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Biomedical subjects

K Ozeki

Publications and source records attributed to K Ozeki.

At least 19 recordsLinked to original sources

Dissolution behavior and in vitro evaluation of sputtered hydroxyapatite films subject to a low temperature hydrothermal treatment.

Hydroxyapatite (HA) was coated onto titanium substrates using radio frequency sputtering. Some of the as-sputtered films were hydrothermally recrystallized at 110 degrees C. In immersion tests, the as-sputtered film completely dissolved after 2 days in a culture medium, whereas the thickness of hydrothermally treated films increased with an increase in immersion period, reaching a thickness of 127% after a period of 4 weeks. The proliferation and alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblast-like cells on the as-sputtered and hydrothermally treated films were investigated, and the cell morphology was also observed using scanning electron microscopy. The proliferation of MC3T3-E1 cells on the as-sputtered films was suppressed, whereas proliferation on the hydrothermally treated films was comparable to that on control and titanium substrate. The suppression of cell proliferation is associated with an increase in pH of the culture medium caused by dissolution of the as-sputtered film. After a 96-h culture time, the ALP activity of the cells on the hydrothermally treated film was higher than that on the control, titanium substrate, and as-sputtered film samples. From scanning electron microscopic observations, it was found that the MC3T3-E1 cells on the hydrothermally treated films were elongated and had established more intricate filopodia networks with each other, which were also observed for MC3T3-E1 cells on the as-sputtered films after a period of 24 h.

Animals↗

Preparation of germanium apatite and its cytotoxicity.

Germanium apatite was synthesized via the solid-state reaction between GeO(2) and (NH(4))(3)PO(4). The synthesized materials were characterized using XRD, and thermal analysis was carried out using TG-DTA. Ge(2)P(2)O(7) was preferentially produced at temperatures between 300-900 degrees C, and at temperatures above 1000 degrees C, germanium apatite (Ge(5)O(PO(4))(6), GeAp) was synthesized. In solubility tests, 0.36% and 0.65% of Ge ions were liberated from GeAp powder in distilled water at 37 and 80 degrees C after four weeks, respectively. A GeAp aqueous solution maintained at 37 degrees C was strongly acidic with a pH=1.67 after four weeks. The growth rate of human adult gingival fibroblast cells in a medium that included GeAp, HA, and GeO(2) was investigated. The growth rate of the cells in a 0.1 mg/ml GeAp medium was almost the same as that in the control. The cell growth was restricted in a 1.0 mg/ml GeAp medium, whereas the cell growth in a pH-adjusted 1.0 mg/ml GeAp medium at pH=7.60 was higher than that in non-adjusted medium at pH = 7.06.

Apatites↗

Effect of a thin HA coating on the stress/strain distribution in bone around dental implants using three-dimensional finite element analysis.

Three-dimensional finite element analysis was performed for thin hydroxyapatite (HA) coated and titanium dental implants to study the effects on stress/strain distribution in the mandible with application of axial and oblique loads. The implants were of screw and cylinder types. With an axial load, the maximum equivalent bone stresses in the titanium implants were 21.5 and 29.0 MPa for the cylinder and screw types respectively, and the stress and strain distributions differed. For the cylinder type, the highest stress was located at the implant base, and for the screw type, it was located at the top edge of the first thread within the cortical bone. For the HA-coated cylinder and screw implants, the maximum equivalent bone stresses were 7.1 and 7.2 MPa respectively. The stress and strain distributions were similar, and the highest stress was located on the upper side of the cortical bone around the implant neck for both implants. Of the implants examined, the screw type HA-coated implant had the most uniform stress distribution in bone.

Anisotropy↗

Bone response to titanium implants coated with thin sputtered HA film subject to hydrothermal treatment and implanted in the canine mandible.

Hydroxyapatite (HA) was coated onto titanium implants using radio frequency magnetron sputtering. The HA films were crystallized in an autoclave tube using low temperature hydrothermal treatment. The average film thickness on the implant was 1.1 microm. HA-coated and pure-titanium implants were inserted into canine mandibles for up to 24 weeks. Forty-eight implants were placed in eight beagles. After 2, 4, 12 and 24 weeks, implants were retrieved and prepared for histological observation, and the HA film thickness was determined using energy-dispersive X-ray spectroscopy. Light microscopy revealed that, after two weeks, the bone response to the HA-coated implants was much better than to the pure titanium implants, and osteoblasts were observed at the bone-implant interface. After four weeks, the screw threads of the HA-coated implants were almost completely covered with bone. The HA film thickness rapidly decreased up to four weeks of implantation, then gently decreased, reaching 0.40+/-0.03 microm at the upper region of the implant after 12 weeks. That indicates that about 80% of the HA film had dissolved after 12 weeks of implantation. The rate of decrease in the HA film thickness was greater with increasing implant depth.

Animals↗

Expression and methylation status of the FHIT gene in acute myeloid leukemia and myelodysplastic syndrome.

To clarify the role of fragile histidine triad (FHIT) in hematological malignancies, we examined the methylation status and the expression level of the FHIT gene in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) cells in comparison with the methylation of the p15(INK4B) gene. The FHIT methylation was found in 13 of 94 (13.8%) AML and 22 of 40 (55.0%) MDS cases, but not in normal mononuclear cells (MNCs). Both the frequency and density of methylation increased in the advanced-stages MDS and the relapsed AML cases. Although FHIT and p15(INK4B) methylations were not correlated in MDS and AML, increased FHIT methylation at the relapse in AML was associated with p15(INK4B) methylation. The median expression level in AML was significantly higher than in normal MNCs, although the median expression level in those with methylation was significantly lower than in those without methylation. Furthermore, the methylation level at relapse was significantly higher than at diagnosis in AML. These results suggested that FHIT methylation was accumulated through the disease progression of MDS and AML, and the role of the FHIT gene as a tumor suppressor seemed different in AML and MDS.

Acid Anhydride Hydrolases↗

Preparation and osteocompatibility of hydroxyapatite coated on titanium from the reaction of sputtered CaO and vaporized P2O5.

Hydroxyapatite (HA) and other calcium phosphates were synthesized on titanium plates by a solid-gas state reaction of sputtered CaO and vaporized P(2)O(5). The calcium phosphates formed were HA, beta-tricalcium phosphate (beta-TCP; Ca(3)(PO(4))(2)), beta-calcium pyrophosphate (beta-PYR; Ca(2)P(2)O(7)), and beta-calcium metaphosphate (beta-MET; Ca(2)(PO(3))(2)). Their formation depended on the ratio of the sputtered CaO and the reacting P(2)O(5). For a mole ratio of CaO/P(2)O(5)=4 (Ca/P=2), an HA film was synthesized. The surface roughness increased by over seven times after the solid-gas state reaction from Ra = 0.16+/-0.02 microm (for the CaO film) to Ra = 1.15+/-0.25 microm (for the reacted film). The synthesized HA film-coated titanium plates and control non-coated titanium plates were implanted in the femora of two dogs for a period of two, four and 12 weeks, and observed using a soft X-ray radiograph and histological sections. New bone formation was observed without any connective tissue at four weeks around the HA film, whereas over the 12 week experimental period, there was no new bone formation around the control and connective tissue was observed over all periods, reaching a thickness of more than 200 microm at 12 weeks.

Animals↗

Inhibition of Ni release from NiTi alloy by hydroxyapatite, alumina, and titanium sputtered coatings.

To alleviate the effects of Ni allergy from NiTi alloy implants, hydroxyapatite (Ca10(PO4)6(OH)2; HA), alumina (Al2O3), or titanium (Ti) was coated onto NiTi alloy plates to form 1-microm thick films using radio frequency magnetron sputtering. The coatings on the plates were characterized using XRD. After the plates had been immersed in physiological saline for periods of one, four, or eight weeks, the concentration of Ni ions released in each solution was detected using a microwave induced plasma mass spectrometer. After eight weeks, the concentration of Ni ions released from the non-coated, the Ti-coated, the HA-coated, and the alumina-coated plates were 238, 19.7, 183, and 106 ppb, respectively. The bonding strength of the Ti film, the HA film, and the alumina film to the NiTi substrate were 3.8 +/- 1.2, 2.6 +/- 0.7, and 3.1 +/- 1.2 MPa, respectively. The non-coated, the HA-coated, the alumina-coated, and the Ti-coated plates were implanted into the femurs of a dog for four weeks for histological observation. In case of the non-coated plates, connective tissue more than 300 microm thick was observed, whereas for the coated plates the thickness of the connective tissue was around 100 microm.

Aluminum Oxide↗

Deterioration in the superelasticity of Ti sputter coated on NiTi orthodontic wire.

To evaluate the deterioration in the superelasticity of Ti-coated NiTi orthodontic wires, titanium was coated onto wires to form a 1-microm thick film using radio frequency magnetron sputtering. In superelasticity cycle tests, the sigma5.0 value (the stress at 5.0% strain) of a Ti-coated wire was lower than that of a non-coated wire by 6.2% after one cycle, and by 10.4% after 20 cycles. In a three point bend tests, the L2.0 value (the load at 2.0 mm deflection) of a Ti-coated wire was 2.55 N, and this was reduced by 39.0% of the L2.0 value of a non-coated wire (4.18 N). After the non-coated and the Ti-coated wires had been immersed in physiological saline for periods of four and eight weeks, the concentration of Ni ions released into each solution was detected using MIP-Mass, and Ni peak intensity of the wires was measured using XPS. After eight weeks, the concentration of Ni ions released from the non-coated and the Ti-coated wires was 657 and 135 ppb, respectively. In a clinical test, no inflammation was observed during the entire attached period of four weeks in both the non-coated and the Ti-coated wire. In the Ti-coated wire, little exfoliation of the Ti-coating was observed using SEM and EDS. It was concluded that Ti-coated NiTi orthodontic wire has much potential for its application, with a low probability of causing any Ni allergy.

Coated Materials, Biocompatible↗

Bone bonding strength of sputtered hydroxyapatite films subjected to a low temperature hydrothermal treatment.

Hydroxyapatite (HA) was coated onto titanium substrates using radio frequency sputtering, and the coated HA films were crystallized in an autoclave at 110 degrees C using a low temperature hydrothermal method. The crystallite size, the Ca/P ratio, and the surface of the films were observed using XRD, EDS, and SEM, respectively. An immersion test was carried out in physiological saline solution, and the film-to-substrate adhesion strength was measured using a pull-out test. From the XRD patterns, a sputtered film subjected to the hydrothermal treatment had crystallized after 24 h, and the crystallite size increased from 38.0+/-8.7 to 81.4+/-19.1 nm. In immersion tests, an as-sputtered film completely dissolved after 1 d, whereas a sputtered film subjected to the hydrothermal treatment survived to 18.7+/-5.8% of the initial film thickness after four weeks. In pull-out tests, the adhesion strength of the sputtered film to the substrate increased from 1.9+/-0.2 to 5.3+/-1.6 MPa after the hydrothermal treatment. A sputtered film subjected to the hydrothermal treatment and a plasma-sprayed coating on titanium columns were implanted in the diaphysis of the femora of six adult dogs, and a pull-out test was carried out after two, four, and 12 weeks. The sputtered film showed higher bone bonding strength than the plasma-sprayed coating at any period.

Animals↗

Isolation of a new lytic enzyme for hiochi bacteria and other lactic acid bacteria.

A microorganism producing a lytic enzyme preparation that could rapidly lyse bacterial cells such as hiochi bacteria and other lactic acid bacteria was screened. The microorganism was identified as Streptomyces fulvissimus. The enzyme produced by this organism lysed boil-denatured cells quicker than intact cells of hiochi bacteria. A mutant strain of S. fulvissimus producing the enzyme exhibiting high activity against intact cells of hiochi bacteria was screened on plates, containing intact cells inactivated with UV irradiation. The optimal pH for lytic activity against intact cells of the hiochi bacterium Lactobacillus casei S-4 was from 3.5 to 4.0, and the optimum temperature was close to 50 degrees C. This enzyme activity was stable between pH 3.5 and pH 8.0 and up to 60 degrees C. The enzyme exhibits N-acetyl glucosaminidase and muramidase activities. The effects of adjusting the pH and using different inducers for enzyme production were investigated. Chitin was the most effective inducer of enzyme production. Intact DNA was easily isolated from the cells of many lactic acid bacteria following lysis with the enzyme. It is thought that this enzyme will be a good biotechnological tool.

Journal Article↗

Push-out strength of hydroxyapatite coated by sputtering technique in bone.

Hydroxyapatite was coated with 1 microm thickness on titanium columns of a length of 10 mm, an outer diameter of 4.0 mm by radio frequency magnetron sputtering. The hydroxyapatite coating titanium columns were implanted in the diaphysis of femora of 3 adult dogs, and push-out test was carried out after 2, 4 and 12 weeks of implantation using a testing machine. The interface of bone/column was observed histologically after the test. At 12 weeks the push-out strengths of coating and non-coating columns were 3.5 and 1 MPa, respectively. Histological observation indicated a formation of thin connective tissue with 5-30 microm thickness at the interface of the bone/column. No inflammation was observed during the implantation periods.

Animals↗

Sleep apnea syndrome in patients with pulmonary thromboembolism.

Seven patients (one man and six women) with a diagnosis of pulmonary thromboembolism (PTE) were examined by polysomnography in order to clarify the relationship between sleep-related breathing disorders and PTE. In the chronic stage of PTE, sleep apnea syndrome (SAS) was recognized in two patients (a man and a woman) among the subjects. Four of the five patients without SAS showed nocturnal hypoxemia. The female predominance was different from ordinary SAS without the disease background of PTE even though PTE is accompanied by hypoxemia and pulmonary hypertension like SAS. A pathophysiological relationship between PTE and SAS was not found in this study.

Aged↗

Molecular cloning and characterization of a transcriptional activator gene, amyR, involved in the amylolytic gene expression in Aspergillus oryzae.

A gene, designated amyR, coding for a transcriptional activator involved in amylolytic gene expression has been cloned from Aspergillus oryzae by screening for a clone that enabled to reverse the reduced expression of the alpha-amylase gene (amyB) promoter. amyR encodes 604 amino acid residues of a putative DNA-binding protein carrying a zinc binuclear cluster motif (Zn(II)2Cys6) belonging to the GAL4 family of transcription factors. The amyR gene disruptants showed a significant restricted growth on starch medium and produced little of the amylolytic enzymes including alpha-amylase and glucoamylase compared with a non-disruptant, indicating that amyR is a transcriptional activator gene involved in starch/maltose-induced efficient expression of the amylolytic genes in A. oryzae. In addition, sequencing analysis found that amyR, agdA (encoding alpha-glucosidase), and amyA (encoding alpha-amylase), are clustered on a 12-kb DNA fragment of the largest chromosome in A. oryzae, and that amyR is about 1.5 kb upstream of agdA and transcribed in the opposite direction. Furthermore, transcriptional analysis revealed that the amyR gene was expressed in the presence of glucose comparable to the level in the presence of maltose, while the amylolytic genes were transcribed at high levels only in the presence of maltose.

Amino Acid Sequence↗

Crystal chemistry of hydroxyapatite deposited on titanium by sputtering technique.

Crystalline hydroxyapatite (HA) powder was coated on titanium substrate by radio frequency (RF) magnetron sputtering. The coating was homogeneous thin film and the thickness was 1microm. Crystallinity of the HA coating was low and Ca/P ratio was high as 3.0. Particle sizes were 40 to approximately 100 nm, and the crystallite size was calculated by 30 to approximately 50 nm using an X-ray diffractometry (XRD) and a transmission electron microscopy (TEM). When the coating was heated at 800 degrees C for 1 h, the low crystalline HA grew up crystalline HA, and a diffraction pattern of CaO appeared. When the coating was immersed in pH 7.4 of bovine serum for 1 week, c-axis of HA increased.

Animals↗

Molecular and enzymic properties of recombinant 1, 2-alpha-mannosidase from Aspergillus saitoi overexpressed in Aspergillus oryzae cells.

For the construction of an overexpression system of the intracellular 1,2-alpha-mannosidase (EC 3.2.1.113) gene (msdS) from Aspergillus saitoi (now designated Aspergillus phoenicis), the N-terminal signal sequence of the gene was replaced with that of the aspergillopepsin I (EC 3.4.23.18) gene (apnS) signal, one of the same strains as described previously. Then the fused 1, 2-alpha-mannosidase gene (f-msdS) was inserted into the NotI site between P-No8142 and T-agdA in the plasmid pNAN 8142 (9.5 kbp) and thus the Aspergillus oryzae expression plasmid pNAN-AM1 (11.2 kbp) was constructed. The fused f-msdS gene has been overexpressed in a transformant A. oryzae niaD AM1 cell. The recombinant enzyme expressed in A. oryzae cells was purified to homogeneity in two steps. The system is capable of making as much as about 320 mg of the enzyme/litre of culture. The recombinant enzyme has activity with methyl-2-O-alpha-d-mannopyranosyl alpha-D-mannopyranoside at pH 5.0, while no activity was determined with methyl-3-O-alpha-D-mannopyranosyl alpha-D-mannopyranoside or methyl-6-O-alpha-D-mannopyranosyl alpha-D-mannopyranoside. The substrate specificity of the enzyme was analysed by using pyridylaminated (PA)-oligomannose-type sugar chains, Man9-6(GlcNAc)2-PA (Man is mannose; GlcNAc is N-acetylglucosamine). The enzyme hydrolysed Man8GlcNAc2-PA (type 'M8A') fastest, and 'M6C' {Manalpha1-3[Manalpha1-2Manalpha1-3(Manalpha1-6) Manalpha1-6]Manbeta1- 4GlcNAcbeta1-4GlcNAc-PA} slowest, among the PA-sugar chains. Molecular-mass values of the enzyme were determined to be 63 kDa by SDS/PAGE and 65 kDa by gel filtration on Superose 12 respectively. The pI value of the enzyme was 4.6. The N-terminal amino acid sequence of the enzyme was GSTQSRADAIKAAFSHAWDGYLQY, and sequence analysis indicated that the signal peptide from apnS gene was removed. The molar absorption coefficient, epsilon, at 280 nm was determined as 91539 M-1.cm-1. Contents of the secondary structure (alpha-helix, beta-structure and the remainder of the enzyme) by far-UV CD determination were about 55, 38 and 7% respectively. The melting temperature, Tm, of the enzyme was 71 degrees C by differential scanning calorimetry. The calorimetric enthalpy, DeltaHcal, of the enzyme was calculated as 13.3 kJ.kg of protein-1. Determination of 1 g-atom of Ca2+/mol of enzyme was performed by atomic-absorption spectrophotometry.

Amino Acid Sequence↗

Granular Cell Tumor of the Breast: A Case Report Describing Dynamic MR Mammography.

A case of granular cell tumor of the breast in a 36-year-old woman is reported. The patient presented with a hard mass,1.5 cm in diameter in the upper-outer quadrant of the left breast. Physical examination and ultrasonography suggested the presence of breast carcinoma. An aspiration biopsy cytology(ABC)specimen wasevaluated as class II, and dynamic magnetic resonance(MR)mammography indicated a benign tumor. Granular cell tumor was finally diagnosed on examination of an excisional biopsy specimen. Granular cell tumor of the breast can mimic breast carcinoma on physical examination, mammography, ultrasonography, and even gross inspection. Dynamic MR mammography has the potential distinguish this condition from carcinoma. Awareness of this disease and prudent use of diagnostic procedures, including MR mammography, will help prevent misdiagnosis and unnecessary surgery.

Journal Article↗

Changes in colonic motility in dogs after a resection of the inferior mesenteric ganglion and plexus.

In a curative resection for advanced sigmoid or rectal cancer, an extensive dissection of the regional lymph nodes is generally required. This often necessitates the removal of the autonomic nerves around the inferior mesenteric artery. The present study was done in an attempt to clarify the influence of a neurectomy around the inferior mesenteric ganglion and plexus on the motility of the colon. In eight dogs, we resected the ganglion and plexus around the inferior mesenteric artery, together with an implantation of strain gauge force transducers in various parts of the colon, and 7-10 days later, colonic motility was examined. The percentage of contractile states and contractile forces increased at both the distal colon in fasting dogs, as well as at the middle colon in the late postprandial period. At the distal colon, contractile forces were noted in the early and late postprandial periods. These contractile abnormalities at the middle and distal colon may thus explain the frequent bowel movements or diarrhea often observed after extensive surgery in patients with sigmoid or rectal cancer.

Animals↗