bluuebunny/crossref_metadata_2025 huggingface dataset:
Salt-Related Knowledge, Attitudes, and Behaviors on Efate Island, Vanuatu
Authors: Sparks Emalie , Paterson Katherine , Santos Joseph Alvin , Trieu Kathy , Hinge Nerida , Tarivonda Len , Snowdon Wendy , Johnson Claire , Webster Jacqui
Published: March 2019

In Vanuatu, mean salt intake exceeds the recommended maximum daily intake, and contributes to the high proportion of deaths attributable to cardiovascular diseases. Understanding salt-related knowledge, attitudes, and behaviors of the Vanuatu population can inform appropriate interventions. This cross-sectional study was conducted as part of the 2016–2017 Vanuatu Salt Survey. In total, 753 participants aged between 18 and 69 years from rural and urban communities on the Island of Efate were included. Demographic and clinical data were collected and a salt-related knowledge, attitudes, and behaviors survey was administered. Knowledge relating to the need to reduce salt consumption was high, but reported behaviors did not reflect this knowledge. A total of 83% of participants agreed that too much salt could cause health problems, and 86% reported that it was “very important” to lower the amount of salt in the diet. However, more than two-thirds of the population reported always/often adding salt to food during cooking/meal preparation and at the table, and always/often consuming processed foods high in salt. Strategic, targeted, and sustained behavior change programs in parallel with interventions to change the food environment to facilitate healthier choices should be key components of a salt reduction program. Actions should implemented as part of a comprehensive strategy to prevent and control non-communicable diseases in Vanuatu.
Tree Species Classification Using Hyperion and Sentinel-2 Data with Machine Learning in South Korea and China
Authors: Lim Joongbin , Kim Kyoung-Min , Jin Ri
Published: March 2019

Remote sensing (RS) has been used to monitor inaccessible regions. It is considered a useful technique for deriving important environmental information from inaccessible regions, especially North Korea. In this study, we aim to develop a tree species classification model based on RS and machine learning techniques, which can be utilized for classification in North Korea. Two study sites were chosen, the Korea National Arboretum (KNA) in South Korea and Mt. Baekdu (MTB; a.k.a., Mt. Changbai in Chinese) in China, located in the border area between North Korea and China, and tree species classifications were examined in both regions. As a preliminary step in developing a classification algorithm that can be applied in North Korea, common coniferous species at both study sites, Korean pine (Pinus koraiensis) and Japanese larch (Larix kaempferi), were chosen as targets for investigation. Hyperion data have been used for tree species classification due to the abundant spectral information acquired from across more than 200 spectral bands (i.e., hyperspectral satellite data). However, it is impossible to acquire recent Hyperion data because the satellite ceased operation in 2017. Recently, Sentinel-2 satellite multispectral imagery has been used in tree species classification. Thus, it is necessary to compare these two kinds of satellite data to determine the possibility of reliably classifying species. Therefore, Hyperion and Sentinel-2 data were employed, along with machine learning techniques, such as random forests (RFs) and support vector machines (SVMs), to classify tree species. Three questions were answered, showing that: (1) RF and SVM are well established in the hyperspectral imagery for tree species classification, (2) Sentinel-2 data can be used to classify tree species with RF and SVM algorithms instead of Hyperion data, and (3) training data that were built in the KNA cannot be used for the tree classification of MTB. Random forests and SVMs showed overall accuracies of 0.60 and 0.51 and kappa values of 0.20 and 0.00, respectively. Moreover, combined training data from the KNA and MTB showed high classification accuracies in both regions; RF and SVM values exhibited accuracies of 0.99 and 0.97 and kappa values of 0.98 and 0.95, respectively.
Finding Visible kNN Objects in the Presence of Obstacles within the User’s View Field †
Authors: Su I-Fang , Chen Ding-Li , Lee Chiang , Chung Yu-Chi
Published: March 2019

In many spatial applications, users are only interested in data objects that are visible to them. Hence, finding visible data objects is an important operation in these real-world spatial applications. This study addressed a new type of spatial query, the View field-aware Visible k Nearest Neighbor (V2-kNN) query. Given the location of a user and his/her view field, a V2-kNN query finds data object p so that p is the nearest neighbor of and visible to the user, where visible means the data object is (1) not hidden by obstacles and (2) inside the view field of the user. Previous works on visible NN queries considered only one of these two factors, but not both. To the best of our knowledge, this work is the first to consider both the effect of obstacles and the restriction of the view field in finding the solutions. To support efficient processing of V2-kNN queries, a grid structure is used to index data objects and obstacles. Pruning heuristics are also designed so that only data objects and obstacles relevant to the final query result are accessed. A comprehensive experimental evaluation using both real and synthetic datasets is performed to verify the effectiveness of the proposed algorithms.
Making the Invisible Visible—Strategies for Visualizing Underground Infrastructures in Immersive Environments
Authors: Ortega Sebastián , Wendel Jochen , Santana José Miguel , Murshed Syed Monjur , Boates Isaac , Trujillo Agustín , Nichersu Alexandru , Suárez José Pablo
Published: March 2019

Visualization of underground infrastructure in an interactive 3D immersive environment is extremely important for efficient management of city’s infrastructure. This paper describes different geometric modelling approaches to illustrate appropriate visualization of such data. A multimodal prototype has been developed by exploiting different algorithms to render these invisible underground objects as part of an urban model. This prototype has been integrated in an immersive geographic information system (GIS), named MultiVis, for handheld iOS and Android devices. As a part of the study, three distinct strategies have been tested; the first is based on the use of transparencies to convey a sense of depth, the second relies on an image-space superposition of “ditches” on top of the rendered frame and the third is a world-space deformation of the elevation model that exposes the underground elements. Furthermore, a comparative user experience analysis of different techniques aimed to the geometrically accurate visualisation of utility networks and other underground facilities are performed and evaluated. It includes a set of user evaluations for different parameters of these techniques, which gives us an insight on how the proposed methods affect the experience and usability for technical and non-technical users.
Olive Tree Biophenols in Inflammatory Bowel Disease: When Bitter is Better
Authors: Larussa Tiziana , Imeneo Maria , Luzza Francesco
Published: March 2019

The current therapeutic scenario for inflammatory bowel diseases (IBD) involves aminosalicylates, corticosteroids, and immunomodulators, but concerns regarding their safety profiles and high costs heavily impact their widespread use. In recent years, the beneficial effects thatbiophenols—from fruit and vegetables—have on human health have been investigated. The antioxidant and anti-inflammatory properties of phenolic fraction, from olive leaves and fruits, have been suggested, and a potential application in gut inflammation has been supported by in vitro and IBD-animal models studies. In the present review, we first introduced the potential therapeutic role of olive tree biophenolsin chronic inflammatory disease. Then, we aimed to describe their most interesting application for gut inflammation, as the results of basic science studies and animal experimental models. Finally, the potential role of olive tree biophenols in the setting of human IBD is discussed.
Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
Authors: Balaguer Francisco de Asís , Mühlethaler Tobias , Estévez-Gallego Juan , Calvo Enrique , Giménez-Abián Juan Francisco , Risinger April L. , Sorensen Erik J. , Vanderwal Christopher D. , Altmann Karl-Heinz , Mooberry Susan L. , Steinmetz Michel O. , Oliva María Ángela , Prota Andrea E. , Díaz J. Fernando
Published: March 2019

It has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, including cyclostreptin, to gain further insight into this mechanism. The crystal structure of cyclostreptin-bound tubulin reveals covalent binding to βHis229, but no stabilization of the M-loop. The capacity of cyclostreptin to induce microtubule assembly compared to other covalent taxane-site agents demonstrates that the induction of tubulin assembly is not strictly dependent on M-loop stabilization. We further demonstrate that most covalent taxane-site ligands are able to partially overcome drug resistance mediated by βIII-tubulin (βIII) overexpression in HeLa cells, and compare their activities to pironetin, an interfacial covalent inhibitor of tubulin assembly that displays invariant growth inhibition in these cells. Our findings suggest a relationship between a diminished interaction of taxane-site ligands with βIII-tubulin and βIII tubulin-mediated drug resistance. This supports the idea that overexpression of βIII increases microtubule dynamicity by counteracting the enhanced microtubule stability promoted by covalent taxane-site binding ligands.
Differential Methylation in APOE (Chr19; Exon Four; from 44,909,188 to 44,909,373/hg38) and Increased Apolipoprotein E Plasma Levels in Subjects with Mild Cognitive Impairment
Authors: Mancera-Páez Oscar , Estrada-Orozco Kelly , Mahecha María Fernanda , Cruz Francy , Bonilla-Vargas Kely , Sandoval Nicolás , Guerrero Esneyder , Salcedo-Tacuma David , Melgarejo Jesús D. , Vega Edwin , Ortega-Rojas Jenny , Román Gustavo C. , Pardo-Turriago Rodrigo , Arboleda Humberto
Published: March 2019

Background: Biomarkers are essential for identification of individuals at high risk of mild cognitive impairment (MCI) for potential prevention of dementia. We investigated DNA methylation in the APOE gene and apolipoprotein E (ApoE) plasma levels as MCI biomarkers in Colombian subjects with MCI and controls. Methods: In total, 100 participants were included (71% women; average age, 70 years; range, 43–91 years). MCI was diagnosed by neuropsychological testing, medical and social history, activities of daily living, cognitive symptoms and neuroimaging. Using multivariate logistic regression models adjusted by age and gender, we examined the risk association of MCI with plasma ApoE and APOE methylation. Results: MCI was diagnosed in 41 subjects (average age, 66.5 ± 9.6 years) and compared with 59 controls. Elevated plasma ApoE and APOE methylation of CpGs 165, 190, and 198 were risk factors for MCI (p < 0.05). Higher CpG-227 methylation correlated with lower risk for MCI (p = 0.002). Only CpG-227 was significantly correlated with plasma ApoE levels (correlation coefficient = −0.665; p = 0.008). Conclusion: Differential APOE methylation and increased plasma ApoE levels were correlated with MCI. These epigenetic patterns require confirmation in larger samples but could potentially be used as biomarkers to identify early stages of MCI.
Deletion of Osteopontin Enhances β2-Adrenergic Receptor-Dependent Anti-Fibrotic Signaling in Cardiomyocytes
Authors: Pollard Celina M. , Desimine Victoria L. , Wertz Shelby L. , Perez Arianna , Parker Barbara M. , Maning Jennifer , McCrink Katie A. , Shehadeh Lina A. , Lymperopoulos Anastasios
Published: March 2019

Cardiac β2-adrenergic receptors (ARs) are known to inhibit collagen production and fibrosis in cardiac fibroblasts and myocytes. The β2AR is a Gs protein-coupled receptor (GPCR) and, upon its activation, stimulates the generation of cyclic 3′,5′-adenosine monophosphate (cAMP). cAMP has two effectors: protein kinase A (PKA) and the exchange protein directly activated by cAMP (Epac). Epac1 has been shown to inhibit cardiac fibroblast activation and fibrosis. Osteopontin (OPN) is a ubiquitous pro-inflammatory cytokine, which also mediates fibrosis in several tissues, including the heart. OPN underlies several cardiovascular pathologies, including atherosclerosis and cardiac adverse remodeling. We found that the cardiotoxic hormone aldosterone transcriptionally upregulates OPN in H9c2 rat cardiac myoblasts—an effect prevented by endogenous β2AR activation. Additionally, CRISPR-mediated OPN deletion enhanced cAMP generation in response to both β1AR and β2AR activation in H9c2 cardiomyocytes, leading to the upregulation of Epac1 protein levels. These effects rendered β2AR stimulation capable of completely abrogating transforming growth factor (TGF)-β-dependent fibrosis in OPN-lacking H9c2 cardiomyocytes. Finally, OPN interacted constitutively with Gαs subunits in H9c2 cardiac cells. Thus, we uncovered a direct inhibitory role of OPN in cardiac β2AR anti-fibrotic signaling via cAMP/Epac1. OPN blockade could be of value in the treatment and/or prevention of cardiac fibrosis.
Antigen Targets for the Development of Immunotherapies in Leukemia
Authors: Bauer Jens , Nelde Annika , Bilich Tatjana , Walz Juliane S.
Published: March 2019

Immunotherapeutic approaches, including allogeneic stem cell transplantation and donor lymphocyte infusion, have significantly improved the prognosis of leukemia patients. Further efforts are now focusing on the development of immunotherapies that are able to target leukemic cells more specifically, comprising monoclonal antibodies, chimeric antigen receptor (CAR) T cells, and dendritic cell- or peptide-based vaccination strategies. One main prerequisite for such antigen-specific approaches is the selection of suitable target structures on leukemic cells. In general, the targets for anti-cancer immunotherapies can be divided into two groups: (1) T-cell epitopes relying on the presentation of peptides via human leukocyte antigen (HLA) molecules and (2) surface structures, which are HLA-independently expressed on cancer cells. This review discusses the most promising tumor antigens as well as the underlying discovery and selection strategies for the development of anti-leukemia immunotherapies.
Normal FGF-21-Serum Levels in Patients with Carnitine Palmitoyltransferase II (CPT II) Deficiency
Authors: Motlagh Scholle Leila , Lehmann Diana , Joshi Pushpa Raj , Zierz Stephan
Published: March 2019

Fibroblast growth factor 21 (FGF-21) is known to be a biomarker for mitochondrial disorders. An upregulation of FGF-21 in serum and muscle of carnitine palmitoyltransferase I (CPT I) and carnitine palmitoyltransferase II (CPT II) knock-out mice has been reported. In human CPT II deficiency, enzyme activity and protein content are normal, but the enzyme is abnormally regulated by malonyl-CoA and is abnormally thermolabile. Citrate synthase (CS) activity is increased in patients with CPT II deficiency. This may indicate a compensatory response to an impaired function of CPT II. In this study, FGF-21 serum levels in patients with CPT II deficiency during attack free intervals and in healthy controls were measured by enzyme linked immunosorbent assay (ELISA). The data showed no significant difference between FGF-21 concentration in the serum of patients with CPT II deficiency and that in the healthy controls. The results of the present work support the hypothesis that in muscle CPT II deficiency, in contrast to the mouse knockout model, mitochondrial fatty acid utilization is not persistently reduced. Thus, FGF-21 does not seem to be a useful biomarker in the diagnosis of CPT II deficiency.
Beyond the Flavour: The Potential Druggability of Chemosensory G Protein-Coupled Receptors
Authors: Di Pizio Antonella , Behrens Maik , Krautwurst Dietmar
Published: March 2019

G protein-coupled receptors (GPCRs) belong to the largest class of drug targets. Approximately half of the members of the human GPCR superfamily are chemosensory receptors, including odorant receptors (ORs), trace amine-associated receptors (TAARs), bitter taste receptors (TAS2Rs), sweet and umami taste receptors (TAS1Rs). Interestingly, these chemosensory GPCRs (csGPCRs) are expressed in several tissues of the body where they are supposed to play a role in biological functions other than chemosensation. Despite their abundance and physiological/pathological relevance, the druggability of csGPCRs has been suggested but not fully characterized. Here, we aim to explore the potential of targeting csGPCRs to treat diseases by reviewing the current knowledge of csGPCRs expressed throughout the body and by analysing the chemical space and the drug-likeness of flavour molecules.
Actin Cytoskeleton as Actor in Upstream and Downstream of Calcium Signaling in Plant Cells
Authors: Qian Dong , Xiang Yun
Published: March 2019

In plant cells, calcium (Ca2+) serves as a versatile intracellular messenger, participating in several fundamental and important biological processes. Recent studies have shown that the actin cytoskeleton is not only an upstream regulator of Ca2+ signaling, but also a downstream regulator. Ca2+ has been shown to regulates actin dynamics and rearrangements via different mechanisms in plants, and on this basis, the upstream signaling encoded within the Ca2+ transient can be decoded. Moreover, actin dynamics have also been proposed to act as an upstream of Ca2+, adjust Ca2+ oscillations, and establish cytosolic Ca2+ ([Ca2+]cyt) gradients in plant cells. In the current review, we focus on the advances in uncovering the relationship between the actin cytoskeleton and calcium in plant cells and summarize our current understanding of this relationship.
PD-1 Ligand Expression in Epithelial Thyroid Cancers: Potential Clinical Implications
Authors: Ulisse Salvatore , Tuccilli Chiara , Sorrenti Salvatore , Antonelli Alessandro , Fallahi Poupak , D’Armiento Eleonora , Catania Antonio , Tartaglia Francesco , Amabile Maria Ida , Giacomelli Laura , Metere Alessio , Cornacchini Nicola , Pironi Daniele , Carbotta Giovanni , Vergine Massimo , Monti Massimo , Baldini Enke
Published: March 2019

The new immunotherapy targeting the programmed cell death 1 (PD-1) receptor and its cognate ligand PD-L1 has renewed hopes of eradicating the most difficult human cancers to treat. Among these, there are the poorly differentiated and anaplastic thyroid cancers, unresponsive to all the therapies currently in use. In the present review we will summarize information regarding the expression of PD-L1 in the different thyroid cancer histotypes, its correlation with clinicopathological features, and its potential prognostic value. Then, we will evaluate the available data indicating the PD-1/PD-L1 axis as a promising target for thyroid cancer therapy.
Exosomes and Hepatocellular Carcinoma: From Bench to Bedside
Authors: Sasaki Reina , Kanda Tatsuo , Yokosuka Osamu , Kato Naoya , Matsuoka Shunichi , Moriyama Mitsuhiko
Published: March 2019

As hepatocellular carcinoma (HCC) usually occurs in the background of cirrhosis, which is an end-stage form of liver diseases, treatment options for advanced HCC are limited, due to poor liver function. The exosome is a nanometer-sized membrane vesicle structure that originates from the endosome. Exosome-mediated transfer of proteins, DNAs and various forms of RNA, such as microRNA (miRNA), long noncoding RNA (lncRNA) and messenger RNA (mRNA), contributes to the development of HCC. Exosomes mediate communication between both HCC and non-HCC cells involved in tumor-associated cells, and several molecules are implicated in exosome biogenesis. Exosomes may be potential diagnostic biomarkers for early-stage HCC. Exosomal proteins, miRNAs and lncRNAs could provide new biomarker information for HCC. Exosomes are also potential targets for the treatment of HCC. Notably, further efforts are required in this field. We reviewed recent literature and demonstrated how useful exosomes are for diagnosing patients with HCC, treating patients with HCC and predicting the prognosis of HCC patients.
Bioengineered Skin Intended for Skin Disease Modeling
Authors: Sarkiri Maria , Fox Stephan C. , Fratila-Apachitei Lidy E. , Zadpoor Amir A.
Published: March 2019

Clinical use of bioengineered skin in reconstructive surgery has been established for more than 30 years. The limitations and ethical considerations regarding the use of animal models have expanded the application of bioengineered skin in the areas of disease modeling and drug screening. These skin models should represent the anatomical and physiological traits of native skin for the efficient replication of normal and pathological skin conditions. In addition, reliability of such models is essential for the conduction of faithful, rapid, and large-scale studies. Therefore, research efforts are focused on automated fabrication methods to replace the traditional manual approaches. This report presents an overview of the skin models applicable to skin disease modeling along with their fabrication methods, and discusses the potential of the currently available options to conform and satisfy the demands for disease modeling and drug screening.
Single-Molecule Imaging and Computational Microscopy Approaches Clarify the Mechanism of the Dimerization and Membrane Interactions of Green Fluorescent Protein
Authors: Wang Xiaohua , Song Kai , Li Yang , Tang Ling , Deng Xin
Published: March 2019

Green fluorescent protein (GFP) is widely used as a biomarker in living systems; however, GFP and its variants are prone to forming low-affinity dimers under physiological conditions. This undesirable tendency is exacerbated when fluorescent proteins (FP) are confined to membranes, fused to naturally-oligomeric proteins, or expressed at high levels in cells. Oligomerization of FPs introduces artifacts into the measurement of subunit stoichiometry, as well as interactions between proteins fused to FPs. Introduction of a single mutation, A206K, has been shown to disrupt hydrophobic interactions in the region responsible for GFP dimerization, thereby contributing to its monomerization. Nevertheless, a detailed understanding of how this single amino acid-dependent inhibition of dimerization in GFP occurs at the atomic level is still lacking. Single-molecule experiments combined with computational microscopy (atomistic molecular dynamics) revealed that the amino group of A206 contributes to GFP dimer formation via a multivalent electrostatic interaction. We further showed that myristoyl modification is an efficient mechanism to promote membrane attachment of GFP. Molecular dynamics-based site-directed mutagenesis has been used to identify the key functional residues in FPs. The data presented here have been utilized as a monomeric control in downstream single-molecule studies, facilitating more accurate stoichiometry quantification of functional protein complexes in living cells.
The Role of Metabotropic Glutamate Receptors in Social Behavior in Rodents
Authors: Zoicas Iulia , Kornhuber Johannes
Published: March 2019

The appropriate display of social behavior is critical for the well-being and survival of an individual. In many psychiatric disorders, including social anxiety disorder, autism spectrum disorders, depression and schizophrenia social behavior is severely impaired. Selective targeting of metabotropic glutamate receptors (mGluRs) has emerged as a novel treatment strategy for these disorders. In this review, we describe some of the behavioral paradigms used to assess different types of social behavior, such as social interaction, social memory, aggressive behavior and sexual behavior. We then focus on the effects of pharmacological modulation of mGluR1-8 on these types of social behavior. Indeed, accumulating evidence indicates beneficial effects of selective ligands of specific mGluRs in ameliorating innate or pharmacologically-induced deficits in social interaction and social memory as well as in reducing aggression in rodents. We emphasize the importance of future studies investigating the role of selective mGluR ligands on different types of social behavior to provide a better understanding of the neural mechanisms involved which, in turn, might promote the development of selective mGluR-targeted tools for the improved treatment of psychiatric disorders associated with social deficits.
Functional Mechanisms Underlying the Antimicrobial Activity of the Oryza sativa Trx-like Protein
Authors: Park Seong-Cheol , Kim Il Ryong , Hwang Jung Eun , Kim Jin-Young , Jung Young Jun , Choi Wonkyun , Lee Yongjae , Jang Mi-Kyeong , Lee Jung Ro
Published: March 2019

Plants are constantly subjected to a variety of environmental stresses and have evolved regulatory responses to overcome unfavorable conditions that might reduce or adversely change a plant’s growth or development. Among these, the regulated production of reactive oxygen species (ROS) as a signaling molecule occurs during plant development and pathogen defense. This study demonstrates the possible antifungal activity of Oryza sativa Tetratricopeptide Domain-containing thioredoxin (OsTDX) protein against various fungal pathogens. The transcription of OsTDX was induced by various environmental stresses known to elicit the generation of ROS in plant cells. OsTDX protein showed potent antifungal activity, with minimum inhibitory concentrations (MICs) against yeast and filamentous fungi ranging between 1.56 and 6.25 and 50 and 100 µg/mL, respectively. The uptake of SYTOX-Green into fungal cells and efflux of calcein from artificial fungus-like liposomes suggest that its killing mechanism involves membrane permeabilization and damage. In addition, irregular blebs and holes apparent on the surfaces of OsTDX-treated fungal cells indicate the membranolytic action of this protein. Our results suggest that the OsTDX protein represents a potentially useful lead for the development of pathogen-resistant plants.
Interactions between Membrane Resistance, GABA-A Receptor Properties, Bicarbonate Dynamics and Cl−-Transport Shape Activity-Dependent Changes of Intracellular Cl− Concentration
Authors: Lombardi Aniello , Jedlicka Peter , Luhmann Heiko J. , Kilb Werner
Published: March 2019

The effects of ionotropic γ-aminobutyric acid receptor (GABA-A, GABAA) activation depends critically on the Cl−-gradient across neuronal membranes. Previous studies demonstrated that the intracellular Cl−-concentration ([Cl−]i) is not stable but shows a considerable amount of activity-dependent plasticity. To characterize how membrane properties and different molecules that are directly or indirectly involved in GABAergic synaptic transmission affect GABA-induced [Cl−]i changes, we performed compartmental modeling in the NEURON environment. These simulations demonstrate that GABA-induced [Cl−]i changes decrease at higher membrane resistance, revealing a sigmoidal dependency between both parameters. Increase in GABAergic conductivity enhances [Cl−]i with a logarithmic dependency, while increasing the decay time of GABAA receptors leads to a nearly linear enhancement of the [Cl−]i changes. Implementing physiological levels of HCO3−-conductivity to GABAA receptors enhances the [Cl−]i changes over a wide range of [Cl−]i, but this effect depends on the stability of the HCO3− gradient and the intracellular pH. Finally, these simulations show that pure diffusional Cl−-elimination from dendrites is slow and that a high activity of Cl−-transport is required to improve the spatiotemporal restriction of GABA-induced [Cl−]i changes. In summary, these simulations revealed a complex interplay between several key factors that influence GABA-induced [Cl]i changes. The results suggest that some of these factors, including high resting [Cl−]i, high input resistance, slow decay time of GABAA receptors and dynamic HCO3− gradient, are specifically adapted in early postnatal neurons to facilitate limited activity-dependent [Cl−]i decreases.
Symmetrical Modification of Minimized Dermaseptins to Extend the Spectrum of Antimicrobials with Endotoxin Neutralization Potency
Authors: Shao Changxuan , Li Weizhong , Tan Peng , Shan Anshan , Dou Xiujing , Ma Deying , Liu Chunyu
Published: March 2019

Antimicrobial peptides (AMPs) have emerged as a promising class of antimicrobial agents that could potentially address the global antibiotic resistance. Generating mirror-like peptides by minimizing dermaseptin family sequences is an effective strategy for designing AMPs. However, the previous research still had some limitations such as lower effectiveness and a narrow spectrum of antibacterial activity. To further expand and hone this strategy, we designed a series of AMPs consisting of the WXMXW-NH2 motif (X represents V, I, F, and W; M represents KAAAKAAAK). The peptides formed α-helices and displayed broad-spectrum antimicrobial activities against eleven types of clinical bacteria including both Gram-negative and Gram-positive bacteria. The optimized peptide WW exhibited high physical rupture by inducing membrane shrinkage, disruption, and lysis. Moreover, WW effectively neutralized endotoxins and inhibited the inflammatory response while having the highest therapeutic index. In conclusion, these results indicated that the peptide WW has potential as a broad-spectrum antimicrobial agent or preservative for overcoming the risk of multidrug resistance in localized or external therapeutic applications.
Caprate Modulates Intestinal Barrier Function in Porcine Peyer’s Patch Follicle-Associated Epithelium
Authors: Radloff Judith , Cornelius Valeria , Markov Alexander G. , Amasheh Salah
Published: March 2019

Background: Many food components influence intestinal epithelial barrier properties and might therefore also affect susceptibility to the development of food allergies. Such allergies are triggered by increased antibody production initiated in Peyer’s patches (PP). Usually, the presentation of antigens in the lumen of the gut to the immune cells of the PP is strongly regulated by the follicle-associated epithelium (FAE) that covers the PP. As the food component caprate has been shown to impede barrier properties in villous epithelium, we hypothesized that caprate also affects the barrier function of the PP FAE, thereby possibly contributing a risk factor for the development of food allergies. Methods: In this study, we have focused on the effects of caprate on the barrier function of PP, employing in vitro and ex vivo experimental setups to investigate functional and molecular barrier properties. Incubation with caprate induced an increase of transepithelial resistance, and a marked increase of permeability for the paracellular marker fluorescein in porcine PP to 180% of control values. These effects are in accordance with changes in the expression levels of the barrier-forming tight junction proteins tricellulin and claudin-5. Conclusions: This barrier-affecting mechanism could be involved in the initial steps of a food allergy, since it might trigger unregulated contact of the gut lumen with antigens.
Obesity-Altered Adipose Stem Cells Promote ER+ Breast Cancer Metastasis through Estrogen Independent Pathways
Authors: Sabol Rachel A. , Beighley Adam , Giacomelli Paulina , Wise Rachel M. , Harrison Mark A. A. , O’Donnnell Ben A. , Sullivan Brianne N. , Lampenfeld Jacob D. , Matossian Margarite D. , Bratton Melyssa R. , Wang Guangdi , Collins-Burow Bridgette M. , Burow Matthew E. , Bunnell Bruce A.
Published: March 2019

Adipose stem cells (ASCs) play an essential role in tumor microenvironments. These cells are altered by obesity (obASCs) and previous studies have shown that obASCs secrete higher levels of leptin. Increased leptin, which upregulates estrogen receptor alpha (ERα) and aromatase, enhances estrogen bioavailability and signaling in estrogen receptor positive (ER+) breast cancer (BC) tumor growth and metastasis. In this study, we evaluate the effect of obASCs on ER+BC outside of the ERα signaling axis using breast cancer models with constitutively active ERα resulting from clinically relevant mutations (Y537S and D538G). We found that while obASCs promote tumor growth and proliferation, it occurs mostly through abrogated estrogen signaling when BC has constitutive ER activity. However, obASCs have a similar promotion of metastasis irrespective of ER status, demonstrating that obASC promotion of metastasis may not be completely estrogen dependent. We found that obASCs upregulate two genes in both ER wild type (WT) and ER mutant (MUT) BC: SERPINE1 and ABCB1. This study demonstrates that obASCs promote metastasis in ER WT and MUT xenografts and an ER MUT patient derived xenograft (PDX) model. However, obASCs promote tumor growth only in ER WT xenografts.
MicroRNA Sequencing Revealed Citrus Adaptation to Long-Term Boron Toxicity through Modulation of Root Development by miR319 and miR171
Authors: Huang Jing-Hao , Lin Xiong-Jie , Zhang Ling-Yuan , Wang Xian-Da , Fan Guo-Cheng , Chen Li-Song
Published: March 2019

Boron (B) toxicity in Citrus is a common physiological disorder leading to reductions in both productivity and quality. Studies on how Citrus roots evade B toxicity may provide new insight into plant tolerance to B toxicity. Here, using Illumina sequencing, differentially expressed microRNAs (miRNAs) were identified in B toxicity-treated Citrus sinensis (tolerant) and C. grandis (intolerant) roots. The results showed that 37 miRNAs in C. grandis and 11 miRNAs in C. sinensis were differentially expressed when exposed to B toxicity. Among them, miR319, miR171, and miR396g-5p were confirmed via 5′-RACE and qRT-PCR to target a myeloblastosis (MYB) transcription factor gene, a SCARECROW-like protein gene, and a cation transporting ATPase gene, respectively. Maintenance of SCARECROW expression in B treated Citrus roots might fulfill stem cell maintenance, quiescent center, and endodermis specification, thus allowing regular root elongation under B-toxic stress. Down-regulation of MYB due to up-regulation of miR319 in B toxicity-treated C. grandis roots might decrease the number of root tips, thereby dramatically changing root system architecture. Our findings suggested that miR319 and miR171 play a pivotal role in Citrus adaptation to long-term B toxicity by targeting MYB and SCARECROW, respectively, both of which are responsible for root growth and development.
Energy Landscapes of Ligand Motion Inside the Tunnel-Like Cavity of Lipid Transfer Proteins: The Case of the Pru p 3 Allergen
Authors: Cuevas-Zuviría Bruno , Garrido-Arandia María , Díaz-Perales Araceli , Pacios Luis F.
Published: March 2019

Allergies are a widespread problem in western countries, affecting a large part of the population, with levels of prevalence increasingly rising due to reasons still not understood. Evidence accumulated in recent years points to an essential role played by ligands of allergen proteins in the sensitization phase of allergies. In this regard, we recently identified the natural ligand of Pru p 3, a lipid transfer protein, a major allergen from peach fruit and a model of food allergy. The ligand of Pru p 3 has been shown to play a key role in the sensitization to peach and to other plant food sources that provoke cross-reactivity in a large proportion of patients allergic to peach. However, the question of which is the binding pose of this ligand in its carrier protein, and how it can be transferred to receptors of the immune system where it develops its function as a coadjuvant was not elucidated. In this work, different molecular dynamics simulations have been considered as starting points to study the properties of the ligand–protein system in solution. Besides, an energy landscape based on collective variables that describe the process of ligand motion within the cavity of Pru p 3 was obtained by using well-tempered metadynamics. The simulations revealed the differences between distinct binding modes, and also revealed important aspects of the motion of the ligand throughout its carrier protein, relevant to its binding–unbinding process. Our findings are potentially interesting for studying protein–ligand systems beyond the specific case of the allergen protein dealt with here.
The Genetic Background of Mice Influences the Effects of Cigarette Smoke on Onset and Severity of Experimental Autoimmune Encephalomyelitis
Authors: Enzmann Gaby , Adelfio Roberto , Godel Aurélie , Haghayegh Jahromi Neda , Tietz Silvia , Burgener Sabrina S. , Deutsch Urban , Wekerle Hartmut , Benarafa Charaf , Engelhardt Britta
Published: March 2019

Multiple sclerosis (MS) is the most common inflammatory disorder of the central nervous system (CNS) in young adults leading to severe disability. Besides genetic traits, environmental factors contribute to MS pathogenesis. Cigarette smoking increases the risk of MS in an HLA-dependent fashion, but the underlying mechanisms remain unknown. Here, we explored the effect of cigarette smoke exposure on spontaneous and induced models of experimental autoimmune encephalomyelitis (EAE) by evaluating clinical disease and, when relevant, blood leukocytes and histopathology. In the relapsing-remitting (RR) transgenic model in SJL/J mice, we observed very low incidence in both smoke-exposed and control groups. In the optico-spinal encephalomyelitis (OSE) double transgenic model in C57BL/6 mice, the early onset of EAE prevented a meaningful evaluation of the effects of cigarette smoke. In EAE models induced by immunization, daily exposure to cigarette smoke caused a delayed onset of EAE followed by a protracted disease course in SJL/J mice. In contrast, cigarette smoke exposure ameliorated the EAE clinical score in C57BL/6J mice. Our exploratory studies therefore show that genetic background influences the effects of cigarette smoke on autoimmune neuroinflammation. Importantly, our findings expose the challenge of identifying an animal model for studying the influence of cigarette smoke in MS.
An Integrative Evaluation Method for the Biological Safety of Down and Feather Materials
Authors: Kawada Toshikatsu , Kuroyanagi Junya , Okazaki Fumiyoshi , Taniguchi Mizuki , Nakayama Hiroko , Suda Narumi , Abiko Souta , Kaneco Satoshi , Nishimura Norihiro , Shimada Yasuhito
Published: March 2019

Background: Down and feather materials have been commonly used and promoted as natural stuffing for warm clothing and bedding. These materials tend to become more allergenic as they become contaminated with microorganisms, in addition to being subjected to several kinds of chemical treatments. The biological or chemical contaminants in these materials pose a major risk to human health, to consumers and manufacturers alike. Here, we report the development of an integrative evaluation method for down and feather materials to assess bacterial contamination and in vivo toxicity. Methods: To assess bacterial contamination, we quantified 16S ribosomal RNA, performed culture tests, and established a conversion formula. To determine in vivo toxicity, we performed a zebrafish embryo toxicity testing (ZFET). Results: Washing the material appropriately decreases the actual number of bacteria in the down and feather samples; in addition, after washing, 16S rRNA sequencing revealed that the bacterial compositions were similar to those in rinse water. The ZFET results showed that even materials with low bacterial contamination showed high toxicity or high teratogenicity, probably because of the presence of unknown chemical additives. Conclusions: We established an integrative evaluation method for down and feather safety, based on bacterial contamination with in vivo toxicity testing.
Oenanthe Javanica Extract Protects Mouse Skin from UVB Radiation via Attenuating Collagen Disruption and Inflammation
Authors: Her Young , Shin Bich-Na , Lee Yun Lyul , Park Joon Ha , Kim Dae Won , Kim Ki Seob , Kim Hyunjung , Song Minah , Kim Jong-Dai , Won Moo-Ho , Ahn Ji Hyeon
Published: March 2019

In recent years, the use of botanical agents to prevent skin damage from solar ultraviolet (UV) irradiation has received considerable attention. Oenanthe javanica is known to exert anti-inflammatory and antioxidant activities. This study investigated photoprotective properties of an Oenanthe javanica extract (OJE) against UVB-induced skin damage in ICR mice. The extent of skin damage was evaluated in three groups: control mice with no UVB, UVB-exposed mice treated with vehicle (saline), and UVB-exposed mice treated with 1% extract. Photoprotective properties were assessed in the dorsal skin using hematoxylin and eosin staining, Masson trichrome staining, immunohistochemical staining, quantitative real-time polymerase chain reaction, and western blotting to analyze the epidermal thickness, collagen expression, and mRNA and protein levels of type I collagen, type III collagen, and interstitial collagenases, including matrix metalloproteinase (MMP)-1 and MMP-3. In addition, tumor necrosis factor (TNF)-α and cyclooxygenase (COX)-2 protein levels were also assessed. In the UVB-exposed mice treated with extract, UV-induced epidermal damage was significantly ameliorated. In this group, productions of collagen types I and III were increased, and expressions of MMP-1 and MMP-3 were decreased. In addition, TNF-α and COX-2 expressions were reduced. Based on these findings, we conclude that OJE displays photoprotective effects against UVB-induced collagen disruption and inflammation and suggest that Oenanthe javanica can be used as a natural product for the treatment of photodamaged skin.
Synthesis of New GABAA Receptor Modulator with Pyrazolo[1,5-a]quinazoline (PQ) Scaffold
Authors: Guerrini Gabriella , Vergelli Claudia , Cantini Niccolò , Giovannoni Maria Paola , Daniele Simona , Mascia Maria Paola , Martini Claudia , Crocetti Letizia
Published: March 2019

We previously published a series of 8-methoxypirazolo[1,5-a]quinazolines (PQs) and their 4,5-dihydro derivatives (4,5(H)PQ) bearing the (hetero)arylalkylester group at position 3 as ligands at the γ-aminobutyric type A (GABAA) subtype receptor. Continuing the study in this field, we report here the design and synthesis of 3-(hetero)arylpyrazolo[1,5-a]quinazoline and 3-(hetero)aroylpyrazolo[1,5-a]quinazoline 8-methoxy substituted as interesting analogs of the above (hetero)arylalkylester, in which the shortening or the removal of the linker between the 3-(hetero)aryl ring and the PQ was performed. Only compounds that are able to inhibit radioligand binding by more than 80% at 10 μM have been selected for electrophysiological studies on recombinant α1β2γ2L GABAA receptors. Some compounds show a promising profile. For example, compounds 6a and 6b are able to modulate the GABAAR in an opposite manner, since 6b enhances and 6a reduces the variation of the chlorine current, suggesting that they act as a partial agonist and an inverse partial agonist, respectively. The most potent derivative was 3-(4-methoxyphenylcarbonyl)-8-methoxy-4,5-dihydropyrazolo[1,5-a] quinazoline 11d, which reaches a maximal activity at 1 μM (+54%), and it enhances the chlorine current at ≥0.01 μM. Finally, compound 6g, acting as a null modulator at α1β2γ2L, shows the ability to antagonize the full agonist diazepam and the potentiation of CGS 9895 on the new α+/β− ‘non-traditional’ benzodiazepine site.
Protective Effects of Fermented Oyster Extract against RANKL-Induced Osteoclastogenesis through Scavenging ROS Generation in RAW 264.7 Cells
Authors: Jeong Jin-Woo , Choi Sung Hyun , Han Min Ho , Kim Gi-Young , Park Cheol , Hong Su Hyun , Lee Bae-Jin , Park Eui Kyun , Kim Sung Ok , Leem Sun-Hee , Jeon You-Jin , Choi Yung Hyun
Published: March 2019

Excessive bone resorption by osteoclasts causes bone loss-related diseases and reactive oxygen species (ROS) act as second messengers in intercellular signaling pathways during osteoclast differentiation. In this study, we explored the protective effects of fermented oyster extract (FO) against receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation in murine monocyte/macrophage RAW 264.7 cells. Our results showed that FO markedly inhibited RANKL-induced activation of tartrate-resistant acid phosphatase and formation of F-actin ring structure. Mechanistically, FO has been shown to down-regulate RANKL-induced expression of osteoclast-specific markers by blocking the nuclear translocation of NF-κB and the transcriptional activation of nuclear factor of activated T cells c1 (NFATc1) and c-Fos. Furthermore, FO markedly diminished ROS production by RANKL stimulation, which was associated with blocking the expression of nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) and its regulatory subunit Rac-1. However, a small interfering RNA (siRNA) targeting NOX1 suppressed RANKL-induced expression of osteoclast-specific markers and production of ROS and attenuated osteoclast differentiation as in the FO treatment group. Collectively, our findings suggest that FO has anti-osteoclastogenic potential by inactivating the NF-κB-mediated NFATc1 and c-Fos signaling pathways and inhibiting ROS generation, followed by suppression of osteoclast-specific genes. Although further studies are needed to demonstrate efficacy in in vivo animal models, FO may be used as an effective alternative agent for the prevention and treatment of osteoclastogenic bone diseases.
Recent Advances in Protein Kinase Activity Analysis Based on Nanomaterials
Authors: Yan Zhiyong , Deng Pingye , Liu Yang
Published: March 2019

Protein phosphorylation regulated by protein kinases, as well as their dephosphorylation, is one of the most common post-translational modifications, and plays important roles in physiological activities, such as intracellular signal communications, gene transcription, cell proliferation and apoptosis. Over-expression of protein kinases is closely associated with various diseases. Consequently, accurate detection of protein kinases activities and their relevant inhibitors screening is critically important, not only to the biochemical research, but also to the clinical diagnosis and therapy. Nanomaterials, taking advantage of large surface areas, as well as excellent electrical, catalytic, magnetic and optical properties, have been utilized as target concentrators, recognition components, signal transducer or amplification elements in protein kinase related assays. This review summarizes the recent representative works to highlight the applications of nanomaterials in different biosensor technologies for protein kinases activities detection and their inhibitors screening. First, different nanomaterials developed for phosphoprotein/phosphopeptide enrichment and phosphate recognition are introduced. Next, representative works are selected that mainly focus on the utilization of nanomaterials as signal transducer or amplification elements in various protein kinases sensing platforms, such as electrochemical, colorimetric, fluorescent, and mass spectroscopy-based approaches. Finally, the major challenges and perspectives of nanomaterials being applied in protein kinases related assays are discussed.