bluuebunny/crossref_metadata_2025 huggingface dataset:
Coniferyl aldehyde alleviates LPS-induced WI-38 cell apoptosis and inflammation injury via JAK2–STAT1 pathway in acute pneumonia
Authors: He Yichun , Li Qin , Zhou Weijun , Gu Yanhong , Jiang Yu
Published: September 2021

Pneumonia is a kind of inflammatory disease characterized by pathogen infection of lower respiratory track. Lipopolysaccharide (LPS) is the main bioactive component of Gram-negative bacteria responsible for inflammatory response. Recently, coniferyl aldehyde (CA) has been reported to play a crucial role because of its anti-inflammatory activity. However, the effect and mechanisms of CA in ameliorating symptoms of acute pneumonia remain unknown. Evaluating and identifying the value and exploring the mechanisms of CA on LPS-mediated WI-38 apoptosis and inflammation were the aims of this study. Here, CCK-8 cell viability assay was applied on WI-38 after treatment with or without LPS at different doses of CA to verify that CA can increase LPS-induced cell viability. Then, quantitative polymerase chain reaction (qPCR) and enzyme-linked-immunosorbent serologic assays (ELISA) suggested that LPS treatment dramatically decreased the expression level of IL-10 (anti-inflammatory factor) while strikingly increasing the expression levels of IL-1β, IL-6, and TNF-α (tumor necrosis factor-α; proinflammatory factor) whereas CA treatment attenuates LPS-induced inflammation of WI-38. Further, flow cytometry and Western blot assay verified that LPS treatment dramatically promoted apoptosis of WI-38 cells, while administration of CA notably inhibited apoptosis of WI-38 cells. Moreover, the Western blot assay hinted that CA could inactivate LPS-induced JAK2–STAT1 signaling pathway. These findings indicated that CA could alleviate LPS-mediated WI-38 apoptosis and inflammation injury through JAK2–STAT1 pathway in acute pneumonia.
KCNQ1OT1 promotes the proliferation and migration of psoriatic keratinocytes by regulating miR-183-3p/GAB1
Authors: Liu Ting , Duan Xi , He Jia , Yang Chuan
Published: September 2021

Background: Differentially expressed long non-coding RNAs (lncRNA) have been reported to be involved in the proliferation and migration of keratinocyte. Potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 (KCNQ1OT1) was implicated in the pathogenesis of various diseases, including cancer, sepsis, diabetic cardiomyopathy, and atherosclerosis. In this study, the influence of KCNQ1OT1 on the proliferation and migration of psoriatic keratinocytes was explained.
Methods: Cultured human keratinocyte cell line (HaCaT) was incubated with tumor necrosis factor-α (TNF-α). Cell viability and migration were assessed by MTT assay and wound healing, respectively. Target miRNA of KCNQ1OT1 was identified by luciferase activity and RNA immunoprecipitation (RIP) assays.
Results: KCNQ1OT1 was up-regulated in TNF-α-treated HaCaT cell line, and knockdown of KCNQ1OT1 reduced viability and suppressed the migration of TNF-α-treated HaCaT cell line. KCNQ1OT1 was bound to microRNA-183-3p (miR-183-3p) and negatively regulated its expression. Over-expression of growth factor receptor binding 2-associated binding protein 1 (GAB1) counteracted with the suppressive effects of KCNQ1OT1-induced silence on the viability and migration of TNF-α-treated HaCaT cells.
Conclusion: KCNQ1OT1 silence suppressed the proliferation and migration of TNF-α-treated HaCaT cells through regulation of miR-183-3p/GAB1.
PRMT5 promotes inflammation of cigarette smoke extract-induced bronchial epithelial cells by up-regulation of CXCL10
Authors: Ju Junfang , He Yingchun
Published: September 2021

Background: Chronic obstructive pulmonary disease (COPD) is related to inflammation and obstruction of the lungs and airways. Protein arginine methyltransferase 5 (PRMT5) that pro-motes arginine methylation of histones is associated with inflammation of endothelial cell and is implicated in lung branching morphogenesis and progression of lung cancer. The mechanism of PRMT5 in inflammatory response of COPD was explored in this study.
Methods: Human bronchial epithelial cells, 16HBE, were treated with cigarette smoke extract for 24 h to establish cell model of COPD. Cell viability was examined by MTT assay. Western blot and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays were used to explore expression of PRMT5. Expression of Interleukin (IL)-6, IL-8, tumor necrosis factor-α (TNF-α), and IL-1β were investigated by enzyme-linked-ithe mmunosorbent serologic assay.
Results: Cigarette smoke extract treatment induced cytotoxity of 16HBE with reduced cell viability. PRMT5 was enhanced in cigarette smoke extract-induced 16HBE. Knockdown of PRMT5 increased cell viability of cigarette smoke extract-induced 16HBE, and attenuated cigarette smoke extract-induced increase of IL-6, IL-8, TNF-α, and IL-1β. Up-regulation of C-X-C Motif Chemokine 10 (CXCL10) in cigarette smoke extract-induced 16HBE was restored by knockdown of PRMT5. Over-expression of CXCL10 counteracted with the suppressive effect of PRMT5 silence on expression of IL-6, IL-8, TNF-α, and IL-1β. Moreover, PRMT5 silence-induced increase of cell viability in cigarette smoke extract-induced 16HBE was reversed by over-expression of CXCL10.
Conclusion: Knockdown of PRMT5 promoted cell viability of cigarette smoke extract-induced 16HBE, and reduced inflammation through down-regulation of CXCL10.
Presentation of the Dossier - The royalty in the ancient Eastern world: Textual and image narratives
Authors: Santos João Batista Ribeiro , Pozzer Katia Maria Paim
Published: September 2021

The idea of organizing a dossier on Eastern Antiquity has haunted us for some years. As scholars of Eastern history and archeology, it is our duty to provoke debate and reflection on the ancient Near East in its various temporalities and regions. Thus emerged the thematic proposal on royalty in the ancient oriental world.
Análise espacial do fluxo origem-destino das internações por síndrome respiratória aguda grave por COVID-19 na região metropolitana do Rio de Janeiro
Authors: Cavalcante João Roberto , Xavier Diego Ricardo , Santos Cleber Vinicius Brito dos , Pungartnik Paula Cristina , Guimarães Raphael Mendonça
Published: September 2021

RESUMO: Este estudo analisou o fluxo intermunicipal das internações por síndrome respiratória aguda grave por COVID-19 na região metropolitana do Rio de Janeiro. Foram identificadas 12.676 internações intermunicipais por COVID-19 envolvendo o município do Rio de Janeiro. Dessas, 11.288 (89,0%) eram de residentes na região metropolitana, 87% de residentes em outros municípios da mesma região e internados na capital do estado, e 13% eram residentes da capital internados em outros municípios da região. Há correlação negativa entre a distância dos municípios e o fluxo origem-destino (r=0,62, p<0,001). O município do Rio de Janeiro importa mais internações por síndrome respiratória aguda grave por COVID-19 do que exporta. Este estudo evidenciou a importância das redes de atendimento para casos mais graves, os quais necessitem, principalmente, de atenção especializada.
Desigualdade na atenção pré-hospitalar no Brasil: Análise da eficiência e suficiência da cobertura do SAMU 192
Authors: Malvestio Marisa Aparecida Amaro , Sousa Regina Márcia Cardoso de
Published: September 2021

Objetivo : Analisar a evolução e o cenário de cobertura da atenção pré-hospitalar no Brasil entre 2015 e 2019, conforme dimensões de eficiência e suficiência, na totalidade dos municípios, segundo macrorregiões, população coberta, estratégia de cobertura, área geográfica, coberta disponível disponível e recursos móveis distribuídos. Método : Estudo censitário, observacional, abordagem quantitativa, descritiva e exploratória, realizado com dados sobre o Serviço de Atendimento Móvel de Urgência (SAMU 192) fornecido pelo Ministério da Saúde, complementados com área geográfica e população, por município. Resultados: No período, a cobertura cresceu 5,4%, alcançou 85,0% dos cidadãos em 3750 municípios (67,3%), com avanço (0,6%) e predomínio (83,0%) da modalidade de Suporte Básico de Vida. Há variações na eficiência de cobertura entre regiões (entre 77,2% na região norte e 94,6% na região sul) com desigualdade na composição das superiores de resposta (entre 10,5% de equipes de suporte avançado de vida na região norte à 18,3% na região sul), acentuadamente maior entre capitais e municípios do interior. Capitais e municípios com 10 mil a 50 mil hab, concentram recursos do SAMU 192. A face mais vulnerável da cobertura está nos 1938 municípios cobertos regionalmente e nos 1820 municípios sem cobertura (51,8% com <10 mil hab). Conclusão: A eficiência na cobertura é coerente com uma direção de integralidade do sistema de saúde, entretanto, uma expansão exígua e a persistência de cidadãos descobertos e um cenário de desigualdade de recursos e informação de resposta nos locais diferentes regiões, corrompem a diretriz de igualdade no acesso à saúde (suficiência).
Pulp mill wastewater management using an advanced oxidation process and activated sludge
Authors: Davarnejad Reza , Abbasi Vajihe , Izadi Ali
Published: September 2021

In this study, the electro-Fenton process (with response surface methodology as an experiments design and optimisation technique) was applied to evaluate the effects of five independent variables on the chemical oxygen demand (COD) and colour removal from paper pulp wastewater (PW). The parameters were statistically optimised under response surface methodology. The optimum conditions for 98.2% of COD and 96.8% of colour removal experimentally were at a reaction time of 30 min, current density of 79.9 mA/cm2, pH of 6.9, hydrogen peroxide (H2O2)/PW volume ratio of 1.7 ml/l, hydrogen peroxide (H2O2)/ferrous ions (Fe2+) molar ratio of 1.6, electrode consumption of 0.1 (g/l) and energy consumption of 1.3 (kWh/m3). In the next step, the filtration process was carried out on the treated effluent obtained at the optimal conditions. Then, this was mixed with activated sludge in a ratio of 1 : 10; 99.9% of COD and colour removal was found after 150 and 60 min, respectively.
Rates of bubble diffusion and turbulence dissipation in highly aerated hydraulic jumps
Authors: Bai Ruidi , Wang Hang , Liu Shanjun
Published: September 2021

Air–water mixing associated with high turbulence is a common hydraulic phenomenon, where the transport of entrained air bubbles and the dissipation of turbulent energy are key processes affecting the air–water mass transfer. Herein hydraulic jumps were selected as a seminal case of self-air-entrainment in transitional water flow, and the air–water flow properties were investigated at high Froude numbers from 10.5 to 13.5. The analysis of air concentration distributions and bubble size spectra enabled deviation of the rates of bubble diffusion and turbulence dissipation. The study was facilitated with re-analyses of earlier experimental data for smaller Froude numbers between 3.8 and 10. The spatial evolution of the diffusion and dissipation rates suggested that the diffusion of air bubbles and dissipation of turbulent structures were not independent processes, and, at high Froude numbers, the mix of air and water was subject to re-aeration of the turbulent shear layer by the roller.
Reviewer Acknowledgements 2021
Authors: Terrazas Teresa
Published: September 2021

Reviewer Acknowledgements 2021
MAGNETIC FORCE MICROSCOPE IMAGE EVALUATION OF MAGNETIC IRON OXIDE FLOPPY DISC WITH VARIOUS LIFT HEIGHTS
Authors: Shabrina Nanda , Mardiyanto Mardiyanto , Rivai Abu Khalid
Published: September 2021

Magnetic Force Microscope Image Evaluation of Magnetic Iron Oxide Floppy Disc with Various Lift Heights. One of the advantages of Scanning Probe Microscopes (SPM) or better known as the Atomic Force Microscope (AFM) is its ability to "see" in detail at the level of atoms and molecules, so as to improve understanding of how a system works and leads to new discoveries in areas such as life science, materials science, electrochemistry, polymer science, biophysics, nanotechnology and biotechnology. To understand about the MFM mode, amplitude, phase and topographic image, a piece of a flop-py disk as references sample was used in various lift heights. In this paper presented the principles of AFM work and explain the necessary components of such equipment. The MFM images were taken in floating operation method at different heights namely 30, 40 and 60 nm between the surface sample and Co-Cr cantilever tip. Result showed the distance between the cantilever tip to the sample surface influenced the image quality. The best amplitude image could be taken in 60 nm distance.
EFFECT OF CRYSTALLINITY TO OVERPOTENTIAL ON Ni₃Fe ALLOY AS ELECTROCATALYST IN HYDROGEN EVOLUTION REACTION
Authors: Mu'minah Qonita , Rochliadi Achmad , Patah Aep
Published: September 2021

EFFECT OF CRYSTALLINITY TO OVERPOTENTIAL ON Ni3Fe ALLOY AS ELECTROCATALYST IN HYDROGEN EVOLUTION REACTION. Ni-Fe alloys can be used as electrocatalyst for the hydrogen evolution reaction (HER) in an alkaline solution. HER consumed highly energy and overpotential driven. The overpotential value corresponding to the electron transfer in reaction can be affected either by metal combination or alloy as a cathode. Ni₃Fe  alloy had been successfully synthesized by the electrodeposition method using direct-current (DC) on a 304 L type stainless steel substrate. The modified Watts bath deposition was used NiCl2·6H2O and FeCl3·6H2O as precursors of the alloy. The optimum conditions of the reaction were obtained at pH of the solution is 2.20±0.02 with 25 mA/cm² current density at 55 °C for 160 minutes. Ni₃Fe alloy was characterized by Powder X-ray Diffraction (PXRD), Energy-Dispersive X-ray Spectroscopy (EDX), and Scanning Electron Microscopy (SEM). The electrocatalytic property of Ni3Fe alloy was electrochemically measured in 1 M KOH solution by polarization method using a Tafel plot with a scanning rate of 1 mV/s. As a result, the mass ratio of Ni²+ /Fe³+ in bath deposition influenced the electrocatalytic property of Ni₃Fe alloy. Ni₃Fe alloy with a higher crystallinity lowered the overpotential value of HER up to 67% compared to Ni metal.
EFFECT OF ULTRASONIC TREATMENT ON THE CONSOLIDATION OF ODS STEEL FeCrY2O3 PROCESSING WITH CAPSULATED SINTERING PROCESS.
Authors: Silalahi Marzuki , Bandriyana Bernardus , Bayu Harum Andriadi , Sugeng Bambang , Salam Rohmad
Published: September 2021

EFFECT OF ULTRASONIC TREATMENT ON THE CONSOLIDATION OF ODS STEEL FeCrY2O3 PROCESSING WITH CAPSULATED SINTERING PROCESS. A new method on the synthesis of ODS (Oxide Dispersion Strengthened) steel for advanced nuclear material was performed by ultrasonic treatment to improve the consolidation process. The raw material of Fe, Cr and Y2O3powder with the composition of Fe-15 wt% Cr, 0.5 wt% and Ytria (Y2O3)  as disperzoid were  processed by the powder metallurgy method with the main process of pre-alloying, iso-compaction and sintering process. The pre-alloying process was carried out by mixing the alloying elements using ultrasonically treatment at frquency of 20 kHz with variation of  40, 50 and 60 % amplitude. Iso-compaction process was done using the load of  800 psi to obtain a pellet-shaped sample, then continued by the sintering process for consolidation. The sintering process was done in two stages, 1-step sintering and 2-steps sintering, using the heating furnace by putting the sample in a quartz capsule to prevent oxidation attack. Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy (SEM-EDS), X-ray Diffraction (XRD), and Micro-hardness tests were carried out to analyze the microstructure and phase formation in relation to the consolidation process. The highest hardness occurred in the addition of treatment with an amplitude of 60% which produces a micro structure with the most fine grain. For 1-step sintering process, the highest hardness of 134.51 VHN obtained at 40 % amplitude. The hardness of the alloy depends on the size of the grain boundary associated with the difficulty of the dislocation movement.
BIOREDUCTION AND CHARACTERIZATION OF SILVER NANOPARTICLES FROM OIL PALM EMPTY FRUIT BUNCH
Authors: Suhartati Siti , Saputra Iwan Syahjoko , Rahmi Dwinna , Yulizar Yoki , Sudirman Sudirman
Published: September 2021

BIOREDUCTION AND CHARACTERIZATION OF SILVER NANOPARTICLES FROM OIL PALM EMPTY FRUIT BUNCH (OPEFB). The synthesis of silver nanoparticles was successfully carried out by extracting oil palm empty fruit bunch. The precursor used was silver nitrate (AgNO3) with a concentration of 9x10-4 M and 5 wt% of the oil palm empty fruit bunch extract. OPEFB acted as a capping agent in the synthesis of silver nanoparticles. The bioreduction method Ag+ to Ag0 produced a silver nanoparticle colloid in brown color. The results of the UV-Vis spectrophotometer showed the silver nanoparticles colloids spectrum at a wavelength of 420 nm with an absorbance value of 0.5. FTIR shows the reduction and shift of absorption peak in the hydroxyl functional group (-OH) at wavenumbers of 3323 cm-1 and the presence of absorption peaks at 560 cm-1. While, XRD pattern showed the specific crystallinity peaks of silver nanoparticles at 2θ: 33.24°; 39.98°; 61.23°; dan 79.13° respectively with the face-centered cubic crystal structure (FCC) and crystallite size of 15 nm. PSA analysis showed two specific peaks with an average size distribution silver nanoparticles of 43.5 nm and a PDI value of 0.4. Analysis of TEM shows the average particle size of 20 nm with a spherical particle shape.
THE EFFECT OF CALCINATION TEMPERATURE TO THE COMPOSITE CHARACTERISTICS OF TiO2SiO2 NANOPARTICLE
Authors: Ananda Della Dwi , Maharani Dina Kartika
Published: September 2021

THE EFFECT OF CALCINATION TEMPERATURE TO THE COMPOSITECHARACTERISTICS OF TiO2SiO2 NANOPARTICLE. Self-cleaning material is a material that utilizes the photocatalytic property to degrade organic and inorganic compounds with the help of UV light. One of the materials that have good photocatalytic property is TiO2, the photocatalytic property causes TiO2 to be amphiphilic: becomes hydrophilic when there is light and becomes hydrophobic when there is no light. The photocatalytic property of TiO2 can be improved with the addition of buffer material such as SiO2. TiO2SiO2 nanoparticle was synthesized using a sol-gel method with Titanium (IV) Isopropoxide (TTIP) precursors for TiO2 and Tetraethyl Orthosilicate (TEOS) precursors for SiO2 and followed by a variation of calcination temperature of 400 °C, 450 °C, 500 °C, and 550 °C for 2 hours. TiO2SiO2 composite was synthesized using composition TiO2 sol 75% and SiO2 25%. The result of the synthesis TiO2SiO2 composite was characterized by Fourier Transform Infra-Red (FT-IR) instrument to determine the functional groups in the composites and X-Ray Diffraction (XRD) instruments to determine the phase, crystallite size and degree of crystallinity in the composite. The purpose of this research is to synthesized TiO2SiO2 nanoparticle as a self cleaning agent with variation of the calcination temperature, to obtain composite characteristics that can support self cleaning. The self-cleaning ability was based on a produced composite characteristic of TiO2SiO2. The result of FTIR characterization showed that at calcination temperature of 400 °C, 450 °C, 500 °C, and 550 °C there was a Ti-OSi bond at the peak of 948.91 cm-1, 950.77 cm-1, 941.13 cm-1, 942.13 cm-1. The result of XRD characterization showed that at the temperature of 400 °C had the best characteristics, the 75.27% anatase phase and brookite phase 24.72%. Calcinations temperature 400 °C had best degree of crystallinity of 91.66%.
SANS STUDIES ON THE BOVINE SERUM ALBUMIN DENATURATION IN THE PRESENCE OF SDS
Authors: Rahayu Dwi , Patriati Arum , Suparno Nadi , Putra Edy Giri Rachman
Published: September 2021

The effect of the presence of sodium dodecyl sulfate (SDS) on the denaturation of bovine serum albumin (BSA) has been studied using 36 m small-angle neutron scattering (SANS) BATAN spectrometer (SMARTer). The neutron scattering data reduction used the Graphical Reduction and Analysis SANS Program (GRASP) software, and the fitting process used the IGOR SANS Analysis software. The denaturation process was identified by observing the changes BSA globular structure. The experimental results showed the addition of SDS at low concentrations (2 mM, 5 mM, 10 mM) into BSA solution at pH 7 do not cause a significant change in the size of the BSA globular structure. The SANS scattering profile of BSA fitted with the triaxial ellipsoid model, a simple shape approach for protein globular structure. The fitting result showed the semi-axis B for BSA in the addition of 2 mM, 5 mM, 10 mM SDS were 33.8 Å, 33.8 Å, and 37.8 Å, respectively. While the semi-axis A and semi-axis C were constant for those three variations at 14.6 Å and 32.2 Å, respectively. In higher addition of SDS, the globular structure of BSA unfolded into flexible cylinder structure with the radius of 14.4 Å and length of 83.5 Å. The denaturation of BSA was clearly showed by the addition of 40 mM SDS. The structure of BSA in this condition fitted to fractal structure with fractal dimension of 1.1, the block radius of 16.7 Å and the correlation length of 42.5 Å. These results indicated that the addition of SDS at low concentrations has not caused the denaturation of BSA. Meanwhile, the addition of SDS at high concentrations made BSA to unfold that lead to the denaturation of BSA.
A SYNERGISTIC ABSORPTION AND PLASMONIC EFFECT OF SiO2@Au@TiO2 IN TiO2 PHOTOANODE FOR DYE-SENSITIZED SOLAR CELLS
Authors: Pakpahan Matius Nata , Hartanto Aldi , Gultom Yonatan Davidson , Fadhilah Nur , Risanti Doty Dewi
Published: September 2021

A SYNERGISTIC ABSORPTION AND PLASMONIC EFFECT OF SiO2@Au@TiO2 IN A TiO2 PHOTOANODE FOR DYE-SENSITIZED SOLAR CELLS. A method for increasing the visible-light harvesting of a TiO2 anatase photoanode in dye-sensitized solar cells by incorporating plasmonic nanostructures was developed. Sidoarjo mud as the SiO2 source was used to successfully synthesized core/multishell SiO2@Au@TiO2, with varying amounts of Au (60, 90, and 120 mL). In addition, the core/multishell fractions in TiO2 paste were varied, i.e., 0.5%, 1%, and 5%. The UV–Vis spectrum shows that a more ripple spectrum at higher wavelengths is obtained with increasing Au content, as suggested by the presence of large Au nanoparticles; however, a similar value of efficiency is observed for all sample variations studied compared to a pure TiO2 photoanode. The incident photon-to-current efficiency reveals that all photoanodes containing the core/multishell SiO2@Au@TiO2 studied show somewhat broader and enhanced spectra for all studied wavelengths compared to the pure TiO2 photoanode, resulting from the synergistic effect between plasmonic nanostructures and the presence of silica that boost the absorption to higher wavelengths.
SYNTHESIS POLYMER MATRIX COMPOSITE EPOXY FeNdB-Cu FOR RADAR ABSORBER COATING APPLICATION
Authors: Latifah Sri Mulyati , Prajitno Djoko Hadi , Syukron Muhammad Fadhil
Published: September 2021

SYNTHESIS POLYMER MATRIX COMPOSITE EPOXY FeNdB-Cu FOR RADAR ABSORBER COATING APPLICATION. Radar (Radio Detection and Ranging) is a technology using the principle of electromagnetic for detecting, measuring, and mapping target coordinates. So stealth technology was created to tackle these threats, one of which is the radar absorber coating method (RAC). This research aims to identify the effect addition of Cu to Fe-Nd-B radar absorber material as polymer matrix composite (PMC). The processes were done by powder metallurgy method, started by milling and mechanical alloying process using planetary ball mill for 60 minutes 1000 rpm, 25 kg/mm2 of compaction load, and sintering at the temperature 1000 oC for 3 hours. The coating process was carried out conventional method as much as 1 layer, the variation of resin and magnetic powder are (95:5), (90:10), and (85:5) with the addition of Cu 1%wt. Based on the characterization the lowest RL was obtained at the composition (85:15) with the value 32.08 dB at the frequency of 10.36 GHz, magnetic characterization after doping addition coercivity 0.096 kOe and Saturation 5.22 kG. PMC hardness was raising following to addition of magnetic powder with the 29.2 HD Shore D, and adhesivity value was decreased following to addition of magnetic powder to 0.5 MPa.
Protocols for Northern Analysis of Exosome Substrates and Other Noncoding RNAs v1
Authors: Houseley Jonathan , Cruz Cristina
Published: September 2021

Over the past decade a plethora of noncoding RNAs (ncRNAs) have been identified, initiating an explosion in RNA research. Although RNA sequencing methods provide unsurpassed insights into ncRNA distribution and expression, detailed information on structure and processing are harder to extract from sequence data. In contrast, northern blotting methods provide uniquely detailed insights into complex RNA populations but are rarely employed outside specialist RNA research groups. Such techniques are generally considered difficult for nonspecialists, which is unfortunate as substantial technical advances in the past few decades have solved the major challenges. Here we present simple, reproducible and highly robust protocols for separating glyoxylated RNA on agarose gels and heat denatured RNA on polyacrylamide–urea gels using standard laboratory electrophoresis equipment. We also provide reliable transfer and hybridization protocols that do not require optimization for most applications. Together, these should allow any molecular biology lab to elucidate the structure and processing of ncRNAs of interest.
Identifying and Validating Tankyrase Binders and Substrates: A Candidate Approach v1
Authors: Pollock Katie , Ranes Michael , Collins Ian , Guettler Sebastian
Published: September 2021

The poly(ADP-ribose)polymerase (PARP) enzyme tankyrase (TNKS/ARTD5, TNKS2/ARTD6) uses its ankyrin repeat clusters (ARCs) to recognize degenerate peptide motifs in a wide range of proteins, thereby recruiting such proteins and their complexes for scaffolding and/or poly(ADP-ribosyl)ation. Here, we provide guidance for predicting putative tankyrase-binding motifs, based on the previously delineated peptide sequence rules and existing structural information. We present a general method for the expression and purification of tankyrase ARCs from Escherichia coli and outline a fluorescence polarization assay to quantitatively assess direct ARC–TBM peptide interactions. We provide a basic protocol for evaluating binding and poly(ADP-ribosyl)ation of full-length candidate interacting proteins by full-length tankyrase in mammalian cells.
Small-Volume Flow Cytometry-Based Multiplex Analysis of the Activity of Small GTPases v1
Authors: Simons Peter , Bondu Virginie , Wandinger-Ness Angela , Buranda Tione
Published: September 2021

Small, monomeric guanine triphosphate hydrolases (GTPases) are ubiquitous cellular integrators of signaling. A signal activates the GTPase, which then binds to an effector molecule to relay a signal inside the cell. The GTPase effector trap flow cytometry assay (G-Trap) utilizes bead-based protein immobilization and dual-color flow cytometry to rapidly and quantitatively measure GTPase activity status in cell or tissue lysates. Beginning with commercial cytoplex bead sets that are color-coded with graded fluorescence intensities of a red (700nm) wavelength, the bead sets are derivatized to display glutathione on the surface through a detailed protocol described here. A different glutathione-S-transferase-effector protein (GST-effector protein) can then beattached to the surface of each set. For the assay, users can incubate bead sets individually or in a multiplex format with lysates for rapid, selective capture of active, GTP-bound GTPases from a single sample. After that, flow cytometry is used to identify the bead-borneGTPase based on red bead intensity, and the amount of active GTPase per bead is detected using monoclonal antibodies conjugated to a green fluorophore or via labeled secondary antibodies. Three examples are provided to illustrate the efficacy of the effector-functionalized beads for measuring the activation of at least five GTPases in a single lysate from fewer than 50,000 cells.
3.1 Preparation of Monocellular Lung Suspension from a Mouse Lung Lobe v1
Authors: Graves Helen , Evans Steven , Fauler Michael , Frick Manfred , A. Moschos Sterghios
Published: September 2021

The clinical potential of DNA and RNA-targeting therapeutics for airways disease has been hampered by the poor translation of promising drug candidates from cell culture to in vivo models and the clinic. For example, classical preclinical approaches routinely report 20–60% target knockdown effects in the lung, where 1 or 2 log effects are observed in isolated cell cultures in vitro. Preparation of monocellular suspensions of tissues by mechanoenzymatic disruption followed by cell sorting (TDCS) after in vivo drug dosing, however, can offer pharmacokinetic and pharmacodynamic insights on the effects of drugs to precise cell subpopulations. Moreover, this can be reliably achieved with up to 66% fewer animals than standard in vivo pharmacology approaches due to lower data variance afforded through analytics on defined, viable cell numbers. Here we describe the TDCS methodology for the isolation of total lung epithelia, lung macrophages, and epithelium/macrophage-depleted cell fractions from mouse lungs using a two-stage sorting process of immunomagnetic bead separation followed by flow cytometric sorting using fluorescent antibodies against well-established surface markers such as F4/80, CD11b, and CD326. Validated antibodies for additional cell types and markers are also provided.
3.3 Lentivirus Production and Transduction v1
Authors: J. Sauter Evelyn , K. Kutsche Lisa , D. Klapper Simon , Busskamp Volker
Published: September 2021

This is part 3.3 of the "Induced Neurons for the Study of Neurodegenerative and Neurodevelopmental Disorders" collection of protocols. Collection Abstract: Patient-derived or genomically modified human induced pluripotent stem cells (iPSCs) offer the opportunity to study neurodevelopmental and neurodegenerative disorders. Overexpression of certain neurogenic transcription factors (TFs) in iPSCs can induce efficient differentiation into homogeneous populations of the disease-relevant neuronal cell types. Here we provide protocols for genomic manipulations of iPSCs by CRISPR/Cas9. We also introduce two methods, based on lentiviral delivery and the piggyBac transposon system, to stably integrate neurogenic TFs into human iPSCs. Furthermore, we describe the TF-mediated neuronal differentiation and maturation in combination with astrocyte cocultures.
Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDER v1
Authors: Karina Brinkman Eva , van Steensel Bas
Published: September 2021

Current genome editing tools enable targeted mutagenesis of selected DNA sequences in many species. However, the efficiency and the type of introduced mutations by the genome editing method are largely dependent on the target site. As a consequence, the outcome of the editing operation is difficult to predict. Therefore, a quick assay to quantify the frequency of mutations is vital for a proper assessment of genome editing actions. We developed two methods that are rapid, cost-effective, and readily applicable: (1) TIDE, which can accurately identify and quantify insertions and deletions (indels) that arise after introduction of double strand breaks (DSBs); (2) TIDER, which is suited for template-mediated editing events including point mutations. Both methods only require a set of PCR reactions and standard Sanger sequencing runs. The sequence traces are analyzed by the TIDE or TIDER algorithm (available at https://tide.nki.nl or https://deskgen.com). The routine is easy, fast, and provides much more detailed information than current enzyme-based assays. TIDE and TIDER accelerate testing and designing of DSB-based genome editing strategies.
Binding Relations and Their Implications for Word Order in Arabic
Authors: Alenazy Mamdouh Ayed
Published: September 2021

This study aims at investigating the distribution of the possessive pronouns in Modern Standard Arabic. It shows that when the possessive pronouns are used as reflexives they have implications for the word order. The different positions occupied by the objects are determined by the presence of these pronouns and the binding relations within the c-commanding domain. Building on the basic assumptions of Binding Theory, possessive pronouns are best treated as normal pronominal elements which are subject to condition B. However, when they are used as anaphoric elements in certain contexts, they have to be c-commanded by their antecedents. Depending on the derivational level at which c-command relation is established between the reflexive possessive pronoun and its antecedent, movement of the possessive pronoun along with the phrase containing is optional in certain structures or, in other structures, the pronoun becomes frozen in the position in which it is base-generated.
Effects of Teaching Approach Based on Three-dimensional Grammar on Grade Eight Students’ Acquisition of English Phrasal Verbs
Authors: Tao Mingxing , Wang Jiaxin
Published: September 2021

Phrasal verbs, an essential but complex aspect of English vocabulary, are difficult for Chinese EFL learners to acquire, whose first language lacks of phrasal verbs. This quasi-experiment explores the effectiveness of the teaching approach based on three-dimensional grammar on the acquisition of English phrasal verbs by Chinese middle school students compared with that based on conceptual metaphor theory. The finding is that there is a significant difference between the effect of three-dimensional grammar teaching and that of conceptual metaphor teaching, and the former is of great benefit for learners to acquire English phrasal verbs.
A Study of the Reflection of Naturalism in the Heroine in Sister Carrie
Authors: Zeng Xiu
Published: September 2021

Sister Carrie is one of the greatest works composed by Theodore Dreiser, one of the representatives of naturalists in American literature at the beginning of the 20th century. Among numerous works of Theodore Dreiser, Sister Carrie enjoys a quite high literary status, and also meets with different comments after its publication. The novel mainly tells the personal experiences of a rural beautiful girl Carrie in the big city Chicago. Driven by natural desires and urban environment, she changes her social status and original values in the end. Naturalism is a scientific and literary approach employed here to depict the characters in the novel where a person's fate is decided or predetermined by impersonal forces of nature and environment beyond human control. The novel is an experiment where the author could discover and analyze the natural forces and survival laws that influence personal behaviors, emotion and fate. This paper introduces the origin, development and characteristics of naturalism in American literature, studies the effect of human weakness on their life, and makes clear the influences of environment on people’s way of thinking and way of life.
Comic Function in the Animated Ecodiscourse (Case Study of “Zootopia”)
Authors: Ivchenko Natalia
Published: September 2021

This study focuses on the role of comic functions in exposing and challenging hidden ideologies, intentions, potential significance and other phenomena behind the animated ecological discourse of the film "Zootopia". The paper consistently considers two views on the ecology of animated discourse and comic functions that are used to uncover violation and establish an eco-friendly relation by means of language forms. The material of the English-language animated discourse of the film "Zootopia" examines cooperation of verbal and nonverbal modes, exposes problems and encourages solving them as well as promotes establishment of sustainable relationships between humans themselves, humans and nature as well as its phenomena.
REVIEW OF THEORETICAL APPROACHES TO DEFINING THE ESSENCE OF THE DIGITAL ECONOMY
Authors:   Siberian Federal University, Kravchenko Ekaterina
Published: September 2021

In the presented article, theoretical approaches to defining the essence of the digital economy were considered. The author proposed his own interpretation of the term "digital economy" on the basis of the presented combined approaches, also highlighted the classification of definitions of the digital economy.
A NEW LOOK AT THE SYSTEM OF BALANCED INDICATORS (IN RELATION TO THE BUSINESS ENTITIES OF THE HOSPITALITY INDUSTRY)
Authors:   Siberian Federal University, Maksimenko Irina
Published: September 2021

Currently, business entities of the hospitality industry, being in the conditions of post-pandemic market compression, strive to ensure their survival by transforming their own business models. The process of any strategic transformation should be accompanied by a measurement of its effectiveness. Traditional performance indicators in the hospitality industry are financial aspects, such as net income, return on investment, and return on assets. However, the focus on finance seriously limits the ability to recognize intangible factors. So, Professors Robert Kaplan and David Norton developed a strategic management tool called the Balanced Scorecard (BSC). BSC enables business entities to visualize mission, vision, and strategy into a comprehensive set of performance indicators and provide a framework for strategic measurement and management. This is made possible by the fact that BSC complements traditional financial indicators with other three key business perspectives that are useful for evaluating performance: customers, internal business processes, training, and growth. However, three decades after the research of Norton and Kaplan, their model needs to be improved taking into account the current realities, both the economy in general and the hospitality industry in particular. The strategic map of balanced indicators proposed in this paper is formed in two dimensions. The first of them presents four thematic areas (supplemented by the classic BSC perspectives) that form the basis of the map: the financial perspective in terms of the cost component; the customer perspective in terms of online reputation; the perspective of business processes from the point of view of innovation; the perspective of training and growth from the point of view of employees. In the second dimension, each of these perspectives is concretized through the interests of four different stakeholders: managers, business partners, guests, and academic experts in the hospitality industry. As a result, the structure of the strategic map of performance indicators in the hospitality industry covers a two-dimensional panorama – one in the thematic dimension, the other from the point of view of various stakeholders.
REMOTE SALES: COMPREHENSIVE ASSESSMENT OF INFORMATION FOR BUYERS AND QUALITY OF SOLD PRODUCTS
Authors:   Siberian Federal University, Demakova Evgeniya
Published: September 2021

The theoretical basis of the study was publications that study the premises of the customer experience as the main determinant of satisfaction and loyalty of users of online food delivery services. The aim of the study is to analyze the specifics of the formation of loyalty of Russian buyers by assessing the convenience of using the information attributes of the seller and the quality of the products sold. Our hypothesis lies in the assumption that at the current stage for Russia of the transition from single to massive online purchases, there are great risks of unjustified enthusiasm for the organizational aspects of remote interactions and underestimation of consumer values traditional for offline purchases. At the first stage of the study, an expert assessment of the completeness, accessibility and ease of use of information on the activities of the most popular online delivery services was carried out. Further, according to standard methods, an examination of the quality and safety of caviar and salted fish of the salmon family, purchased from the studied delivery services, was carried out. We have proposed a new model for shaping the online shopping experience. Its first difference is in complementing the list of prerequisites of the shopping experience with independent reviews of product quality and safety. The second feature of the model we propose is predicting consumer satisfaction with interactions with sellers by means of a multi-criteria expert assessment of information about the activities of online stores for buyers. The results of such an assessment for the four studied delivery services turned out to be equally high in terms of the following criteria: information about the online store, the seller, payment and delivery terms. The results of a study of information about goods provided to consumers turned out to be noticeably worse. The highest risks of reduced customer satisfaction of the studied online delivery services were shown by the results of the examination of the quality and safety of the products sold. Two of the six salmon roe samples tested and most of the red fish samples did not pass the microbiological safety test. The results of the study showed that for distance shopping service providers, the struggle for a buyer is shifting towards providing information about products and objectively confirming their quality.
Estimation of State Variables in the Ćuk Converter Mathematical Model with Partially Unknown Parameters
Authors: Bobtsov A. A. , Ortega R. , Nikolaev N. A. , Slita O. V. , Kozachek O. A. , Wang C.
Published: September 2021

In this paper the solution was proposed for the state variables estimation problem in the mathematical model of the DC switch-mode power converter built according to the Ćuk scheme. Pulse converters are one of the main components of most modern electrical devices and the circuit proposed by Slobodan Ćuk in the 70s of the 20th century is still relevant and demanded. Traditionally, PI (proportional-integral) controllers or proportional-integral adaptive control algorithm (PI-PBC), based on passification methods and superior to standard PI controllers in accuracy, are used as the control algorithm for power converters. However, you need to know the entire vector of the state variables of the converter to build a PI-PBC controller, and moreover, all its parameters must be accurately known. Unfortunately, in practice, such assumptions are not fulfilled, since parametric drifting is possible, and measurements of the converter’s state require additional sensors, which in some cases does not justify itself. Thus, there is a need to develop additional observers or estimators that allow obtaining data on all variables of the converter, as well as its parameters. The solution is based on the GPEBO method (generalized parameter estimation-based observers). The problem was solved under assumption that only the output signal (the output voltage of the converter) is measurable and some of the mathematical model parameters are unknown. An important aspect of the observer design is the development of an algorithm for unknown parameters and the state vector of a mathematical model estimation that ensures convergence in a finite time. Finite-time convergence is extremely important when designing observers since transients in pulse converters occur very quickly.
Methodology of Automated Monitoring and Control of Pump Complex Operation Modes in Conditions of Cavitation Occurrence
Authors: Lutov A. G. , Novozhenin M. B.
Published: September 2021

The results of solving the problem of increasing the efficiency of an automated pumping complex for pumping liquids in conditions of non-stationary hydraulic processes, such as cavitation, are presented. The difficulty of determining the conditions for the occurrence of cavitation is associated with a large number of parameters, the mutual correlation of which is difficult to determine. It is shown that the methods used in practice in these conditions for monitoring and controlling pumping complexes based on centrifugal pumps and adjacent pipelines have significant disadvantages or solve the problem only partially. A mathematical model of the pump complex operation for operational control of the parameters of cavitation modes based on the similarity of the modes of operation of the centrifugal pump and the movement of the piston through the pipeline is presented, which simplifies the procedure for determining the presence of cavitation. A criterion for determining the efficiency of the pump complex operation mode is proposed based on an integral assessment of the difference between experimental and model data. A methodology for controlling the modes of operation of the pumping complex in the conditions of cavitation is formed. Due to the complexity of the direct calculation of the cavitation volume, a neural network model was proposed, trained based on experimental data. The structure, algorithms and software of the automated control and control system are developed using neural network models and a case-based approach to quickly determine the conditions for the occurrence of cavitation and correct the operating modes of the pumping complex. Decisions based on case — based reasoning are offered to the operator in the form of a "control effect-expected result" pair. The practical implementation of the automated system for monitoring and controlling the operating modes of the pumping complex is carried out in the AppDesigner package of the Matlab mathematical package. The use of the developed automated monitoring and control system provides an increase (restoration) of the pump complex performance in the conditions of cavitation, prevents the destruction of its elements, increases the service life, reduces operating costs and equipment repair costs.
Study of thermal conductivity coefficient of basalt fiber insulation at various temperature conditions
Authors: Bazukova E.R. , Vankov Yu.V. , Gaponenko S.O. , Smirnov N.N.
Published: September 2021

Industrial thermal insulation fibrous materials (mineral wool and fiberglass products) are mainly used for thermal protection of the systems transporting high-temperature heat carriers. The data known in the scientific literature about the effect of insulation density and operating temperature conditions on the thermophysical properties of materials made of basalt fiber differ significantly. Taking these data into account let us improve the accuracy of thermal calculations and the validity of design solutions for thermal protection of industrial networks up to 60 %. The aim of the study is to obtain data on the change of the heat-shielding properties of insulating materials made of basalt fiber of various densities, depending on the temperature conditions of operation. An experimental study of the thermophysical properties of several samples of basalt fiber insulation has been carried out on a laboratory bench. The density of a constant heat flux passing through the sample has been determined by the additional wall method. The experimental unit consists of a cylindrical metal tube with a built-in electric heater, on the top of which a heat-insulating basalt fiber of various densities is placed. The temperature on the pipe surface is changed in the range from 50 to 350 оC. The dependence of the thermal conductivity coefficient of thermal insulation basalt fiber samples on its density and operating temperature of the material is obtained. The research results show that thermal conductivity coefficient of basalt fiber insulation increases with temperature increase of the pipeline surface. The lower the value of the material density is, the higher the increment of the thermal conductivity coefficient is. The obtained results enrich the data about the dependence of the thermal conductivity of fibrous heat-insulating materials on density and temperature (for cases of operation at high temperatures). Considering the density of the examined materials made of basalt fiber when determining the thermal conductivity makes it possible to increase the accuracy of calculations up to 20 % and temperature modes of operation up to 60 %. The results obtained can be applied to find the thickness of the thermal insulation of pipelines and heat losses in the networks.