bluuebunny/crossref_metadata_2025 huggingface dataset:
RBM20 Regulates CaV1.2 Surface Expression by Promoting Exon 9* Inclusion of CACNA1C in Neonatal Rat Cardiomyocytes
Authors: Morinaga Akihito , Ito Jumpei , Niimi Tomoaki , Maturana Andrés D.
Published: November 2019

The CACNA1C gene encodes for the CaV1.2 protein, which is the pore subunit of cardiac l-type voltage-gated calcium (Ca2+) channels (l-channels). Through alternative splicing, CACNA1C encodes for various CaV1.2 isoforms with different electrophysiological properties. Splice variants of CaV1.2 are differentially expressed during heart development or pathologies. The molecular mechanisms of CACNA1C alternative splicing still remain incompletely understood. RNA sequencing analysis has suggested that CACNA1C is a potential target of the splicing factor RNA-binding protein motif 20 (RBM20). Here, we aimed at elucidating the role of RBM20 in the regulation of CACNA1C alternative splicing. We found that in neonatal rat cardiomyocytes (NRCMs), RBM20 overexpression promoted the inclusion of CACNA1C’s exon 9*, whereas the skipping of exon 9* occurred upon RBM20 siRNA knockdown. The splicing of other known alternative exons was not altered by RBM20. RNA immunoprecipitation suggested that RBM20 binds to introns flanking exon 9*. Functionally, in NRCMs, RBM20 overexpression decreased l-type Ca2+ currents, whereas RBM20 siRNA knockdown increased l-type Ca2+ currents. Finally, we found that RBM20 overexpression reduced CaV1.2 membrane surface expression in NRCMs. Taken together, our results suggest that RBM20 specifically regulates the inclusion of exon 9* in CACNA1C mRNA, resulting in reduced cell-surface membrane expression of l-channels in cardiomyocytes.
In Silico Study of Rett Syndrome Treatment-Related Genes, MECP2, CDKL5, and FOXG1, by Evolutionary Classification and Disordered Region Assessment
Authors: Fahmi Muhamad , Yasui Gen , Seki Kaito , Katayama Syouichi , Kaneko-Kawano Takako , Inazu Tetsuya , Kubota Yukihiko , Ito Masahiro
Published: November 2019

Rett syndrome (RTT), a neurodevelopmental disorder, is mainly caused by mutations in methyl CpG-binding protein 2 (MECP2), which has multiple functions such as binding to methylated DNA or interacting with a transcriptional co-repressor complex. It has been established that alterations in cyclin-dependent kinase-like 5 (CDKL5) or forkhead box protein G1 (FOXG1) correspond to distinct neurodevelopmental disorders, given that a series of studies have indicated that RTT is also caused by alterations in either one of these genes. We investigated the evolution and molecular features of MeCP2, CDKL5, and FOXG1 and their binding partners using phylogenetic profiling to gain a better understanding of their similarities. We also predicted the structural order–disorder propensity and assessed the evolutionary rates per site of MeCP2, CDKL5, and FOXG1 to investigate the relationships between disordered structure and other related properties with RTT. Here, we provide insight to the structural characteristics, evolution and interaction landscapes of those three proteins. We also uncovered the disordered structure properties and evolution of those proteins which may provide valuable information for the development of therapeutic strategies of RTT.
Lipid Related Genes Altered in NASH Connect Inflammation in Liver Pathogenesis Progression to HCC: A Canonical Pathway
Authors: Desterke Christophe , Chiappini Franck
Published: November 2019

Nonalcoholic steatohepatitis (NASH) is becoming a public health problem worldwide. While the number of research studies on NASH progression rises every year, sometime their findings are controversial. To identify the most important and commonly described findings related to NASH progression, we used an original bioinformatics, integrative, text-mining approach that combines PubMed database querying and available gene expression omnibus dataset. We have identified a signature of 25 genes that are commonly found to be dysregulated during steatosis progression to NASH and cancer. These genes are implicated in lipid metabolism, insulin resistance, inflammation, and cancer. They are functionally connected, forming the basis necessary for steatosis progression to NASH and further progression to hepatocellular carcinoma (HCC). We also show that five of the identified genes have genome alterations present in HCC patients. The patients with these genes associated to genome alteration are associated with a poor prognosis. In conclusion, using an integrative literature- and data-mining approach, we have identified and described a canonical pathway underlying progression of NASH. Other parameters (e.g., polymorphisms) can be added to this pathway that also contribute to the progression of the disease to cancer. This work improved our understanding of the molecular basis of NASH progression and will help to develop new therapeutic approaches.
Tilivalline- and Tilimycin-Independent Effects of Klebsiella oxytoca on Tight Junction-Mediated Intestinal Barrier Impairment
Authors: Hering Nina A. , Fromm Anja , Bücker Roland , Gorkiewicz Gregor , Zechner Ellen , Högenauer Christoph , Fromm Michael , Schulzke Jörg-Dieter , Troeger Hanno
Published: November 2019

Klebsiella oxytoca causes antibiotic-associated hemorrhagic colitis and diarrhea. This was attributed largely to its secreted cytotoxins tilivalline and tilimycin, inductors of epithelial apoptosis. To study whether Klebsiella oxytoca exerts further barrier effects, T84 monolayers were challenged with bacterial supernatants derived from tilivalline/tilimycin-producing AHC6 or its isogeneic tilivalline/tilimycin-deficient strain Mut-89. Both preparations decreased transepithelial resistance, enhanced fluorescein and FITC-dextran-4kDa permeabilities, and reduced expression of barrier-forming tight junction proteins claudin-5 and -8. Laser scanning microscopy indicated redistribution of both claudins off the tight junction region in T84 monolayers as well as in colon crypts of mice infected with AHC6 or Mut-89, indicating that these effects are tilivalline/tilimycin-independent. Furthermore, claudin-1 was affected, but only in a tilivalline/tilimycin-dependent manner. In conclusion, Klebsiella oxytoca induced intestinal barrier impairment by two mechanisms: the tilivalline/tilimycin-dependent one, acting by increasing cellular apoptosis and a tilivalline/tilimycin-independent one, acting by weakening the paracellular pathway through the tight junction proteins claudin-5 and -8.
Cytocompatibility of Titanium, Zirconia and Modified PEEK after Surface Treatment Using UV Light or Non-Thermal Plasma
Authors: Guo Linna , Smeets Ralf , Kluwe Lan , Hartjen Philip , Barbeck Mike , Cacaci Claudio , Gosau Martin , Henningsen Anders
Published: November 2019

A number of modifications have been developed in order to enhance surface cytocompatibility for prosthetic support of dental implants. Among them, ultraviolet (UV) light and non-thermal plasma (NTP) treatment are promising methods. The objective of this study was to compare the effects of UV light and NTP on machined titanium, zirconia and modified polyetheretherketone (PEEK, BioHPP) surfaces in vitro. Machined samples of titanium, zirconia and BioHPP were treated by UV light and NTP of argon or oxygen for 12 min each. Non-treated disks were set as controls. A mouse fibroblast and a human gingival fibroblast cell line were used for in vitro experiments. After 2, 24 and 48 h of incubation, the attachment, viability and cytotoxicity of cells on surfaces were assessed. Results: Titanium, zirconia and BioHPP surfaces treated by UV light and oxygen plasma were more favorable to the early attachment of soft-tissue cells than non-treated surfaces, and the number of cells on those treated surfaces was significantly increased after 2, 24 and 48 h of incubation (p < 0.05). However, the effects of argon plasma treatment on the cytocompatibility of soft tissue cells varied with the type of cells and the treated material. UV light and oxygen plasma treatments may improve the attachment of fibroblast cells on machined titanium, zirconia and PEEK surfaces, that are materials for prosthetic support of dental implants.
Knockdown of β-N-acetylglucosaminidase 2 Impairs Molting and Wing Development in Lasioderma serricorne (Fabricius)
Authors: Yang Wen-Jia , Xu Kang-Kang , Yan Xin , Li Can
Published: November 2019

β-N-acetylglucosaminidases (NAGs) are carbohydrate enzymes that degrade chitin oligosaccharides into N-acetylglucosamine monomers. This process is important for chitin degradation during insect development and metamorphosis. We identified and evaluated a β-N-acetylglucosaminidase 2 gene (LsNAG2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The full-length open reading frame of LsNAG2 was 1776 bp and encoded a 591 amino acid protein. The glycoside hydrolase family 20 (GH20) catalytic domain and an additional GH20b domain of the LsNAG2 protein were highly conserved. Phylogenetic analysis revealed that LsNAG2 clustered with the group II NAGs. Quantitative real-time PCR analyses showed that LsNAG2 was expressed in all developmental stages and was most highly expressed in the late larval and late pupal stages. In the larval stage, LsNAG2 was predominantly expressed in the integument. Knockdown of LsNAG2 in fifth instar larvae disrupted larval–pupal molting and reduced the expression of four chitin synthesis genes (trehalase 1 (LsTRE1), UDP-N-acetylglucosamine pyrophosphorylase 1 and 2 (LsUAP1 and LsUAP2), and chitin synthase 1 (LsCHS1)). In late pupae, LsNAG2 depletion resulted in abnormal adult eclosion and wing deformities. The expression of five wing development-related genes (teashirt (LsTSH), vestigial (LsVG), wingless (LsWG), ventral veins lacking (LsVVL), and distal-less (LsDLL)) significantly declined in the LsNAG2-depleted beetles. These findings suggest that LsNAG2 is important for successful molting and wing development of L. serricorne.
Special Issue ”Instruments and Methods for Cyclotron Produced Radioisotopes”
Authors: Braccini Saverio , Alves Francisco
Published: November 2019

The 17th Workshop on Targets and Target Chemistry (WTTC17) was held in Coimbra (Portugal) on 27–31 August 2018. A few months before, the 13th Workshop of the European Cyclotron Network (CYCLEUR) took place in Lisbon (Portugal) on 23–24 November 2017. These two events reassembled major experts in the field of radioisotope production, targets, target chemistry and cyclotrons. In the last few years, significant advances have been obtained in these fields with direct implications for science and society. Instruments and methods, originally developed for nuclear and particle physics, played a crucial role and remarkable developments are on-going. The production of novel radioisotopes for both diagnostics and therapy is expected to produce a breakthrough in nuclear medicine in the next years, paving the way towards theranostics and personalized medicine. This Special Issue presents a collection of original scientific contributions on the latest developments on instruments and methods for medical and research cyclotrons as well as on target and target chemistry for the production of radioisotopes.
The Association between SOCS1−1656G>A Polymorphism, Insulin Resistance and Obesity in Nonalcoholic Fatty Liver Disease (NAFLD) Patients
Authors: Kempinska-Podhorodecka Agnieszka , Wunsch Ewa , Milkiewicz Piotr , Stachowska Ewa , Milkiewicz Malgorzata
Published: November 2019

Suppressor of cytokine signaling (SOCS) proteins prevent uncontrolled cytokine signaling and appear to play a role in the pathological processes behind obesity and insulin resistance. The polymorphism of the SOCS1 gene (rs243330, −1656G>A) is associated with obesity and glucose sensitivity. To estimate the effect of this SOCS1 gene polymorphism on nonalcoholic fatty liver disease (NAFLD) susceptibility, we performed a study on 138 patients with ultrasound-confirmed NAFLD and 1000 healthy blood donors. The relationship between the SOCS1−1656G>A polymorphism and serum biochemical parameters in NAFLD was additionally investigated. The SOCS1 variant was genotyped using a dedicated TaqMan assay. The frequency of rs243330 polymorphism did not differ between patients and controls. However, in a cohort of obese individuals (BMI ≥ 30 kg/m2) the occurrence of the G allele of the SOCS1−1656G>A polymorphism was strongly associated with NAFLD (odds ratio (OR) 1.6; 95% CI,1.1–2.5; p = 0.009), and carriers of the AA genotype have lower risk of developing NAFLD (OR 0.4; 95% CI, 0.2–0.7; p = 0.004). Overweight NAFLD patients who were carriers of GG genotypes had significantly lower levels of homeostasis model assessment of insulin resistance (HOMA-IR) values (p = 0.03 vs. AA), and the obese GG homozygotes had lower serum concertation of triglyceride (GG vs. AA; p = 0.02). Serum liver enzyme activities were not modified by the presence of SOCS1 risk variants. In conclusion, the observed phenotype of overweight NAFLD patients with non-elevated levels of TG and HOMA-IR, which is associated with genetic variants of SOCS1, provides a rationale for further research on the pathophysiology of fatty liver disease.
Fatigue Life for Different Stress Concentration Factors for Stainless Steel 1.4301
Authors: Strzelecki Przemysław , Mazurkiewicz Adam , Musiał Janusz , Tomaszewski Tomasz , Słomion Małgorzata
Published: November 2019

This paper presents the results of the static tensile and fatigue life tests under rotating bending of round 1.4301 (AISI 304) steel samples. The fatigue tests were carried out on smooth and notched samples with three different rounding angles with a shape factor of 1.4, 2 and 2.6. A fatigue life was determined for samples with different shape factors subject to identical loads. The results showed that the scatter of fatigue test results decreases with an increase in shape factor. To evaluate the cracking properties (cracking mode and mechanism), microstructure and fractographic tests of the fractured samples were carried out.
Analytical Method for Predicting Early Age Thermal Effects in Thick Foundation Slabs
Authors: Klemczak Barbara
Published: November 2019

Prediction of hydration temperature and induced stresses in mass foundation slabs, due to the hydration effects is a difficult task. The complexity of this issue is compounded by transient and non-linear thermo-mechanical phenomena as well by a significant number of contributing technological and material factors that affect the early-age volume changes. This is a probable reason for the limited number of simple analytical methods allowing for the estimation of these effects. This work presents a new proposal in the discussed field. The submitted analytical method for determining the hydration temperature rise, its differentials at a cross-section and induced thermal stresses in mass concrete foundation slabs considers the majority of important technological and material factors, such as the initial temperature of the concrete, the ambient temperature, the thermal properties of the concrete and the heat exchange conditions on the slab surfaces. In stress analysis, both self-balanced and restraint stresses are calculated. Finally, the method is validated in FE analysis conducted for the slabs with various heights and made of different types of cements, as well as by the thermal measurements from the construction site. Due to the limited number of methods allowing for the analytical estimation of the early age thermo-mechanical effects in slabs, this new proposal can be useful in the assessment of these effects.
Investigation of Copper Alloying in a TNTZ-Cux Alloy
Authors: Fowler Lee , Janse Van Vuuren Arno , Goosen William , Engqvist Håkan , Öhman-Mägi Caroline , Norgren Susanne
Published: November 2019

Alloying copper into pure titanium has recently allowed the development of antibacterial alloys. The alloying of biocompatible elements (Nb, Ta and Zr) into pure titanium has also achieved higher strengths for a new alloy of Ti-1.6 wt.% Nb-10 wt.% Ta-1.7 wt.% Zr (TNTZ), where strength was closer to Ti-6Al-4V and higher than grade 4 titanium. In the present study, as a first step towards development of a novel antibacterial material with higher strength, the existing TNTZ was alloyed with copper to investigate the resultant microstructural changes and properties. The initial design and modelling of the alloy system was performed using the calculation of phase diagrams (CALPHAD) methods, to predict the phase transformations in the alloy. Following predictions, the alloys were produced using arc melting with appropriate heat treatments. The alloys were characterized using energy dispersive X-ray spectroscopy in scanning transmission electron microscopy (STEM-EDS) with transmission Kikuchi diffraction (TKD). The manufactured alloys had a three-phased crystal structure that was found in the alloys with 3 wt.% Cu and higher, in line with the modelled alloy predictions. The phases included the α-Ti (HCP-Ti) with some Ta present in the crystal, Ti2Cu, and a bright phase with Ti, Cu and Ta in the crystal. The Ti2Cu crystals tended to precipitate in the grain boundaries of the α-Ti phase and bright phase. The hardness of the alloys increased with increased Cu addition, as did the presence of the Ti2Cu phase. Further studies to optimize the alloy could result in a suitable material for dental implants.
In Vitro Evaluation of the Biological Effects of ACTIVA Kids BioACTIVE Restorative, Ionolux, and Riva Light Cure on Human Dental Pulp Stem Cells
Authors: López-García Sergio , Pecci-Lloret María P. , Pecci-Lloret Miguel R. , Oñate-Sánchez Ricardo E. , García-Bernal David , Castelo-Baz Pablo , Rodríguez-Lozano Francisco Javier , Guerrero-Gironés Julia
Published: November 2019

This study aimed to analyze the biological effects of three new bioactive materials on cell survival, migration, morphology, and attachment in vitro. ACTIVA Kids BioACTIVE Restorative (Pulpdent, Watertown, MA, USA) (Activa), Ionolux (Voco, Cuxhaven, Germany), and Riva Light Cure UV (SDI, Bayswater, Australia) (Riva) were handled and conditioned with a serum-free culture medium. Stem cells from human dental pulp (hDPSCs) were exposed to material extracts, and metabolic activity, cell migration, and cell morphology were evaluated. Cell adhesion to the different materials was analyzed by scanning electron microscopy (SEM). The chemical composition of the materials was evaluated by energy-dispersive X-ray (EDX). One-way analysis of variance followed by a Tukey test was performed (p < 0.05). Ionolux promoted a drastic reduction in metabolic activity and wound closure compared to the control (p < 0.05), whereas Activa induced adequate metabolic activity and cell migration. Moreover, SEM and immunofluorescence analysis showed abundant cells exposed to Activa. The materials showed different surface morphologies, and EDX spectra exhibited different peaks of C, O, Si, S, Ca, and F ions in glass ionomer cements. The results showed that Activa induced cell migration, cell attachment, and cell viability to a greater extent than Riva and Ionolux.
Direct Integration of Boundary Value Problems Using the Block Method via the Shooting Technique Combined with Steffensen’s Strategy
Authors: Mohd Nasir Nadirah , Abdul Majid Zanariah , Ismail Fudziah , Bachok Norfifah
Published: November 2019

This study is intended to evaluate numerically the solution of second order boundary value problems (BVPs) subject to mixed boundary conditions using a direct method. The mixed set of boundary conditions is subsumed under Type 1: mixed boundary conditions of Dirichlet and Robin and Type 2: mixed boundary conditions of Robin and Neumann. The direct integration procedure will compute the solutions at two values concurrently within a block with a fixed step size. The shooting technique adapted to the derivative free Steffensen method is employed as the iterative strategy to generate the new initial estimates. Four numerical examples are given to measure the efficiency and effectiveness of the developed numerical scheme of order six. The computational comparison indicates that the proposed method gives favorably competitive performance compared to the existing method in terms of accuracy, total function calls, and time saving.
A New Optimal Family of Schröder’s Method for Multiple Zeros
Authors: Behl Ramandeep , Alsolami Arwa Jeza , Pansera Bruno Antonio , Al-Hamdan Waleed M. , Salimi Mehdi , Ferrara Massimiliano
Published: November 2019

Here, we suggest a high-order optimal variant/modification of Schröder’s method for obtaining the multiple zeros of nonlinear uni-variate functions. Based on quadratically convergent Schröder’s method, we derive the new family of fourth -order multi-point methods having optimal convergence order. Additionally, we discuss the theoretical convergence order and the properties of the new scheme. The main finding of the present work is that one can develop several new and some classical existing methods by adjusting one of the parameters. Numerical results are given to illustrate the execution of our multi-point methods. We observed that our schemes are equally competent to other existing methods.
A New Scheme Using Cubic B-Spline to Solve Non-Linear Differential Equations Arising in Visco-Elastic Flows and Hydrodynamic Stability Problems
Authors: Tassaddiq Asifa , Khalid Aasma , Naeem Muhammad Nawaz , Ghaffar Abdul , Khan Faheem , Karim Samsul Ariffin Abdul , Nisar Kottakkaran Sooppy
Published: November 2019

This study deals with the numerical solution of the non-linear differential equations (DEs) arising in the study of hydrodynamics and hydro-magnetic stability problems using a new cubic B-spline scheme (CBS). The main idea is that we have modified the boundary value problems (BVPs) to produce a new system of linear equations. The algorithm developed here is not only for the approximation solutions of the 10th order BVPs but also estimate from 1st derivative to 10th derivative of the exact solution as well. Some examples are illustrated to show the feasibility and competence of the proposed scheme.
Biotechnological Potential of Bacteria Isolated from the Sea Cucumber Holothuria leucospilota and Stichopus vastus from Lampung, Indonesia
Authors: Wibowo Joko T. , Kellermann Matthias Y. , Versluis Dennis , Putra Masteria Y. , Murniasih Tutik , Mohr Kathrin I. , Wink Joachim , Engelmann Michael , Praditya Dimas F. , Steinmann Eike , Schupp Peter J.
Published: November 2019

In order to minimize re-discovery of already known anti-infective compounds, we focused our screening approach on understudied, almost untapped marine environments including marine invertebrates and their associated bacteria. Therefore, two sea cucumber species, Holothuria leucospilota and Stichopus vastus, were collected from Lampung (Indonesia), and 127 bacterial strains were identified by partial 16S rRNA-gene sequencing analysis and compared with the NCBI database. In addition, the overall bacterial diversity from tissue samples of the sea cucumbers H. leucospilota and S. vastus was analyzed using the cultivation-independent Illumina MiSEQ analysis. Selected bacterial isolates were grown to high densities and the extracted biomass was tested against a selection of bacteria and fungi as well as the hepatitis C virus (HCV). Identification of putative bioactive bacterial-derived compounds were performed by analyzing the accurate mass of the precursor/parent ions (MS1) as well as product/daughter ions (MS2) using high resolution mass spectrometry (HRMS) analysis of all active fractions. With this attempt we were able to identify 23 putatively known and two previously unidentified precursor ions. Moreover, through 16S rRNA-gene sequencing we were able to identify putatively novel bacterial species from the phyla Actinobacteria, Proteobacteria and also Firmicutes. Our findings suggest that sea cucumbers like H. leucospilota and S. vastus are promising sources for the isolation of novel bacterial species that produce compounds with potentially high biotechnological potential.
Requirement of lmo1930, a Gene in the Menaquinone Biosynthesis Operon, for Esculin Hydrolysis and Lithium Chloride Tolerance in Listeria monocytogenes
Authors: Parsons Cameron , Jahanafroozi Midya , Kathariou Sophia
Published: November 2019

Listeria monocytogenes is a foodborne pathogen that is widely distributed in nature, having been isolated from a variety of sources such as soil, water, plant matter, and animals. In addition, L. monocytogenes is often detected in the regular sampling of food and food processing environments. The most common method for detecting L. monocytogenes is the use of selective enrichments. Both lithium chloride and esculin, in combination with ferric ammonium citrate, are utilized in several of the most commonly-employed selective enrichment schemes for L. monocytogenes. Here we report that transposon-based inactivation of lmo1930, one of the genes in the menaquinone biosynthesis operon, via transposon mutagenesis severely impaired the ability of L. monocytogenes to grow in the presence of lithium chloride or hydrolyze esculin, and conferred reduced growth and colony size. All phenotypes were restored upon genetic complementation. Thus, strains of L. monocytogenes with mutations leading to inactivation of lmo1930 may evade many commonly-used selective enrichment protocols employed in the detection of L. monocytogenes.
Dislocated Boardgames: Design Potentials for Remote Tangible Play
Authors: Maurer Bernhard , Fuchsberger Verena
Published: November 2019

Conventional digital and remote forms of play lack the physicality associated with analog play. Research on the materiality of boardgames has highlighted the inherent material aspects to this analog form of play and how these are relevant for the design of digital play. In this work, we analyze the inherent material qualities and related experiences of boardgames, and speculate how these might shift in remote manifestations. Based on that, we depict three lenses of designing for remote tangible play: physicality, agency, and time. These lenses present leverage points for future designs and illustrate how the digital and the physical can complement each other following alternative notions of hybrid digital–physical play. Based on that, we illustrate the related design space and discuss how boardgame qualities can be translated to the remote space, as well as how their characteristics might change. Thereby, we shed light on related design challenges and reflect on how designing for shared physicality can enrich dislocated play by applying these lenses.
Hybrid Au/Si Disk-Shaped Nanoresonators on Gold Film for Amplified SERS Chemical Sensing
Authors: Barbillon Grégory , Ivanov Andrey , Sarychev Andrey K.
Published: November 2019

We present here the amplification of the surface-enhanced Raman scattering (SERS) signal of nanodisks on a gold film for SERS sensing of small molecules (thiophenol) with an excellent sensitivity. The enhancement is achieved by adding a silicon underlayer for the composition of the nanodisks. We experimentally investigated the sensitivity of the suggested Au/Si disk-shaped nanoresonators for chemical sensing by SERS. We achieved values of enhancement factors of 5 × 10 7 − 6 × 10 7 for thiophenol sensing. Moreover, we remarked that the enhancement factor (EF) values reached experimentally behave qualitatively as those evaluated with the E 4 model.
Curcumin Ameliorates Nonalcoholic Fatty Liver Disease through Inhibition of O-GlcNAcylation
Authors: Lee Da Eun , Lee Su Jin , Kim Su Ji , Lee Hyun-Shik , Kwon Oh-Shin
Published: November 2019

The cause of progression to non-alcoholic fatty liver disease (NAFLD) is not fully understood. In the present study, we aimed to investigate how curcumin, a natural phytopolyphenol pigment, ameliorates NAFLD. Initially, we demonstrated that curcumin dramatically suppresses fat accumulation and hepatic injury induced in methionine and choline-deficient (MCD) diet mice. The severity of hepatic inflammation was alleviated by curcumin treatment. To identify the proteins involved in the pathogenesis of NAFLD, we also characterized the hepatic proteome in MCD diet mice. As a result of two-dimensional proteomic analysis, it was confirmed that thirteen proteins including antioxidant protein were differentially expressed in hepatic steatosis. However, the difference in expression was markedly improved by curcumin treatment. Interestingly, eight of the identified proteins are known to undergo O-GlcNAcylation modification. Thus, we further focused on elucidating how the regulation of O-linked β-N-acetylglucosamine (O-GlcNAc) modification is associated with the progression of hepatic steatosis leading to hepatitis in MCD diet mice. In parallel with lipid accumulation and inflammation, the MCD diet significantly up-regulated hexosamine biosynthetic pathway (HBP) and O-GlcNAc transferase (OGT) via ER stress. Curcumin treatment alleviates the severity of hepatic steatosis by relieving the dependence of O-GlcNAcylation on nuclear factor-κB (NF-κB) in inflammation signaling. Conversely, the expressions of superoxide dismutase 1 (SOD1) and SIRT1 were significantly upregulated by curcumin treatment. In conclusion, curcumin inhibits O-GlcNAcylation pathway, leading to antioxidant responses in non-alcoholic steatohepatitis (NASH) mice. Therefore, curcumin will be a promising therapeutic agent for diseases involving hyper-O-GlcNAcylation, including cancer.
Frequency of Trying to Lose Weight and Its Association with Children’s Weight Perception and Dietary Intake (NHANES 2011–2012)
Authors: Brown Taiya , Overcash Francine , Reicks Marla
Published: November 2019

Weight loss practices and frequency among children and adolescents can impact overall diet quality. We used cross-sectional U.S. National Health and Nutrition Examination Survey data (NHANES) 2011–2012 from 1303 children and adolescents (8–15 years) to examine associations between frequency of trying to lose weight (never vs. sometimes/a lot) and sociodemographic characteristics, self-perception of weight, and dietary intake. A greater frequency of trying to lose weight was reported by participants with overweight/obesity, those from households with lower annual income and those who perceived they were overweight or obese compared to their counterparts. A high proportion of participants with overweight and obesity considered themselves to be “about the right weight” (76.7 and 42.8%, respectively). Intake data based on one 24-h dietary recall were examined using multivariable regression models adjusted for child and parent/family characteristics. In adjusted analyses, energy and total fat intakes were lower among those trying to lose weight compared to those never trying to lose weight. Intakes of cholesterol, sodium, and refined grains were not different by frequency of trying to lose weight (all p = 0.059–0.074). Weight loss efforts may be related to better nutritional profiles for some children and adolescents.
Leptin, Obesity, and Leptin Resistance: Where Are We 25 Years Later?
Authors: Izquierdo Andrea G. , Crujeiras Ana B. , Casanueva Felipe F. , Carreira Marcos C.
Published: November 2019

Leptin, a hormone that is capable of effectively reducing food intake and body weight, was initially considered for use in the treatment of obesity. However, obese subjects have since been found to have high levels of circulating leptin and to be insensitive to the exogenous administration of leptin. The inability of leptin to exert its anorexigenic effects in obese individuals, and therefore, the lack of clinical utility of leptin in obesity, is defined as leptin resistance. This phenomenon has not yet been adequately characterized. Elucidation of the molecular mechanisms underlying leptin resistance is of vital importance for the application of leptin as an effective treatment for obesity. Leptin must cross the blood–brain barrier (BBB) to reach the hypothalamus and exert its anorexigenic functions. The mechanisms involved in leptin transportation across the blood–brain barrier continue to be unclear, thereby preventing the clinical application of leptin in the treatment of obesity. In recent years, new strategies have been developed to recover the response to leptin in obesity. We have summarized these strategies in this review.
Deriving the Dietary Approaches to Stop Hypertension (DASH) Score in Women from Seven Pregnancy Cohorts from the European ALPHABET Consortium
Authors: Aubert Adrien M. , Forhan Anne , de Lauzon-Guillain Blandine , Chen Ling-Wei , Polanska Kinga , Hanke Wojciech , Jankowska Agnieszka , Mensink-Bout Sara M. , Duijts Liesbeth , Suderman Matthew , Relton Caroline L. , Crozier Sarah R. , Harvey Nicholas C. , Cooper Cyrus , McAuliffe Fionnuala M. , Kelleher Cecily C. , Phillips Catherine M. , Heude Barbara , Bernard Jonathan Y.
Published: November 2019

The ALPHABET consortium aims to examine the interplays between maternal diet quality, epigenetics and offspring health in seven pregnancy/birth cohorts from five European countries. We aimed to use the Dietary Approaches to Stop Hypertension (DASH) score to assess diet quality, but different versions have been published. To derive a single DASH score allowing cross-country, cross-cohort and cross-period comparison and limiting data heterogeneity within the ALPHABET consortium, we harmonised food frequency questionnaire (FFQ) data collected before and during pregnancy in ≥26,500 women. Although FFQs differed strongly in length and content, we derived a consortium DASH score composed of eight food components by combining the prescriptive original DASH and the DASH described by Fung et al. Statistical issues tied to the nature of the FFQs led us to re-classify two food groups (grains and dairy products). Most DASH food components exhibited pronounced between-cohort variability, including non-full-fat dairy products (median intake ranging from 0.1 to 2.2 servings/day), sugar-sweetened beverages/sweets/added sugars (0.3–1.7 servings/day), fruits (1.1–3.1 servings/day), and vegetables (1.5–3.6 servings/day). We successfully developed a harmonized DASH score adapted to all cohorts being part of the ALPHABET consortium. This methodological work may benefit other research teams in adapting the DASH to their study’s specificities.
The Timing of Water and Beverage Consumption During the Day Among Children and Adults in the United States: Analyses of NHANES 2011–2016 Data
Authors: Vieux Florent , Maillot Matthieu , Rehm Colin D. , Barrios Pamela , Drewnowski Adam
Published: November 2019

Dietary Guidelines for Americans 2015–20 recommend replacing sugar sweetened beverages (SSBs) with plain water in order to promote adequate hydration while reducing added sugar intake. This study explored how water intakes from water, beverages, and foods are distributed across the day. The dietary intake data for 7453 children (4–18 y) and 15,263 adults (>19 y) came from the National Health and Nutrition Examination Survey (NHANES 2011–2016). Water was categorized as tap or bottled. Beverages were assigned to 15 categories. Water intakes (in mL/d) from water, beverages, and food moisture showed significant differences by age group, meal occasion, and time of day. Plain water was consumed in the morning, mostly in the course of a morning snack and between 06:00 and 12:00. Milk and juices were consumed at breakfast whereas SSBs were mostly consumed at lunch, dinner, and in the afternoon. Children consumed milk and juices, mostly in the morning. Adults consumed coffee and tea in the morning, SSBs in the afternoon, and alcohol in the evening. Relatively little drinking water was consumed with lunch or after 21:00. Dietary strategies to replace caloric beverages with plain water need to build on existing drinking habits by age group and meal type.
Reduced Oxidative Stress and Enhanced FGF21 Formation in Livers of Endurance-Exercised Rats with Diet-Induced NASH
Authors: Henkel Janin , Buchheim-Dieckow Katja , Castro José P. , Laeger Thomas , Wardelmann Kristina , Kleinridders André , Jöhrens Korinna , Püschel Gerhard P.
Published: November 2019

Non-alcoholic fatty liver diseases (NAFLD) including the severe form with steatohepatitis (NASH) are highly prevalent ailments to which no approved pharmacological treatment exists. Dietary intervention aiming at 10% weight reduction is efficient but fails due to low compliance. Increase in physical activity is an alternative that improved NAFLD even in the absence of weight reduction. The underlying mechanisms are unclear and cannot be studied in humans. Here, a rat NAFLD model was developed that reproduces many facets of the diet-induced NAFLD in humans. The impact of endurance exercise was studied in this model. Male Wistar rats received control chow or a NASH-inducing diet rich in fat, cholesterol, and fructose. Both diet groups were subdivided into a sedentary and an endurance exercise group. Animals receiving the NASH-inducing diet gained more body weight, got glucose intolerant and developed a liver pathology with steatosis, hepatocyte hypertrophy, inflammation and fibrosis typical of NAFLD or NASH. Contrary to expectations, endurance exercise did not improve the NASH activity score and even enhanced hepatic inflammation. However, endurance exercise attenuated the hepatic cholesterol overload and the ensuing severe oxidative stress. In addition, exercise improved glucose tolerance possibly in part by induction of hepatic FGF21 production.
Dietary and Physical Activity Behaviors in Women with Polycystic Ovary Syndrome per the New International Evidence-Based Guideline
Authors: Lin Annie W. , Kazemi Maryam , Jarrett Brittany Y. , Vanden Brink Heidi , Hoeger Kathleen M. , Spandorfer Steven D. , Lujan Marla E.
Published: November 2019

Lifestyle modifications are recommended as first-line therapy in polycystic ovary syndrome (PCOS). However, usual dietary and physical activity (PA) behaviors of women with PCOS remain uncertain, likely owing to controversy in diagnostic criteria. Our objective was to contrast the usual dietary and PA behaviors of women with PCOS (n = 80) diagnosed by the 2018 International Evidence-based Guideline for the Assessment and Management of PCOS to that of controls (n = 44). Study outcomes were dietary intake, diet quality (Healthy Eating Index-2015), and PA (questionnaire, waist-worn accelerometers). Women with PCOS met the acceptable macronutrient distribution ranges for carbohydrate, fat, and protein, but did not meet the recommended dietary reference intakes for vitamin D (mean (95% confidence interval); 6 (5–7) μg/d), vitamin B9 (275 (252–298) μg/d), total fiber (24 (22–26) g/d), or sodium (4.0 (3.6–4.4) g/d). Women with PCOS also met the US recommendations for PA. No differences were detected in dietary intake, diet quality, or PA levels between groups (p ≥ 0.11). In conclusion, women with and without PCOS have comparable dietary and PA behaviors. A lack of unique targets for dietary or PA interventions supports the position of the new guideline to foster healthy lifestyle recommendations for the management of PCOS.
Plant-Based Diets in the Reduction of Body Fat: Physiological Effects and Biochemical Insights
Authors: Najjar Rami S. , Feresin Rafaela G.
Published: November 2019

Obesity affects over one-third of Americans and increases the risk of cardiovascular disease and type II diabetes. Interventional trials have consistently demonstrated that consumption of plant-based diets reduces body fat in overweight and obese subjects, even when controlling for energy intake. Nonetheless, the mechanisms underlying this effect have not been well-defined. This review discusses six major dietary mechanisms that may lead to reduced body fat. These include (1) reduced caloric density, (2) improved gut microbiota symbiosis, (3) increased insulin sensitivity, (4) reduced trimethylamine-N-oxide (TMAO), (5) activation of peroxisome proliferator-activated receptors (PPARs), and (6) over-expression of mitochondrial uncoupling proteins. Collectively, these factors improve satiety and increase energy expenditure leading to reduced body weight.
C60 Fullerene as an Effective Nanoplatform of Alkaloid Berberine Delivery into Leukemic Cells
Authors: Grebinyk Anna , Prylutska Svitlana , Buchelnikov Anatoliy , Tverdokhleb Nina , Grebinyk Sergii , Evstigneev Maxim , Matyshevska Olga , Cherepanov Vsevolod , Prylutskyy Yuriy , Yashchuk Valeriy , Naumovets Anton , Ritter Uwe , Dandekar Thomas , Frohme Marcus
Published: November 2019

A herbal alkaloid Berberine (Ber), used for centuries in Ayurvedic, Chinese, Middle-Eastern, and native American folk medicines, is nowadays proved to function as a safe anticancer agent. Yet, its poor water solubility, stability, and bioavailability hinder clinical application. In this study, we have explored a nanosized carbon nanoparticle—C60 fullerene (C60)—for optimized Ber delivery into leukemic cells. Water dispersions of noncovalent C60-Ber nanocomplexes in the 1:2, 1:1, and 2:1 molar ratios were prepared. UV–Vis spectroscopy, dynamic light scattering (DLS), and atomic force microscopy (AFM) evidenced a complexation of the Ber cation with the negatively charged C60 molecule. The computer simulation showed that π-stacking dominates in Ber and C60 binding in an aqueous solution. Complexation with C60 was found to promote Ber intracellular uptake. By increasing C60 concentration, the C60-Ber nanocomplexes exhibited higher antiproliferative potential towards CCRF-CEM cells, in accordance with the following order: free Ber < 1:2 < 1:1 < 2:1 (the most toxic). The activation of caspase 3/7 and accumulation in the sub-G1 phase of CCRF-CEM cells treated with C60-Ber nanocomplexes evidenced apoptosis induction. Thus, this study indicates that the fast and easy noncovalent complexation of alkaloid Ber with C60 improved its in vitro efficiency against cancer cells.
IVIVC Assessment of Two Mouse Brain Endothelial Cell Models for Drug Screening
Authors: Puscas Ina , Bernard-Patrzynski Florian , Jutras Martin , Lécuyer Marc-André , Bourbonnière Lyne , Prat Alexandre , Leclair Grégoire , Roullin V. Gaëlle
Published: November 2019

Since most preclinical drug permeability assays across the blood-brain barrier (BBB) are still evaluated in rodents, we compared an in vitro mouse primary endothelial cell model to the mouse b.End3 and the acellular parallel artificial membrane permeability assay (PAMPA) models for drug screening purposes. The mRNA expression of key feature membrane proteins of primary and bEnd.3 mouse brain endothelial cells were compared. Transwell® monolayer models were further characterized in terms of tightness and integrity. The in vitro in vivo correlation (IVIVC) was obtained by the correlation of the in vitro permeability data with log BB values obtained in mice for seven drugs. The mouse primary model showed higher monolayer integrity and levels of mRNA expression of BBB tight junction (TJ) proteins and membrane transporters (MBRT), especially for the efflux transporter Pgp. The IVIVC and drug ranking underlined the superiority of the primary model (r2 = 0.765) when compared to the PAMPA-BBB (r2 = 0.391) and bEnd.3 cell line (r2 = 0.019) models. The primary monolayer mouse model came out as a simple and reliable candidate for the prediction of drug permeability across the BBB. This model encompasses a rapid set-up, a fair reproduction of BBB tissue characteristics, and an accurate drug screening.
Advances in Lipid and Metal Nanoparticles for Antimicrobial Peptide Delivery
Authors: Makowski Marcin , Silva Ítala C. , Pais do Amaral Constança , Gonçalves Sónia , Santos Nuno C.
Published: November 2019

Antimicrobial peptides (AMPs) have been described as excellent candidates to overcome antibiotic resistance. Frequently, AMPs exhibit a wide therapeutic window, with low cytotoxicity and broad-spectrum antimicrobial activity against a variety of pathogens. In addition, some AMPs are also able to modulate the immune response, decreasing potential harmful effects such as sepsis. Despite these benefits, only a few formulations have successfully reached clinics. A common flaw in the druggability of AMPs is their poor pharmacokinetics, common to several peptide drugs, as they may be degraded by a myriad of proteases inside the organism. The combination of AMPs with carrier nanoparticles to improve delivery may enhance their half-life, decreasing the dosage and thus, reducing production costs and eventual toxicity. Here, we present the most recent advances in lipid and metal nanodevices for AMP delivery, with a special focus on metal nanoparticles and liposome formulations.
Preparation, Characterization, and In Vivo Pharmacokinetic Study of the Supercritical Fluid-Processed Liposomal Amphotericin B
Authors: Lim Chang-baek , Abuzar Sharif Md , Karn Pankaj Ranjan , Cho Wonkyung , Park Hee Jun , Cho Cheong-Weon , Hwang Sung-Joo
Published: November 2019

Here, we aimed to prepare and optimize liposomal amphotericin B (AmB) while using the supercritical fluid of carbon dioxide (SCF-CO2) method and investigate the characteristics and pharmacokinetics of the SCF-CO2-processed liposomal AmB. Liposomes containing phospholipids, ascorbic acid (vit C), and cholesterol were prepared by the SCF-CO2 method at an optimized pressure and temperature; conventional liposomes were also prepared using the thin film hydration method and then compared with the SCF-CO2-processed-liposomes. The optimized formulation was evaluated by in vitro hemolysis tests on rat erythrocytes and in vivo pharmacokinetics after intravenous administration to Sprague-Dawley rats and compared with a marketed AmB micellar formulation, Fungizone®, and a liposomal formulation, AmBisome®. The results of the characterization studies demonstrated that the SCF-CO2-processed-liposomes were spherical particles with an average particle size of 137 nm (after homogenization) and drug encapsulation efficiency (EE) was about 90%. After freeze-drying, mean particle size, EE, and zeta potential were not significantly changed. The stability study of the liposomes showed that liposomal AmB that was prepared by the SCF method was stable over time. In vivo pharmacokinetics revealed that the SCF-CO2-processed-liposomes were bioequivalent to AmBisome®; the hemolytic test depicted less hematotoxicity than Fungizone®. Therefore, this method could serve as a potential alternative for preparing liposomal AmB for industrial applications.
Biofunctionalized Polyelectrolyte Microcapsules Encoded with Fluorescent Semiconductor Nanocrystals for Highly Specific Targeting and Imaging of Cancer Cells
Authors: Nifontova Galina , Kalenichenko Daria , Baryshnikova Maria , Ramos Gomes Fernanda , Alves Frauke , Karaulov Alexander , Nabiev Igor , Sukhanova Alyona
Published: November 2019

Fluorescent semiconductor nanocrystals or quantum dots (QDs) are characterized by unique optical properties, including a high photostability, wide absorption spectrum, and narrow, symmetric fluorescence spectrum. This makes them attractive fluorescent nanolabels for the optical encoding of microcarriers intended for targeted drug delivery, diagnosis, and imaging of transport processes on the body, cellular, and subcellular levels. Incorporation of QDs into carriers in the form of polyelectrolyte microcapsules through layer-by-layer adsorption of oppositely charged polyelectrolyte polymers yields microcapsules with a bright fluorescence signal and adaptable size, structure, and surface characteristics without using organic solvents. The easily modifiable surface of the microcapsules allows for its subsequent functionalization with capture molecules, such as antibodies, which ensures specific and selective interaction with cells, including tumor cells, with the use of the bioconjugation technique developed here. We obtained stable water-soluble nanolabels based on QDs whose surface was modified with polyethylene glycol (PEG) derivatives and determined their colloidal and optical characteristics. The obtained nanocrystals were used to encode polyelectrolyte microcapsules optically. The microcapsule surface was modified with humanized monoclonal antibodies (Abs) recognizing a cancer marker, epidermal growth factor receptor (EGFR). The possibility of effective, specific, and selective delivery of the microcapsules to tumor cells expressing EGFR has been demonstrated. The results show that the QD-encoded polyelectrolyte microcapsules functionalized with monoclonal Abs against EGFR can be used for targeted imaging and diagnosis.