ALBERT

Description: Waterborne diseases have emerged as global health problems and their rapid and sensitive detection in environmental water samples is of great importance. Bacterial identification and enumeration in water samples is significant as it helps to maintain safe drinking water for public consumption. Culture-based methods are laborious, time-consuming, and yield false-positive results, whereas viable but nonculturable (VBNCs) microorganisms cannot be recovered. Hence, numerous methods have been developed for rapid detection and quantification of waterborne pathogenic bacteria in water. These rapid methods can be classified into nucleic acid-based, immunology-based, and biosensor-based detection methods. This review summarizes the principle and current state of rapid methods for the monitoring and detection of waterborne bacterial pathogens. Rapid methods outlined are polymerase chain reaction (PCR), digital droplet PCR, real-time PCR, multiplex PCR, DNA microarray, Next-generation sequencing (pyrosequencing, Illumina technology and genomics), and fluorescence in situ hybridization that are categorized as nucleic acid-based methods. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence are classified into immunology-based methods. Optical, electrochemical, and mass-based biosensors are grouped into biosensor-based methods. Overall, these methods are sensitive, specific, time-effective, and important in prevention and diagnosis of waterborne bacterial diseases. Waterborne diseases pose constant threats to public health. Therefore, public healthcare needs rapid, sensitive, and accurate detection of pathogens for the diagnosis and treatment.

Description: Testicular germ cell tumors (TGCTs) are frequent solid malignant tumors and cause of death in men between 20–40 years of age. Genetic and environmental factors play an important role in the origin and development of TGCTs. Although the majority of TGCTs are responsive to chemotherapy, about 20% of patient presents incomplete response or tumors relapse. In addition, the current treatments cause acute toxicity and several chronic collateral effects, including sterility. The present mini-review collectively summarize the most recent findings on the new discovered molecular biomarkers such as tyrosine kinases, HMGAs, Aurora B kinase, and GPR30 receptor predictive of TGCTs and as emerging new possible molecular targets for therapeutic strategies. This article is protected by copyright. All rights reserved

Description: ABSTRACT Atrogin-1, an E3 ligase present in skeletal, cardiac and smooth muscle, down-regulates myocardin protein during skeletal muscle differentiation. Myocardin, the master regulator of smooth muscle cell (SMC) differentiation, induces expression of smooth muscle marker genes through its association with serum response factor (SRF), which binds to the CArG box in the promoter. Myocardin undergoes ubiquitylation and proteasomal degradation. Evidence suggests that proteasomal degradation of myocardin is critical for myocardin to exert its transcriptional activity, but there is no report about the E3 ligase responsible for myocardin ubiquitylation and subsequent transactivation. Here, we showed that overexpression of atrogin-1 increased contractility of cultured SMCs and mouse aortic tissues in organ culture. Overexpression of dominant-negative myocardin attenuated the increase in SMC contractility induced by atrogin-1. Atrogin-1 overexpression increased expression of the SM contractile markers while downregulated expression of myocardin protein but not mRNA. Atrogin-1 also ubiquitylated myocardin for proteasomal degradation in vascular SMCs. Deletion studies showed that atrogin-1 directly interacted with myocardin through its amino acids 284-345. Immunostaining studies showed nuclear localization of atrogin-1, myocardin and the Rpt6 subunit of the 26S proteasome. Atrogin-1 overexpression not only resulted in degradation of myocardin but also increased recruitment of RNA Polymerase II onto the promoters of myocardin target genes. In summary, our results have revealed the roles for atrogin-1 in the regulation of smooth muscle contractility through enhancement of myocardin ubiquitylation/degradation and its transcriptional activity. This article is protected by copyright. All rights reserved

Description: Cholesterol is an important component for cell physiology. It regulates the fluidity of cell membranes and determines the physical and biochemical properties of proteins. In the central nervous system, cholesterol controls synapse formation and function and supports the saltatory conduction of action potential. In recent years, the role of cholesterol in the brain has caught the attention of several research groups since a breakdown of cholesterol metabolism has been associated with different neurodevelopmental and neurodegenerative diseases, and interestingly also with psychiatric conditions. The aim of this review is to summarize the current knowledge about the connection between cholesterol dysregulation and various neurologic and psychiatric disorders based on clinical and preclinical studies. This article is protected by copyright. All rights reserved

Description: The study of cellular senescence and proliferative lifespan is becoming increasingly important because of the promises of autologous cell therapy, the need for model systems for tissue disease and the implication of senescent cell phenotypes in organismal disease states such as sarcopenia, diabetes and various cancers, among others. Here, we explain the concepts of proliferative cellular lifespan and cellular senescence, and we present factors that have been shown to mediate cellular lifespan positively or negatively. We review much recent literature and present potential molecular mechanisms by which lifespan mediation occurs, drawing from the fields of telomere biology, metabolism, NAD + and sirtuin biology, growth factor signaling and oxygen and antioxidants. We conclude that cellular lifespan and senescence are complex concepts that are governed by multiple independent and interdependent pathways, and that greater understanding of these pathways, their interactions and their convergence upon specific cellular phenotypes may lead to viable therapies for tissue regeneration and treatment of age-related pathologies, which are caused by or exacerbated by senescent cells in vivo. Replicative cellular lifespan is regulated by myriad cellular factors and processes, including telomeres, oxygen, DNA damage signaling, growth factors and metabolism. In this review, we will explain some of the molecular means by which these and other factors mediate cellular lifespan.

Description: Uncoupling proteins (UCPs) regulate mitochondrial function, and thus cellular metabolism. Angiotensin-converting enzyme (ACE) is the central component of endocrine and local tissue renin–angiotensin systems (RAS), which also regulate diverse aspects of whole-body metabolism and mitochondrial function (partly through altering mitochondrial UCP expression). We show that ACE expression also appears to be regulated by mitochondrial UCPs. In genetic analysis of two unrelated populations ( healthy young UK men and Scandinavian diabetic patients ) serum ACE (sACE) activity was significantly higher amongst UCP3-55C (rather than T) and UCP2 I (rather than D) allele carriers. RNA interference against UCP2 in human umbilical vein endothelial cells reduced UCP2 mRNA sixfold ( P  〈 0·01) whilst increasing ACE expression within a physiological range (〈1·8-fold at 48 h; P  〈 0·01). Our findings suggest novel hypotheses. Firstly, cellular feedback regulation may occur between UCPs and ACE. Secondly, cellular UCP regulation of sACE suggests a novel means of crosstalk between (and mutual regulation of) cellular and endocrine metabolism. This might partly explain the reduced risk of developing diabetes and metabolic syndrome with RAS antagonists and offer insight into the origins of cardiovascular disease in which UCPs and ACE both play a role. Uncoupling proteins (UCPs) regulate mitochondrial function, and thus cellular metabolism. Angiotensin-converting enzyme (ACE) is the central component of endocrine and local tissue renin–angiotensin systems, which also regulate diverse aspects of whole-body metabolism and mitochondrial function. We demonstrate that ACE expression appears to be regulated by mitochondrial UCPs.

Description: Positive transcription elongation factor (P-TEFb) plays an important role in host cell and viral gene expression. Many viruses, including Herpes Simplex Virus 1, have evolved strategies to hijack this key factor via their own regulatory proteins. The central role of P-TEFb in viral life cycles raises the possibility that Cdk9 inhibitors might be useful antiviral agents. See article “P-TEFb goes viral” by Justyna Zaborowska, Nur F. Isa and Shona Murphy in this issue.

Description: Tumors are often viewed as unique entities with specific behaviors. However, tumors are a mixture of differentially evolved subpopulations of cells in constant Darwinian evolution, selecting the fittest clone and allowing it to outgrow the rest. As in the natural environment, the niche defines the properties the fittest clones must possess. Therefore, there can be multiple fit clones because of the various microenvironments inside a single tumor. Hypoxia is considered to be a major feature of the tumor microenvironment and is a potential contributor to the cancer stem cell (CSC) phenotype and its enhanced tumorigenicity. The acidic microenvironment around hypoxic cells is accompanied by the activation of a subset of proteases that contribute to metastasis. Because of aberrant angiogenesis and the inaccessibility of their locations, hypoxic cells are less likely to accumulate therapeutic concentrations of chemotherapeutics that can lead to therapeutic resistance. Therefore, the targeting of the hypoxic CSC niche in combination with chemotherapy may provide a promising strategy for eradicating CSCs. In this review, we examine the cancer stem cell hypothesis and its relationship to the microenvironment, specifically to hypoxia and the subsequent metabolic switch and how they shape tumor behavior. Tumors are a mixture of differentially evolved subpopulations of cells in constant evolution. As in the natural environment, the niche defines the properties the fittest clones must possess. Therefore, there can be multiple fit clones because of the various microenvironments inside a single tumor. Hypoxia is considered to be a major feature of the tumor microenvironment and is a potential contributor to the cancer stem cell phenotype and its enhanced tumorigenicity.

Description: Publication date: Available online 18 July 2016 Source: Journal of Asian Ceramic Societies Author(s): J. Raj Mohamed, L. Amalraj The present work investigates the effect of precursor concentration ( m c ) on the structural, optical, morphological and electrical conductivity properties of In 2 S 3 thin films grown on amorphous glass substrates by nebulized spray pyrolysis (NSP) technique. The mixed phase of cubic and tetragonal structure of In 2 S 3 thin films at higher concentration has been observed by X-ray diffraction pattern. The reduced strain by increasing the precursor concentration increased the average crystallite from 17.8 to 28.9 nm. The energy dispersive analysis by X-ray (EDAX) studies confirmed the presence of In and S. The transmittance, optical direct band gap energy, Urbach energy and skin depth of In 2 S 3 films have been analyzed by optical absorption spectra. The better conductivity and mobility noticed at m c = 0.15 M are explained by carrier concentration and crystallite. Better optical and electrical conductivity behaviour of In 2 S 3 thin film sample proposes for effective solar cell fabrication.

Description: Eukaryotic gene expression is extensively controlled at the level of mRNA stability and the mechanisms underlying this regulation are markedly different from their archaeal and bacterial counterparts. We propose that two such mechanisms, nonsense-mediated decay (NMD) and motif-specific transcript destabilization by CCCH-type zinc finger RNA-binding proteins, originated as a part of cellular defense against RNA pathogens. These branches of the mRNA turnover pathway might have been used by primeval eukaryotes alongside RNA interference to distinguish their own messages from those of RNA viruses and retrotransposable elements. We further hypothesize that the subsequent advent of “professional” innate and adaptive immunity systems allowed NMD and the motif-triggered mechanisms to be efficiently repurposed for regulation of endogenous cellular transcripts. This scenario explains the rapid emergence of archetypical mRNA destabilization pathways in eukaryotes and argues that other aspects of post-transcriptional gene regulation in this lineage might have been derived through a similar exaptation route. mRNA turnover in eukaryotes is remarkably different from its prokaryotic counterparts and possible reasons underlying this divergence remain unclear. Here we propose that eukaryotic mRNA destabilization pathways evolved as a part of host defense against RNA pathogens and were subsequently repurposed for post-transcriptional regulation of cell-encoded genes.

Description: Photodynamic therapy (PDT) is a non-thermal technique for inducing tumour damage following administration of a light-activated photosensitizing drug (PS). In a previous work we found that PDT induces cytoskeleton changes in HB4a-Ras cells (human mammary breast carcinoma HB4a cells transfected with the RAS oncogene). In the present work we have studied the migratory and invasive features and the expression of proteins related to these processes on HB4a-Ras cells after 3 successive cycles of PDT using different PSs: 5-aminolevulinic acid (ALA), Verteporfin (Verte), m -tetrahydroxyphenylchlorin ( m -THPC) and Merocyanine 540 (MC). A slight (1.25- to -2 fold) degree of resistance was acquired in cell populations subjected to the three successive PDT treatments. However, complete cell killing was achieved after a light dose increase. Regardless of the PS employed, all the PDT-treated populations had shorter stress fibres than the untreated control HB4a-Ras cells, and the number of dorsal stress fibres was decreased in the PDT-treated populations. E-Cadherin distribution, which was already aberrant in HB4a-Ras cells, became even more diffuse in the PDT-treated populations, though its expression was increased in some of them. The strong migratory and invasive ability of HB4a-Ras cells in vitro was impaired in all the PDT-treated populations, with a behaviour that was similar to the parental non-tumoral HB4a cells. MMP-2 and MMP-9 metalloproteinase activities were also impaired in the PDT-treated populations. The evidence presented herein suggests that the cells surviving PDT would be less metastatic than the initial population. These findings encourage the use of PDT in combination with other treatments such as intraoperative or post-surgery therapeutic procedures. This article is protected by copyright. All rights reserved

Description: ABSTRACT Nestin (+) -cardiomyocytes were identified in the ischemically damaged human/rodent heart, albeit the cellular source and signaling events implicated in the appearance of the intermediate filament protein remained undefined. Expression of the enhanced green fluorescent protein (EGFP) driven by the second intron of the nestin gene identified a subpopulation of EGFP/nestin (+) cells that differentiated to a vascular phenotype in the peri-infarct/infarct region of post-MI mice albeit the transgene was not detected in nestin (+) -cardiomyocytes. α-MHC-driven expression of the reporter mCherry was detected in troponin-T (+) - and nestin (+) -cardiomyocytes in the peri-infarct/infarct region of post-MI mice. However, the cell cycle re-entry of nestin/mCherry (+) -cardiomyocytes was not observed. Nestin staining was identified in a paucity of neonatal rat ventricular cardiomyocytes (NNVM). Exposure to phorbol 12, 13-dibutyrate (PDBu) induced NNVM hypertrophy but did not promote nestin expression or Brdu incorporation. PDBu treatment of NNVMs phosphorylated p38 MAPK and HSP27 and HSP27 phosphorylation was abrogated by the p38 MAPK inhibitor SB203580. PDBu/SB203580 co-treatment significantly increased the percentage of NNVMs that expressed nestin and incorporated Brdu. In the heart of embryonic 10.5 day mice, nestin was detected in cycling troponin-T (+) -cardiomyocytes. Nestin was also detected in embryonic rat ventricular cardiomyocytes and depletion of the intermediate filament protein attenuated cell cycle re-entry. Thus, nestin expressed by pre-existing cardiomyocytes following ischemic damage recapitulated in part an embryonic phenotype and may provide the requisite phenotype to initiate cell cycle re-entry. However, the overt activation of the p38 MAPK pathway post-MI may in part limit the appearance and inhibit the cell cycle re-entry of nestin (+) -cardiomyocytes. This article is protected by copyright. All rights reserved

Description: Soma to germline inheritance of extrachromosomal genetic information. Non-Mendelian transgenerational inheritance is a growingly recognized phenomenon, yet still elusive in its molecular nature . On pages 726–733 of this issue, Corrado Spadafora proposes a model, whereby extrachromosomal genetic information released form somatic cells can cross the Weismann barrier and become internalized in epididymal spermatozoa, which thereafter mediate the acquisition of new traits in the offspring at fertilization. The sperm endogenous reverse transcriptase (RT) plays a key role in developmental control. Sperm cells therefore act as recipients, and at the same time transgenerational vectors, of somatically derived genetic information, which they pass to the next generation in a non chromosomally-integrated form, yet with the potential to modify the fate of the developing embryos.

Description: Beet Armyworm, Spodoptera exigua is a herbivorous moth and a serious pest of many economically important plants, which are used as food sources. Because of rigorous standards of food quality, usage of synthetic insecticides in crop protection, against pests, is limited. Solanaceae plant extracts may be a relatively cheap source of efficient natural insecticides that can limit usage of synthetic substances. Their biological activity is not fully known. In particular, ultrastructural studies, using transmission electron microscopy, are not usual. In the present article we describe the effects of sublethal concentrations of tomato and potato leaf extracts against S. exigua . Acute lethal effects were not observed. Both extracts exerted similar effects within midgut and fat body cells. Midgut cells were not significantly altered while fat body cells showed prominent swelling of nuclear envelope and endoplasmic reticulum, vacuolization of mitochondria and fusion of fat droplets. These changes were much more intensive within groups exposed to potato than tomato extracts at highest concentration at least. Light microscopy was used to observe and document developmental alterations of S. exigua exposed to potato and tomato leaf extracts. Potato leaf extracts significantly decreased hatching success and caused morphological malformations of imagoes. Among them, malformations of wings were the most prominent. Interestingly, these effects were not observed within populations exposed to tomato extracts at highest concentration at least.

Description: Dense (~98.5%), lithium aluminum silicate glass-ceramics were obtained via the sinter-crystallization of glass particle compacts at relatively low temperatures, that is, 790–875°C. The effect of P 2 O 5 on the glass-ceramics' sinter-crystallization behavior was evaluated. We found that P 2 O 5 does not modify the surface crystallization mechanism but instead delays the crystallization kinetics, which facilitates viscous flow sintering. Our glass-ceramics had virgilite (Li x Al x Si 3-x O 6 ; 0.5 〈 x 〈 1), a crystal size 〈1 μm, and a linear thermal expansion coefficient of 2.1 × 10 −6 °C −1 in the temperature range 40–500°C. The overall heat treatment to obtain these GCs was quite short, at ~25 min.

Description: The effect of postgrowth thermal annealing on ZnO/ZnCr 2 O 4 nanocomposites synthesized by hydrothermal method has been thoroughly investigated. XRD data have clearly revealed the transformation of spinel cubic phase of ZnCr 2 O 4 to wurtzite phase ZnO and indicated the incorporation of Cr in ZnO lattice with annealing at high temperatures. Photoluminescence spectra have shown a strong dependence of emission on annealing temperatures. This work demonstrated the unique and simple route to fabricate Cr-doped ZnO and tuning of the luminescence with annealing temperature. Thus, the work has immense potential for optoelectronic and photovoltaic applications.

Description: Alkaptonuria (AKU) is an ultra-rare autosomal genetic disorder caused by a defect in the activity of the enzyme homogentisate 1,2-dioxygenase (HGD) that leads to the accumulation of homogentisic acid (HGA) and its oxidized product, benzoquinone acetic acid (BQA), in the connective tissues causing a pigmentation called “ochronosis”. The consequent progressive formation of ochronotic aggregates generate a severe condition of oxidative stress and inflammation in all the affected areas. Experimental evidences have also proved the presence of serum amyloid A (SAA) in several AKU tissues and it allowed classifying AKU as a secondary amyloidosis. Although AKU is a multisystemic disease, the most affected system is the osteoarticular one and articular cartilage is the most damaged tissue. In this work, we have analyzed for the first time the cytoskeleton of AKU chondrocytes by means of immunofluorescence staining. We have shown the presence of SAA within AKU chondrocytes and finally we have demonstrated the co-localization of SAA with three cytoskeletal proteins: actin, vimentin and β-tubulin. Furthermore, in order to observe the ultrastructural features of AKU chondrocytes we have performed TEM analysis, focusing on the Golgi apparatus structure and, to demonstrate that pigmented areas in AKU cartilage are correspondent to areas of oxidation, 4-HNE presence has been evaluated by means of immunofluorescence. This article is protected by copyright. All rights reserved

Description: In this report, we present an analysis of several recycling protocols based on labeling of membrane proteins with specific monoclonal antibodies (mAbs). We analyzed recycling of membrane proteins that are internalized by clathrin-dependent endocytosis, represented by the transferrin receptor, and by clathrin-independent endocytosis, represented by the Major Histocompatibility Class I molecules. Cell surface membrane proteins were labeled with mAbs and recycling of mAb:protein complexes was determined by several approaches. Our study demonstrates that direct and indirect detection of recycled mAb:protein complexes at the cell surface underestimate the recycling pool, especially for clathrin-dependent membrane proteins that are rapidly reinternalized after recycling. Recycling protocols based on the capture of recycled mAb:protein complexes require the use of the Alexa Fluor 488 conjugated secondary antibodies or FITC-conjugated secondary antibodies in combination with inhibitors of endosomal acidification and degradation. Finally, protocols based on the capture of recycled proteins that are labeled with Alexa Fluor 488 conjugated primary antibodies and quenching of fluorescence by the anti-Alexa Fluor 488 displayed the same quantitative assessment of recycling as the antibody-capture protocols. This article is protected by copyright. All rights reserved

Description: ABSTRACT Mechanotransduction is a key process by which cells perceive extracellular mechanical cues / intercellular physical interactions and transform them into intracellular biochemical signals. This physiological process is crucial during bone development and bone remodeling throughout childhood and adult life, whereas several aberrations during this process have emerged as a distinct pathogenic molecular entity in bone maladies and tumor formation. The present review focuses on recent advances regarding the mechanobiology of osteosarcoma, the most common type of bone cancer. Special emphasis is given on the mechano-responsive signal transduction pathways underlying osteosarcoma pathology and on specific mechanosensitive molecules engaged in osteosarcoma development. This article is protected by copyright. All rights reserved

Description: ABSTRACT Protandim and 6-gingerol, two potent nutraceuticals, have been shown to decrease free radicals production through enhancing endogenous antioxidant enzymes. In this study, we evaluated the effects of these products on the expression of different factors involved in osteoarthritis (OA) process. Human OA chondrocytes were treated with 1 ng/ml IL-1β in the presence or absence of protandim (0-10 μg/ml) or 6-gingerol (0-10 μM). OA was induced surgically in mice by destabilization of the medial meniscus (DMM). The animals were treated weekly with an intraarticular injection of 10 μl of vehicle or protandim (10 μg/ml) for 8 weeks. Sham-operated mice served as controls. In vitro , we demonstrated that protandim and 6-gingerol preserve cell viability and mitochondrial metabolism and prevented 4-hydroxynonenal (HNE)-induced cell mortality. They activated Nrf2 transcription factor, abolished IL-1β-induced NO, PGE 2 , MMP-13, and HNE production as well as IL-β − induced GSTA4-4 down-regulation. Nrf2 overexpression reduced IL-1β-induced HNE and MMP-13 as well as IL-1β-induced GSTA4-4 down-regulation. Nrf2 knockdown following siRNA transfection abolished protandim protection against oxidative stress and catabolism. The activation of MAPK and NF-κB by IL-1β was not affected by 6-gingerol. In vivo , we observed that Nrf2 and GSTA4-4 expression was significantly lower in OA cartilage from humans and mice compared to normal controls. Interestingly, protandim administration reduced OA score in DMM mice. Altogether, our data indicate that protandim and 6-gingerol are essential in preserving cartilage and abolishing a number of factors known to be involved in OA pathogenesis. This article is protected by copyright. All rights reserved

Description: In the kidney, vitamin C is reabsorbed from the glomerular ultrafiltrate by sodium-vitamin C cotransporter isoform 1 (SVCT1) located in the brush border membrane of the proximal tubules. Although we know that vitamin C levels decrease with age, the adaptive physiological mechanisms used by the kidney for vitamin C reabsorption during aging remain unknown. In this study, we used an animal model of accelerated senescence (SAMP8 mice) to define the morphological alterations and aging-induced changes in the expression of vitamin C transporters in renal tissue. Aging induced significant morphological changes, such as periglomerular lymphocytic infiltrate and glomerular congestion, in the kidneys of SAMP8 mice, although no increase in collagen deposits was observed using 2-photon microscopy analysis and second harmonic generation. The most characteristic histological alteration was the dilation of intracellular spaces in the basolateral region of proximal tubule epithelial cells. Furthermore, a combination of laser microdissection, qRT-PCR and immunohistochemical analyses allowed us to determine that SVCT1 expression specifically increased in the proximal tubules from the outer strip of the outer medulla (segment S3) and cortex (segment S2) during aging and that these tubules also express GLUT1. We conclude that aging modulates vitamin C transporter expression and that renal over-expression of SVCT1 enhances vitamin C reabsorption in aged animals that may synthesize less vitamin C. This article is protected by copyright. All rights reserved

Description: ABSTRACT Epithelial morphogenesis in the mammary gland proceeds as a consequence of complex cell behaviors including apoptotic cell death and epithelial-mesenchymal transition (EMT); the extracellular matrix (ECM) protein laminin is crucially involved. Syntaxins mediate intracellular vesicular fusion, yet certain plasmalemmal members have been shown to possess latent extracellular functions. In this study, the extracellular subpopulation of syntaxin-4, extruded in response to the induction of differentiation or apoptosis in mammary epithelial cells, was detected. Using a tetracycline-repressive transcriptional system and clonal mammary epithelial cells, SCp2, we found that the expression of cell surface syntaxin-4 elicits EMT-like cell behaviors. Intriguingly, these cells did not up-regulate key transcription factors associated with the canonical EMT such as snail, slug , or twist , and repressed translation of E-cadherin. Concurrently, the cells completely evaded the cellular aggregation/rounding triggered by a potent EMT blocker laminin-111. We found that the recombinant form of syntaxin-4 not only bound to laminin but also latched onto the glycosaminoglycan (GAG) side chains of syndecan-1, a laminin receptor that mediates epithelial morphogenesis. Thus, temporal extracellular extrusion of syntaxin-4 emerged as a novel regulatory element for laminin-induced mammary epithelial cell behaviors. This article is protected by copyright. All rights reserved

Description: Glial cell line-derived neurotrophic factor (GDNF) is known to mediate multiple biological activities such as promotion of cell motility and proliferation, and morphogenesis. However, little is known about its effects on periodontal ligament (PDL) cells. Recently, we reported that GDNF expression is increased in wounded rat PDL tissue and human PDL cells (HPDLCs) treated with proinflammatory cytokines. Here, we investigated the associated expression of GDNF and the proinflammatory cytokine interleukin-1 beta (IL-1β) in wounded PDL tissue, and whether HPDLCs secrete GDNF which affects neurocytic differentiation. Rat PDL cells near the wounded area showed intense immunoreactions against an anti-GDNF antibody, where immunoreactivity was also increased against an anti-IL-1β antibody. Compared with untreated cells, HPDLCs treated with IL-1β or tumor necrosis factor-alpha showed an increase in the secretion of GDNF protein. Conditioned medium of IL-1β-treated HPDLCs (IL-1β-CM) increased neurite outgrowth of PC12 rat adrenal pheochromocytoma cells. The expression levels of two neural regeneration-associated genes, growth-associated protein-43 (Gap-43) and small proline-rich repeat protein 1A (Sprr1A), were also upregulated in IL-1β-CM-treated PC12 cells. These stimulatory effects of IL-1β-CM were significantly inhibited by a neutralizing antibody against GDNF. In addition, U0126, a MEK inhibitor, inhibited GDNF-induced neurite outgrowth of PC12 cells. These findings suggest that an increase of GDNF in wounded PDL tissue might play an important role in neural regeneration probably via the MEK/ERK signaling pathway. This article is protected by copyright. All rights reserved

Description: Electrochemotherapy (ECT) is a medical strategy that allows an increased efficacy of chemotherapy agents after the application of permeabilizing electric pulses having appropriate characteristics (form, voltage, frequency). In the past ten years, the clinical efficacy of this therapeutic approach in several spontaneous models of tumors in animals has been shown. Moreover, some of the molecular and cellular mechanisms responsible for this phenomenon have been elucidated. Our group has been deeply involved in the development of new ECT protocols for companion animals, implementing the use of the technique as first line treatment, and evaluating different chemotherapy agents in laboratory animals as well as pets. This article summarizes the most important advances in veterinary ECT, including the development of novel equipment, therapeutic protocols and their translation to humans. This article is protected by copyright. All rights reserved

Description: Analyzing the temperature evolution in pressureless mold-assisted flash sintering, we found the same onset condition as in standard flash sintering: When sample's DC or AC Joule heating replaces environment's radiation heating as the dominant power input term, thermal runaway ensues. Various serial and parallel components connected to the sample, including the mold, insulation, and punches, can affect Joule heating and conduction heat loss, thus play an important role in successful mold-assisted flash sintering.

Description: This work reports the processing steps of Al 2 O 3 (1–5 vol%) nanoparticulate ( d V.50 = 13 nm) LZS glass–ceramic matrix (19.58Li 2 O·11.10ZrO 2 ·69.32SiO 2 , mol%, d v.50 = 3.5 μm) composites for production of multilayered materials with thermal expansion gradients obtained by tape casting. Suspensions were prepared in water to solids contents ranging from 40 to 47 vol% using ammonium polyacrylate as a deflocculant, and an acrylic copolymer and polyvinyl alcohol as binders. Optimum performance was achieved by sonication and controlling the rheological properties for every step of the process. To prepare the composites, different concentrations (1, 2.5 and 5 vol%) of nanoalumina were added to fresh, as-prepared LZS suspensions, by changing the solid contents as required to maintain similar viscosities. Green tapes with high uniformity, without macroscopic defects and easy to handle were sintered to relative densities between 89% and 94%. Dense and homogeneous laminates with gradual composition with increasing concentrations of alumina were obtained.

Description: A new lead-free perovskite solid solution (1− x )BaTiO 3 – x Bi(Mg 1/2 Zr 1/2 )O 3 with morphotropic phase boundary (MPB) has been developed, and its structural and dielectric properties have been investigated. Rietveld structural analysis of the X-ray diffraction data suggest a composition-dependent tetragonal ( P 4 mm ) to cubic ( ) phase transition with an intermediate, phase coexistence region, demarcating the MPB. The compositions with x ≤ 0.05 are tetragonal in the P 4 mm space group and the compositions with x ≥ 0.25 are cubic in the space group. Coexistence of monoclinic phase (space group Cm ) with tetragonal/cubic phase (space group P 4 mm / ) is observed in the MPB region for the compositions with 0.10 ≤ x ≤ 0.22. The temperature dependence of permittivity exhibits a nonrelaxor type diffuse phase transition for all the compositions across the MPB.

Description: The amorphous silica (a-SiO 2 ) and germania (a-GeO 2 ) have a wide range of applications in glass industry. Based on a previously constructed near-perfect continuous random network model with 1296 atoms and periodic boundary conditions, we extend our study to amorphous Si 1− x Ge x O 2 models of homogeneous random substitution of Si by Ge with x ranging from 0 to 1. We have calculated the structural, electronic, mechanical, and optical properties for the series by using the first-principles density functional theory methods. The x -dependence of the variations in the properties is analyzed and critically compared with available experimental data. The mass density, volume, total bond order density, bulk mechanical properties, and refractive index are found to vary linearly as a function of x . For x = 0.5, we have also constructed six different kinds of particle immersion models to test the effect of inclusion of spherical particles of one glass of different sizes in the medium of the other glass on their physical properties. It is shown that particle sizes do affect the properties of particle immersion. Our calculations provide deep insight on the properties of mixture and nanocomposites of a-SiO 2 and a-GeO 2 glasses.

Description: Hertzian testing is applied to obtain flaw distributions in two fusion-drawn glasses and two glass-ceramics. A tungsten carbide sphere (diameter either 1.0, 2.5, or 5.0 mm) was used to produce surface cracks (ring cracks and cone cracks). Two theoretical approaches were employed to describe the data. Both approaches are only descriptive for very high strength materials in which the surface flaw sizes are small (e.g., 〈1 μm). In the first, a Weibull distribution for strength was assumed, and an expression for the probability of fracture was derived based on the stress field around the indent contact area. The unique aspect of this is that the stress field used includes material that has been “probed” at loads below the fracture load. A Weibull plot with this expression shows a slope of m + 2, where m is the conventional Weibull modulus. For the four different materials, the Weibull modulus varied between 8.0 for β-quartz glass-ceramic to 14.2 for fusion drawn alumni silicate glass. The second theoretical approach employs a modification of the method of Poloniecki and Wilshaw (the PW Method) to describe the distributions of very small flaws. The modification removes the need to bin the flaw distribution data. The modified PW Method revealed distinct differences in the flaw distributions between the four materials. These differences are consistent with the different Weibull moduli determined by ranking the different materials according to flaw size. However, Hertzian testing only probes relatively small flaw sizes and thus may differ from typical tensile or bending tests; nevertheless, the method should be applicable for extremely high strength materials.

Description: A strain-induced structural phase transition and its subsequent effect on the piezoelectric properties of BZT-BCT– x CeO 2 ( x  = 0–0.1 wt%) ceramics have been reported in this manuscript. BZT-BCT– x  wt% CeO 2 lead-free piezoceramics were successfully been prepared using sol–gel method. X-ray diffraction spectra of BZT-BCT– x  wt% CeO 2 ( x  = 0–0.10 wt%) ceramics showed a purely single-phase perovskite structure and a change in crystal structure between tetragonal and rhombohedral phases with increasing CeO 2 content. The strain-induced structural phase transition has resulted in excellent piezoelectric properties, d 33  = 673 pC/N, k p ~ 56%, and T c  = 110°C for BZT-BCT–0.08 wt% CeO 2 ceramics. Greatly improved temperature stability of the piezoelectric properties has been observed over a temperature range between 20°C and 90°C. The ferroelectric–paraelectric transition temperature, T c obtained in this study is 17°C higher than those reported earlier.

Description: The oxygen nonstoichiometry of large oxygen-deficient Ruddlesden–Popper oxides La x Sr 3− x Fe 2 O 7−δ (LSFO7- x ) ( x  = 0, 0.25, 0.5) was measured by the high-temperature gravimetry and the coulometric titration. In the composition series, the P (O 2 ) dependencies exhibited typical plateaus at δ = (2−[ ])/2. Meanwhile, La 0.5 Sr 2.5 Fe 2 O 7−δ showed the smallest oxygen nonstoichiometry and was the most thermochemically stable compound against P (O 2 ), temperature, and the La content. Based on the defect equilibrium model and the statistical thermodynamic calculation derived oxygen nonstoichiometric data, the substitution of La for Sr-site can promote the forward reaction of oxygen incorporation, the backward reaction of the disproportionation of the charge carriers, and oxygen redistribution between the O1 and O3 sites, resulting in the reduction of oxygen-deficient and the lower decomposition P (O 2 ). The obtained thermodynamic quantities of the partial molar enthalpy of oxygen, , and the partial molar entropy of oxygen, , calculated from the statistical thermodynamic calculation are in good agreement with those using the Gibbs–Helmholtz equation.

Description: Microstress in reaction-bonded silicon carbide (RBSiC) has been measured using piezo-Raman spectroscopy. Compressive microstresses as high as 2 GPa exist in the silicon phase and tensile microstresses as high as 2.3 GPa exist in the SiC phase of RBSiC. This is much larger than expected for thermoelastic microstress from coefficient of thermal expansion mismatch would provide. Instead the microstresses arise from the crystallization of liquid silicon. During the reaction bonding process, not all of the silicon reacts to form SiC and there is liquid free silicon. The phase transformation of the free silicon from liquid to solid has a large volume expansion, which results in large residual microstress within the silicon and SiC phases of RBSiC.

Description: The single crystal solid-state conversion of fluorapatite-type Sr 5 (PO 4 ) 3 F (Sr-FAP) has been achieved by spark plasma sintering with the assistance of NaF additive. NaF was determined to act as both a sintering aid and impurity solute along the grain boundaries (GBs), controlling both the space charge and GB migration rate. Postsintering isothermal annealing was performed to examine the effect of DC electric field on grain growth. From the space charge potential determined from impedance spectra measurements, in combination with the theoretical contribution of space charge to grain-boundary energy reduction, it was concluded that the magnitude of the space charge in Sr-FAP is temperature dependent. As such, a moderate decrease in polycrystalline GB driving force is the main cause for the increased single crystal migration length that was observed in this study.

Description: Surface ultrastructure of the gills of the angler catfish Chaca chaca was investigated to unravel the adaptive modifications associated with the feeding ecology of the fish. The fish is often found in mud or in soft substrates where they remain buried both for protection and to feed. Gill rakers present on the gill arch in most fish species are absent in this fish. The absence of gill rakers are associated with the feeding habit of the fish and is considered to facilitate the swallowing of captured prey smoothly without any hindrance. Highly corrugated surface of the gill arch and gill filaments could be associated to retain water/mucus to prevent dessicassion of the fish. Papillae like epithelial protuberances each bearing a taste bud at its summit toward the pharyngeal side of the gill arch is associated with the sorting of the food. Large number of mucous goblet cells on the gill arch epithelium are considered to secret copious mucus to lubricate the prey for easy swallowing. In C. chaca the gill septa between gill filaments are reduced. This could enhance the flexibility and permit the free movement of the gill filaments. Extensive secondary lamellae and infrequent mucous goblet cells on secondary lamellae are associated to increase the surface area to enhance efficiency of gaseous exchange.

Description: Phase relationships of the CaO–ZnO system in equilibrium with air ( = 0.21 atm) have been studied for the first time using the equilibration/quenching/EPMA technique in the temperature range from 1000°C to 1600°C. In this study, the mutual solubility boundaries of the CaO and ZnO phases have been determined. Both CaO and ZnO have obvious solubilities in each other. The maximum solubility of ZnO in solid CaO phase is 23.21 at.% at 1535°C, and ZnO solid phase can take up to 4.81 at.% CaO at 1535°C. The eutectic point and parts of the liquidus lines have also been determined. The eutectic point was found to be at 1535°C ± 2°C and 0.6612 mole fraction of ZnO. No compound was found in this system, and there were two primary phases: halite-structured CaO (lime) and wurtzite-structured ZnO (zincite). Results from this study were compared with the CaO–ZnO phase diagram calculated by MTDATA 5.10 and its MTOX 8.1 database. The difference was significant, and the thermodynamic description of the system requires a reassessment.

Description: In the present research, different adhesive techniques were used to set up fillings with composite resins. After the application of etch and rinse or self etch adhesive technique, marginal adaptation of composite fillings was estimated by the length of margins without gaps, and by the microretention of resin in enamel and dentin. The study material consisted of 40 extracted teeth. Twenty Class V cavities were treated with 35% phosphorous acid and restored after rinsing by Adper Single Bond 2 and Filtek Ultimate - ASB/FU 3M ESPE composite system. The remaining 20 cavities were restored by Adper Easy One - AEO/FU 3M ESPE composite system. Marginal adaptation of composite fillings was examined using a scanning electron microscope (SEM). The etch and rinse adhesive technique showed a significantly higher percentage of margin length without gaps (in enamel: 92.5%, in dentin: 57.3%), compared with the self-etch technique with lower percentage of margin length without gaps, in enamel 70.4% ( p  〈 .001), and in dentin-22.6% ( p  〈 .05). In the first technique, microretention was composed of adhesive and hybrid layers as well as resin tugs in interprismatic spaces of enamel, while the dentin microretention was composed of adhesive and hybrid layers with resin tugs in dentin canals. In the second technique, resin tugs were rarely seen and a microgap was dominant along the border of restoration margins. The SEM analysis showed a better marginal adaptation of composite resin to enamel and dentin with better microretention when the etch and rinse adhesive procedure was applied.

Description: The purpose of this study was to compare total-etch, self-etch, and selective etching techniques on the marginal microleakage of Class V composite restorations prepared by Er:YAG laser and bur. Class V cavities prepared on both buccal and lingual surfaces of 30 premolars by Er:YAG laser or bur and divided into six groups. The occlusal margins were in enamel, and the cervical margins were in cementum. Group-1: bur preparation(bp)+Adper Single Bond 2 (ASB)+Filtek Z550 (FZ); Group-2: laser preparation(lp)+(ASB)+(FZ); Group-3: bp + Clearfil S3 Bond Plus (CSBP)+(FZ); Group-4: lp+(CSBP) (FZ); Group-5: bp + acid etching+(CSBP)+(FZ); Group-6: lp + acid etching+(CSBP)+(FZ). All teeth were stored in distilled water at 37°C for 24 hr, and then thermocycled 1000 times (5–55°C). Five teeth from each group were chosen for the microleakage investigation, and two teeth for the scanning electron microscope evaluation. Teeth which were prepared for the microleakage test were immersed in .5% methylene blue dye for 24 hr. After immersion, the teeth were sectioned and observed under a stereomicroscope for dye penetration. Data were analyzed using Kruskal–Wallis and Mann–Whitney U tests ( p  〈 .05). More microleakage was observed in the cervical regions compared to the occlusal regions in Groups 3, 5, and 6, respectively ( p  〈 .05). There is no statistically significant difference in Groups 1, 2, and 4, in terms of cervical regions versus occlusal regions ( p  〉 .05). No significant differences were observed among any groups in terms of occlusal and cervical surfaces, separately ( p  〉 .05). Different etching techniques did not influence microleakage of Class V restorations prepared by Er:YAG laser and bur.

Description: The method of pulsed electric current sintering (PECS) has been used to obtain dense boron carbide (B 4 C) and B 4 C-based composite materials containing tungsten boride (W 2 B 5 ). To elucidate the role of the sintering additives and the mechanism of reactive densification, three types of materials have been obtained by PECS at 1850°C and 1900°C: “pure” B 4 C, B 4 C doped with 10 wt% W 2 B 5 , and B 4 C doped with 10 wt% tungsten carbide (WC). X-ray diffraction and X-ray photoelectron spectroscopy have been used to determine crystallite size, phase changes, and the peculiarities of the chemical bonds of the densified materials. Structural and mechanical properties of the materials have been investigated using scanning electron microscopy, optical microscopy, ultrasound velocity measurements, and hardness tests. The electrochemical impedance spectra have been used to investigate the electrical properties of the PECS-ed materials.

Description: Forsterite powders with controllable morphology were synthesized using oxides as raw materials in NaCl–KCl molten salt media. The effects of MgO/SiO 2 ratio, calcining temperature, and salt/oxide ratio on the phase composition and morphology of the powders are investigated. The results indicate that single-phase forsterite powders can be synthesized from a mixture of MgO and SiO 2 with a MgO/SiO 2 molar ratio of 2:1.3 at 700°C. With the increase in calcining temperature, the powders obtained changes from an irregular to a columnar morphology. In addition, the morphology of the forsterite powders produced can also be controlled by altering the salt/oxide ratio.

Description: This work investigated the effect of MnO 2 addition on the phase structure, microstructure, and electrical properties of AgSbO 3 -modified (Li,K,Na)(Nb,Ta)O 3 (abbreviated as LKNNT-AS) lead-free piezoelectric ceramics with an optimized composition endowed with a state of two-phase coexistence. A small amount (0.1 wt%) of MnO 2 can significantly further enhance the piezoelectric property of LKNNT-AS ceramics, whose piezoelectric constant d 33 and converse piezoelectric coefficient d 33 * as well as planar electromechanical coupling factor k p reach 363 pC/N, 543 pm/V, and 0.49, respectively. The temperature stability of piezoelectricity in MnO 2 -modified LKNNT-AS samples also improved, which is associated with the more uniform and better thermally stable ferroelectric domains that were revealed by piezoresponse force microscopy.

Description: Protein–membrane interactions are still an important topic of investigation. One of the suitable experimental techniques used by the scientific community to address such question is atomic force microscopy. In a previous work, we have reported that the binding mechanism between the cytolytic and antimicrobial protein (Cyt2Aa2) and lipid/cholesterol bilayers was concentration-dependent, leading to either the formation of holes in the bilayer or aggregates. Here we study such binding mechanism as a function of time at low protein concentrations (10 µg/mL). We demonstrate that although holes are formed during the first stages of the protein–lipid interaction, a reparation process due to molecular mobility in the bilayer leads to a homogenous and isotropic protein–lipid/cholesterol layer within 3 hr. The combination of imaging, force spectroscopy, and phase contrast delivered information about topography dynamics (molecular mobility), layer thickness, and mechanical properties of the protein–lipid/cholesterol system. These results highlight the importance of the observation time in (such type of) protein–lipid interactions (at low protein concentrations).

Description: Atomic force microscopy (AFM) is an enormous tool to observe nature in highest resolution and understand fundamental processes like friction and tribology on the nanoscale. Atomic resolution in highest quality was possible only in well-controlled environments like ultrahigh vacuum (UHV) or controlled buffer environments (liquid conditions) and more specified for long-term high-resolution analysis at low temperatures (∼4 K) in UHV where drift is nearly completely absent. Atomic resolution in these environments is possible and is widely used. However, in uncontrolled environments like air, with all its pollutants and aerosols, unspecified thin liquid films as thin as a single molecular water-layer of 200 pm or thicker condensation films with thicknesses up to hundred nanometer, have been a problem for highest resolution since the invention of the AFM. The goal of true atomic resolution on hydrophilic as well as hydrophobic samples was reached recently. In this manuscript we want to review the concept of ambient AFM with atomic resolution. The reader will be introduced to the phenomenology in ambient conditions and the problems will be explained and analyzed while a method for scan parameter optimization will be explained. Recently developed concepts and techniques how to reach atomic resolution in air and ultra-thin liquid films will be shown and explained in detail, using several examples.

Description: The conventional approach to developing disease-modifying treatments for neurodegenerative conditions has been to identify drivers of pathology and inhibit such pathways. Here we discuss the possibility that the efficacy of such approaches may be increasingly attenuated as disease progresses. This is based on experiments using mouse models of spinocerebellar ataxia type 1 and Huntington's disease (HD), where expression of the dominantly acting mutations could be switched off, as well as studies in human HD, which suggest that the primary genetic driver of age-of-onset of disease is a much weaker determinant of disease progression in affected individuals. The idea that one may approach a point in the disease course where such rational therapeutic strategies based on targets which determine onset of disease have minimal efficacy, suggests that one needs to consider other approaches to therapies and clinical trial design, including initiation of therapies in presymptomatic individuals. Different factors may determine the onset of a neurodegenerative disease versus its progression. Thus, treatments aiming to slow progression based on targets which determine onset may have less efficacy with increased disease duration.

Description: The visualization of taxonomically diagnostic features of individual pollen grains can be a challenge for many ecologically and phylogenetically important pollen types. The resolution of traditional optical microscopy is limited by the diffraction of light (250 nm), while high resolution tools such as electron microscopy are limited by laborious preparation and imaging workflows. Airyscan confocal superresolution and structured illumination superresolution (SR-SIM) microscopy are powerful new tools for the study of nanoscale pollen morphology and three-dimensional structure that can overcome these basic limitations. This study demonstrates their utility in capturing morphological details below the diffraction limit of light. Using three distinct pollen morphotypes ( Croton hirtus, Dactylis glomerata , and Helianthus sp.) and contrast-enhancing fluorescent staining, we were able to assess the effectiveness of the Airyscan and SR-SIM. We further demonstrate that these new superresolution methods can be easily applied to the study of fossil pollen material.

Description: Segmentation of objects from a noisy and complex image is still a challenging task that needs to be addressed. This article proposed a new method to detect and segment nuclei to determine whether they are malignant or not (determination of the region of interest, noise removal, enhance the image, candidate detection is employed on the centroid transform to evaluate the centroid of each object, the level set [LS] is applied to segment the nuclei). The proposed method consists of three main stages: preprocessing, seed detection, and segmentation. Preprocessing stage involves the preparation of the image conditions to ensure that they meet the segmentation requirements. Seed detection detects the seed point to be used in the segmentation stage, which refers to the process of segmenting the nuclei using the LS method. In this research work, 58 H&E breast cancer images from the UCSB Bio-Segmentation Benchmark dataset are evaluated. The proposed method reveals the high performance and accuracy in comparison to the techniques reported in literature. The experimental results are also harmonized with the ground truth images.

Description: Herpes simplex viruses can cause uncommon systemic complications as acute liver failure or urinary tract dysfunctions. Diphenyl diselenide, (PhSe) 2 , a classical studied organic selenium compound, has a novel antiviral action against HSV-2 infection and well-known antioxidant and anti-inflammatory properties. This study aimed to investigate if (PhSe) 2 reduces oxidative stress and systemic toxicity caused by HSV-2 infection in mice. Adult BALB/c mice were pre-treated with (PhSe) 2 (5 mg kg −1 /day, intragastric, i.g.) during 5 days; at day 6 mice were infected with HSV-2 (10 µl-10 5 PFU/ml −1 ) and post-treated with (PhSe) 2 for more 5 days. At day 11, they were killed and samples of liver and kidney were obtained to determine: reactive species (RS); malondialdehyde (MDA) and non-protein thiols (NPSH) levels; the activities of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT). The activities of adenosine deaminase (ADA), Na + /K + -ATPase (liver and kidney); alanine aminotransferase (ALT), aspartate aminotransferase (AST) and the levels of urea (plasma) were determined as markers of hepatic and renal toxicity. The results revealed that (PhSe) 2 treatment was effective against the increase of renal and hepatic oxidative stress in infected mice and also normalized hepatic and renal ADA activity. It recovered the activity of Na + /K + - and was not effective against the increase in urea levels in infected mice. Different from (PhSe) 2 , acyclovir (positive control), caused an increase in ADA activity and a decrease in hepatic CAT activity. Considering the interest of alternative therapies to treat HSV-2 infections and secondary complications, (PhSe) 2 become a notable candidate. This article is protected by copyright. All rights reserved

Description: ABSTRACT Conjugated linoleic acid (CLA) has been reported to improve muscle hypertrophy, steroidogenesis, physical activity, and endurance capacity in mice, although the molecular mechanisms of its actions are not completely understood. The aim of the present study was to identify whether CLA alters the expression of any of the peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) isoforms, and to evaluate the possible existence of fibre-type-specific hypertrophy in the gastrocnemius and plantaris muscles. Mice were randomly assigned to one of four groups: placebo sedentary, CLA sedentary, placebo trained, or CLA trained. The CLA groups were gavaged with 35 µl per day of Tonalin® FFA 80 food supplement containing CLA throughout the 6-week experimental period, whereas the placebo groups were gavaged with 35 µl sunflower oil each day. Each administered dose of CLA corresponded to approximately 0.7 g/kg or 0.5%, of the dietary daily intake. Trained groups ran 5 days per week on a Rota-Rod for 6 weeks at increasing speeds and durations. Mice were sacrificed by cervical dislocation and hind limb posterior muscle groups were dissected and used for histological and molecular analyses. Endurance training stimulated mitochondrial biogenesis by PGC1α isoforms (tot, α1, α2 and α3) but CLA supplementation did not stimulate PGC1α isoforms or mitochondrial biogenesis in trained or sedentary mice. In the plantaris muscle, CLA supplementation induced a fibre-type-specific hypertrophy of type IIx muscle fibres, which was associated with increased capillary density and was different from the fibre-type-specific hypertrophy induced by endurance exercise (of type I and IIb muscle fibres). This article is protected by copyright. All rights reserved

Description: Diclofenac is the most commonly used phenylacetic acid derivative non-steroidal anti-inflammatory drug (NSAID) that demonstrates significant analgesic, antipyretic, and anti-inflammatory effects. Several epidemiological studies have demonstrated anti-proliferative activity of NSAIDs and examined their apoptotic induction effects in different cancer cell lines. However, the precise molecular mechanisms by which these pharmacological agents induce apoptosis and exert anti-carcinogenic properties are not well known. Here, we have observed that diclofenac treatment induces proteasome malfunction and promotes accumulation of different critical proteasome substrates, including few pro-apoptotic proteins in cells. Exposure of diclofenac consequently elevates aggregation of various ubiquitylated misfolded proteins. Finally, we have shown that diclofenac treatment promotes apoptosis in cells, which could be because of mitochondrial membrane depolarization and cytochrome c release into cytosol. This study suggests possible beneficial insights of NSAIDs-induced apoptosis that may improve our existing knowledge in anti-proliferative interspecific strategies development. This article is protected by copyright. All rights reserved

Description: Diabetic macular edema (DME), characterized by an increase of thickness in the eye macular area, is due to breakdown of the blood-retinal barrier (BRB). Hypoxia plays a key role in the progression of this pathology by activating the hypoxia-inducible factors. In the last years, various studies have put their attention on the role of pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) in retinal dysfunction. However, until now, no study has investigated their protective role against the harmful combined effect of both hyperglycemia and hypoxia on outer BRB. Therefore, in the present study, we have analyzed the role of these peptides on permeability, restoration of tight junctions expression and inhibition of hyperglycemia/hypoxia-induced apoptosis, in an experimental in vitro model of outer BRB. Our results have demonstrated that the peptides' treatment have restored the integrity of outer BRB induced by cell exposure to hyperglycemia/hypoxia. Their effect is mediated through the activation of phosphoinositide 3 kinase (PI3K)/Akt and mammalian mitogen activated protein kinase/Erk kinase (MAPK/ERK) signaling pathways. In conclusion, our study further clarifies the mechanism through which PACAP and VIP perform the beneficial effect on retinal damage induced by hyperglycemic/hypoxic insult, responsible of DME progression. This article is protected by copyright. All rights reserved

Description: Crystalline pure NaTi 2 (PO 4 ) 3 (NTP) powder was synthesized at 700°C using a simple and low energy, hybrid inorganic–organic, steric entrapment method. Sodium nitrite (NaNO 2 ) and ammonium phosphate dibasic ((NH 4 ) 2 HPO 4 ) dissolved in water, whereas titanium (IV) isopropoxide (Ti[OCH(CH 3 ) 2 ] 4 ) hydrolyzed in water. Ethylene glycol (HOCH 2 CH 2 OH) was used as a polymeric entrapper and hydrolysis of the Ti source was hindered by its dissolution in isopropyl alcohol. The resulting NTP powder was characterized by thermogravimetric analysis/differential thermal analysis, X-ray diffractometry, scanning electron microscopy, specific surface area by Brunauer–Emmett–Teller nitrogen absorption, and particle size analysis. Furthermore, C, H, N were measured by the classical Pregl-Dumas method. The thermal expansion behavior in all { hkl } pole directions was also determined by in situ high-temperature X-ray diffraction using synchrotron radiation and was found to be in agreement with other published studies.

Description: In this paper, by applying the differential quadrature (DQ) method, a semi analytical model has been developed for atomic force microscope cantilever, and then by using the interfacial forces between the cantilever tip and imaged surfaces, a 2D model has been extracted for imaging nano-sized fine samples. By employing the present model, several simple and standard samples have been imaged, and finally the effects of the microcantilever's structural damping and its stiffness on the imaging results have been investigated. It has been observed that, through the control of damping, the quality of the acquired images is considerably improved. It has also been shown that the self-softening and self-hardening properties of cantilever have serious effects on the obtained images. The present model can be used to study the effects of different parameters on the process of imaging small-scale samples. Also, as one of its most important applications, this model can be used in common multiscale models for simulating and predicting the effects of large and small fields on each other.

Description: The synthesis of scandium nitride (ScN) nanoscale crystals by dissolving N 2 into In-Sc melts is demonstrated for the first time. The crystallization mechanism of ScN from In-Sc melts is investigated. In the N 2 pressure of 0.3 MPa, the ScN yield increases with the Sc concentration in flux and the growth temperature within the range 900–1100°C, achieving a maximum value of about 70% at temperatures above 1100°C. Scanning electron microscopy evidences the growth of round-shaped ScN crystals with increasing average size from 48 to 860 nm in the temperature range of 900–1300°C. This study shows that Sc effectively promotes the dissolution of N 2 in the In-Sc melt, and In-Sc is a promising flux for the synthesis of ScN nanocrystals.

Description: Coiled-coils are found in proteins throughout all three kingdoms of life. Coiled-coil domains of some proteins are almost invariant in sequence and length, betraying a structural and functional role for amino acids along the entire length of the coiled-coil. Other coiled-coils are divergent in sequence, but conserved in length, thereby functioning as molecular spacers. In this capacity, coiled-coil proteins influence the architecture of organelles such as centrioles and the Golgi, as well as permit the tethering of transport vesicles. Specialized coiled-coils, such as those found in motor proteins, are capable of propagating conformational changes along their length that regulate cargo binding and motor processivity. Coiled-coil domains have also been identified in enzymes, where they function as molecular rulers, positioning catalytic activities at fixed distances. Finally, while coiled-coils have been extensively discussed for their potential to nucleate and scaffold large macromolecular complexes, structural evidence to substantiate this claim is relatively scarce. Supercoiling of α-helices gives rise to coiled-coil structures of theoretically infinite length. The tunable length and mechanical properties of coiled-coils make them suitable for a wide variety of functions in the cell including molecular rulers or spacers, molecular tethers, and for communicating information along their length.

Description: Interleukin (IL)-10–expressing B cells play a critical role in the immune homeostasis in the body; its regulation has not been fully understood. Micro-RNA (miR)-17-92 cluster has strong regulation in the immunity. This study tests a hypothesis that miR-17-92 cluster suppresses IL-10 expression in B cells. In this study, peripheral B cells were collected from patients with allergic rhinitis (AR). The B cells were treated with specific allergens, dust mite extracts, in the culture. The expressions of miR-17-92 cluster and IL-10 in the culture were assessed by real-time quantitative reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. The results showed that the levels of miR-19a, but not the rest of the 5 members (miR-17, miR-18a, miR-19b, miR-20a, and miR-92a), were significantly higher in peripheral B cells from AR patients as than in B cells from healthy participants. Exposure of B cells from AR patients to specific allergen, dust mite extracts, significantly increased the levels if miR-19a and suppressed the expression of IL-10 in B cells. The levels of histone deacetylase 11 and acetylated H3K9 were higher, and the RNA polymerase II and c-Maf (the IL-10 transcription factor) were lower, at the IL-10 promoter locus. In conclusion, miR-19a mediates the allergen-specific immune response–decreased IL-10 expression in B cells.

Description: Publication date: Available online 11 July 2016 Source: Journal of Asian Ceramic Societies Author(s): Chaouki Sadik, Omar Moudden, Abdselam El Bouari, Iz-Eddine El Amrani One of the most important elements of furnaces, boilers and other heating units is the structure (lining), usually made of silica–alumina, basic or special refractories. The basic refractories are materials that are increasingly in demand and whose manufacturing involves necessarily the use of MgO and CaO. In this article, the description and characterization of magnesite (MgCO 3 ) and dolomite (Mg,Ca(CO 3 ) 2 ) and their contribution in industrial ceramics-refractories have been reviewed.

Description: Observing adaptive evolution is difficult. In the fossil record, phenotypic evolution happens much more slowly than in artificial selection experiments or in experimental evolution. Yet measures of selection on phenotypic traits, with high heritabilities, suggest that phenotypic evolution should also be rapid in the wild, and this discrepancy often remains even after accounting for correlations between different traits (i.e. making predictions using the multivariate version of the breeder's equation). Are fitness correlations with quantitative traits adequate measures of selection in the wild? We should instead view fitnesses as average properties of genotypes, while acknowledging that they can be environment-dependent. Populations will tend to remain at fitness equilibria, once these are attained, and phenotypes will then be stable. Thus, studying the causes of adaptive change at a genotypic rather than phenotypic level may reveal that, typically, it is occurring too slowly to be easily observed. Measured phenotypic evolutionary rates are high under artificial selection and in changed environments, but are low when environments are constant and are very low over paleontological time. Additive genetic variance (heritability) and apparent selection predict high rates of phenotypic evolution, which is not seen. Why?

Description: Apolipoprotein A-I (ApoA-I) is a key component of High Density Lipoproteins which possess anti-atherosclerotic and anti-inflammatory properties. Insulin is a crucial mediator of the glucose and lipid metabolism that has been implicated in atherosclerotic and inflammatory processes. Important mediators of insulin signaling such as Liver X Receptors (LXRs) and Forkhead Box A2 (FOXA2) are known to regulate apoA-I expression in liver. Forkhead Box O1 (FOXO1) is a well-known target of insulin signaling and a key mediator of oxidative stress response. Low doses of insulin were shown to activate apoA-I expression in human hepatoma HepG2 cells. However, the detailed mechanisms for these processes are still unknown. We studied the possible involvement of FOXO1, FOXA2, LXRα and LXRβ transcription factors in the insulin-mediated regulation of apoA-I expression. Treatment of HepG2 cells with high doses of insulin (48 hours, 100 nM) suppresses apoA-I gene expression. siRNAs against FOXO1, FOXA2, LXRβ or LXRα abrogated this effect. FOXO1 forms a complex with LXRβ and insulin treatment impairs FOXO1/LXRβ complex binding to hepatic enhancer and triggers its nuclear export. Insulin as well as LXR ligand TO901317 enhance the interaction between FOXA2, LXRα and hepatic enhancer. These data suggest that high doses of insulin downregulate apoA-I gene expression in HepG2 cells through redistribution of FOXO1/LXRβ complex, FOXA2 and LXRα on hepatic enhancer of apoA-I gene. This article is protected by copyright. All rights reserved

Description: ABSTRACT Vimentin (Vim), a cytoskeletal intermediate filament, is part of a naturally occurring reversible program, the Epithelial-Mesenchymal Transition (EMT), which converts epithelial cells into mesenchymal-like derivatives. Based on previous results showing that epithelial cells co-express Vim and keratin (Krt) as part of a cytoskeletal network which confers them a highly motile phenotype, we explored the role of Vim in rabbit corneal epithelial cells or RCE1(5T5) cells, an established model of corneal epithelial differentiation. Vim and keratin filaments were co-expressed in cells localized at the proliferative/migratory rim of the growing colonies, but not in basal cells from the center of the colonies nor at suprabasal cell layers. Flow cytometry and qPCR demonstrated that there was a decrease in Krt + /Vim + cell number and ΔNp63α expression when cells reached confluence and formed a 4-5 layered epithelium, while there was a concomitant increase of both Pax-6 expression and Krt + /Vim - cells. Inhibition of cell proliferation with mitomycin C did not modify cell motility nor the expression of Vim. We studied the distribution and expression of α6 integrin, a protein also involved in cell migration. The results demonstrated that α6 integrin had a distribution which was, in part, co-linear with Vim at the proliferative/migratory rim of cell colonies, suggesting an indirect interaction between these proteins. Immunoprecipitation and immunostaining assays indicated that plectin might be mediating such interaction. These data suggest that Vim expression in corneal epithelium is found in a cell population composed of highly motile cells with a Vim + /Krt + /ΔNp63α + /Pax-6 low /α6 integrin + phenotype. This article is protected by copyright. All rights reserved

Description: DNA methylation has emerged as a crucial regulator of chondrocyte dedifferentiation, which severely compromises the outcome of autologous chondrocyte implantation (ACI) treatment for cartilage defects. However, the full -scale DNA methylation profiling in chondrocyte dedifferentiation remains to be determined. Here, we performed a genome-wide DNA methylation profiling of dedifferentiated chondrocytes in monolayer culture and chondrocytes treated with DNA methylation inhibitor 5-azacytidine (5-AzaC). This research revealed that the general methylation level of CpG was increased while the COL-1A1 promoter methylation level was decreased during the chondrocyte dedifferentiation. 5-AzaC could reduce general methylation levels and reverse the chondrocyte dedifferentiation. Surprisingly, the DNA methylation level of COL-1A1 promoter was increased after 5-AzaC treatment. The COL-1A1 expression level was increased while that of SOX-9 was decreased during the chondrocyte dedifferentiation. 5-AzaC treatment up-regulated the SOX-9 expression while down-regulated the COL-1A1 promoter activity and gene expression. Taken together, these results suggested that differential regulation of the DNA methylation level of cartilage-specific genes might contribute to the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation. This article is protected by copyright. All rights reserved

Description: Angiogenesis is associated with changes in endothelial cell (EC) proliferation and tube formation, controlled by extracellular receptor-activated kinase (ERK)/mitogen activated protein kinase (MAPK) and Akt signaling. Important regulators of these systems include hormones acting on G-protein-coupled receptors, such as beta 2-adrenoceptors (β2-ARs). In central nervous system (CNS) trauma, the importance of β2-AR modulation has been highlighted, although the effects on revascularization remain unclear. Vascular protection and revascularization are, however, key to support regeneration. We have investigated the angiogenic capacity of the specific β2-AR agonist terbutaline on ECs derived from the CNS, namely bEnd.3-cells. As angiogenesis is a multistep process involving increased proliferation and tube formation of ECs, we investigated the effects of terbutaline on these processes. We show that terbutaline significantly induced bEnd.3 tube formation in a matrigel in vitro assay. Moreover, administration of specific inhibitors of ERK and Akt signaling both inhibited terbutaline-induced tube formation. The proliferation rate of the ECs was not affected. In order to investigate the general effects of terbutaline in an organotypic system, we have used the chick chorioallantoic membrane (CAM)-assay. Most importantly, terbutaline increased the number of blood vessels in this in ovo setting. Although we observed a positive trend, the systemic administration of terbutaline did not significantly improve the functional outcome, nor did it affect revascularization in our spinal cord injury model. In conclusion, these data indicate that terbutaline is promising to stimulate blood vessel formation, underscoring the importance of further research into the angiotherapeutic relevance of terbutaline and β2-AR signaling after CNS-trauma. This article is protected by copyright. All rights reserved

Description: Extracellular ATP and other nucleotides induce autocrine and/or paracrine purinergic signalling via activation of the P2 receptors on the cell surface, which represents one of the most common signalling mechanisms. Mesenchymal stem cells (MSC) are a type of multipotent adult stem cells that have many promising applications in regenerative medicine. There is increasing evidence to show that extracellular nucleotides regulate MSC functions and P2 receptor-mediated purinergic signalling plays an important role in such functional regulation. P2 receptors comprise ligand-gated ion channel P2X receptors and G-protein-coupled P2Y receptors. In this review, we provide an overview of the current understanding with respect to expression of the P2X and P2Y receptors in MSC and their roles in mediating extracellular nucleotide regulation of MSC proliferation, migration and differentiation. This article is protected by copyright. All rights reserved

Description: ABSTRACT Chronic inflammation and metabolic reprogramming have been proposed as hallmarks of cancer development. Currently, many of the functional clues between these two phenomena are studied under the integrative view of functional stroma-epithelia interaction. It has been proposed that stromal cells, due to their abundance and avidity for glucose, are able to modify the metabolic behavior of an entire solid tumor”. In the present study, using a mammary stromal cell line derived from healthy tissue subjected to long-term culture in low (5 mM) or high (25 mM) glucose, we found that the hyperglycemic condition favors the establishment of a pro-inflammatory and pro-oxidant environment characterized by the induction of the COX-2/PGE2 axis. In this condition, epithelial migration was stimulated. Moreover, we also found that stromal-derived PGE2, acting as a stimulator of IL-1 epithelial expression was one of the factors that promote the acquisition of motile properties by epithelial cells and the maintenance of a COX-2/PGE2-dependent inflammatory condition. Overall, our work provides experimental evidence that glucose stimulates a tumor inflammatory environment that, as a result of a functional cross-talk between stroma and epithelia, may be responsible for tumor progression. This article is protected by copyright. All rights reserved

Description: Wnt/β-catenin signals are important regulators of embryonic and adult stem cell self-renewal and differentiation and play causative roles in tumorigenesis. Purified recombinant Wnt3a protein, or Wnt3a-conditioned culture medium, has been widely used to study canonical Wnt signaling in vitro or ex vivo. To study the role of Wnt3a in embryogenesis and cancer models, we developed a Cre recombinase activatable Rosa26 Wnt3a allele, in which a Wnt3a cDNA was inserted into the Rosa26 locus to allow for conditional, spatiotemporally defined expression of Wnt3a ligand for gain-of-function (GOF) studies in mice. To validate this reagent, we ectopically overexpressed Wnt3a in early embryonic progenitors using the T-Cre transgene. This resulted in up-regulated expression of a β-catenin/Tcf-Lef reporter and of the universal Wnt/β-catenin pathway target genes, Axin2 and Sp5 . Importantly, T-Cre; Rosa26 Wnt3a mutants have expanded presomitic mesoderm (PSM) and compromised somitogenesis and closely resemble previously studied T-Cre; Ctnnb1 ex3 (β-catenin GOF ) mutants. These data indicate that the exogenously expressed Wnt3a stimulates the Wnt/β-catenin signaling pathway, as expected. The Rosa26 Wnt3a mouse line should prove to be an invaluable tool to study the function of Wnt3a in vivo . This article is protected by copyright. All rights reserved.

Description: Endometrioid adenocarcinomas represent 80% of endometrial carcinomas 1 . Molecular features, including microsatellite instability, mutations of the PTEN, PIK3CA, K-Ras and β-catenin genes 1 and dysregulations in sncRNAs (small non coding RNAs) are described for this disease. However, mechanisms and molecules that determine cell survival and response/resistance to therapy in different subtypes of this tumour are not fully clarified 1 . This article is protected by copyright. All rights reserved

Description: Cyanobacteria, such as the toxin producer Microcystis aeruginosa , are predicted to be favored by global warming both directly, through elevated water temperatures, and indirectly, through factors such as prolonged stratification of waterbodies. M. aeruginosa is able to produce the hepatotoxin microcystin, which causes great concern in freshwater management worldwide. However, little is known about the expression of microcystin synthesis genes in response to climate change-related factors. In this study, a new RT-qPCR assay employing four reference genes ( GAPDH , gltA, rpoC1, and rpoD ) was developed to assess the expression of two target genes (the microcystin synthesis genes mcyB and mcyD ). This assay was used to investigate changes in mcyB and mcyD expression in response to selected environmental factors associated with global warming. A 10°C rise in temperature significantly increased mcyB expression, but not mcyD expression. Neither mixing nor the addition of microcystin-LR (10 μg L −1 or 60 μg L −1 ) significantly altered mcyB and mcyD expression. The expression levels of mcyB and mcyD were correlated but not identical. Cyanobacteria, such as the toxin producer Microcystis aeruginosa , are predicted to be favored by global warming. In this study, a new RT-qPCR assay was used to investigate the changes in mcyB and mcyD expression, genes responsible for the biosynthesis of the cyanotoxin microcystin, in response to selected environmental factors associated with global warming.

Description: Three-dimensional optical super-resolution imaging is capable of providing 3D visualization of cellular structures in nanoscale detail. The past decade has witnessed the blossoming of 3D super-resolution imaging technologies. In this review, we comprehensively discuss and compare the imaging depth, resolution enhancement, and imaging speed of the existing 3D super-resolution imaging techniques.