Search

Simple Summary Hyperactivation of the canonical Wnt and inactivation of the Hippo pathway are well-known genetic backgrounds for familial adenomatosis polyposis (FAP) and colorectal cancer (CRC), although the reciprocal regulation between those pathways is not yet clear. In this study, we found that Axin2, a bona fide downstream target of canonical Wnt, activates the Hippo pathway in APC-mutated CRC, limiting the therapeutic potential of niclosamide on advanced CRC through the inactivation of the Hippo pathway. To overcome the limitation, we combined niclosamide with AMPK activator metformin to activate Hippo and found that this combination synergistically suppressed canonical Wnt and activated Hippo in APC-mutated CRC. Using patient-derived cancer organoid and an APC-MIN mice model, we found the combinatory approach to be effective for APC-mutated CRC. Our results provide not only the reciprocal link between Wnt and Hippo in APC-mutated CRC, but they also provide an effective therapeutic approach with clinically available drugs for FAP and CRC patients. Abstract The Wnt and Hippo pathways are tightly coordinated and understanding their reciprocal regulation may provide a novel therapeutic strategy for cancer. Anti-helminthic niclosamide is an effective inhibitor of Wnt and is now in a phase II trial for advanced colorectal cancer (CRC) patients. We found that Axin2, an authentic target gene of canonical Wnt, acts as aYAP phosphorylation activator in APC-mutated CRC. While niclosamide effectively suppresses Wnt, it also inhibits Hippo, limiting its therapeutic potential for CRC. To overcome this limitation, we utilized metformin, a clinically available AMPK activator. This combinatory approach not only suppresses canonical Wnt activity, but also inhibits YAP activity in CRC cancer cells and in patient-derived cancer organoid through the suppression of cancer stemness. Further, combinatory oral administration suppressed in vivo tumorigenesis and the cancer progression of APC-MIN mice models. Our observations provide not only a reciprocal link between Wnt and Hippo, but also clinically available novel therapeutics that are able to target Wnt and YAP in APC-mutated CRC.

Purpose Wnt and mammalian target of rapamycin (mTOR) are major molecular signaling pathways associated with the development and progression of tumor, as well as the maintenance and proliferation of cancer stem cells (CSCs), in colorectal cancer (CRC). Identifying patients at risk of poor prognosis is important to determining whether to add adjuvant treatment in stage II CRC and thus improve survival. In the present study, we evaluated the prognostic value of Wnt, mTOR, and CSC markers as survival predictors in stage II CRC. Materials and methods We identified 148 cases of stage II CRC and acquired their tumor tissue. Tissue microarrays for immunohistochemical staining were constructed, and the expressions of CD166, CD44, EphB2, β-catenin, pS6 were evaluated using immunohistochemical staining. Results The expressions of CD166 (p=0.045) and pS6 (p=0.045) and co-expression of pS6/CD166 (p=0.005), pS6/CD44 (p=0.042), and pS6/CD44/CD166 (p=0.013) were negatively correlated with cancer-specific survival. Cox proportional hazard analysis showed the combination of CD166/pS6 [hazard ratio, 9.42; 95% confidence interval, 2.36-37.59; p=0.002] to be the most significant predictor related with decreased cancer-specific survival. In addition, co-expression of CD44/CD166 (p=0.017), CD166/β-catenin (p=0.036), CD44/β-catenin (p=0.001), and CD44/CD166/β-catenin (p=0.001) were significant factors associated with liver metastasis. Conclusion Specific combinations of CSC markers and β-catenin/mTOR signaling could be a significant predictor of poor survival in stage II CRC.

To obtain an enhanced population of cardiomyocytes from differentiating mouse embryonic stem (ES) cells, we confirmed the role of noggin treatment during the cardiac differentiation of mouse ES cells. ES cells were cultured in ES medium containing both noggin and LIF for 3 days on the mouse embryonic fibroblast feeder layer, followed by dissociated and suspension culture without LIF to form the embryoid body (EB). The next day, noggin was eliminated and EBs were cultured continuously. Noggin treated ES cells showed a relatively rapid increase of cardiac marker genes, while the vehicle (PBS) treated group showed no significant cardiac marker expression at 4 days after the EB formation. Furthermore, Noggin treated ES cells showed 68.00±9.16% spontaneous beating EBs at 12 days after the EB formation. To develop a more efficient cardiomyocyte differentiation method, we tested several known cardiogenic reagents (ascorbic acid, 5’-Azacytidine, LiCl, oxytocin, FGF2 and PDGF-BB) after noggin induction or we cultured noggin treated ES cells on various extracellular matrixes (collagen, fibronectin and Matrigel). Quantitative RT-PCR and immunocytochemistry results showed a significantly increased cardiac differentiation rate in the FGF2 treated group. Differentiation on the collagen extracellular matrix (ECM) could slightly increase the cardiac differentiation efficiency. These results show the possibilities for the establishment of selective differentiation conditions for the cardiac differentiation of mouse ES cells.

Optimum injection molding condition for a box mold was searched by the Response Surface Analysis(RSA) with the aid of process monitoring system(PMS). Process variables on the control panel of the injection molding machine such as barrel temperatures, screw speed profile, holding pressures, etc. cannot guarantee the uniformity of the material variables directly related with the state of the product in the mold cavity. In order to make sure the state of the resin in the cavity, pressures and temperatures in the cavity, runner and nozzle were monitored in the experiment with the PMS. To accomplish the consistency of the molding process, dependent variables such as the switchover point and holding time were searched with the PMS. With a proper objective function about deflection of the box-type product, the optimum injection molding condition was obtained.

During the injection molding process an excessive packing can occur in the smaller volume cavity because of volumetric difference of the family-mold. It causes warpage by increased residual stress in the product and flesh by over packing. In this study, we used a variable-runner system for the filling balance of the cavities by changing the cross-sectional area of a runner, and confirmed the filling imbalance by temperature and pressure sensors. We carried out experiments to examine the influence of types of resins such as LDPE/ABS/PA6,6 on the filling balancing of the system, in order to help mold designers, who can easily adopt the variable-runner system to their design. We also examined filling imbalance in the system with CAE analysis.

Dental impression materials are used to register or reproduce the form and relationship of the teeth and oral tissues. They should not be torn when removing from the wet mouth after setting. Nowadays, silica is widely used as filler to overcome the low mechanical strength of the dental impression materials. The purpose of this study was to synthesize high strength glass in the system of MgO-CaO-Al2O3-SiO2 and investigate its usefulness according to ISO standard after mixing with addition silicone. Commercial products, Contrast, Examix, Express, and Perfect-F were selected as control group. When the prepared glass filler was introduced in addition silicone impression material, tear strength was drastically increased significantly than that of the control group, keeping the consistency. All experimental groups showed higher tear stength than that of control groups. As the amount of filler content increases, tear strength was increased. Strain in compression, recovery from deformation, and linear dimensional change were satisfied the ISO standard either all the experimental or control groups. Therefore, calcium-substituted magnesium aluminosilicate glass in the system of 12.5MgO-17.5CaO-20Al2O3-50SiO2 is expected the useful filler in the light body of addition silicone impression materials.

Seven ferromanganese crusts from the northwest intertropical Pacific seamounts were analyzed for photomicroscopic growth structures, microprobe chemistry, and ages based on Co-chronometer growth rate. The crusts on the Marshall Islands seamounts are thick and are divided into phosphatized lower older and nonphosphatized upper younger growth generations: the older crust consists of compact laminations and columns impregnated with carbonate fluoapatite (CFA), whereas the younger crust is characterized by porous botryoids and columns of δ-MnO2 and Fe oxyhydroxide. The crusts on the Federated States of Micronesia (FSM) and Palau Islands seamounts are thin and are often incorporated with inorganic opal-A in the uppermost part, comprising the younger generation. Some crusts show scours and fractures. Although the growth of crusts has been often interrupted by mass failure of slope sediments, the crusts on the Marshall Islands seamounts are estimated to have grown at rate of about 3 mm/Ma since the middle Eocene and to have been phosphatized in the late Oligocene during the host seamounts were located beneath the equatorial zone of high productivity. Prolonged infiltration of the oxygen minimum zone (OMZ) water into shallower water older crusts redistributed crust composition by precipitating CFA, enriching subsequent amounts of Mn and Ni, and removing some Co. The younger crust has formed at slower rate (about 2 mm/Ma) under the stronger influence of bottom-water circulation in the north of the equatorial zone, concentrating abundant Co. In the uppermost part of some crusts, siliceous skeletons transform with burial to inorganic opal-A and Si-rich Fe oxyhydroxide, suggesting that biosilica diagenesis can enhance crust growth.

In the southern part of the Korea Deep Ocean Study (KODOS) area, the western part of the Clarion-Clipperton fracture zones, abundant nodules occur in the Oligocene to Miocene radiolarian-bearing smectitic clay. The nodules are generally concentrated in patchy pavements (more than 5 kg/m2) on the hilly seafloor with sediment waves. They are either rare or abundant on the seamounts and are generally rare in the abyssal plain. The nodules can be classified into three types based on distinct surface texture. The S-type nodules occur in intensely bioturbated sediment of the abyssal hill and seamount areas. In the abyssal plain, the R-type nodules are dominant, embedded within the topmost Quaternary sediment layer. In the southeastern part where Pliocene sediment occurs, they are in an incipient stage of growth. The S-type nodules are rich in Fe and Co in columnar structures of δ-MnO2, whereas the R-types contain abundant Mn, Ni, Cu, and Zn in dendritic and massive 10A-manganate layers. The T-type nodules occur together with either the R- or S-type, showing intermediate composition. Regardless of the type and shape, the bottoms of nodules contain more abundant Mn, Ni, Cu, Zn, and 10A-manganate than the tops. As the Mn Fe ratio increases in the nodules, the abundance of Ni, Cu, and Zn increases up to a constant value (about 11% of Mn) southward. In the underlying sediment, however, it decreases northward. Manganese and these minor metals are concentrated in the lowermost Tertiary sediment as much as 2 to 3 times than those in the topmost sediment layer which is intensely bioturbated. Iron and Co are relatively uniform downcore. Manganese, Ni, and Cu are abundant in micronodules, whereas Fe is concentrated in smectite. Redistributional processes of bottom currents seem to be most responsible for the (re)generation and concentration of nodules in patchy pavements, by excavating buried nodules. These processes result in an inverse relation of nodule abundance to the thickness of uppermost sedimentary layer which comprises the radiolarian ooze of Clipperton Formation. In the seamount areas, sediment slumping has also contributed to the concentration of nodules. The fragmentation of old nodules also enhances the nodule (re)generation. Activities of benthic animals cause the nodules to be lifted upward onto the seafloor mostly during the quiescent periods of bottom currents. The growth of nodules by resedimentation processes results in the large discrepancy in composition not only between the types but also the tops and bottoms, controlled by the burial rates of nodules within the topmost sediment layer where the diagenesis takes place. Diagenetic growth of nodules occurs by utilizing Mn and minor metals remobilized from micronodules during biomixing and decomposition of organic materials within Tertiary sediment. Nodule growth is more active southward toward the equator. Latitudinal variations of nodule size, distribution, abundance, and composition result from the change in surface water productivity and the timing of nodule nucleation as the Pacific plate migrates northwestward.

We present, in the form of an overview, some of the results of our study on the miniaturization and integration of micro/nano-photonic devices for optical printed circuit board (O-PCB) and VLSI photonic applications. The micro- and nano-photonic devices are designed to perform the functions of transporting, switching, wavelength filtering, and routing optical signals on flat modular boards or substrates. We have designed micron-scale devices and nanoscale photonic crystal devices of diverse functions for VLSI applications and have fabricated some of them using embossing or imprinting techniques. We also designed and fabricated novel subwavelength nano-scale plasmonic devices. We have constructed and assembled O-PCBs using polymer-based waveguide arrays and circuits and have examined their information handling performances. We have used near-field scanning optical microscopic (NSOM) technique to characterize the fabricated micro/nano-scale photonic materials and devices

We report on the design and fabrication of what we call "optical printed circuit board (O-PCB)" by way of interconnecting and integrating micro/nano-scale optical waveguide circuits and miniaturized photonic devices. Planar optical waveguide circuits were fabricated out of polymer materials using thermal and UV embossing techniques. Micro/nano-scale photonic devices include arrayed waveguide gratings, directional couplers, beam splitters, filters, multimode interference devices, micro-ring resonator devices, photonic crystal devices and plasmonic devices. The O-PCBs were assembled by integrating these devices for the functions of transporting, switching, routing and distributing optical signals on flat modular boards. Potential use of O-PCBs for telecommunications, computers, transportation systems, and VLSI photonic circuits are discussed.

We present a low-cost C/S-band WDM-PON employing colorless uncooled bidirectional transceivers and SOA-based broadband light sources. Colorless operations over 32 DWDM channels are demonstrated from -20 to 80/spl deg/C in 155-Mb/s bidirectional transmissions over 25 km.

A current regulator for the resonant inverter, which chooses the nearest available voltage vector to the predicted desired voltage vector is proposed. It is proved that the proposed regulator minimizes RMS current error. The performance of the regulator is tested by computer simulations and compared with that of a conventional pulse density modulation (PDM) regulator. The effects of the parameter and estimation error in the back EMF are considered in the computer simulation. The implementation of the regulator using a digital signal processor and its experimental results are described. The computer simulation and experimental results reveal improved responses compared with a conventional PDM regulator. >

This paper reports a method for measuring magnetostriction, Young's modulus of a substrate or film, and the /spl Delta/E effect with one apparatus. A substrate with a thin magnetic film deposited on it is in parallel with and cantilevered above a metal plate electrode, and they form a capacitive cell. The cantilever deflects due to its own weight and applied electric and magnetic fields. The small change of capacitance caused by this deflection is measured by a sensitive capacitance bridge. Young's modulus, magnetostriction, and /spl Delta/E effect can be calculated by a theoretical analysis with the measured deflection data. >

Some of the important characteristics required for field-oriented AC drives are a fast run-stop capability and good speed regulation regardless of sudden load change. The properties of a PDF (pseudo derivative feedback) controller are analyzed, and some key features are described compared with the PI (proportional plus integral) controller. In order to get robust speed control performance against variation of load inertia, a load disturbance observer is also applied to the PDF controller, and experimental results are presented. Nominal load inertia is measured by using a digital-signal-processor-based inverter self-tuning function, and parameters needed for the controller can be calculated to get good speed control performance. >

A global proteomics approach by 2-D gel electrophoresis and mass spectrometry was used to analyze the differential protein patterns of three placentae obtained after postnatal death of fetuses from SCNT of Korean Native cattle and three normal placentae obtained after birth of AI fetuses, as previously reported (Kim et al. 2004 Reprod. Fertil. Develop. 16, 145). One differentially up-regulated proteins in SCNT placenta was identified as TIMP-2 protein that is related to extracellular matrix degradation and tissue remodeling processes. To ensure that the identified protein was truly up-regulated in SCNT placenta, a small portion of the protein lysates were subjected to Western blotting with the antibodies against TIMP-2 and β-actin protein. Band images were scanned with a GT-6000 Scanner (Epson-Seiko, Suwa, Nagano, Japan) and densitometric analyses were performed using NIH Image (version 1.56). Proteins were normalized against the levels of β-actin protein. Indeed, Western blot analysis revealed a significant increase in TIMP-2 protein level (about 5-fold) in SCNT placenta compared with normal. Epigenetic programming of the genome by DNA methylation and histone modification ensures proper gene expression during development. To examine how the TIMP-2 gene was programmed to be active in SCNT placenta, we first investigated the status of DNA methylation in the promoter region of the TIMP-2 gene. Sodium bisulfite mapping revealed that most CpG sites in SCNT placenta cells were unmethylated in agreement with expression of TIMP-2 compared with those in normal placenta cells, suggesting that TIMP-2 gene programming by hypomethylation of CpG sites in the promoter region was responsible for the expression in SCNT placenta. We then examined histone modification of the nucleosome in the coding region of the TIMP-2 gene of SCNT placenta cells and normal placenta cells. Chromatin immunoprecipitation (ChIP) assay revealed that H3-K9/K14 acetylation of the TIMP-2 locus in SCNT placenta cells was about 2-fold higher than in normal placenta cells. Therefore, it is postulated that the SCNT placenta at the end of gestation seems to be unusual in tissue remodeling via improper expression of TIMP-2 by abnormal epigenetic modification, and that may affect placental abnormality such as enlarged and dysfunctional placentae as reported in cloned bovine and mouse.

Antibiotics are commonly added to mammalian oocyte maturation media, but their effects on oocytes maturation have not been examined thoroughly. Goat follicular oocytes were used to investigate whether penicillin, streptomycin or gentamycin affect maturational competence of oocytes and subsequent parthenogenetic activation potential in vitro. Cumulus-oocyte complexes collected from a local abattoir were matured for 24h in five treatments, and matured oocytes were cultured for 48h in five treatments after parthenogenetic activation by treatment with ionomycin, followed by immediate exposure to 6-diethlaminopurine; (1) Control: TCM-199 medium with no antibiotics, (2) TCM-199 with 100IU/mL−1 penicillin (P-4687, Sigma, St. Louis, MO, USA), (3) TCM-199 with 50μgmL−1 streptomycin (S-1277, Sigma), (4) TCM-199 with 50μgmL−1 gentamycin (G-1264, Sigma) and (5) TCM-199 with both 100IUmL−1 penicillin and 50μgmL−1 streptomycin. Maturation rates at 24h post-in vitro maturation and parthenogenetic cleavage development at 48h post-activation were evaluated. Data were analyzed by ANOVA and Student’s t-test. Penicillin and gentamicin treatment groups did not affect maturation rates and percentages of cleavage to 2–4 cell stage at 48h post-chemical oocyte activation. However, when streptomycin was present in the maturation medium, the percentages of matured oocytes at 24h post-in vitro maturation of immature goat oocytes were significantly lower than those from the other groups. However, among the five treatments, there was no significant difference in cleavage rates of matured oocytes at 48h post-activation (Table 1). Therefore, streptomycin did interfere with the maturation of immature goat oocytes, but did not affect the subsequent development of matured goat oocytes. The mechanism by which streptomycin affects the maturation of goat follicular oocytes needs to be investigated further. We conclude that streptomycin in oocyte maturation medium can be detrimental during in vitro maturation of goat follicular oocytes. Table 1 Effect of antibiotics on maturational competence of goat follicular oocytes and subsequent parthenogenetic activation potential in vitro

Practical application of animal cloning by somatic cell nuclear transfer (SCNT) has been hampered by extremely low success rate. Most clones die before birth and survivors frequently display abnormalities. It is speculated that epigenetic reprogramming is somehow defective in reconstituted embryos (Reik W et al., 2003 Theriogenology 59 21–32; Han YM et al., 2003 Theriogenology 59, 33–44). It is likely that placental anomalies are directly or indirectly responsible for the death of cloned fetus and neonates. To address this question, we analyzed protein patterns of two placentae obtained after postnatal death of fetuses from SCNT of Korean Native Cattle and two normal placentae obtained after birth of AI fetuses. Global proteomics approach was employed by using 2-D gel electrophoresis and mass spectrometry to separate the different placenta proteins. Proteins within an isoelectric point range of 4.0 to 7.0 and a molecular weight range of 20–100kDa were analyzed by means of 2-D gel electrophoresis with three replications of each sample. The stained gels were scanned and calibrated at an optical resolution of 63.5μm/pixel using a GS-710 (Bio-Rad Laboratories, Hercules, CA, USA). Approximately 480 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis by Malanie III (Swiss Institute for Bioinformatics, Geneva, Switzerland) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, at least 15 protein spots were identified as regulated differentially. Using MALDI-TOF-MS (PerSeptive Biosystems, Framinham, MA, USA), 10 spots were identified as up-regulated proteins in SCNT placentae including BPLP-I, Rho GDI 2, osteoclast stimulating factors, SM22, 60S Acidic Ribosomal and Protein P2, whereas five spots were down-regulated proteins such as Peroxiredoxin 2. Mass spectrometry with sequencing was used to further analyze the uncharacterized proteins. Most identified proteins in this analysis appeared to be related to cell proliferation and differentiation, fetal growth and development or metabolism. Further, specific functions of proteins in placenta have been investigated at the molecular levels during pregnancy.