ALBERT

Beschreibung: New, commensurate members of the fluorite-related Bi 3 Nb 1− x Ta x O 7 family were synthesized and their crystal structures, microstructures, and microwave (MW) dielectric properties were characterized. The incorporation of Ta into the tetragonal Bi 3 Nb 1− x Ta x O 7 solid solution was found to gradually affect the density and the MW dielectric properties. The materials sintered at 870°C exhibited relative permittivities in the range k ′ = 86–72, Q  ×  f values from 793 to 1189 GHz and a positive temperature coefficient of resonant frequency from 88 to 12 ppm/K. The formation of the members of the fluorite-related solid solution along the Bi 3 Nb 1− x Ta x O 7 composition depends on a phase transition, and thus their properties are compared within the compositional range. The correlations between their MW dielectric properties, compositions, crystal structures, and processing parameters were discussed in detail. Optimization of MW properties can be achieved by utilizing the ability of the Bi 3 Nb 1− x Ta x O 7 solid solution that it undergoes a phase transformation from cubic to tetragonal structure which are both characterized by unique properties, under certain synthesis conditions.

Beschreibung: Transparent novel glass-ceramics containing Sr 2 YbF 7 :Er 3+ nanocrystals were successfully fabricated by melt-quenching technique. Their structural and up-conversion luminescent properties were systemically investigated by XRD, HRTEM, and a series of spectroscopy methods. The temperature-dependent up-conversion spectra prove that 2 H 11/2 and 4 S 3/2 levels of Er 3+ are thermally coupled energy levels (TCEL). Consequently, the 2 H 11/2 4 I 15/2 and 4 S 3/2 4 I 15/2 emissions of Er 3+ in Sr 2 YbF 7 :Er 3+ glass-ceramics can be used as optical thermometry based on fluorescence intensity ratio (FIR) technique. Combined with low phonon energy and high thermal stability, Er 3+ ions in Sr 2 YbF 7 glass-ceramics present broad operating temperature range (300–500 K), large energy gap of TCEL (786 cm −1 ) and high theoretical maximum value of relative sensitivity (62.14 × 10 −4  K −1 at 560 K), which suggests that Sr 2 YbF 7 :Er 3+ glass-ceramics may be excellent candidates for optical temperature sensors.

Beschreibung: World Health Organization reports that methicillin-resistant Staphylococcus aureus (MRSA) is the origin of higher proportion of hospital acquired infections. In order to combat the effect of MRSA infection, an ideal drug should stimulate the allosteric exposure of active site, prompting penicillin binding proteins (PBP2a) to bind with that particular compound. Ceftaroline shows high binding affinity towards PBP2a and also confers resistance against degrading enzymes. Recently, two amino acid alterations in the allosteric site of PBP2a, asparagine (N) to lysine (K) at position 146 and glutamic acid (E) to lysine at position 150 are reported to confer resistance against ceftaroline resulting in the rise of ceftaroline-resistant MRSA strains. The present study focuses on the identification of potential ligands that can effectively bind with allosteric site of PBP2a, that leads to the access of active site and entry of a β-lactam antibiotic for effective inhibition. The results obtained from our study will be useful for designing effective compounds with potential therapeutic effects against ceftaroline resistant MRSA strains. This article is protected by copyright. All rights reserved

Beschreibung: The human protein kinase X gene (PRKX) and cAMP-dependent protein kinase (PKA) are both c-AMP-dependent serine/threonine protein kinases within the protein kinase AGC subgroup. Of all the protein kinases in this group, PRKX is the least studied. PRKX has been isolated from patients with chondrodysplasia punctate and is involved in numerous processes, including sexual differentiation and fertilization, normal kidney development, and autosomal dominant polycystic kidney disease (ADPKD), blood maturation, neural development and angiogenesis in vitro. Although the role of PRKX in development and disease has been reported recently, the underlying mechanism of PRKX activity is largely unknown. In addition, based on the expression pattern of PRKX and the extensive role of PKA in disease and development, PRKX might have additional crucial functions that have not been addressed in the literature. In this review, we summarize the characteristics and developmental functions of PRKX that have been reported by recent studies. In particular, we elucidate the structural and functional differences between PRKX and PKA, as well as the possible roles of PRKX in development and related diseases. Finally, we propose future studies that could lead to important discoveries of more PRKX functions and the underlying mechanisms involved. This article is protected by copyright. All rights reserved

Beschreibung: The endocannabinoid system is the target of the main psychoactive component of the plant Cannabis sativa , the Δ 9 -tetrahydrocannabinol (THC). This system is composed by the cannabinoid receptors, the endogenous ligands, and the enzymes involved in their metabolic processes, which works both centrally and peripherally to regulate a plethora of physiological functions. This review aims at explaining how the site-specific actions of the endocannabinoid system impact on memory and feeding behavior through the cannabinoid receptors 1 (CB 1 R). Centrally, CB 1 R is widely distributed in many brain regions, different cell types (e.g. neuronal or glial cells) and intracellular compartments (e.g. mitochondria). Interestingly, cellular and molecular effects are differentially mediated by CB 1 R according to their cell-type localization (e.g. glutamatergic or GABAergic neurons). Thus, understanding the cellular and subcellular function of CB 1 R will provide new insights and aid the design of new compounds in cannabinoid-based medicine. The widespread localization of CB1 receptors in different brain regions (e.g. hippocampus, hypothalamus, and cortex), cell types (e.g. GABAergic and glutamatergic neurons), and subcellular domains (e.g. plasma membrane and mitochondria) allows the endocannabinoid system to control different behaviors (e.g. memory and food intake) in a multimodal and versatile fashion. Also watch the Video Abstract .

Beschreibung: Several key transcription factors regulate cell growth, survival, and differentiation during neural crest and melanoblast development in the embryo, and these same pathways may be reactivated in tumors arising from the progenitors of these cells. The transcription factors PAX3 and FOXD3 have essential roles in melanoblasts and melanoma. In this study, we define a regulatory pathway where FOXD3 promotes the expression of PAX3. Both factors are expressed in melanoma cells and there is a positive correlation between the transcript levels of PAX3 and FOXD3. The PAX3 gene contains two FOX binding motifs within highly conserved enhancer regulatory elements that are essential for neural crest development. FOXD3 binds to both of these motifs in vitro but only one of these sites is preferentially utilized in melanoma cells. Overexpression of FOXD3 upregulates PAX3 levels while inhibition of FOXD3 function does not alter PAX3 protein levels, supporting that FOXD3 is sufficient but not necessary to drive PAX3 expression in melanoma cells. Here, we identify a molecular pathway where FOXD3 upregulates PAX3 expression and therefore contributes to melanoma progression. This article is protected by copyright. All rights reserved

Beschreibung: Metabolic networks are significantly altered in neoplastic cells. This altered metabolic program leads to increased glycolysis and lipogenesis and decreased dependence on oxidative phosphorylation and oxygen consumption. Despite their limited mitochondrial respiration, cancer cells, nonetheless, derive sufficient energy from alternative carbon sources and metabolic pathways to maintain cell proliferation. They do so, in part, by utilizing fatty acids, amino acids, ketone bodies and acetate, in addition to glucose. The alternative pathways used in the metabolism of these carbon sources provide opportunities for therapeutic manipulation. Acetate, in particular, has garnered increased attention in the context of cancer as both an epigenetic regulator of posttranslational protein modification, and as a carbon source for cancer cell biomass accumulation. However, to date, the data have not provided a clear understanding of the precise roles that protein acetylation and acetate oxidation play in carcinogenesis, cancer progression or treatment. This review highlights some of the major issues, discrepancies and opportunities associated with the manipulation of acetate metabolism and acetylation-based signaling in cancer development and treatment. This article is protected by copyright. All rights reserved

Beschreibung: Direct application of histone-deacetylase-inhibitors (HDACis) to dental pulp cells (DPCs) induces chromatin changes, promoting gene expression and cellular-reparative events. We have previously demonstrated that HDACis (Valproic acid, Trichostatin A) increase mineralization in dental papillae-derived cell-lines and primary DPCs by stimulation of dentinogenic gene expression. Here, we investigated novel genes regulated by the HDACi, suberoylanilide hydroxamic acid (SAHA), to identify new pathways contributing to DPC differentiation. SAHA significantly compromised DPC viability only at relatively high concentrations (5 μM); while low concentrations (1 μM) SAHA did not increase apoptosis. HDACi-exposure for 24 h induced mineralization-per-cell dose-dependently after 2 weeks; however, constant 14d SAHA-exposure inhibited mineralization. Microarray analysis (24 h and 14d) of SAHA exposed cultures highlighted that 764 transcripts showed a significant 〉2.0-fold change at 24 h, which reduced to 36 genes at 14d. 59% of genes were down-regulated at 24 h and 36% at 14d, respectively. Pathway analysis indicated SAHA increased expression of members of the matrix metalloproteinase (MMP) family. Furthermore, SAHA-supplementation increased MMP-13 protein expression (7d, 14 d) and enzyme activity (48 h, 14d). Selective MMP-13-inhibition (MMP-13i) dose-dependently accelerated mineralization in both SAHA-treated and non-treated cultures. MMP-13i-supplementation promoted expression of several mineralization-associated markers, however, HDACi-induced cell migration and wound healing were impaired. Data demonstrate that short-term low-dose SAHA-exposure promotes mineralization in DPCs by modulating gene pathways and tissue proteases. MMP-13i further increased mineralization-associated events, but decreased HDACi cell migration indicating a specific role for MMP-13 in pulpal repair processes. Pharmacological inhibition of HDAC and MMP may provide novel insights into pulpal repair processes with significant translational benefit. This article is protected by copyright. All rights reserved

Beschreibung: Adipogenesis comprises a complex network of signaling pathways and transcriptional cascades; the GSK3β-C/EBPβ- srebf1a axis is a critical signaling pathway at early stages leading to the expression of PPARγ2, the master regulator of adipose differentiation. Previous work has demonstrated that retinoic acid inhibits adipogenesis affecting different signaling pathways. Here, we evaluated the anti-adipogenic effect of retinoic acid on the adipogenic transcriptional cascade, and the expression of adipogenic genes cebpb , srebf1a , srebf1c , pparg2 , and cebpa . Our results demonstrate that retinoic acid blocks adipose differentiation during commitment, returning cells to an apparent non-committed state, since they have to be newly induced to adipose conversion after the retinoid is removed from the culture medium. Retinoic acid down regulates the expression of the adipogenic genes, srebf1a, srebf1c , pparg2 , and cebpa . Retinoic acid did not down regulate the expression of cebpb , but it inhibited C/EBPβ phosphorylation at Thr188, a critical step for the progression of the adipogenic program. We also found that RA inhibition of adipogenesis did not increase the expression of dlk1 , the gene encoding for Pref1, a well-known anti-adipogenic transcription factor. This article is protected by copyright. All rights reserved

Beschreibung: ABSTRACT Platelets are important in hemostasis, but also detect particles and pathogens in the circulation. Phagocytic and endocytic activities of platelets are widely recognized, however, receptors and mechanisms involved remain poorly understood. We previously demonstrated that platelets internalize and store phospholipid microvesicles enriched in human tissue factor (TF + MVs) and that platelet-associated TF enhances thrombus formation at sites of vascular damage. Here we investigate the mechanisms implied in the interactions of TF + MVs with platelets and the effects of specific inhibitory strategies. Aggregometry and electron microscopy were used to assess platelet activation and TF + MVs uptake. Cytoskeletal assembly and activation of phosphoinositide 3-kinase (PI3K) and RhoA were analyzed by western blot and ELISA. Exposure of platelets to TF + MVs caused reversible platelet aggregation, actin polymerization and association of contractile proteins to the cytoskeleton being maximal at 1 min. The same kinetics were observed for activation of PI3K and translocation of RhoA to the cytoskeleton. Inhibitory strategies to block glycoprotein IIb-IIIa (GPIIb-IIIa), scavenger receptor CD36, serotonin transporter (SERT) and PI3K, fully prevented platelet aggregation by TF + MVs. Ultrastructural techniques revealed that uptake of TF + MVs was efficiently prevented by anti-CD36 and SERT inhibitor, but only moderately interfered by GPIIb-IIIa blockade. We conclude that internalization of TF + MVs by platelets occurs independently of receptors related to their main hemostatic function (GPIIb-IIIa), involves the scavenger receptor CD36, SERT and engages PI3-Kinase activation and cytoskeletal assembly. CD36 and SERT appear as potential therapeutic targets to interfere with the association of TF + MVs with platelets and possibly downregulate their prothrombotic phenotype. This article is protected by copyright. All rights reserved

Beschreibung: Deficiency in the retinoblastoma protein (Rb) favors leanness and a healthy metabolic profile in mice largely attributed to activation of oxidative metabolism in white and brown adipose tissues. Less is known about Rb modulation of skeletal muscle metabolism. This was studied here by transiently knocking down Rb expression in differentiated C2C12 myotubes using small interfering RNAs. Compared with control cells transfected with non-targeting RNAs, myotubes silenced for Rb (by 80–90%) had increased expression of genes related to fatty acid uptake and oxidation such as Cd36 and Cpt1b (by 61% and 42%, respectively), increased Mitofusin 2 protein content (∼2.5-fold increase), increased mitochondrial to nuclear DNA ratio (by 48%), increased oxygen consumption (by 65%) and decreased intracellular lipid accumulation. Rb silenced myotubes also displayed up-regulated levels of glucose transporter type 4 expression (∼5-fold increase), increased basal glucose uptake, and enhanced insulin-induced Akt phosphorylation. Interestingly, exercise in mice led to increased Rb phosphorylation (inactivation) in skeletal muscle as evidenced by immunohistochemistry analysis. In conclusion, the silencing of Rb enhances mitochondrial oxidative metabolism and fatty acid and glucose disposal in skeletal myotubes, and changes in Rb status may contribute to muscle physiological adaptation to exercise. This article is protected by copyright. All rights reserved

Beschreibung: Human pancreatic and prostate cancers metastasize along nerve axons during perineural invasion. The extracellular matrix laminin class of proteins is an abundant component of both myelinated and non-myelinated nerves. Analysis of human pancreatic and prostate tissue revealed both perineural and endoneural invasion with Schwann cells surrounded or disrupted by tumor, respectively. Tumor and nerve cell co-culture conditions were used to determine if myelinating or non-myelinating Schwann cell (S16 and S16Y, respectively) phenotype was equally likely to promote integrin-dependent cancer cell invasion and migration on laminin. Conditioned medium from S16 cells increased tumor cell (DU145, PC3, and CFPAC1) invasion into laminin approximately 1.3 to 2.0 fold compared to fetal bovine serum (FBS) treated cells. Integrin function (e.g., ITGA6p formation) increased up to 1.5 fold in prostate (DU145, PC3, RWPE-1) and pancreatic (CFPAC1) cells, and invasion was dependent on ITGA6p formation and ITGB1 as determined by function-blocking antibodies. In contrast, conditioned medium isolated from S16Y cells (non-myelinating phenotype) decreased constitutive levels of ITGA6p in the tumor cells by 50% compared to untreated cells and decreased ITGA6p formation 3.0 fold compared to S16 treated cells. Flow cytometry and western blot analysis revealed loss of ITGA6p formation as reversible and independent of overall loss of ITGA6 expression. These results suggest that the myelinating phenotype of Schwann cells within the tumor microenvironment increased integrin-dependent tumor invasion on laminin. This article is protected by copyright. All rights reserved

Beschreibung: Accurate information on the interactions between water and silica is critical to the understanding of its properties including mechanical strength under stress and long-term chemical durability of silica and silicate glasses. In this study, interactions between water and nanoporous amorphous silica models were investigated using density functional theory (DFT) based ab initio molecular dynamics (AIMD) simulations which accurately describe bond breakage and formation as well as chemical reactions. AIMD simulations up to 30 ps were performed for systems containing water and nanoporous silica with a wide range of porosities (31%–67%). Partial removal of defects, such as two-membered rings, was observed during the AIMD runs whereas more reactive coordination defects were removed during the initial geometry optimization. The limited two-membered ring removal can be attributed to restricted water-defect movement or the increased stability of rings located on concave surfaces. Two-membered ring removal mechanisms included the formation of an overcoordinated silicon (Si 5 ) intermediate defect from the dynamic simulations. Si 5 defects continued to develop throughout the simulations, indicating a thermodynamic drive for two-membered ring removal which is kinetically limited. Changes in the electronic structures, such as atomic charges, and bond length-bond angle correlation functions were monitored during the hydroxylation process.

Beschreibung: Amorphous calcium polyphosphate (ACPP), an inorganic polymer ceramic, has shown promise as a drug delivery matrix following a repeat gelling protocol. This study described a simple method of preparing ACPP hydrogel in the presence of an excess volume of water. The increased water availability accelerates water molecule ingress and microstructural transformation of ACPP hydrogels. The impact of some experimental settings (soaking time, temperature, stirring, and ACPP particle size) on the physiochemical and rheological natures of ACPP hydrogel were investigated and from which possible hydrogel formation mechanisms were inferred. We believe that the formation of ACPP hydrogel is through the mechanisms of intermolecular ionic interaction and entanglement of polyphosphate chains. The potential application of ACPP hydrogel as a ceramic matrix for sustained drug release warrants further investigation.

Beschreibung: Melting gels are hybrid gels that have the ability to soften and flow at around 100°C for some combinations of mono- and di-substituted alkoxysiloxanes, where substitutions are either all aromatic or all aliphatic. In this study, melting gels were prepared using phenyltriethoxysilane (PhTES) and dimethyldiethoxysilane (DMDES), meaning both an aromatic and aliphatic substitution. Differential scanning calorimetry was performed to identify glass-transition temperatures, and thermal gravimetric analysis coupled with differential thermal analysis (TGA-DTA) was performed to measure weight loss. The glass-transition temperatures ( T g ) ranged from −61°C to +5.6°C, which are between the values in the methyl only system, where all T g values are less than 0°C, and those values in the phenyl only system, where T g values are greater than 0°C. The T g decreased with an increase in the DMDES fraction. Below 450°C, the gels lost little weight, but around 600°C there was a drop in weight. This temperature is lower than the temperature for gels prepared with only aromatic substitutions, but higher than that for gels prepared with only aliphatic substitutions. Final heat treatment was carried out at 150°C for the gel with 80%PhTES-20%DMDES (in mol%), and the consolidation temperature increased with increasing DMDES content to 205°C for the gel with 50%PhTES-50%DMDES. After this heat treatment, the melting gels no longer soften.

Beschreibung: Direct integration of nanostructures into macroscopic substrates is very important for their practical applications. In this work, we report a simple method that can be introduced for the Sn-catalyzed growth of alumina nanowires on ceramic substrates such as porous disk, monolith, and foam. Our study focuses on the role of the Sn catalysts in the formation mechanisms governing nanowire growth. Using the proposed approach, hair- or grass-like tufts of 20 nm diameter nanowires grow on the surface of the ~3 μm diameter Sn particles, in a tip growth mechanism. The nanowires of α-phased polycrystalline structure grow and are packed via a complex process involving batch-by-batch, branching, and amalgamation growth. The detailed observations reveal that the Sn catalyst is key to tailoring the growth patterns of the nanowires. In addition, cathodoluminescence studies highlight the potential optical applications of the alumina nanowires.

Beschreibung: Flash sintering is a nonlinear phenomenon characterized by a sharp increase in the conductivity of the sample and concomitant rapid densification under an electric field in low temperatures in a matter of seconds. Since it is a transient phenomenon, the power dissipation on the sample is not uniform during the process. Thus, a transient analysis is needed to estimate the temperature of the sample during flash sintering due to Joule heating. In this work, the Finite Element Method on a coupled electrothermal nonlinear analysis was used in order to obtain the specimen temperature of 8YSZ after 5 s of flashing. The results agree with the experimental data obtained by the flashing of dense samples and with previous literature.

Beschreibung: Temperature-dependent in-situ Raman spectroscopy is used to investigate the phase transformation of zinc metastannate (ZnSnO 3 ) to zinc orthostannate (Zn 2 SnO 4 ) induced upon annealing in the ambient. ZnSnO 3 microcubes (MCs) were synthesized at room temperature using a simple aqueous synthesis process, followed by characterization using electron microscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and thermogravimetric analysis (TGA). Annealing of the ZnSnO 3 MCs was carried out up to 1000°C, while recording the Raman spectra in-situ at regular intervals. Phase transformation from metastannate to orthostannate was found to begin around 500°C with an activation energy of ~0.965 eV followed by the recrystallization into the inverse spinel orthostannate phase at ~750°C. Results from this study provide detailed understanding of the phase transformation behavior of perovskite ZnSnO 3 to inverse spinel Zn 2 SnO 4 upon thermal annealing.

Beschreibung: Here, we demonstrate the relationship between glass network topological structure and the chemical state of embedded lanthanide ions. It is revealed that a more dispersed state of lanthanide ions is shown in more constrained 3D rigid network, which delivers valuable information toward homogeneous doping in glasses from the perspective of glass topological structure. The results are believed to be of great significances in the development of advanced optoelectronic devices like high-power laser, efficient fiber amplifier, smaller integrated photonic circuit, etc.

Beschreibung: The mechanofusion process, a dry particle coating route, has been successfully applied to coat micrometric SiC particles with submicrometric Ni filaments. In a first step, the mechanofusion parameters were optimized to form a continuous Ni coating onto SiC particles. In a second step, the Ni-coated SiC particles were sintered by hot isostatic pressing. The temperature and pressure cycles were determined to ensure a good densification of the material. Such a densification process leads to the formation of a δ-Ni 2 Si bilayer at the SiC/Ni interface; the inner δ-Ni 2 Si layer in contact with SiC being more rich in carbon than the one in contact with the matrix. From X-ray diffraction, wavelength-dispersive X-ray spectrometry and scanning electron microscopy characterizations, a mechanism is proposed to explain the microstructure of the end-product.

Beschreibung: MDA-9/Syntenin is a small PDZ domain containing scaffolding protein with diverse array of function regulating membrane trafficking, cell adhesion, neural and synaptic development, ubiquitination and exosome biogenesis. An appreciable number of studies also established a pivotal role of MDA-9/Syntenin in cancer development and progression. In this review, we will discuss the dynamic role of MDA-9/Syntenin in regulating normal and abnormal fate of various cellular processes. This article is protected by copyright. All rights reserved

Beschreibung: Because early-stage hepatocellular carcinoma (HCC) is difficult to diagnose using the existing techniques, identifying better biomarkers would likely improve the patients' prognoses. We performed a systematic review and meta-analysis of published studies to appraise the utility of microRNAs (miRNAs) for the early diagnosis of HCC. Pertinent literature was collected from the Medline, Embase, and Chinese National Knowledge Infrastructure databases. We analyzed 50 studies that included 3423 cases of HCC, 2403 chronic hepatic disease (CH) patients, and 1887 healthy controls in 16 articles. Summary receiver operating characteristic analyses of all miRNAs showed an area under the curve (AUC) of 0.82, with 75.8% sensitivity and 75.0% specificity in discriminating patients with HCC from healthy controls. miR-21 and miR-122 individually distinguished patients with HCC from healthy controls, with an AUC of 0.88 for miR-21 and 0.77 for miR-122. The sensitivity and specificity for miR-21 were 86.6% and 79.5%, respectively; those for miR-122 were 68.0% and 73.3%. We conclude that circulating miRNAs, particularly miR-21 and miR-122, are promising biomarkers for the early diagnosis of HCC. This article is protected by copyright. All rights reserved

Beschreibung: Here, we review genomic target site selection during retroviral integration as a multistep process in which specific biases are introduced at each level. The first asymmetries are introduced when the virus takes a specific route into the nucleus. Next, by co-opting distinct host cofactors, the integration machinery is guided to particular chromatin contexts. As the viral integrase captures a local target nucleosome, specific contacts introduce fine-grained biases in the integration site distribution. In vivo, the established population of proviruses is subject to both positive and negative selection, thereby continuously reshaping the integration site distribution. By affecting stochastic proviral expression as well as the mutagenic potential of the virus, integration site choice may be an inherent part of the evolutionary strategies used by different retroviruses to maximise reproductive success. Retroviral integration is a multistep process moulded by nuclear entry, host cofactors and target recognition by the viral integrase. At each level, biases are introduced and the resulting distribution is reshaped by host selection processes. By affecting gene expression, site selection represents a selectable part of the retroviral evolutionary strategy.

Beschreibung: N 6 -methyladenine (6mA) is one of the most abundant types of DNA methylation, and plays an important role in bacteria; however, its roles in higher eukaryotes, such as plants, insects, and mammals, have been considered less important. Recent studies highlight that 6mA does indeed occur, and that it plays an important role in eukaryotes, such as worm, fly, and green algae, and thus the regulation of 6mA has emerged as a novel epigenetic mechanism in higher eukaryotes. Despite this intriguing development, a number of important issues regarding its biological roles are yet to be addressed. In this review, we focus on the 5mC and 6mA modifications in terms of their production, distribution, and the erasure of 6mA in higher eukaryotes including mammals. We perform an analysis of the potential functions of 6mA, hence widening understanding of this new epigenetic mark in higher eukaryotes, and suggesting future studies in this field. Like 5mC, 6mA functions as a potential epigenetic mark in higher eukaryotes. The expression of target genes can be modulated via dynamic and reversible pattern of DNA methylation in a variety of biological processes.

Beschreibung: The common methods for synthesizing calcium phosphates include solid-state reaction, hydrothermal, sol–gel, and wet precipitation. The purpose of this study was to prepare various calcium phosphate bioceramics through a continuous reactor equipped with a static mixer. The precursors, amorphous calcium phosphate (ACP), and poor crystal hydroxyapatite (PC-HAp) were prepared using calcium and phosphate ion-containing solutions by adjusting the pH and the [Ca]/[P] input ratio. The phase transformation from precursors to HAp, alpha-tricalcium phosphates (α-TCP), beta-tricalcium phosphate (β-TCP), or biphasic calcium phosphate (BCP) was dependent on the sintering temperature.

Beschreibung: Rotate and activate . On pages 959–967 , Ichiro Maruyama proposes a “rotation model” for the activation of transmembrane cell-surface receptors. Following this model, receptors exist in a dimeric form prior to ligand binding. The extracellular domain (ECD) has a rotationally flexible structure while the intracellular domain (ICD) is relatively stable. Ligand binding stabilizes the flexible ECD and induces conformational changes of the domains. This in turn induces or allows a rotation of the transmembrane domains leading to a rearrangement of the ICD, hence making the ICD flexible for activation of/interaction with cytoplasmic proteins.

Beschreibung: A complex network of transcription factors regulates specification of neural crest cells at early neurula stage by stabilizing neural crest identity and activating neural crest effector genes so that distinct subpopulations evolve. In this network c-myc acts on top of the gene hierarchy controlling snail2, AP2 and prohibitin1 ( phb1 ) expression. While snail2 and AP2 are well studied neural crest specifier genes little is known about the role of phb1 in this process. To identify phb1 regulated genes we analyzed the transcriptome of neural crest explants of phb1 morphant Xenopus embryos. Among 147 phb1 regulated genes we identified the membrane associated protein-tyrosine phosphatase PRP4A3 ( prl3 ) and the atypical cadherin and Wnt-PCP component van gogh like1 ( vangl1 ). Gain of function, loss of function and epistasis experiments allowed us to allocate both genes in the neural crest specification network between phb1 and twist . Interestingly, both, vangl1 and prl3 regulate only a small subset of neural crest marker genes. The identification of two membrane associated proteins as novel neural crest specifiers indicates that in addition to gene regulation by combinatory effects of transcription factors also post-translational modifications ( prl3 ) and cell-cell adhesion and/or regulation of cell-polarity ( vangl1 ) specify the identity of neural crest cell populations. This article is protected by copyright. All rights reserved.

Beschreibung: We have synthesized a novel derivative of Digitoxin, termed “MonoD”, which demonstrates cytotoxic effects in lung cancer cells with much higher potency as compared to Digitoxin. Our data show that within one hour of MonoD treatment, H460 cells showed increased oxidative stress, increased formation of autophagic vacuoles and increased expression of pro-autophagic markers Beclin-1 and LC3-II. Cells pretreated with MnTBAP, a superoxide scavenger not only lowered superoxide production, but also had lower levels of LC3-II and Beclin-1. Prolonged treatment with MonoD induced apoptosis in lung cancer cells. We investigated MonoD-dependent regulation of Akt and Bcl2, proteins that are known regulators of both autophagy and apoptosis. Molecular and pharmacologic inhibitors of Bcl2 and Akt, when combined with MonoD, led to higher expression of LC3-II and Beclin-1 as compared to MonoD alone, suggesting a repressive effect for these proteins in MonoD-dependent autophagy. Pretreatment of cells with an autophagy inhibitor repressed the apoptotic potential of MonoD, confirming that early autophagic flux is important to drive apoptosis. Therapeutic entities such as MonoD that target multiple pathways such as autophagy and apoptosis may prove advantageous over current therapies that have unimodal basis for action and may drive sustained tumor regression, which is highly desirable. This article is protected by copyright. All rights reserved

Beschreibung: Multiple doping is widely used to improve the performance of a material, including its electrical transport, mechanical, and photovoltaic properties. In this paper, Sn–Se dual-doped Li 10 GeP 2 S 12 (LGPS, thio-LISICON II analogue) electrolytes were synthesized via ball milling and sintering and compared with those Sn or Se single-doped. Successful Sn and/or Se substitution expanded the unit cell and formed units, which were verified by X-ray powder diffraction, energy-dispersive X-ray spectroscopy, and Raman spectroscopy. In contrast to the limited benefits of Se single doping and the negative effects of Sn single doping, Sn–Se dual doping demonstrated up to 53% enhancement in ionic conductivity. More importantly, Sn–Se dual-doped LGPS showed an extremely low activation energy of 16 kJ/mol, which is one of the lowest known values for lithium ion conductors; as well as one of the widest electrochemical windows of 8 V. Sn–Se dual-doped LGPS is a promising electrolyte for advanced all-solid-state batteries.

Beschreibung: Profilin (Pfn1) regulates cytoskeletal reorganization and migration, but its role in osteoblasts is not known. BMPs (bone morphogenetic proteins) aree a multifunctional cytokine involved in osteoblastic differentiation and promote bone regeneration and repair. Although several molecules are known to modulate BMP signaling, mechanisms that determine the levels of BMP action in osteoblastic function are still incompletely understood. We therefore examine the expression of Pfn1 in osteoblasts and its role in BMP-induced differentiation in osteoblasts. In osteoblastic MC3T3-E1(MC) cells, Pfn1 mRNA is expressed constitutively and its expression levels are declined during the culture in a time dependent manner in contrast to the increase in alkaline phosphatase activity revealing that Pfn1 expression is down regulated along with differentiation. To test the effects of osteoblastic differentiation on Pfn1expression further, MC cells are treated with BMP. BMP treatment suppresses the levels of Pfn1 mRNA. This suppressive effect of BMP is time dependent and further down regulation of Pfn1 mRNA levels is observed when the BMP treatment is continued for a longer period of time. Pfn1mRNA knock down (KD) by siRNAs enhances BMP-induced increase in alkaline phosphatase (Alp) activity in MC cells. To analyze the regulatory mechanism, Alp mRNA levels are examined and Pfn1 KD enhances the BMP-induced increase in the levels of Alp mRNA expression. Furthermore, Pfn1 KD enhances BMP-induced transcriptional expression of luciferase reporter activity via BMP response element in osteoblasts. These data indicate that Pfn1 is a novel target of BMP and suppresses BMP-induced differentiation of osteoblasts at least in part via transcriptional event. This article is protected by copyright. All rights reserved

Beschreibung: Ellis-van Creveld (EvC) syndrome (OMIM 225500) is an autosomal recessive disease characterized with chondrodysplastic dwarfism in association with abnormalities in oral cavity. Ciliary proteins EVC and EVC2 have been identified as causative genes and they play an important role on Hedgehog signal transduction. We have also identified a causative gene LIMBIN for bovine chondrodysplastic dwarfism ( bcd ) that is later identified as the bovine ortholog of EVC2 . Here, we report generation of conventional and conditional mutant Evc2 / Limbin alleles that mimics mutations found in EvC patients and bcd cattle. Resulted homozygous mice showed no ciliary localization of EVC2 and EVC and displayed reduced Hedgehog signaling activity in association with skeletal and oral defects similar to the EvC patients. Cartilage-specific disruption of Evc2 / Limbin resulted in similar but milder skeletal defects, whereas osteoblast-specific disruption did not cause overt changes in skeletal system. Neural crest-specific disruption of Evc2 / Limbin resulted in defective incisor growth similar to that seen in conventional knockouts; however, differentiation of amelobolasts was relatively normal in the conditional knockouts. These results showcased functions of EVC2/LIMBIN during formation of mineralized tissues. Availability of the conditional allele for this gene should facilitate further detailed analyses of the role of EVC2/LIMBIN in pathogenesis of EvC syndrome. This article is protected by copyright. All rights reserved.

Beschreibung: Polycrystalline Cd 1− x Ba x O (0 ≤  x  ≤ 0.08) ceramics were synthesized via conventional solid-state reaction method, and the effect of Ba 2+ doping on the microstructure as well as the thermoelectric transport properties of the samples were investigated. It was found that doping of Ba 2+ can inhibit the grain growth of CdO, resulting in a considerable reduction in grain size. Moreover, with the increase in Ba 2+ doping content, both the electrical conductivity and the thermal conductivity of Cd 1− x Ba x O decreased, whereas the Seebeck coefficient increased. A high ZT value of 0.47 was achieved for Cd 0.99 Ba 0.01 O at 1000 K, 38% higher than the undoped CdO, mostly due to reduction of the thermal conductivity.

Beschreibung: It has long been thought that transmembrane cell-surface receptors, such as receptor tyrosine kinases and cytokine receptors, among others, are activated by ligand binding through ligand-induced dimerization of the receptors. However, there is growing evidence that prior to ligand binding, various transmembrane receptors have a preformed, yet inactive, dimeric structure on the cell surface. Various studies also demonstrate that during transmembrane signaling, ligand binding to the extracellular domain of receptor dimers induces a rotation of transmembrane domains, followed by rearrangement and/or activation of intracellular domains. The paper here describes transmembrane cell-surface receptors that are known or proposed to exist in dimeric form prior to ligand binding, and discusses how these preformed dimers are activated by ligand binding. “Rotation model” for a molecular mechanism underlying ligand-induced activation of preformed, cell-surface receptor dimers. Ligand binding induces conformational changes of the extracellular domains that cause rotations of the transmembrane domains. The transmembrane domain rotations dissociate and rearrange the intracellular domain dimers for activation and/or interaction with other cytoplasmic proteins.

Beschreibung: To overcome the drug resistance phenomenon induced by Imatibib (IM), in clinical practice, are often used second generation of tyrosine kinase inhibitors as Nilotinib (NIL) a such potent inhibitor of the BCR/ABL kinase and Dasatinib (DAS) a inhibitor of BCR/ABL kinase and inhibitor SrC family kinase. In this study we evaluated the in vivo effect of DAS, NIL and IM on intracellular calcium concentration, oxidative stress and apoptosis in peripheral blood leukocytes of 45 newly diagnosed patients with chronic myeloid leukaemia (CML-PBM). Our data demonstrated that treatment with DAS and NIL showed an higher modulating potential than IM on intracellular calcium concentration by inhibiting the thapsigargin, a sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) inhibitor and Lithium (Li) an inositol 1,4,5-triphosphate (InsP3) receptor inhibitor activities. Moreover our data demonstrated that NIL and DAS have significantly increased apoptosis more than IM by involving both intracellular calcium signaling as well as oxidative stress. The acquisition of the oxidative stress and calcium channels receptors values data could help the hematologist to modulate and improve the treatment of chronic myeloid leukaemia (CML) pathology. This article is protected by copyright. All rights reserved

Beschreibung: Fucoidan, a natural component of seaweeds, is reported to have immunomodulatory and anti-tumor effects. The mechanisms underpinning these activities remain poorly understood. In this study, the cytotoxicity and anti-tumor activities of fucoidan were investigated in acute myeloid leukemia (AML) cells. The human AML cell lines NB4, KG1a, HL60 and K562 were treated with fucoidan and cell cycle, cell proliferation and expression of apoptotic pathways molecules were analyzed. Fucoidan suppressed the proliferation and induced apoptosis through the intrinsic and extrinsic pathways in the acute promyelocytic leukemia (APL) cell lines NB4 and HL60, but not in KG1a and K562 cells. In NB4 cells, apoptosis was caspase-dependent as it was significantly attenuated by pre-treatment with a pan-caspase inhibitor. P21/WAF1/CIP1 was significantly up-regulated leading to cell cycle arrest. Fucoidan decreased the activation of ERK1/2 and down-regulated the activation of AKT through hypo-phosphorylation of Thr(308) residue but not Ser(473). In vivo , a xenograft model using the NB4 cells was employed. Mice were fed with fucoidan and tumor growth was measured following inoculation with NB4 cells. Subsequently, splenic natural killer (NK) cell cytotoxic activity was also examined. Oral doses of fucoidan significantly delayed tumor growth in the xenograft model and increased cytolytic activity of NK cells. Taken together, these data suggest that the selective inhibitory effect of fucoidan on APL cells and its protective effect against APL development in mice warrant further investigation of fucoidan as a useful agent in treatment of certain types of leukemia. This article is protected by copyright. All rights reserved

Beschreibung: One of the major features of neurodegenerative disease is the selective vulnerability of different neuronal populations that are affected in a progressive and often stereotyped manner. Despite the susceptible neuronal population varies between diseases, oxidative stress is implicated as the major pathogenic process in all of them. Natural Extract of Castanea sativa Mill . bark (ENC), recently characterized in its phenolic composition, acts as antioxidant and cardioprotective agent. Its neuroprotettive properties, however, have never been investigated. The aim of this study was to assess neuroprotection of ENC in in vitro models of oxidative-stress-mediate injury. Human neuroblastoma SH-SY5Y cells treated with glutamate (50 mM for 24h) or hydrogen peroxide (25 µM for 1h followed by 24 with medium) were used. The results showed that the addition of ENC (1-50 µg/ml) to cell medium before the neuronal damage provided neuroprotection in both experimental models used, while its addition after the injury was ineffective. In conclusion, the present results suggest that ENC could be a valuable support as dietary supplement, combining beneficial preventive neuroprotettive effects with a high antioxidant activity. This article is protected by copyright. All rights reserved

Beschreibung: PKR-like ER-resident kinase (PERK) phosphorylates eukaryotic translation initiation factor 2 alpha (eIF2α) under endoplasmic reticulum (ER) stress; this results in repression of general translation and induction of specific gene expression, such as activating transcription factor 4 (ATF4). We previously showed that, upon ER stress, transducin (beta)-like 2 (TBL2) was an ER-localized transmembrane protein and interacted with PERK and that TBL2 was involved in ATF4 expression and cell survival. Here, we show that TBL2 is able to associate with ATF4 mRNA and regulate its translation. The RNA-immunoprecipitation analysis using several TBL2 deletion mutants revealed that the WD40 domain was essential for association with ATF4 mRNA. Importantly, suppression of TBL2 by knockdown or overexpression of the TBL2 mutant with a defective WD40 domain diminished ATF4 induction at the translational level. Thus, our findings indicate that, under ER stress, TBL2 participates in ATF4 translation through its association with the mRNA. This article is protected by copyright. All rights reserved

Beschreibung: Toll-like receptor 2 (TLR2)-mediated signaling cascades and gene regulation are mainly involved in diseases such as immunity and inflammation. In this study microarray analysis was performed using bone marrow-derived macrophages (BMDM) and Raw 264.7 cells to identify novel proteins involved in the TLR2-mediated cellular response. We found that pleckstrin homology-like domain family, member 1 (PHLDA1) is a novel gene up-regulated by TLR2 stimulation and determined the unique signaling pathway for its expression. Treatment with TLR2 agonist Pam 3 CSK 4 increased mRNA, protein, and fluorescence staining of PHLDA1. Induction of PHLDA1 by TLR2 stimulation disappeared from TLR2 KO mice-derived BMDM. Among janus kinase (JAK) family members, JAK2 was involved in TLR2-stimulated PHLDA1 expression. Signal transducer and activator of transcription 3 (STAT3) also participated in PHLDA1 expression downstream of the JAK2. Interestingly, ERK1/2 was an intermediate between JAK2 and STAT3. In silico analysis revealed the presence of highly conserved γ-activated sites within mouse PHLDA1 promoter and confirmed the JAK2-STAT3 pathway is important to Pam 3 CSK 4 -induced PHLDA1 transcription. These findings suggest that the JAK2-ERK1/2-STAT3 pathway is an important signaling pathway for PHLDA1 expression and that these proteins may play a critical role in eliciting TLR2-mediated immune and inflammatory response. This article is protected by copyright. All rights reserved

Beschreibung: Autophagy is a catabolic cellular mechanism involving lysosomal degradation of unwanted cellular components. Interaction between Beclin-1 and Bcl-2 proteins is known to play a critical role in the initiation of autophagy. We report that malignantly transformed lung epithelial cells are resistant to autophagy, and express lower basal levels of autophagic proteins Beclin-1 and LC3-II as compared to non-tumorigenic cells. Additionally, increased levels of nitric oxide (NO) and Bcl-2 were observed in transformed cells. NO was found to negatively regulate autophagy initiation and autophagic flux by nitrosylating Bcl-2 and stabilizing its interaction with Beclin-1, resulting in inhibition of Beclin-1 activity. An increase in the apoptotic initiator caspase-9 and the apoptosis and autophagy-associated kinase p38/MAPK in both cell types indicated possible autophagy-apoptosis crosstalk. Pre-treatments with ABT-737 (Bcl-2 inhibitor) and aminoguanidine (NO inhibitor), and transfection with a non-nitrosylable Bcl-2 cysteine double-mutant plasmid resulted in increased autophagic flux (LC3-II/p62 upregulation) corresponding with decreased S -nitrocysteine expression, thus corroborating the regulatory role of Bcl-2 S -nitrosylation in autophagy. In conclusion, our study reveals a novel mechanism of autophagy resistance via post-translational modification of Bcl-2 protein by NO, which may be critical in driving cellular tumorigenesis. This article is protected by copyright. All rights reserved

Beschreibung: Virtual Reality (VR), a computer-generated virtual environment, has been increasingly used in the entertainment world becoming a very new evolving field, but VR technology has also found a variety of applications in the biomedical field. VR can offer to subjects a safe environment within which to carry on different interventions ranging from the rehabilitation of discharged patients directly at home, to the support of hospitalized patients during different procedures and also of oncological inpatient subjects. VR appears as a promising tool for support and monitoring treatments in cancer patients influencing psychological and physiological functions. The aim of this systematic review is to provide an overview of all the studies that used VR intervention on cancer patients and analyze their main findings. Nineteen studies across nearly a thousand articles were identified that explored effects of VR interventions on cancer patients. Although these studies varied greatly in setting and design, this review identified some overarching themes. Results found that VR improved patients' emotional well-being, and diminished cancer related psychological symptoms. The studies explored various relevant variables including different types of settings (i.e. during chemotherapy, during pain procedures, during hospitalization). Here, we point to the need of a global and multi-disciplinary approach aimed at analyzing the effects of VR taking advantage of the new technology systems like bio sensors as well as electroencephalogram monitoring pre-during and after intervention. Devoting more attention to bio physiological variables, standardized procedures, extending duration to longitudinal studies and adjusting for motion sickness related to VR treatment need to become standard of this research field. This article is protected by copyright. All rights reserved

Beschreibung: A vast network of cellular circadian clocks regulates 24-hour rhythms of behavior and physiology in mammals. Complex environments are characterized by multiple, and often conflicting time signals demanding flexible mechanisms of adaptation of endogenous rhythms to external time. Traditionally this process of circadian entrainment has been conceptualized in a hierarchical scheme with a light-reset master pacemaker residing in the hypothalamus that subsequently aligns subordinate peripheral clocks with each other and with external time. Here we review new experiments using conditional mouse genetics suggesting that resetting of the circadian system occurs in a more “federated” and tissue-specific fashion, which allows for increased noise resistance and plasticity of circadian timekeeping under natural conditions. A network of cellular circadian clocks adapts physiology to the 24-hour day cycle. Traditionally clock entrainment has been conceptualized in a hierarchical scheme with a light-reset SCN pacemaker that subsequently aligns subordinate peripheral clocks. New experiments suggest that resetting of the circadian system occurs in a more “federated” fashion allowing for increased noise resistance and plasticity of circadian timekeeping under complex natural conditions.

Beschreibung: hiCLIP (RNA hybrid and individual-nucleotide resolution ultraviolet cross-linking and immunoprecipitation), is a novel technique developed by Sugimoto et al. (2015). Here, the use of different adaptors permits a controlled ligation of the two strands of a RNA duplex allowing the identification of each arm in the duplex upon sequencing. The authors chose a notoriously difficult to study double-stranded RNA-binding protein (dsRBP) termed Staufen1, a mammalian homolog of Drosophila Staufen involved in mRNA localization and translational control. Using hiCLIP, they discovered a dominance of intramolecular RNA duplexes compared to the total RNA duplexes identified. Importantly, the authors discovered two different types of intramolecular duplexes in the cell: highly translated mRNAs with long-range duplexes in their 3′-UTRs and poorly translated mRNAs with duplexes in their coding region. In conclusion, the authors establish hiCLIP as an important novel technique for the identification of RNA secondary structures that serve as in vivo binding sites for dsRBPs. The hiCLIP technique has allowed Sugimoto et al. (2015) to unravel Staufen 1 (Stau1) function in mRNA translational regulation, showing that transcript translation is dependent on the location of intramolecular double-stranded duplexes recognized by Stau1.

Beschreibung: Alzheimer's disease (AD) is the most common cause of dementia, and there is currently no cure. The “β-amyloid cascade hypothesis” of AD is the basis of current understanding of AD pathogenesis and drug discovery. However, no AD models have fully validated this hypothesis. We recently developed a human stem cell culture model of AD by cultivating genetically modified human neural stem cells in a three-dimensional (3D) cell culture system. These cells were able to recapitulate key events of AD pathology including β-amyloid plaques and neurofibrillary tangles. In this review, we will discuss the progress and current limitations of AD mouse models and human stem cell models as well as explore the breakthroughs of 3D cell culture systems. We will also share our perspective on the potential of dish models of neurodegenerative diseases for studying pathogenic cascades and therapeutic drug discovery. Recently, we recapitulated key events of Alzheimer's disease pathogenesis in a 3D human stem cells culture system. This model enhances beta-amyloid accumulation and neurofibrillary tau tangles (NFT), providing a powerful cellular model for Alzheimer's disease. In this review, we discuss the current progress of modeling neurodegenerative diseases in 3D cultures.

Beschreibung: In this study, aluminum foil was bonded directly to alumina substrate at a temperature higher than 1000°C in an inert atmosphere. A platinum marker was used to locate the original interface after bonding. With the help of the Pt marker, the microstructure analysis revealed that the aluminum foil oxidized first, and the newly formed alumina was subsequently bonded to the alumina substrate. The bonding strength increased with increases in bonding temperature until 1100°C. The strong interface also led to high thermal conductivity.

Beschreibung: A study of phase transition, microstructure, and dielectric properties of Ba 0.7 Sr 0.3 Ti 1– x Ni x O 3 (BSTN) ceramics prepared by slow-injection solgel technique with x ranging from 0 to 1 mol% is reported in this article. The as-prepared BSTN material was calcined at 800 and 1000°C and subsequently sintered at 1100 and 1200°C, respectively. The optimized condition was found to be Ba 0.7 Sr 0.3 TiO 3 doped with 1 mol% nickel calcined at 1000°C and sintered at 1200°C having the lowest dielectric loss of 0.02 with a dielectric constant of 1603 which was measured at a frequency of 1 kHz at room temperature.

Beschreibung: The Gene Expression Database (GXD) is an extensive and freely available community resource of mouse developmental expression data. GXD curates and integrates expression data from the literature, via electronic data submissions, and by collaborations with large-scale projects. As an integral component of the Mouse Genome Informatics (MGI) Resource, GXD combines expression data with genetic, functional, phenotypic and disease-related data, and provides tools for the research community to search for and analyze expression data in this larger context. Recent enhancements include: an interactive browser to navigate the mouse developmental anatomy and find expression data for specific anatomical structures; the capability to search for expression data of genes located in specific genomic regions, supporting the identification of disease candidate genes; a summary displaying all the expression images that meet specified search criteria; interactive matrix views that provide overviews of spatio-temporal expression patterns (Tissue x Stage Matrix) and enable the comparison of expression patterns between genes (Tissue x Gene Matrix); data zoom and filter utilities to iteratively refine summary displays and data sets; and gene-based links to expression data from other model organisms, such as chicken, Xenopus and zebrafish, fostering comparative expression analysis for species that are highly relevant for developmental research. This article is protected by copyright. All rights reserved.

Beschreibung: Although the mechanism which regulates transcription in the 5'-UTR of the mu opioid receptor (MOR) gene (OPRM1) in lymphocytes has been well studied, a question remains as to whether there is post-transcriptional regulation of MOR gene OPRM1 gene in lymphocytes. In this study, we describe both the role played by miRNAs and the impact of SIVmac239 infection on post-transcriptional regulation of MOR gene OPRM1 gene in CEM x174 cells. Our results show that miR-16 is able to bind the target site in the range of 8699-8719 nt from the stop coden in MOR gene MOR-1 mRNA 3'-UTR and suppress the expression of MOR OPRM1 gene. Mutation of this target site reduces the effect of miR-16. Morphine (1 µM) inhibites the expression of miR-16, and this effect is reversed by the antagonist naloxone. Thus, morphine may up-regulate MOR receptor level by both stimulating MOR OPRM1 gene transcription and stabilizing its mRNA. SIVmac239 infection results in an apparent elevation of miR-16 and gradual reduction of MOR OPRM1 gene expression. The inverse correlation of elevated miR-16 and reduced MOR OPRM1 gene expression under viral loading confirmed the effect of SIVmac239 on post-transcriptional regulation of MOR OPRM1 gene in lymphocytes. We conclude that miR-16 is a primary factor in post-transcriptional regulation of MOR OPRM1 gene. SIVmac239 upregulates miR-16 levels and consequently suppresses MOR OPRM1 gene expression. This finding will be helpful for full understanding of the regulatory mechanism of MOR OPRM1 gene in lymphocytes, as well as the synergistic mechanism of HIV infection and morphine addiction in the pathogenesis of AIDS.

Beschreibung: Structural stability of Oldenlandia affinis cyclotide, kalata B1 of native (1NB1) and two mutants 2F2I ([P20D, V21K] kB1) and 2F2J ([W19K, P20N, V21K] kB1) was investigated. Single model analysis showed high number of intra-molecular interactions followed by more proportion of beta sheet contents in [P20D, V21K] kB1 as compared to that of native and the other mutant of kalata B1. Further, the modern conformational sampling approach, an alternate to classical molecular dynamics was introduced, which revealed that the [P20D, V21K] kB1 was identified as structurally stable one, substantiated by various structural events viz., root mean square deviation, root mean square fluctuation and angular deviation by Ramachandran plot. Moreover, the statistically validated contours of polar surface area, hydrogen bond distribution and the distance of disulfide bridges also supported the priority of [P20D, V21K] kB1 with respect to stability. From this work, it is proposed that the [P20D, V21K] kB1 (2F2I) could be the best template for scaffolding peptide based drug design.

Beschreibung: Inhibition of metabolic features which distinguish cancer cells from their non-malignant counterparts is a promising approach to cancer treatment. Energy support for drug extrusion in multidrug resistance (MDR) is a potential target for metabolic inhibition. Two major sources of ATP-based metabolic energy are partial (glycolysis) and complete (mitochondrial oxidative phosphorylation) oxidation of metabolic fuels. In cancer cells, the balance between them tends to be shifted towards glycolysis; this shift is considered to be characteristic of the cancer metabolic phenotype. Numerous earlier studies, conducted with cells cultured in a monolayer (2-D model), suggested inhibition of glycolytic ATP production as an efficient tool to suppress MDR in cancer cells. Yet, more recent work challenged the appropriateness of the 2-D model for such studies and suggested that a more clinically relevant approach would utilize a more advanced cellular model such as a 3-D model. Here we show that the transition from the 2-D model (cultured monolayer) to a 3-D model (cultured spheroids) introduces essential changes into the concept of energetic suppression of MDR. The 3-D cell organization leads to the formation of a discrete cell subpopulation (not formed in the 2-D model) with elevated MDR transport capacity. This subpopulation has a specific metabolic phenotype (mixed glycolytic/oxidative MDR support) different from that of cells cultured in the 2-D model. Finally, the shift to the oxidative phenotype becomes greater when the spheroids are grown under conditions of lactic acidosis that are typical for solid tumors. The potential clinical significance of these findings is discussed.

Beschreibung: We investigated the effects of exogenous sodium pyruvate (SP) on adipocyte differentiation, lipid accumulation, and the mRNA expression levels of adipogenesis-related genes in 3T3-L1 pre-adipocytes. Differentiation of pre-adipocytes was induced by MDI (3-isobutyl-1-methylxanthine: IBMX, dexamethasone: DEX, and insulin), in the presence or absence of SP. Adipogenesis was stimulated by SP in a concentration-dependent manner. SP also induced the expression of genes encoding aP2, GLUT4, and adiponectin, but had no effect on cell proliferation. Exogenous glucose did not promote adipogenesis or lipid accumulation. 2-deoxy-D-glucose inhibited adipogenesis initiated by MDI, but failed to influence the effects of SP on adipogenesis, whereas 3-bromopyruvate inhibited adipogenesis regardless of whether SP was present. The pro-adipogenic properties of SP were limited to the early events of adipogenesis. To determine whether SP mimics the adipogenic action of dexamethasone or insulin, we examined the effects of SP on adipogenesis with combinations of IBMX, DEX, and insulin. SP did not improve incomplete lipid accumulation observed in cells grown under IBMX-, DEX-, or insulin-free conditions. Insulin-stimulated ERK1/2 phosphorylation was diminished by SP, while phosphorylation of Akt was increased, correlating with increased glucose uptake in response to insulin. We also observed that SP stimulated immediate early expression of C/EBPβ and C/EBPδ. The PPARγ antagonist GW9662 inhibited adipogenesis. Our findings highlight the adipogenic function of exogenous SP by stimulating early events of adipogenesis.

Beschreibung: Diabetic nephropathy is characterized by inordinate secretion of extracellular matrix (ECM) proteins from mesangial cells (MCs), which is tightly associated with excessive activation of TGF-β signaling. The forkhead transcription factor O1 (FoxO1) protects mesangial cells from hyperglycemia-induced oxidative stress, which may be involved in ameliorating the redundant secretion of ECM proteins under high glucose conditions. Here we reported that high glucose elevated the level of p-Akt to attenuate endogenous FoxO1 bioactivities in MCs, accompanied with decreases in the mRNA expressions of catalase (CAT) and superoxide dismutase 2 (SOD2). Meanwhile, the expression of major ECM proteins-FN and Col I- increased under high glucose condition, in consistent with the activation of TGF-β/Smad signaling. By contrast, overexpression of nucleus-localized FoxO1 (insensitive to Akt phosphorylation) directly up-regulated the expressions of anti-oxidative enzymes, accompanied with inactivation of TGF-β/Smad3 pathway, as well as decreases of extracellular matrix proteins. Moreover, similar to those MCs overexpressed of nucleus-localized FoxO1 in high glucose conditions, MCs with down-regulation of FoxO1 by small interference-RNA under normal glucose conditions showed increased FN level and activated TGF-β/Smad3 pathway. Our findings link the anti-oxidative activity of FoxO1 and the TGF-β-induced secretion of ECM proteins, indicating the novel role of FoxO1 in protecting MCs under high glucose conditions.

Beschreibung: RNA binding proteins (RBPs) are key factors for the regulation of gene expression by binding to cis elements, i.e. short sequence motifs in RNAs. Recent studies demonstrate that cooperative binding of multiple RBPs is important for the sequence-specific recognition of RNA and thereby enables the regulation of diverse biological activities by a limited set of RBPs. Cross-linking immuno-precipitation (CLIP) and other recently developed high-throughput methods provide comprehensive, genome-wide maps of protein-RNA interactions in the cell. Structural biology gives detailed insights into molecular mechanisms and principles of RNA recognition by RBPs, but has so far focused on single RNA binding proteins and often on single RNA binding domains. The combination of high-throughput methods and detailed structural biology studies is expected to greatly advance our understanding of the code for protein-RNA recognition in gene regulation, as we review in this article. Multi-protein-RNA networks play important roles in post-transcriptional regulation of gene expression. Deciphering the underlying protein-RNA recognition code will greatly benefit from combining large-scale quantitative methods with integrated structural biology.

Beschreibung: A phase-pure and high-brightness blue phosphor, Eu 2+ doped SrMgAl 10 O 17 (SAM), was synthesized through a hybrid urea-sol combustion route. The structure, photoluminescence spectra, and thermal stability of the SAM were investigated in this work. The phosphor had a homogeneous and rod-like microstructure, showing a broad emission band centered at 465 nm under the 330 nm excitation. The concentration for luminescence quenching of SAM: Eu 2+ occurred at 10 mol%, which doubled that of the phosphor prepared by the conventional combustion method. Compared with the traditional combustion method, the hybrid route led to improvements in luminescence by 52.2%, external quantum efficiency by 16.2%, and thermal stability by 8.8% at 435 K. The blue phosphor prepared by the new method could thus be potentially used with near ultraviolet light-emitting diodes.

Beschreibung: Fabry disease (FD) is a hereditary X-linked metabolic lysosomal storage disorder due to insufficient amounts or a complete lack of the lysosomal enzyme α -galactosidase A ( α  − GalA). The loss of α -GalA activity leads to an abnormal accumulation of globotriaosylceramide (Gb3) in lysosomes and other cellular components of different tissues and cell types, affecting the cell function. However, whether these biochemical alterations also modify functional processes associated to the cell mitotic ability is still unknown. The goal of the present study was to characterize lineages of human dermal fibroblasts (HDFs) of FD patients and healthy controls focusing on Gb3 accumulation, expression of chloride channels that regulate proliferation, and proliferative activity. The biochemical and functional analyses indicate the existence of quantitative differences in some but not all the parameters of cytoskeletal organization, proliferation and differentiation processes. This article is protected by copyright. All rights reserved

Beschreibung: Chromosomes are not only carriers of the genetic material, but also actively regulate the assembly of complex intracellular architectures. During mitosis, chromosome-induced microtubule polymerisation ensures spindle assembly in cells without centrosomes and plays a supportive role in centrosome-containing cells. Chromosomal signals also mediate post-mitotic nuclear envelope (NE) re-formation. Recent studies using novel approaches to manipulate histones in oocytes, where functions can be analysed in the absence of transcription, have established that nucleosomes, but not DNA alone, mediate the chromosomal regulation of spindle assembly and NE formation. Both processes require the generation of RanGTP by RCC1 recruited to nucleosomes but nucleosomes also acquire cell cycle stage specific regulators, Aurora B in mitosis and ELYS, the initiator of nuclear pore complex assembly, at mitotic exit. Here, we review the mechanisms by which nucleosomes control assembly and functions of the spindle and the NE, and discuss their implications for genome maintenance. Chromosomes act as reaction platforms for spindle assembly and nuclear envelope formation. Both processes depend on nucleosomes, which induce spindles by recruiting RCC1 and Aurora B in mitosis, and nuclear envelopes by recruiting RCC1 and ELYS in interphase. Here, we review these mechanisms, and discuss their implications for genome maintenance.

Beschreibung: In the organelles of plants and mammals, recent evidence suggests that genomic instability stems in large part from template switching events taking place during DNA replication. Although more than one mechanism may be responsible for this, some similarities exist between the different proposed models. These can be separated into two main categories, depending on whether they involve a single-strand-switching or a reciprocal-strand-switching event. Single-strand-switching events lead to intermediates containing Y junctions, whereas reciprocal-strand-switching creates Holliday junctions. Common features in all the described models include replication stress, fork stalling and the presence of inverted repeats, but no single element appears to be required in all cases. We review the field, and examine the ideas that several mechanisms may take place in any given genome, and that the presence of palindromes or inverted repeats in certain regions may favor specific rearrangements. Short-range inversions are a major component of genomic instability in the organelles of Arabidopsis thaliana and humans. Here, we review proposed replication-based mechanisms for the formation of these rearrangements. We identify common characteristics of the mechanisms and examine their impact on organelle genomes.

Beschreibung: Mucopolysaccharidosis type I (MPS I) is a rare autosomal recessive disease caused by alpha-L-iduronidase deficiency in which heparan and dermatan sulfate degradation is compromised. Besides primary lysosomal glycosaminoglycan accumulation, further changes in cellular functions have also been described in several murine MPS models. Herein, we evaluated alterations in hematopoiesis and its implications on the production of mature progeny in a MPS I murine model. Despite the significant increase in hematopoietic stem cells, a reduction in common myeloid progenitors and granulocyte-macrophage progenitor cells was observed in Idua -/- mice bone marrow. Furthermore, no alterations in number, viability nor activation of cell death mechanisms were observed in Idua -/- mice mature macrophages but they presented higher sensitivity to apoptotic induction after staurosporine treatment. In addition, changes in Ca 2+ signaling and a reduction in phagocytosis ability were also found. In summary, our results revealed significant intracellular changes in mature Idua -/- macrophages related to alterations in Idua -/- mice hematopoiesis, revealing a disruption in cell homeostasis. These results provide new insights into physiopathology of MPS I. This article is protected by copyright. All rights reserved

Beschreibung: Crystalline argon oxygen decarburization slag, in powdery form, was investigated for its hydration potential by alkali activation and curing at 80°C. Na-silicate and K-silicate of the same modulus were used as activators. Isothermal calorimetry at 80°C indicated exothermic reactions in the slag pastes. When the slag mortars were cured under steam at 80°C appreciable gain in compressive strength was measured. This was attributed to C–S–H which was detected in TG, FTIR, and 29 Si NMR analyses. Upon hydration at 90 d, the amount of crystalline phases decreased, whereas the XRD amorphous content in the slag increased. Electron microscopy showed the formation of different morphologies of reaction products depending on the alkaline activator employed. Presence of reaction rims around the crystalline phases with a major presence of Ca, Si, and O in the reacted matrix was observed in elemental maps.

Beschreibung: The (1− x )BiFeO 3 - x BaTiO 3 (with x  = 0.1, 0.2, 0.3, and 0.4) ceramics were fabricated successfully by solid-state reaction method. Single-phase perovskite was obtained in all ceramics, as confirmed by XRD technique. It was observed that 0.7BiFeO 3 –0.3BaTiO 3 was the morphotropic phase boundary (MPB) between rhombohedral and cubic phases, as also revealed from ferroelectric and magnetic properties. The simulated and experimental X-Ray Absorption Spectroscopy (XAS) study revealed that BT in 0.75BF-0.25BT is possibly taken a rhombohedral structure. Furthermore, the rounded ferroelectric hysteresis loops observed for 0.9BiFeO 3 –0.1BaTiO 3 and 0.8BiFeO 3 –0.2BaTiO 3 compositions could be attributed to their microstructure and surface charge effects and electron transfer between Fe 3+ and Fe 2+ ions. It was also found that high dielectric constant of 0.9BiFeO 3 –0.1BaTiO 3 composition was a result of grain and grain-boundary effects, as observed in SEM micrographs. In addition, a strong signature of dielectric relaxation behavior was observed in this ceramic system with the activation energy 0.467 eV obtained from the Arrhenius' law. Finally, the local structure investigation with XAS technique provided additional information to better understand the electric and magnetic properties in the BF-BT ceramic system.

Beschreibung: Inflammatory responses are essential for the clearance of pathogens and the repair of injured tissues; however, if these responses are not properly controlled chronic inflammation can occur. Chronic inflammation is now recognized as a contributing factor to many age-associated diseases including metabolic disorders, arthritis, neurodegeneration, and cardiovascular disease. Due to the connection between chronic inflammation and these diseases, it is essential to understand underlying mechanisms behind this process. In this review, factors that contribute to chronic inflammation are discussed. Further, we emphasize the emerging roles of microRNAs (miRNAs) and other noncoding RNAs (ncRNA) in regulating chronic inflammatory states, making them important future diagnostic markers and therapeutic targets. Copyright Line: © 2015 The Authors BioEssays Published by Wiley-VCH Verlag GmbH & Co. KGaA. Although immune responses are necessary for proper clearance of pathogens and tissue repair, these responses can become dysregulated resulting in a chronic inflammatory state. Chronic inflammation is a contributing factor to many age-associated diseases. Recently, noncoding RNAs have been shown to regulate chronic inflammation and are emerging as potential therapeutic targets.

Beschreibung: ABSTRACT RFX transcription factors are key regulators of ciliogenesis in vertebrates. In Xenopus and zebrafish embryos, knockdown of Rfx2 causes defects in neural tube closure and in left-right axis patterning. To determine the essential role of the Rfx2 gene in mammalian development, we generated Rfx2 -deficient mice using an embryonic stem cell clone containing a lacZ gene trap reporter inserted into the first intron of the Rfx2 gene. We found that the Rfx2 lacZ reporter is expressed in ciliated tissues during mouse development including the node, the floor plate and the dorsal neural tube. However, mice homozygous for the Rfx2 gene trap mutation did not have defects in neural tube closure or in organ situs . The gene trap insertion appears to create a null allele as Rfx2 mRNA was not detected in Rfx2 gt/gt embryos. Although Rfx2 -deficient mice do not have an obvious embryonic phenotype, we found that Rfx2 gt/gt males are infertile due to a defect in spermatid maturation at or before the round and elongating spermatid stage. Our results indicate that Rfx2 is not essential for embryonic development in the mouse but is required for spermatogenesis. This article is protected by copyright. All rights reserved.

Beschreibung: The neural-epidermal boundary tissues include the neural crest and preplacodal ectoderm (PPE) as primordial constituents. The PPE region is essential for the development of various sensory and endocrine organs, such as the anterior lobe of the pituitary, olfactory epithelium, lens, trigeminal ganglion, and otic vesicles. During gastrulation, a neural region is induced in ectodermal cells that interacts with mesendodermal tissue and responds to several secreted factors. Among them, inhibition of bone morphogenetic protein (BMP) in the presumptive neuroectoderm is essential for the induction of neural regions, and formation of a Wnt and fibroblast growth factor (FGF) signaling gradient along the midline determines anterior-posterior patterning. In this study, we attempted to specifically induce PPE cells from undifferentiated Xenopus cells by regulating BMP, Wnt and FGF signaling. We showed that the proper level of BMP inhibition with an injection of truncated BMP receptor or treatment with a chemical antagonist triggered the expression of PPE genes. In addition, by varying the amount of injected chordin , we optimized specific expression of the PPE genes. PPE gene expression increased by adding an appropriate dose of a FGF receptor antagonist. Furthermore, co-injection with either wnt8 or the Wnt inhibitor dkk-1 altered the expression levels of several region-specific genes according to the injected dose. We specifically induced PPE cell differentiation in animal cap cells from early-stage Xenopus embryos by modulating BMP, Wnt and FGF signaling. This is not the first research on placode induction, but our simple method could potentially be applied to mammalian stem cell systems. This article is protected by copyright. All rights reserved.

Beschreibung: Spherical monodispersed yttrium-doped barium zirconate (BaY 0.1 Zr 0.9 O 3 ; BYZ) particles were successfully prepared in single step without the requirement of calcination process by the sonochemical method in highly basic aqueous solution. The stoichiometric solution of BaCl 2 .2H 2 O, ZrOCl 2 .8H 2 O, and YCl 4 .6H 2 O was precipitated in a 20  M NaOH under high-intensity ultrasonic irradiation (20 kHz, 150 W/cm 2 ) for 15, 30, and 60 min. As-prepared powders were identified by XRD, FT-IR, and Raman spectroscopy as cubic perovskite BYZ. The microstructure examined by SEM and TEM showed spherical-shaped BYZ particles formed by aggregation of primary nanocrystals, and this unique morphology was induced by the effects of the ultrasonication and the strong alkaline environment. BYZ powders prepared under ultrasonication for 60 min had narrow-sized distribution with the average particle size of 267 ± 26 nm and the specific surface area of 40.2 m 2 /g. BYZ ceramics sintered in the air at 1550°C for 20 h showed good densification (95%) and consisted of large grain size (7.67 ± 2.79 μm).

Beschreibung: In this study, ZnS powders with homogeneous morphology were synthesized using a colloidal processing method. Vacuum hot pressing was subsequently applied to consolidate the ZnS powders into infrared transparent ceramics (77.3% transmittance at wavelengths of 6.74 and 9.29 μm). The phase composition of the sintered ZnS suggests the presence of wurtzite as a minor phase in addition to the primary sphalerite phase, and microstructural analysis indicates that the ceramics are highly densified. It has been found that the VHP-sintered ZnS ceramics exhibit blue (450 nm) and green (530 nm) luminescence, which is due to the formation of zinc vacancies and sulfur interstitials, respectively, during the sintering process.

Beschreibung: In this study, tribological investigations were carried out on ZTA ceramics with 17 vol% Y-TZP and different stabilizer contents (1, 1.5, and 2 mol% yttria) to analyze the influence of zirconia transformation on wear properties. Samples were tested in a linearly reciprocating ball on flat setup with different applied loads. Raising the fracture toughness by transformation toughening, microcracking, and residual stresses improves the wear resistance only at transition load but increases the wear at high loads. Higher yttria content of 2 mol% and lower zirconia grain size and thus low transformability, decreases fracture toughness but increases the wear resistance at high loads. Therefore the adjustment of stabilizer content on zirconia volume fraction in ZTA plays a decisive role in tribological applications.

Beschreibung: Highly (100)-oriented 0.38Bi(Ni 1/2 Hf 1/2 )O 3 -0.62PbTiO 3 relaxor-ferroelectric films were fabricated on Pt(111)/Ti/SiO 2 /Si(111) substrates by introducing a lead oxide seeding layer. A moderate relative permittivity , a low dissipation factor (tan δ 〈 5%), and strong relaxor-like behavior (γ = 0.74) over a broad temperature region were observed. The energy storage density of approximately 45.1 ± 2.3 J/cm 3 was achieved for films with (100) preferential orientation, which is much higher than the value ~33.5 ± 1.7 J/cm 3 obtained from films with random orientation. Furthermore, the PbO-seeded films are more capable of providing larger piezoelectric response (~113 ± 10 pm/V) compared to the films without seeds (~85 ± 8 pm/V). These excellent features indicate that the highly (100)-oriented 0.38Bi(Ni 1/2 Hf 1/2 )O 3 -0.62PbTiO 3 films could be promising candidates for applications in high-energy storage capacitors, high-performance MEMS devices, and particularly for potential applications in the next-generation integrated multifunctional piezoelectric energy harvesting and storage system.

Beschreibung: Uniformly dispersed TiC nanoparticle strengthened In 4 Se 2.65 composites have been fabricated by a combined process of mechanical alloying (MA) and hot pressing (HP) successfully. Due to the good electrical conductivity and the extra phonon scattering effect of the TiC nanoinclusions, the electrical resistivity and thermal conductivity decrease with the TiC content up to 0.8 wt%, and a maximum ZT of 0.98 at 723 K was achieved in the sample with 0.8 wt% TiC. Taking account of the measurement uncertainly, the enhancement of ZT value by TiC nanoinclusions is less obvious. On the other hand, the mechanical performance of In 4 Se 2.65 can be effectively improved by TiC nanoinclusions due to the dispersive strengthening effect of the nanoinclusions , and the flexural strength of the sample with 0.8 wt% TiC is improved to 73 MPa, which is over 40% higher than that of the pristine sample.

Beschreibung: The radiation damage response of Ti 3 SiC 2 heated from 120°C to 850°C during 700 keV Si + irradiation has been investigated. The samples were analyzed using glancing incidence X-ray diffraction, Rutherford backscattering spectrometry, Raman spectroscopy, and scanning electron microscopy. For the sample at 120°C, irradiation results in a buildup of a heterogeneous surface and the formation of TiC x . Irradiation at 200°C results in maximum microstrain, a maximum in the c lattice parameter, and the appearance of a β phase in addition to the normal α phase of Ti 3 SiC 2. A minimum in the observed damage level near the surface was seen for irradiation at a sample temperature of 300°C but the damaged phase increases at higher temperatures. Differences between the present work and a previous C irradiation study have been ascribed to the enhanced Si defect transport at low temperatures.

Beschreibung: The effect of targeted expression of an anabolic isoform of basic fibroblast growth factor (FGF2) in osteoblastic lineage on tibial fracture healing was assessed in mice. Closed fracture of the tibiae was performed in Col3.6-18kDa Fgf2 -IRES-GFPsaph mice in which a 3.6 kb fragment of type I collagen promoter (Col3.6) drives the expression of only the 18kD isoform of FGF2 (18kDa Fgf2/ LMW) with green fluorescent protein-sapphire (GFPsaph) as well as Vector mice (Col3.6-IRES-GFPsaph, Vector) that did not harbor the FGF2 transgene. Radiographic, micro-CT, DEXA and histologic analysis of fracture healing of tibiae harvested at 3, 10 and 20 days showed a smaller fracture callus but accelerated fracture healing in LMWTg compared with Vector mice. At post fracture day 3, FGF receptor 3 and Sox 9 mRNA were significantly increased in LMWTg compared with Vector. Accelerated fracture healing was associated with higher FGF receptor 1, platelet derived growth factors B, C and D, type X collagen, vascular endothelial cell growth factor, matrix metalloproteinase 9, tartrate resistant acid phosphatase, cathepsin K, runt-related transcription factor-2, Osterix and Osteocalcin and lower Sox9, and type 2 collagen expression at 10 days post fracture. We postulate that overexpression of LMW FGF2 accelerated the fracture healing process due to its effects on factors that are important in chondrocyte and osteoblast differentiation and vascular invasion. This article is protected by copyright. All rights reserved

Beschreibung: Single Nucleotide Polymorphisms in FTO intron 1 have been associated with obesity risk, leading to the hypothesis that FTO is the obesity-related gene. However, other studies have shown that the FTO gene is part of the regulatory domain of the neighboring IRX3 gene and that enhancers in FTO intron 1 regulate IRX3 . While Irx3 activity was shown to be necessary in the hypothalamus for the metabolic function of Irx3 in mouse, no enhancers with hypothalamic activity have been demonstrated in the risk-associated region within FTO . In order to identify potential enhancers at the human FTO locus in vivo , we tested regulatory activity in FTO intron 1 using BAC transgenesis in zebrafish. A minimal gata2 promoter-GFP cassette was inserted 1.3 kb upstream of the obesity associated SNP rs9939609 in a human FTO BAC plasmid. In addition to the previously identified expression domains in notochord and kidney, human FTO BAC:GFP transgenic zebrafish larvae expressed GFP in the ventral posterior tuberculum, the posterior hypothalamus and the anterior brainstem, which are also expression domains of zebrafish irx3a . In contrast, an in-frame insertion of a GFP cassette at the FTO start codon resulted in weak ubiquitous GFP expression indicating that the promoter of FTO does likely not react to enhancers located in the obesity risk-associated region. This article is protected by copyright. All rights reserved.

Beschreibung: TCERG1 was characterized previously as a repressor of the transcription factor C/EBPα through a mechanism that involved relocalization of TCERG1 from nuclear speckles to pericentromeric regions. The inhibitory activity as well as the relocalization activity has been demonstrated to lie in the amino terminal half of the protein, which contains several discrete motifs including an imperfect glutamine-alanine (QA) repeat. In the present study, we showed that deletion of this domain completely abrogated the ability of TCERG1 to inhibit the growth arrest activity of C/EBPα. Moreover, the QA repeat deletion mutant of TCERG1 lost the ability to be relocalized from nuclear speckles to pericentromeric regions, and caused an increase in the average size of individual speckles. We also showed that deletion of the QA repeat abrogated the complex formation between TCERG1 and C/EBPα. Examination of mutants with varying numbers of QA repeats indicated that a minimal number of repeats are required for inhibitory activity as well as relocalization ability. These data contribute to our overall understanding of how TCERG1 can have gene-specific effects in addition to its more general roles in coordinating transcription elongation and splicing. This article is protected by copyright. All rights reserved

Beschreibung: The small ubiquitin-like modifier SUMO regulates many aspects of cellular physiology to maintain cell homeostasis, both under normal conditions and during cell stress. Components of the transcriptional apparatus and chromatin are among the most prominent SUMO substrates. The prevailing view is that SUMO serves to repress transcription. However, as we will discuss in this review, this model needs to be refined, because recent studies have revealed that SUMO can also have profound positive effects on transcription. SUMO targets a number of transcription factors, and the current dogma is that sumoylation of transcription factors generally inhibits the transcription process. In this review, we nuance this dogma by discussing recent findings that reveal SUMO as an activator of transcription of pro-growth genes in yeast and mammalian cells.

Beschreibung: The temperature distribution in copper and martensitic steel spheres has been investigated for the initial stage of field-activated sintering (FAST)/spark plasma sintering (SPS) using capacitor discharges (CD) with applied voltages from one to 15 V as model experiments. At first, the evolution of the contact resistance between the spheres has been studied. The results show the reduction in the contact resistance after discharge with increasing electrical load, yet no significant dependence on the length or number of the discharge pulses. Thereby the initial resistance is only decreased distinctly if at least a certain minimal voltage was applied. Subsequently, the melting of thin coatings of different metals on copper spheres has been studied and the occurrence of molten phase and its melting point were assigned to the corresponding discharge current. Extrapolation from the currents necessary to melt the coating layers in the CD experiments to lower values typical for FAST was used to estimate the contact overtemperature in the latter case. Resulting values for copper range from 0.05 K for normal heating with 100 K/min to 5 K for maximum current output.

Beschreibung: In this study, we report on the microstructure of SiO 2 -coated Al-doped ZnO nanoparticles densified by spark plasma sintering(SPS), using a multiscale approach. Our observations show that it is possible to successfully prepare dense pellets while keeping the nanostructure with well-defined Si-rich grain boundaries. Although a very limited partial solubility of Si in the ZnO matrix has been observed, Si is mostly concentrated at the grain boundaries. More surprisingly, we evidenced some areas with nanoscale inhomogeneity of the Al concentration, which can locally strongly exceed the average composition of the matrix. It could explain the apparent discrepancy observed in the literature between the simultaneous presence of ZnAl 2 O 4 in Al-doped ZnO, which should be the signature of the doping level exceeding the solubility limit, and the concentration of carriers that still depends on the nominal Al concentration in ZnO even in the presence of ZnAl 2 O 4 .

Beschreibung: G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2. GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblasts and myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts the receptor was present in nucleus, mitochondria and endoplasmic reticulum, in C2C12 myotubes it was additionally found in cytoplasm. Using trypan blue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, we demonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H 2 O 2 as E2. However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of the molecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptotic conditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15 antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps of C2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments in undifferentiated and differentiated C2C12 cells and mediates E2 actions. This article is protected by copyright. All rights reserved

Beschreibung: Chondrogenesis subtends the development of most skeletal elements and involves mesenchymal cell condensations differentiating into growth plate chondrocytes that proliferate, undergo hypertrophy and are replaced by bone. In the pediatric disorder Hereditary Multiple Exostoses, however, chondrogenesis occurs also at ectopic sites and causes formation of benign cartilaginous tumors –exostoses- near the growth plates. No treatment is currently available to prevent or reverse exostosis formation. Here, we asked whether chondrogenesis could be stopped by targeting the hedgehog pathway, one of its major regulators. Micromass cultures of limb mesenchymal cells were treated with increasing amounts of the hedgehog inhibitor HhAntag or vehicle. The drug effectively blocked chondrogenesis and did so in a dose-dependent manner as monitored by: alcian blue-positive cartilage nodule formation; gene expression of cartilage marker genes; and reporter activity in Gli1-LacZ cell cultures. HhAntag blocked chondrogenesis even when the cultures were co-treated with bone morphogenetic protein 2 (rhBMP-2), a strong pro-chondrogenic factor. Immunoblots showed that HhAntag action included modulation of canonical (pSmad1/5/8) and non-canonical (pp38) BMP signaling. In cultures co-treated with HhAntag plus rhBMP-2, there was a surprising strong up-regulation of pp38 levels. Implantation of rhBMP-2-coated beads near metacarpal elements in cultured forelimb explants induced formation of ectopic cartilage that however, was counteracted by HhAntag co-treatment. Collectively, our data indicate that HhAntag inhibits not only hedgehog signaling, but also modulates canonical and non-canonical BMP signaling and blocks basal and rhBMP2-stimulated chondrogenesis, thus representing a potentially powerful drug-based strategy to counter ectopic cartilage growth or induce its involution. This article is protected by copyright. All rights reserved

Beschreibung: Chd5 is an essential factor for neuronal differentiation and spermatogenesis, and known as a tumor suppressor. H3K27me3 and H3K4un are modifications recognized by Chd5; however, it remains unclear how Chd5 remodels chromatin structure. We completely disrupted the Chd5 locus using the CRISPR-Cas9 system to generate a 52 kbp long deletion, and analyzed Chd5 function in mouse embryonic stem cells. Our findings show that Chd5 represses murine endogenous retrovirus-L (MuERV-L/MERVL), an endogenous retrovirus-derived retrotransposon, by regulating H3K27me3 and H3.1/H3.2 function. This article is protected by copyright. All rights reserved

Beschreibung: The effect of octylamine flow rate on the structure and morphology of CdSe quantum dots synthesized in a microreactor was studied. The flow rate of octylamine was varied from 0.005 ml/min to 0.030 ml/min, and the optical properties of the synthesized particles were analyzed by UV–vis and photoluminescence spectroscopy. The particle size of the quantum dots was found to increase with an increasing octylamine flow rate. Further, UV–vis and photoluminescence bands were found to be red-shifted with an increasing flow rate. We determined that, by controlling octylamine flow rate, the particle size of the quantum dots could be controlled. This method will help to determine the optimal octylamine flow conditions for synthesizing nanoparticles for use in a diverse range of applications.

Beschreibung: A novel Ce 3+ -doped Lu 3 MgAl 3 SiO 12 (LCMAS) phosphor was successfully synthesized by a conventional solid-state reaction. The crystal structure of the LCMAS was characterized by Rietveld refinement. The broad emission spectra of the LCMAS were observed at 550 nm, and the temperature dependence on photoluminescence properties of the LCMAS was investigated. Also, the activation energy for thermal quenching was determined by Arrhenius fitting. The experimental results clearly indicate that the LCMAS phosphor has a great potential for the down-conversion of yellow phosphor into white light-emitting diodes.

Beschreibung: Use of a liquid feedstock in thermal spraying (an alternative to the conventional solid powder feedstock) is receiving an increasing level of interest due to its capability to produce the advanced submicrometer/nanostructured coatings. Suspension plasma spraying (SPS) and solution precursor plasma spraying (SPPS) are those advanced thermal spraying techniques which help to feed this liquid feedstock. These techniques have shown to produce better performance thermal barrier coatings (TBCs) than conventional thermal spraying. In this work, a comparative study was performed between SPS- and SPPS-sprayed TBCs which then were also compared with the conventional atmospheric plasma-sprayed (APS) TBCs. Experimental characterization included SEM, porosity analysis using weight difference by water infiltration, thermal conductivity measurements using laser flash analysis, and lifetime assessment using thermo-cyclic fatigue test. It was concluded that SPS coatings can produce a microstructure with columnar type features (intermediary between the columnar and vertically cracked microstructure), whereas SPPS can produce vertically cracked microstructure. It was also shown that SPS coatings with particle size in suspension ( D 50 ) 〈3 μm were highly porous with lower thermal conductivity than SPPS and APS coatings. Furthermore, SPS coatings have also shown a relatively better thermal cyclic fatigue lifetime than SPPS.

Beschreibung: ABSTRACT Cholangiocarcinoma (CCAs) may be defined as tumors that derived from the biliary tree with the differentiation in the biliary epithelial cells. This tumor is malignant, extremely aggressive with a poor prognosis. It can be treated surgically and its pathogenesis is poorly understood. The tumor microenvironment (TME) is a very important factor in the regulation of tumor angiogenesis, invasion, and metastasis. Besides cancer stem cells (CSCs) can modulate tumor growth, stroma formation and migratory capability. The initial stage of tumorigenesis is characterized by genetic mutations and epigenetic alterations due to intrinsic factors which lead to the generation of oncogenes thus inducing tumorigenesis. CSCs may result from precancerous stem cells, cell de-differentiation, normal stem cells or an epithelial-mesenchymal transition (EMT). CSCs have been found in the cancer niche, and EMT may occur early within the tumor microenvironment. Previous studies have demonstrated evidence of cholangiocarcinoma stem cells (CD133, CD24, EpCAM, CD44, and others) and the presence of these markers has been associated with malignant potential. The interaction between TME and cholangiocarcinoma stem cells via signaling mediators may create an environment that accommodates tumor growth, yielding resistance to cytotoxic insults (chemotherarapeutic). While progress has been made in the understanding of the mechanisms, the interactions in the tumorigenic process still remain a major challenge. Our review, addresses recent concepts of TME-CSCs interaction and will emphasize the importance of early detection with the use of novel diagnostic mechanisms such as CCA-CSC biomarkers and the importance of tumor stroma to define new treatments. This article is protected by copyright. All rights reserved

Beschreibung: In this research, p-type Bi 2 Te 3 –75% Sb 2 Te 3 thermoelectric alloy powders were produced by gas atomization and subsequently sintered by hot pressing at different temperatures. The grain growth of the hot-pressed samples was observed with increasing sintering temperature from 380°C to 460°C. The compressive strength increased with increasing hot-pressing temperature due to the high relative density of bulk samples obtained at high temperatures. The effect of sintering temperature on thermoelectric (TE) properties was studied. The maximum power factor 3.48 mW/mK 2 was obtained for the sample hot pressed at 420°C due to the resulting high electrical conductivity and enhanced Seebeck coefficient values.

Beschreibung: Previous studies have shown that promyelocytic leukemia zinc finger (PLZF), chemokine (C-X-C motif) receptor 4 (CXCR4) and mir146a were associated with the self-renewal of mouse spermatogonial stem cells (SSCs); however, there is little information on their effects on the fate of livestock SSCs. Here, we have identified a regulatory pathway in dairy goat mGSCs, involving PLZF, mir146a and the SDF-1 receptor CXCR4. PLZF overexpression downregulated mir146a and simultaneously upregulated the expression of CXCR4 protein, whereas PLZF knockdown (siPLZF) induced the specifically opposite effects. The in vitro assays demonstrated that PLZF specifically interacts with and suppresses the mir146a promoter, and mir146a targets CXCR4 to impede its translation. The levels of ERK1/2 phosphorylation in the mGSCs overexpressed CXCR4 and PLZF were upregulated, respectively, whereas mir146a expression was decreased and CXCR4 protein was increased. Mir146a overexpression and siPLZF impaired mGSC proliferation and differentiation, however, Mir146a knockdown induced the opposite effects. The effects of PLZF and mir146a were mediated regulation by mir146a and CXCR4, respectively. Overexpression of CXCR4 or addition of CXCL12 in cultures of dairy goat mGSCs resulted in the upregulation of their signaling, and the phosphorylation of ERK1/2 was increased. Collectively, these findings indicate that PLZF is an important transcription factor in the regulation of the expression of CXCR4 to promote dairy goat mGSC proliferation by targeting mir146a. This article is protected by copyright. All rights reserved

Beschreibung: Bacterial lipopolysaccharide (LPS) is the most important contributing factor in pathogenesis of bacterial infection in male accessory glands; and it has shown to inhibit testicular steroidogenesis and induce apoptosis. The present study demonstrates that LPS causes mitochondrial dysfunction via suppression of sirtuin 4 (SIRT4); which in turn affects Leydig cell function by modulating steroidogenesis and apoptosis. LC-540 Leydig cells treated with LPS (10µg/ml) showed impaired steroidogenesis and increased cellular apoptosis. The mRNA and protein expression of SIRT4 were decreased in LPS treated cells when compared to controls. The obtained data suggest that the c-Jun N-terminal kinase (JNK) activation suppresses SIRT4 expression in LPS treated Leydig cells. Furthermore, the overexpression of SIRT4 prevented LPS induced impaired steroidogenesis and cellular apoptosis by improving mitochondrial function. These findings provide valuable information that SIRT4 regulates LPS mediated Leydig cell dysfunction. This article is protected by copyright. All rights reserved

Beschreibung: Ketamine, a dissociative anesthetic, is misused and abused worldwide as an illegal recreational drug. In addition to its neuropathic toxicity, ketamine abuse has numerous effects, including renal failure; however, the underlying mechanism is poorly understood. The process called epithelial phenotypic changes (EPCs) causes the loss of cell-cell adhesion and cell polarity in renal diseases, as well as the acquisition of migratory and invasive properties. Madin-Darby canine kidney cells, an in vitro cell model, were subjected to experimental manipulation to investigate whether ketamine could promote EPCs. Our data showed that ketamine dramatically decreased transepithelial electrical resistance and increased paracellular permeability and junction disruption, which were coupled to decreased levels of apical junctional proteins (ZO-1, Occludin and E-cadherin). Consistent with the downregulation of epithelial markers, the mesenchymal markers N-cadherin, Fibronectin and Vimentin were markedly upregulated following ketamine stimulation. Of the E-cadherin repressor complexes tested, the mRNA levels of Snail, Slug, Twist, and ZEB1 were elevated. Moreover, ketamine significantly enhanced migration and invasion. Ketamine-mediated changes were at least partly caused by the inhibition of GSK-3β activity through Ser-9 phosphorylation by the PI3K/Akt pathway. Inhibiting PI3K/Akt with LY294002 reactivated GSK-3β and suppressed ketamine-enhanced permeability, EPCs and motility. These findings were recapitulated by the inactivation of GSK-3β using the inhibitor 3F8. Taken together, these results provide evidence that ketamine induces renal distal tubular EPCs through the downregulation of several junction proteins, the upregulation of mesenchymal markers, the activation of Akt, and the inactivation of GSK-3β. This article is protected by copyright. All rights reserved

Beschreibung: CCN2/connective tissue growth factor (CTGF) is a multifunctional molecule that promotes harmonized development and regeneration of cartilage through its matricellular interaction with a variety of extracellular biomolecules. Thus, deficiency in CCN2 supply profoundly affects a variety of cellular activities including basic metabolism. A previous study showed that the expression of a number of ribosomal protein genes was markedly enhanced in Ccn2 -null chondrocytes. Therefore, in this study, we analyzed the impact of CCN2 on amino acid and protein metabolism in chondrocytes. Comparative metabolome analysis of the amino acids in Ccn2 -null and wild type mouse chondrocytes revealed stable decreases in the cellular levels of all of the essential amino acids. Unexpectedly, uptake of such amino acids was rather enhanced in Ccn2 -null chondrocytes, and the addition of exogenous CCN2 to human chondrocytic cells resulted in decreased amino acid uptake. However, as expected, amino acid consumption by protein synthesis was also accelerated in Ccn2 -null chondrocytes. Furthermore, we newly found that expression of 2 genes encoding 2 glycolytic enzymes, as well as the previously reported Eno 1 gene, was repressed in those cells. Considering the impaired glycolysis and retained mitochondrial membrane potential in Ccn2 -null chondrocytes, these findings suggest that Ccn2 deficiency induces amino acid shortage in chondrocytes by accelerated amino acid consumption through protein synthesis and acquisition of aerobic energy. Interestingly, CCN2 was found to capture such free amino acids in vitro . Under physiological conditions, CCN2 may be regulating the levels of free amino acids in the extracellular matrix of cartilage. This article is protected by copyright. All rights reserved

Beschreibung: ABSTRACT Some cord blood banks freeze entire pieces of UC (mixed cord, MC) which after post-thaw yields mixed heterogeneous populations of mesenchymal stem cells (MSCs) from all its microanatomical compartments. Freezing of such entire tissues results in sub-optimal post-thaw cell recovery because of poor cryoprotectant diffusion and intracellular ice-formation, heat and water transport issues and damage to intercellular junctions. To develop a simple method of harvesting pure homogeneous MSCs for cord blood banks we compared the post-thaw behavior of three groups of frozen UC tissues (i) freshly harvested WJ without cell separation, (ii) MSCs isolated from WJ (WJSC) and (iii) MC. WJ and WJSC produced high post-thaw cell survival rates (93.52 ± 6.12% to 90.83 ± 4.51%) and epithelioid monolayers within 24h in primary culture whereas post-thaw MC explants showed slow growth with mixed epithelioid and fibroblastic cell outgrowths after several days. Viability and proliferation rates of post-thawed WJ and hWJSC were significantly greater than MC. Post-thaw WJ and WJSC produced significantly greater CD24 + and CD108 + fluorescence intensities and significantly lower CD40 + contaminants. Post-thaw WJ and WJSC produced significantly lesser annexin-V-positive and sub-G1 cells and greater degrees of osteogenic and chondrogenic differentiation compared to MC. qRT-PCR analysis of post-thaw MC showed significant decreases in anti-apoptotic gene expression (SURVIVIN, BCL2) and increases in pro-apoptotic (BAX) and cell cycle regulator genes (P53, P21, ROCK 1) compared to WJ and WJSC. We conclude that freezing of fresh WJ is a simple and reliable method of generating large numbers of clinically utilizable MSCs for cell-based therapies. This article is protected by copyright. All rights reserved

Beschreibung: Curcumin-sensitized perovskite Bi 0.5 Na 0.5 TiO 3 (BNT) was synthesized for visible light photocatalytic decolorization of rhodamine 6G (Rh6G). Cube-shaped perovskite BNT was synthesized by solid-state route, and curcumin sensitization was performed using hydrothermal method. The surface sensitization of curcumin was analyzed by UV–Vis absorption spectra and FTIR. The degradation kinetics of Rh6G over Cur-BNT were investigated and discussed through first-order Langmuir–Hinshelwood kinetic model. The effect of catalyst dose on photocatalytic decolorization process was investigated. Photocatalysis was performed with different scavengers to investigate the role of active hole and radical species and photocatalytic degradation mechanism of Rh6G.

Beschreibung: The Wnt ligands are a family of secreted signaling proteins which play key roles in a number of cellular processes under physiological and pathological conditions. Wnts bind to their membrane receptors and initiate a signaling cascade which leads to the nuclear localization and transcriptional activity of β-catenin. The development of purified recombinant Wnt ligands has greatly aided in our understanding of Wnt signaling and its functions in development and disease. In the current study, we identified non-Wnt related signaling activities which were present in commercially available preparations of recombinant Wnt3a. Specifically, we found that treatment of cultured fibroblasts with recombinant Wnt3a induced immediate activation of TGF-β and BMP signaling and this activity appeared to be independent of the Wnt ligand itself. Therefore, while purified recombinant Wnt ligands continue to be a useful tool for studying this signaling pathway, one must exercise a degree of caution when analyzing the results of experiments that utilize purified recombinant Wnt ligands. This article is protected by copyright. All rights reserved

Beschreibung: We sought to explore the effects of doxorubicin on inflammatory profiles and energy metabolism in the hypothalamus of rats. To investigate these effects, we formed two groups: a control (C) group and a Doxorubicin (DOXO) group. Sixteen rats were randomly assigned to either the control (C) or DOXO groups. The hypothalamus was collected. The levels of interleukin (IL)-1β, IL-6, IL-10, TNF-α and energy metabolism (malate dehydrogenase, complex I and III activities) were analysed in the hypothalamus. The DOXO group exhibited a decreased body weight (p 〈 0.01). Hypothalamic malate dehydrogenase activity was reduced when compared with control (p 〈 0.05). In addition, pro-inflammatory cytokine levels were unchanged. Therefore, our results demonstrate that doxorubicin leads to an impairment of \hypothalamic energy metabolism, but do not affect the inflammatory pathway. Copyright © 2015 John Wiley & Sons, Ltd. Significance paragraph The hypothalamus is a central organ that regulates a great number of functions, such as food intake, temperature and energy expenditure, among others. Doxorubicin can lead to deep anorexia and metabolic chaos; thus, we observed the effect of this chemotherapeutic drug on the inflammation and metabolism in rats after the administration of doxorubicin in order to understand the central effect in the hypothalamus. Drug treatment by doxorubicin is used as a cancer therapy; however the use of this drug may cause harmful alterations to the metabolism. Thus, further investigations are needed on the impact of drug therapy over the long term.