[Diagnostic efficacy of WT1 expression in asymptomatic acute promyelocytic leukemia].

We report a case of early asymptomatic acute promyelocytic leukemia (APL) with leukopenia as the only hematologic abnormality. A 55-year-old woman was referred to our hospital with leukopenia (white blood cell [WBC] count of 1,500/µl with 36% neutrophils), which was incidentally determined during an annual medical checkup. Two months before the presentation, her WBC was 3,400/µl with 60% neutrophils. A WBC count was 1,200/µl with 40% neutrophils. Immature myeloid cells were not observed. Her hemoglobin level and platelet count were normal. Moreover, no clinical or laboratory evidence was suggestive of disseminated intravascular coagulation or infection. The peripheral blood WT1 mRNA level was increased to 26,000 copies/µg RNA. The bone marrow aspirate smear revealed 40% myeloperoxidase-positive promyelocytes with occasional Auer rods and faggots; however, circulating leukemia cells were not revealed by cell morphology or flow cytometry analysis. Quantitative reverse-transcription polymerase chain reaction analysis revealed WT1 and PML-RARA fusion transcripts in both the peripheral blood and bone marrow samples. Thus, the determination of peripheral blood WT1 expression may be sufficiently sensitive for detecting a small number of circulating APL cells.

12α-Hydroxylated bile acid enhances accumulation of adiponectin and immunoglobulin A in the rat ileum.

We previously reported that dietary supplementation with cholic acid (CA), the primary 12α-hydroxylated (12αOH) bile acid (BA), reduces plasma adiponectin concentration in rats. The aim of this study was to examine the distribution of adiponectin in the body of CA-fed rats and its influence on mucosal immunoglobulin A concentration in the intestine. Rats were fed a diet supplemented with or without CA (0.5 g CA/kg diet) for 13 weeks. A reduction in plasma adiponectin level was observed from week 3. At the end of the experiment, the CA diet reduced plasma adiponectin concentration both in the portal and aortic plasma. Accumulation of adiponectin was accompanied by an increase in cadherin-13 mRNA expression in the ileal mucosa of CA-fed rats. No increase was observed in adiponectin mRNA expression in the ileal and adipose tissues of the CA-fed rats. Immunoglobulin A concentration in the ileal mucosa was elevated in the CA-fed rats and was correlated with the ileal adiponectin concentration. 12αOH BAs may modulate mucosal immune response that are involved in the accumulation of adiponectin in the ileum.

12α-Hydroxylated bile acid induces hepatic steatosis with dysbiosis in rats.

There is an increasing need to explore the mechanism of the progression of non-alcoholic fatty liver disease. Steroid metabolism is closely linked to hepatic steatosis and steroids are excreted as bile acids (BAs). Here, we demonstrated that feeding WKAH/HkmSlc inbred rats a diet supplemented with cholic acid (CA) at 0.5 g/kg for 13 weeks induced simple steatosis without obesity. Liver triglyceride and cholesterol levels were increased accompanied by mild elevation of aminotransferase activities. There were no signs of inflammation, insulin resistance, oxidative stress, or fibrosis. CA supplementation increased levels of CA and taurocholic acid (TCA) in enterohepatic circulation and deoxycholic acid (DCA) levels in cecum with an increased ratio of 12α-hydroxylated BAs to non-12α-hydroxylated BAs. Analyses of hepatic gene expression revealed no apparent feedback control of BA and cholesterol biosynthesis. CA feeding induced dysbiosis in cecal microbiota with enrichment of DCA producers, which underlines the increased cecal DCA levels. The mechanism of steatosis was increased expression of Srebp1 (positive regulator of liver lipogenesis) through activation of the liver X receptor by increased oxysterols in the CA-fed rats, especially 4ß-hydroxycholesterol (4ßOH) formed by upregulated expression of hepatic Cyp3a2, responsible for 4ßOH formation. Multiple regression analyses identified portal TCA and cecal DCA as positive predictors for liver 4ßOH levels. The possible mechanisms linking these predictors and upregulated expression of Cyp3a2 are discussed. Overall, our observations highlight the role of 12α-hydroxylated BAs in triggering liver lipogenesis and allow us to explore the mechanisms of hepatic steatosis onset, focusing on cholesterol and BA metabolism.

A low coefficient of variation in hepatic triglyceride concentration in an inbred rat strain.

BACKGROUND: Inbred strains are characterized by less genetic variation, which suggests usefulness of inbred strains for evaluations of various parameters. In this study, experimental reproducibility in several parameters was compared between an outbred Wistar rat and Wistar King A Hokkaido (WKAH/HkmSlc) rat, the inbred strain that is originated from Wistar rats. METHODS: Difference of variations was investigated in parameters of body compositions and liver functions such as body weight, liver weight, liver triglycerides (TG), liver cholesterol and plasma alanine aminotransferase activity (ALT) between WKAH rats and outbred Wistar rats by using the coefficient of variation (CV). RESULTS: There was no difference in the CVs of body weight and relative liver weight between WKAH and Wistar rats. The CVs of body weight and relative liver weight were below 10% in both WKAH and Wistar rats. The CVs of TG, cholesterol, and ALT in Wistar rats were between 30 and 40%, whereas those in WKAH rats were between 10 and 25%. A low CV level of TG was observed in WKAH rats compared to that in Wistar rats regardless of the duration of the experimental period in those rat strains. CONCLUSION: The low CV values in metabolic parameters involved in liver functions in the inbred rats suggested an advantage of using inbred rather than outbred rats for the evaluation of liver lipid metabolism.

In Vitro and In Vivo Evaluation of Starfish Bone-Derived ß-Tricalcium Phosphate as a Bone Substitute Material.

We evaluated starfish-derived ß-tricalcium phosphate (Sf-TCP) obtained by phosphatization of starfish-bone-derived porous calcium carbonate as a potential bone substitute material. The Sf-TCP had a communicating pore structure with a pore size of approximately 10 µm. Although the porosity of Sf-TCP was similar to that of Cerasorb M (CM)-a commercially available ß-TCP bone filler-the specific surface area was roughly three times larger than that of CM. Observation by scanning electron microscopy showed that pores communicated to the inside of the Sf-TCP. Cell growth tests showed that Sf-TCP improved cell proliferation compared with CM. Cells grown on Sf-TCP showed stretched filopodia and adhered; cells migrated both to the surface and into pores. In vivo, vigorous tissue invasion into pores was observed in Sf-TCP, and more fibrous tissue was observed for Sf-TCP than CM. Moreover, capillary formation into pores was observed for Sf-TCP. Thus, Sf-TCP showed excellent biocompatibility in vitro and more vigorous bone formation in vivo, indicating the possible applications of this material as a bone substitute. In addition, our findings suggested that mimicking the microstructure derived from whole organisms may facilitate the development of superior artificial bone.

Effect of temperature on crystallinity of carbonate apatite foam prepared from alpha-tricalcium phosphate by hydrothermal treatment.

The effect of temperature on crystallinity of carbonate apatite (CAp) foam prepared from alpha-tricalcium phosphate (alpha-TCP) foam by hydrothermal treatment was investigated in the present study. The alpha-TCP foams were prepared through a conventional sintering method using polyurethane foam as template. Then, the resultant alpha-TCP foams were hydrothermally treated with Na2CO3 aqueous solution at 100 degrees C, 150 degrees C and 200 degrees C for 72 h. After hydrothermal treatment, the cancellous bone-like macroporous structure of the alpha-TCP foams was maintained. However, microscopic morphology of the foams' frame significantly changed after the 72 h treatment period. The smooth surface of alpha-TCP foam disappeared and the whole surface was covered with plate-like deposits. The plate-like deposits treated at 150 degrees C and 200 degrees C had smooth surface while those treated at 100 degrees C were constructed from spherical particles of approximately 200 nm in diameter. The results of X-ray diffraction and Fourier transform infrared analysis showed that alpha-TCP was completely converted to CAp and the crystallinity of CAp prepared at 100 degrees C was significantly lower than those prepared at 150 degrees C and 200 degrees C. Hydrothermal treatment of alpha-TCP foam at 100 degrees C allowed the formation of low-crystalline CAp foam but complete conversion needs a longer treatment period.

Fabrication of low-crystallinity hydroxyapatite foam based on the setting reaction of alpha-tricalcium phosphate foam.

Low-crystallinity hydroxyapatite (HAP) foam is an ideal material for bone substitutes and scaffolds for bone tissue regeneration, because its interconnected pores provide the space for cell growth and tissue penetration, and its composition induces excellent tissue response and good osteoconductivity. In this study, the feasibility of low-crystallinity HAP foam fabrication was evaluated based on the phase transformation reaction or the so-called dissolution-reprecipitation reaction of alpha-tricalcium phosphate (alpha-TCP) foam granules. When alpha-TCP foam granules were placed in water at 37 degrees C for 1 day, no reaction was observed. However, alpha-TCP foam granules set to form low-crystallinity HAP by treating it hydrothermally at 200 degrees C. The network of fully interconnected pores was retained, and porosity was as high as 82%. Pore size ranged from 50 to 300 mum with an average pore size of 160 mum. Compressive strength was 207 kPa. Although no setting reaction was observed at 37 degrees C, the setting reaction caused by the hydrothermal treatment of alpha-TCP foam granules allowed the fabrication of any shape of low-crystallinity HAP. Therefore, this method may be useful for the fabrication of bone substitutes and scaffolds in bone tissue regeneration.

Effects of liquid phase on basic properties of alpha-tricalcium phosphate-based apatite cement.

Effects of liquid phase on the basic properties of alpha-tricalcuim phosphate (alpha-TCP)-based cement, BIOPEX, were investigated by employing three liquid phases: distilled water, neutral sodium hydrogen phosphate solution, and succinic acid disodium salt solution containing sodium salt of chondroitin sulfate. When mixed with neutral sodium hydrogen phosphate or succinic acid disodium salt solution, the initial setting times of the cement were 19.4 +/- 0.55 and 11.8 +/- 0.45 minutes respectively. These setting times were much shorter than that of distilled water, 88.4 +/- 0.55 minutes. Formation of needle-like crystals typical of apatite was much faster when neutral sodium hydrogen phosphate solution was used, as compared to distilled water or succinic acid disodium salt solution. Moreover, at 24 hours after mixing, the largest amount of apatite was formed when neutral sodium hydrogen phosphate solution was used, whereas use of succinic acid resulted in the least. On the final mechanical strength of the cement, that yielded with neutral sodium hydrogen phosphate solution was the highest. In contrast, lower mechanical strength was observed--especially at the initial stage--when succinic acid sodium salt was used. It was thus concluded that alpha-TCP-based cement allowed accelerated transformation to apatite, and that higher mechanical strength since the initial stage was achieved when neutral sodium hydrogen phosphate solution was used as the liquid phase.

Fabrication of B-type carbonate apatite blocks by the phosphorization of free-molding gypsum-calcite composite.

B-type carbonate apatite (CO3Ap) block may be an ideal artificial bone substitute because it is closer in chemical composition to bone mineral. In the present study, the feasibility to fabricate CO3Ap blocks was investigated using compositional transformation, which was based on the dissolution-precipitation reaction of a gypsum-calcite composite with free-molding behavior. For the compositional change, or phosphorization, gypsum-calcite composites of varying CaCO3 contents were immersed in 1 mol/L (NH4)3PO4 aqueous solution at 100 degrees C for 24 hours. No macroscopic changes were found after the treatment, whereas microscopic change was observed at SEM level. X-ray diffraction, Fourier transform infrared spectroscopy and CHN analysis indicated that the composites were B-type CO3Ap containing approximately 6-7 wt% of CO3, a value similar to that of biological bone apatite. Diametral tensile strength of the CO3Ap block was approximately 1-3 MPa. Based on the results obtained, it was therefore concluded that gypsum-calcite was a good candidate for the fabrication of CO3Ap blocks, coupled with the advantage that the composite can be molded to any shape by virtue of the setting property of gypsum.

Biomimetic deposition of hydroxyapatite on a synthetic polypeptide with beta sheet structure in a solution mimicking body fluid.

Deposition of a hydroxyapatite layer with similar structure to bone mineral is an attractive approach to the fabrication of bioactive coating layers to achieve direct bonding to living bone. To get successful coating of a hydroxyapatite layer on an organic polymer using a biomimetic solution, it is essential to find organic substrates that can effectively induce heterogeneous nucleation of hydroxyapatite after exposure to the body environment. Our previous study showed that sericin, a type of silk protein, has the ability to induce hydroxyapatite nucleation in a biomimetic solution when the sericin has a beta sheet structure. To confirm the effectiveness of the beta sheet structure in hydroxyapatite nucleation, we focused on investigating hydroxyapatite deposition on a synthetic polypeptide with a beta sheet structure in a biomimetic solution. The beta sheet forming polypeptides with and without carboxyl groups, poly(FE)(3)FG, poly(FQ)(3)FG, poly(LE)(3)LG and poly(LQ)(3)LG, were synthesized in this study. All the polypeptides had mainly beta sheet structure. After soaking the polypeptide films in 1.5SBF, which has 1.5 times the inorganic ion concentrations of human blood plasma, hydroxyapatite formed on the surfaces of the polypeptides with carboxyl groups, poly(FE)(3)FG and poly(LE)(3)LG, within 2 days, but not on those without carboxyl groups, poly(FQ)(3)FG and poly(LQ)(3)LG. We confirmed that the beta sheet structure was effective for hydroxyapatite nucleation even in the synthetic polypeptide. This finding is useful for the future design of organic polymers that can effectively induce nucleation of hydroxyapatite.

Comparison of the effects of added alpha- and beta- tricalcium phosphate on the basic properties of apatite cement.

Effects of added alpha-tricalcium phosphate (alpha-TCP) and beta-TCP were investigated to shed light on the setting reaction of apatite cement (AC) consisting of tetracalcium phosphate (TTCP) and dicalcium phosphate anhydrous8 (DCPA). Added beta-TCP showed no reactivity, and thus resulted in extended setting time and decreased mechanical strength. In contrast, alpha-TCP dissolved to supply calcium and phosphate ions after initial apatite crystal formation by the reaction of TTCP and DCPA. Although setting time was delayed because alpha-TCP was involved only in the latter reaction of apatite cement, larger apatite crystals were formed due to its addition. As a result of larger apatite crystal formation, the mechanical strength of alpha-TCP-added apatite cement increased by approximately 30%, as compared to alpha-TCP-free apatite cement.

The development of Ti alloys for dental implant with high corrosion resistance and mechanical strength.

The corrosion behaviors of Ti and Ti-6Al-4V, Ti-6Al-7Nb, Ti-0.5Pt, Ti-6Al-4V-0.5Pt, and Ti-6Al-7Nb-0.5Pt alloys were examined using an electrochemical analyzer in artificial saliva containing 0.1 and 0.2% NaF at a pH of 4.0. The SEM observations revealed that the surfaces of the alloys containing 0.5 wt% Pt were not affected in fluoride-containing environments, whereas the surfaces of Ti, Ti-6A1-4V, and Ti-6Al-7Nb alloys were markedly rough. In artificial saliva containing 0.1% NaF at a pH of 4.0, the amounts of Ti dissolved from the Ti, Ti-6Al-4V, and Ti-6Al-7Nb alloys were about 50 times larger than those of the alloys containing 0.5 wt% Pt. The tensile strengths of the alloys containing 0.5 wt% Pt were equal to or higher than those of pure Ti or the alloys without Pt. The Ti-0.5Pt, Ti-6Al-4V-0.5Pt, and Ti-6Al-7Nb-0.5 alloys are expected to be useful in clinical dentistry as new Ti alloys with high corrosion resistance and mechanical strength.

Heterogeneous nucleation of hydroxyapatite on protein: structural effect of silk sericin.

Acidic proteins play an important role during mineral formation in biological systems, but the mechanism of mineral formation is far from understood. In this paper, we report on the relationship between the structure of a protein and hydroxyapatite deposition under biomimetic conditions. Sericin, a type of silk protein, was adopted as a suitable protein for studying structural effect on hydroxyapatite deposition, since it forms a hydroxyapatite layer on its surface in a metastable calcium phosphate solution, and its structure has been reported. Sericin effectively induced hydroxyapatite nucleation when it has high molecular weight and a beta sheet structure. This indicates that the specific structure of a protein can effectively induce heterogeneous nucleation of hydroxyapatite in a biomimetic solution, i.e. a metastable calcium phosphate solution. This finding is useful in understanding biomineralization, as well as for the design of organic polymers that can effectively induce hydroxyapatite nucleation.

Deposition of bone-like apatite on silk fiber in a solution that mimics extracellular fluid.

The fabrication of apatite-organic polymer hybrids is one of several attractive methods for the development of biomaterials as a substitute for bone. Such materials have both bone-bonding ability and mechanical properties analogous to natural bone. The biomimetic process has focused attention on fabricating such hybrids, where bone-like apatite is deposited on an organic polymer surface in solutions that mimic physiological conditions. In this process, a bone-like apatite layer can be coated onto organic substrates either by using a simulated body fluid (SBF) with ion concentrations nearly equal to those of human extracellular fluid, or by using fluids that are supersaturated with respect to apatite at ambient conditions. In this study, we investigated the ability of natural silk and its related materials to facilitate apatite deposition under biomimetic conditions. Cloths made of raw silk or normal silk fibers were soaked in 1.5SBF, which has 1.5 times the ion concentration of SBF. Sericin film, which is made from an extract of degummed raw silk, was soaked in 1.5SBF. The cloth and the film soaked in 1.5SBF then were characterized by scanning electron microscopic (SEM) observation, energy dispersive X-ray microanalysis (EDX), and thin-film X-ray diffraction (TF-XRD). Apatite deposition was observed on the surface of cloth made from raw silk fiber after it was soaked in 1.5SBF, but it was not observed on cloth made from normal silk fibers. The apatite deposition on the raw silk fiber cloth was accelerated when the fibers were subjected to treatment with CaCl(2) solution at a concentration of at least 1 kmol/m(3) before immersion in 1.5SBF. Apatite deposition also was observed on the sericin film after the film was soaked in 1.5SBF for 7 days. These results indicate that apatite deposition on raw silk cloth is attributable to the catalytic effect of sericin because the surface of raw silk consists of sericin whereas that of normal silk contains fibroin. The deposition of the apatite and its crystal growth are accelerated by the presence of calcium ions on the sericin after treatment with CaCl(2) solution. Thus, sericin on natural silk fiber has the potential to facilitate apatite deposition and can be useful as a polymer material in the fabrication of hybrid materials analogous to bone through biomimetic processes.