears sulfated D-(+)-glucose, exhibited anticoagulant activity. The anticoagulant activity of poly(GEMA)-sulfate in solution was prolonged by increasing the dose or degree of sulfation of the polymer. In this study, to recognize the mechanism of anticoagulant activity of poly(GEMA)-sulfate, we estimated the binding capacity of antithrombin III to thrombin and some in vitro clotting tests in the presence of poly(GEMA)-sulfate. These results revealed that poly(GEMA)-sulfate had an anticoagulant mechanism which differed to that of heparin. We concluded that the anticoagulant activity of poly(GEMA)-sulfate is responsible for inhibiting fibrin network formation by the insoluble ion complex between fibrinogen and poly(GEMA)-sulfate.

Anticoagulants↗
PubMed1996

Anti-cancer agent OK432 induces manganese superoxide dismutase in human granulocytes.

Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme involved in scavenging O2-. OK432, a Streptococcal preparation, has anti-tumor activity and is also known to be a biological-response modifier. In this study, we have examined the regulation of MnSOD gene by OK432 in human granulocytes. Granulocytes had a low activity of MnSOD; OK432 increased MnSOD mRNA levels in a dose-dependent manner and its activity in granulocytes. OK432 also induced the MnSOD mRNA in HL60 promyelocytes, while not affecting the levels of MnSOD mRNA in human fibroblasts. Pre-treatment with either actinomycin D or cycloheximide inhibited the induction of MnSOD mRNA by OK432. OK432 increased the levels of interleukin-1 (IL-1) and tumor-necrosis-factor-alpha (TNFalpha) mRNA in granulocytes. Furthermore, the induction of MnSOD mRNA by OK432 was blocked by anti-IL-1 antibody but not by anti-TNF antibody. These results suggest that the induction of MnSOD mRNA by OK432 is regulated at the transcriptional level, and OK432 induces MnSOD mRNA, at least in part, through production of IL-1 in granulocytes.

Antibodies↗