Nuclear Expression of Pygo2 Correlates with Poorly Differentiated State Involving c-Myc, PCNA and Bcl9 in Myanmar Hepatocellular Carcinoma

In Myanmar, hepatocellular carcinoma (HCC) is commonly seen in young adult and associated with poor prognosis, while the molecular mechanisms that characterize HCC in Myanmar are unknown. As co-activation of Wnt/β-catenin signaling and c-Myc (Myc) are reported to associate with malignancy of HCC, we immunohistochemically investigated the expression of Pygo2 and Bcl9, the co-activators of the Wnt/β-catenin signaling, Myc and PCNA in 60 cases of Myanmar HCC. Pygo2 expression was confirmed by in situ hybridization. The signal intensity was measured by image analyzer and then statistically analyzed. As a result, the expression of Pygo2 was significantly higher in HCC compared to normal liver tissue and the nuclear signal was the most intense in poorly differentiated HCC. Cytoplasmic Bcl9 was expressed in the normal liver tissue but decreased in HCC with the progression of histopathological grade. Myc was significantly higher in poorly differentiated HCC, whereas PCNA labeling index increased with the progression of histopathological grade. Nuclear Pygo2 showed strong correlation with nuclear Myc (P < 0.01) and PCNA (P < 0.001), and inversely correlated with cytoplasmic Bcl9 (P < 0.01). Our results suggested Wnt/β-catenin and Myc signaling is commonly activated in Myanmar HCC and that the correlative upregulation of nuclear Pygo2 and Myc characterizes the malignant features of HCC in Myanmar

Enhanced initial cell responses to chemically modified anodized titanium.

BACKGROUND: Previously, we reported that anodized porous titanium implants have photocatalytic hydrophilicity. However, this effect was not always sufficient for the significant improvement of bone apposition. PURPOSE: The purpose of this study was to improve the photocatalytic properties of porous titanium implants by the fluoride modification of the anodized titanium dioxide (TiO(2)), and to investigate the initial cell response to it. MATERIALS AND METHODS: The ideal concentration of ammonium hydrogen fluoride (NH(4)F-HF(2)) used in this study was determined by a static water contact angle assay. The ideal concentration of NH(4)F-HF(2) was 0.175%, and experimental disks were treated with this concentration. A pluripotent mesenchymal cell line, C2C12, was cultured on the disks in order to investigate cell attachment, morphology, and proliferation. RESULTS: Cell attachment after 30 minutes of culturing was significantly higher for the ultraviolet-irradiated, fluoride-modified anodized TiO(2) (p < .05), and the simultaneous scanning electron microscope observation showed a rather flattened and extended cell morphology. The proliferation rate after 24 hours was also significantly higher for the fluoride-modified anodized TiO(2). CONCLUSION: Fluoride chemical modification enhances the hydrophilic property of the anodized TiO(2) and improves the initial cell response to it

Experimental apparatus at KUR-ISOL to identify isomeric transitions from fission products, and decay spectroscopy of 151Ce

Decay studies on 151Ce have been performed using the on-line isotope separator connected to the Kyoto University Reactor. In addition to conventional γ and conversion electron spectroscopy, β-gated measurements were carried out on mass-separated 151Ce to identify an isomeric transition. From the analysis of the obtained data, the half-life of the isotope was ascertained to be 1.76(6) s and a decay scheme containing six excited levels was constructed for the first time. The excited level at 35.1 keV in 151Pr was found to be a long-lived state (a half-life of approximately 10us or longer)

A bone substitute with high affinity for vitamin D-binding protein―relationship with niche of osteoclasts

The biological activity of osteoblasts and osteoclasts is regulated not only by hormones but also by local growth factors, which are expressed in neighbouring cells or included in bone matrix. Previously, we developed hydroxyapatite (HA) composed of rod-shaped particles using applied hydrothermal methods (HHA), and it revealed mild biodegradability and potent osteoclast homing activity. Here, we compared serum proteins adsorbed to HHA with those adsorbed to conventional HA composed of globular-shaped particles (CHA). The two ceramics adsorbed serum albumin and γ-globulin to similar extents, but affinity for γ-globulin was much greater than that to serum albumin. The chemotactic activity for macrophages of serum proteins adsorbed to HHA was significantly higher than that of serum proteins adsorbed to CHA. Quantitative proteomic analysis of adsorbed serum proteins revealed preferential binding of vitamin D-binding protein (DBP) and complements C3 and C4B with HHA. When implanted with the femur of 8-week-old rats, HHA contained significantly larger amount of DBP than CHA. The biological activity of DBP was analysed and it was found that the chemotactic activity for macrophages was weak. However, DBP-macrophage activating factor, which is generated by the digestion of sugar chains of DBP, stimulated osteoclastogenesis. These results confirm that the microstructure of hydroxyapatite largely affects the affinity for serum proteins, and suggest that DBP preferentially adsorbed to HA composed of rod-shaped particles influences its potent osteoclast homing activity and local bone metabolism

Osteoconductivity of hydrothermally synthesized beta-tricalcium phosphate composed of rod-shaped particles under mechanical unloading

Spherical beta-tricalcium phosphate (β-TCP) granules synthesized using a unique dropping slurry method expressed good osteoconductivity with prominent bone apposition and bioresorbability when implanted into the rat femur (Gonda et al, Key Eng. Mater. 361-363:1013-1016, 2008). The spherical β-TCP granules were implanted into the bone defect created in the distal end of the right femur of each 8-week-old female Wistar rat. To analyze performance of the spherical β-TCP granules as bone substitute in the bone with reduction in osteogenic potential, the right sciatic neurectomy was performed after implantation and the right hind limb was kept unloaded for 2 weeks before euthanization. Four weeks after implantation, some spherical β-TCP granules with resorption in part were surrounded by newly formed bone. Eight and 12 weeks after implantation, most of the residual β-TCP granules were embedded in newly formed bone, and total volume of the implant and newly formed bone was more than the other portions of the bone or the bone of control animals. Osteoclast activity in the implanted area was also higher than the other portions of the bone or the bone of control animals. Replacement of the intraosseous residual β-TCP granules for bone progressed at 12 weeks after implantation compared to those at 8 weeks after implantation. These data suggested that the spherical β-TCP granules stimulated osteogenesis and osteoclast activity of the unloaded bone

Coexpression of Ang1 and Tie2 in Odontoblasts of Mouse Developing and Mature Teeth?A New Insight into Dentinogenesis

Agiopoieten regulates vascular angiogenesis and stabilization, and is reported to promote bone formation by facilitating angiogenesis. To estimate the role of Ang1 in odontogenesis, we explored the distribution of Ang1 and the receptor, Tie2 in the mouse developing and mature first molar of the mandible. At embryonic day 18, when differentiation of odontoblasts begins, immunosignals for Ang1 were intensely detected in the basement membrane and the distal side, which faced the basement membrane of odontoblasts. In situ hybridization revealed that Ang1 was expressed in odontoblasts and ameloblasts facing the basement membrane. Tie2 was localized in the distal side of odontoblasts. After birth, Ang1 was detected in the predentin, whereas both Ang1 and Tie2 were colocalized in odontoblasts and odontoblast processes. These distributions were retained up to 8 weeks. In contrast to odontoblasts, ameloblasts, cementoblasts and osteoblasts expressed Ang1 but did not express Tie2. Colocalization of Ang1 and Tie2 in odontoblasts and selective expression of Tie2 in odontoblasts among cells responsible for calcified tissue formation suggested the involvement of autocrine signals of Ang1-Tie2 in dentinogenesis

Enhanced osseointegration by the chemotactic activity of plasma fibronectin for cellular fibronectin positive cells.

Plasma fibronectin (pFN) is known to regulate cell growth, differentiation or survival of osteoblasts in vitro. It is also speculated to be important for the early phase of osseointegration, however, its actual in vivo behavior is unknown. The objective of this study is to clarify the role of pFN during osseointegration. We developed a titanium ion-plated acrylic implant (Ti-acryl) for thin sectioning without removal of the implant. Either Ti-acryl or pFN-coated Ti-acryl (FN-Ti-acryl) was implanted in the mouse femur. Samples were taken on days 1-7 and on day 14 after the operation, and were decalcified and paraffin embedded. The bone healing process and immunofluorescence localization of pFN and cellular fibronectin (cFN), a marker for fibroblastic cells were examined. Simultaneously, the effect of pFN on chemotaxis, proliferation and differentiation of bone marrow stromal cells (BMSCs) was analyzed in vitro. The in vivo results showed that faster direct bone formation was seen for the FN-Ti-acryl group compared to the Ti-acryl group. The in vitro results showed that pFN significantly promoted BMSCs chemotaxis, however, had no effect on proliferation or differentiation. The results indicate that pFN regulated chemotaxis of osteogenic cells and coating the implant with pFN enhanced earlier osseointegration

The renin–angiotensin system promotes arrhythmogenic substrates and lethal arrhythmias in mice with non-ischaemic cardiomyopathy

[Aims]The progression of pathological left ventricular remodelling leads to cardiac dysfunction and contributes to the occurrence of malignant arrhythmias and sudden cardiac death. The underlying molecular mechanisms remain unclear, however. Our aim was to examine the role of the renin–angiotensin system (RAS) in the mechanism underlying arrhythmogenic cardiac remodelling using a transgenic mouse expressing a cardiac-specific dominant-negative form of neuron-restrictive silencer factor (dnNRSF-Tg). This mouse model exhibits progressive cardiac dysfunction leading to lethal arrhythmias. [Methods and results]Subcutaneous administration of aliskiren, a direct renin inhibitor, significantly suppressed the progression of pathological cardiac remodelling and improved survival among dnNRSF-Tg mice while reducing arrhythmogenicity. Genetic deletion of the angiotensin type 1a receptor (AT1aR) similarly suppressed cardiac remodelling and sudden death. In optical mapping analyses, spontaneous ventricular tachycardia (VT) and fibrillation (VF) initiated by breakthrough-type excitations originating from focal activation sites and maintained by functional re-entry were observed in dnNRSF-Tg hearts. Under constant pacing, dnNRSF-Tg hearts exhibited markedly slowed conduction velocity, which likely contributes to the arrhythmogenic substrate. Aliskiren treatment increased conduction velocity and reduced the incidence of sustained VT. These effects were associated with suppression of cardiac fibrosis and restoration of connexin 43 expression in dnNRSF-Tg ventricles. [Conclusion]Renin inhibition or genetic deletion of AT1aR suppresses pathological cardiac remodelling that leads to the generation of substrates maintaining VT/VF and reduces the occurrence of sudden death in dnNRSF-Tg mice. These findings demonstrate the significant contribution of RAS activation to the progression of arrhythmogenic substrates

Adult phenology of two species of tiger beetles (Carabidae, Cicindelinae) and estimates of population size of Cylindela elisae , in Tottori Sand Dunes, Honshu, Japan in 2016.

2015年の調査に引き続き，2016年の夏季にも鳥取砂丘オアシス周辺で標識再捕により，当地で絶滅が心配されるエリザハンミョウの生息数を推定した。マークできた個体はエリザハンミョウが270（昨年は304），カワラハンミョウが170（昨年は77），コハンミョウが4（昨年は1：昨年調査の報告書である鶴崎ら2016では「コニワハンミョウ」と誤記）であった。Jolly-Seber法によるエリザハンミョウの個体数推定値はもっとも多かった調査日（6月28日と7月16日）でともに約1,460で，2015年の2,300個体よりも少なかった。おそらく2016年の夏季の高気温のため，成虫の出現期は2015年よりも早く推移し，6 月中旬には成虫が出現し，8月下旬には終息した。エリザハンミョウの幼虫の営巣地はオアシス周辺の尻無川右岸の裸地であるが，成虫は左岸のコウボウシバ群落でも見つかった。カワラハンミョウの成虫は6月下旬から10月上旬まで見られ，170個体をマークしたが再捕獲できた個体は皆無であった。コハンミョウは尻無川の近くで4個体マークしたが本種も再捕獲にいたらなかった。昨年の2個体（マークしたのは1個体）に続けての確認で，細々ではあるが，本種は当地で世代を繰り返している可能性が高い。 / Following a survey of the population size of a tiger beetle species, Cylindera elisae(Motschulsky,1859) in the Tottori Sand Dunes, Tottori City, in 2015 (Tsurusaki et al.2016), we estimated population size of　the same population of the same species also in 2016 by using mark-recapture experiments. A total of　270 adults of Cy. elisae , 170 adults of Chaetodera laetescripta (Motschulsky, 1860) and 4 adults of Myriochila　speculifera (Chevrolat, 1865) (This species was　erroneously recorded as Cicindela　transbaicalica japanensis　Chaudoir, 1863 in Tsurusaki et al. 2016) were individually marked during the summer in 2016.　None of　those　adults marked were recaptured for the　two latter species. The highest number of　adults of Cy. elisae estimated　by the Jolly-Seber method was ca. 1,460 recorded on both　28 June and 16 July

Bcl9の異なるエピトープを認識する二つの抗体を用いた免疫組織化学的マッピングによるミャンマー国若年性発症肝細胞がんの特徴付け

B-cell lymphoma 9 (Bcl9) is the core component of Wnt/β-catenin signaling and overexpressed in nuclei of various tumors, including hepatocellular carcinoma (HCC). However, the extent of Bcl9 expression relative to HCC differentiation stage and its functional aspects are poorly understood. In this study, we examined the expression pattern of Bcl9 immunohistochemically, using two anti-Bcl9 antibodies; one was a conventional polyclonal-antibody (anti-Bcl9ABC) against amino acid no.800?900 of human-Bcl9, while the other (anti-Bcl9BIO) was against amino acid no.50?200, covering Pygopus-binding sites of Bcl9. Immunohistochemistry using anti-Bcl9BIO demonstrated distinctive staining in the cytoplasm, while the anti-Bcl9ABC signal was detected in both cytoplasm and nuclei of HCC cells, reflecting different states of Bcl9 function because Pygopus-binding to Bcl9 is essential to exert its function together with β-catenin in nucleus. Quantitative analysis revealed a significantly higher immunohistochemical-score by anti-Bcl9BIO in normal liver comparing various differentiation grades of HCC (P < 0.004), whereas no significant difference was noted with anti-Bcl9ABC. Interestingly, immunohistochemical-score of anti-Bcl9BIO in patients aged < 40 years was significantly lower than that of ? 40 years group (P < 0.01). The results indicated that anti-Bcl9BIO detected cytoplasmic Bcl9, which does not bind to Pygopus suggesting it could be a useful indicator for development of HCC in young Myanmar patients.長崎大学学位論文 学位記番号:博(医歯薬)甲第1129号 学位授与年月日:平成31年3月20日Author: Myat Thu Soe, Yasuaki Shibata, Myo Win Htun, Kuniko Abe, Kyaw Soe, Nay Win Than, Thann Lwin, Myat Phone Kyaw and Takehiko KojiCitation: Acta Histochemica et Cytochemica, 52(1), pp.9-17; 2019Nagasaki University (長崎大学)課程博