bluuebunny/crossref_metadata_2025 huggingface dataset:
Social practice of urban environment quality assessment
Authors: Ivashova V A , Tokareva G V , Agalarova E G , Nadtochiy Yu B , Yushchenko I V
Published: April 2020

The article describes the approaches and results of studying the urban environment quality of Stavropol city – the regional centre of southern Russia. The study is based on a questionnaire survey (carried out on a representative sample stratified in accordance with the territorial division of the urban environment into administrative regions). In total, 1172 people took part in the survey. The data are processed in SPSS (version 21). The random sampling error is about 2.0%. Assessing the level of satisfaction of the population with services in the field of culture, education, medicine, trade, housing, the work of the administration gives an idea of the quality of the urban environment and the directions of its development. According to the results of the survey, among the most acute problems requiring enhanced monitoring and operational decisions, respondents identified the problems of cultural, leisure and development activities for children and youth, and medical care for the population. In addition, some questions cause additional criticism from the respondents: personal security and safety, the quality of work of local police officers, the quality of work of management companies, the condition of sports facilities, the work of clinics, and the maintenance of cleanliness in apartment buildings. Thus, in the development strategy of a comfortable urban environment, these areas should be reflected.
City branding. Features of urban image design
Authors: Karakova T V , Zaslavskaya A Y , Smolenskaya E
Published: April 2020

In the article the authors consider the foundations of the methodology related to the creation of the “urban brand,” as well as methodological approaches to solving the set of environmental problems, which play a significant role in forming an authentic image of the city and increasing attention to the urban environment on the example of a specific conglomerate, including for attracting tourist flows. The novelty of the author’s approach is evident in the integrated consideration of the issue with the inclusion of multifunctional programs and interaction of various disciplinary fields, including issues related to graphic art, urban environment improvement, history, literature, philosophy and architecture. The development of the brand of the modern city is seen by the authors in the implementation of various historical and cultural urban scenarios taking into account socio-economic indicators and the involvement of design technologies capable to create a “new face of the city» including landmark historical spaces and objects that increase the complex status of the place in the city scenario.
Concept of a system for monitoring the dynamics of transport infrastructure development in a megalopolis
Authors: Zavyalov D V
Published: April 2020

Sustainable development of a metropolis and its transport system is an urgent task of the modern world. The measures taken to develop mobility in a large city require regular measurement of citizens’ perceptions of the changes introduced and the use of the data obtained to monitor and manage programs for the development of urban public transport, bicycle and pedestrian infrastructure, public spaces and other areas of improving the comfort of life in a megalopolis. The paper discusses the basic principles and approaches to the formation of a monitoring system of the transport system, presents the results of testing the methodology, and proposes a conceptual model of a monitoring system for the transport infrastructure development to support management decisions.
Calculation algorithm of the building of CLT for the Russian Federation
Authors: Gotcadze A G , Vasin E E , Shabikova E G
Published: April 2020

The design of modern CLT structures in Russia is still under development. This calculating algorithm is proposed for designing of the CLT structures in Russia. The calculation of the CLT was carried out in accordance with the Russian codes of SP 64.13330.2017 Timber structures. The calculation algorithm allows, on the one hand, to automate the design and, on the other hand, to calculate the CLT according to the existing Russian codes of SP 64.13330.2017 Timber structures, and from the third, to obtain some necessary value of compression stress perpendicular to the CLT plane. In our case modelling was done in the Revit software for computer-aided design that implements the principle of building information modelling. For comparison internal stresses (shear, tensile and compressive) with the design resistance accordance with Russian codes “CП” calculation results were obtained from the program Dlubal RFEM (finite element structural analysis program).To obtain new values of compression stress perpendicular to the CLT plane and its subsequent verification, there is a branch in the algorithm.
Establishing relevant regulatory framework for construction cost calculation in BIM-systems
Authors: Mamaeva O A , Burakov S M , Savenkov A N
Published: April 2020

Nowadays Building Informational Modeling (BIM-technologies) adoption into the design, construction and operation of capital construction projects is currently a national priority. Optimization of construction costs is also within the state interest. For these reasons, one of the most important functions of BIM-systems which are currently being developed and implemented is the assessment of construction costs at various stages of the full life cycle of different construction objects. At the moment, this function is not fully implemented in software programs used to create BIM-models of capital construction. This paper focuses upon a structure of construction cost development at different stages of the life cycle of an investment and construction project integrated into BIM-systems (as well as the levels of development of its BIM-model). The researchers also introduce a system of estimated standards and indices relevant for BIM-products which can be used to determine the cost of construction.
Garden space. Morphotypes of private gardens of Jiangnan region
Authors: Tceluiko D S
Published: April 2020

The paper discusses the types of space representation of private gardens in the Jiangnan region. The definition of space is given, its main elements and features are revealed. Private gardens of China are described as one of the examples of complex space organization, the work of researchers studying the garden space is analyzed. The method of uniform distribution of space between elements has been studied in detail. The logic and the possibility of using the method are shown. The example of three private gardens in the Jiangnan region is the breaking of space. The article presents an analysis of space with the identification of the main elements and the use of the Voronoi diagram. Morphotypes of landscape gardening complexes are described. The text sets out the methods for applying this type of representation of space and outlines the directions for further work.
Practical restoration-related experience with creation of basement level and underground parking into and near architectural monuments in down-town of Saint-Petersburg
Authors: Mangushev R A , Osokin A I
Published: April 2020

Geotechnical construction in Saint-Petersburg is governed by the specific features of geological conditions. There is large thickness of soft water saturated soil underlays at the basement monuments and historical buildings. This soft water saturated soil become thixotropic under insignificant dynamic effect (construction action, transport, seasonal freezing and etc). The experience of the renovation, reconstruction old buildings with construction of the deep basements, underground parking in the inner yards or near the architectural monuments in the central part of Saint- Petersburg are given in this article. This paper provides the description of geotechnical solutions, which were realized on the various cultural heritage buildings, such as Catholic Church of St.Catherine on the Nevsky prospect, Taurid Palace, Christmas Church on the Sands, shopping mall «Galeria» on the Ligovsky prospect. The authors analyzed some failures of buildings and facilities as the result of incorrect methods and technologies of the deep excavation.
Historical city center in the XX - XXI centuries: back to pedestrian
Authors: Raikhel Y L , Rozhdestvenskaya E S
Published: April 2020

The article describes the main factors that affect the change of the public space paradigm of city planning throughout the XX – XXI centuries. The authors cover cities of Europe, America, and Russia. The research is theoretically based on the works of Scott McQuire, Jan Gehl, Jeff Speck. The research outcome presents two groups of the factors: those affecting change of the city-panning paradigm towards car drivers in the first half of the XX century, and back towards pedestrian in the second half of the XX century. The authors choose the space planning model of the medieval European town as the benchmark.
Space structure features of the Garden of Cultivation in the context of traditional private gardens of Suzhou
Authors: Tceluiko D S
Published: April 2020

The paper is devoted to the identification of the main architectural features of the Garden Cultivation. The history of the garden was analyzed, the main stages of the formation of the planning structure were revealed. The etymology of the name of the estate, which is related to the functional direction of the object, has been studied. The garden was founded during the Ming Dynasty. Over time, both the planning structure and individual elements of the garden changed. The space of the garden consists of two main areas: residential and garden, north and south. Each of the zones is actively integrated into each other. The garden space is very densely built up, consists of 17 elements (morphotypes) located on 0.37 hectares. Garden of Cultivation is unique landscape object for Suzhou and for China, the features of which begins with the name and end with unusual buildings and decor. However, the educational function of most of the morphotypes makes the overall structure of the plan different (unique) from the “traditional” gardens of this era.
Theory and practice of heritage renovation of landscape and recreational spaces in the city of Voronezh. Renovation project for the Park named after A. Durov
Authors: Yenin A E , Grosheva T I , Salko V N
Published: April 2020

The article is devoted to the problem of renovation of public recreational spaces classified as historic heritage involving the use of systematic approach and analysis. This approach can scientifically justify the place and public significance of a rather specific and relevant type of architectural and urban planning activity consisting in harmonization and interaction between people (population) and landscape recreational environment inhabited by them [1,2]. The purpose of the research is identifying the integrity of historic spaces through analysis of interaction of industrial, household, recreational, and communicational processes in view of their restoration and choosing methods and techniques with specific examples from design practice in Voronezh.
Control and management scheme of air flows in inner space of steel silo during grain storage
Authors: Kechkin I A , Ermolaev V A , Buzetti K D , Romanenko A I , Gurkovskaya E A
Published: April 2020

The design of steel silo does not provide for constant monitoring of the air amount circulating inside the silo. Silos are not equipped with measuring equipment to control air flow. The height of the grain layer near the vertical wall corresponded to 15 meters and 20 meters at the same pressure of 3000 Pa in the lower part of the layer. At a higher layer height, the lower filtration rates and lower air flow are performed. Near the silo wall, the air rate depends on the height of the layer and is the same for silos of different diameters at the same pressure in the lower part. The silo with a large diameter has a greater variation in the values of the filtration rates along the surface of the grain mass. A typical technological scheme for storing grain in large-capacity steel silo was proposed and tested, taking into account the control and management system for air flows in the inner space of the grain mass.
Ionic composition of drinking water and its influence on the value of its zeta potential (ZP)
Authors: Kichigin V I , Atanov N A
Published: April 2020

The authors demonstrate the dependence of ZP on the value of water ionic composition, characterize test water physical and chemical composition and introduce thorough analysis of the obtained results. In order to reveal a complete picture of water quality, they also define such additional parameters as test water pH, temperature, turbidity and alkalinity in the course of the research. The study proves that all studied water parameters correspond to SanPiN 2.1.4.1074-01 Standard “Drinking water. Hygienic requirements for the quality of water in centralized drinking water systems. Quality control. Sanitary rules and regulations”. The paper also presents mathematical dependences between the ζ-potential and the content of anions and cations in the test water with the accuracy of approximation R2 not less than 0.83.
Method for predicting trihalomethanes content in drinking water (by the example of surface type water intake)
Authors: Malkova M A , Vozhdaeva M Y , Kantor E A
Published: April 2020

The search for predicting the concentration of trihalomethanes in drinking water equations of the surface water intake was carried out. The turbidity, chromaticity, permanganate oxidation of the water source and the dose of chlorine are taken as dependent parameters.The listed parameters are an indirect reflection of the gunim substances presence in the water source. These substances are precursors for the formation of trihalomethanes. In addition, it was found an equation by which it is possible to predict the dose of chlorine depending on the unlisted generalized indicators of water quality. The coefficients of determination of the obtained equations vary between 0.78 - 0.96, and the average relative forecast error is 6.5 - 19.3%.
Rainfall parameters for the design of surface water treatment facilities
Authors: Palagin E D , Strelkov A K , Pavluhin A A
Published: April 2020

Summarized data is presented on the maximum rainfall layer, runoff from which should be completely diverted to treatment facilities (runoff from residential areas and industrial enterprises of the first group in the Russian Federation). The calculated values of the precipitation layer are obtained from observations at 238 weather stations located in the Russian Federation, which are necessary for calculating the performance of surface water treatment plants. The results can be used in the design in areas for which these weather stations are representative. The size of the precipitation layer, which ensures the discharge of at least 70% of the annual volume of runoff to sewage treatment plants, for the territory of the Russian Federation is in the range of 2.2–12.8 mm.
Evaluation results of the wastewater treatment system of small settlements in Syria
Authors: Saied M A , Serpokrilov N S
Published: April 2020

The results of the environmental evaluation show the deterioration of water quality in the main river basins of Syria as a result of untreated domestic wastewater or insufficiently treated wastewater, commercial and industrial wastewater from certain activities. The sewage system, especially in small settlements, suffers from several problems such as: long sewer lines, errors in the design of treatment plants and the choice of treatment method, lack of qualified personnel, frequent power outages, large fluctuations in the flow and concentrations of wastewater pollution. 98% of treatment plants in small settlements operate with extended aeration activated sludge, which does not meet their requirements and does not provide high efficiency of wastewater treatment. One of the recommended solutions for the development of a wastewater treatment system in small settlements in Syria is considered rotating biological contactors RBC, which are the best option and most meet their requirements.
The impact of the factors of oil-containing wastewater formation of Bezenchuk oil extraction plant
Authors: Strelkov A K , Teplykh S Yu , Bystranova A O
Published: April 2020

Currently, there is a boom of oil and fat industry enterprises. The article studies regularities of changes in wastewater composition, processes of wastewater formation of Bezenchuk oil extraction plant, as well as wastewater chemical and physical parameters.

Waste water of industrial enterprises contains a large amount of organic contaminants: hydrocarbon oils, vegetable and animal fats, surfactants, etc. High contaminants concentration at the enterprises makes the treatment difficult being a complex technical and scientific task. Wastewater treatment problem in food industry is largely related to the extraction of fats. Flotation methods of cleaning with the use of reagents are considered; their advantages and disadvantages are revealed.
Features of the preparation of calcium-containing raw materials in the production of ceramic bricks
Authors: Guryeva V A , Doroshin A V , Dubineckij V V
Published: April 2020

The paper considers possibility of using low-melting loam, drilling waste and ash and slag waste of thermal power plants in the production of wall ceramics by the method of semi-dry pressing. The physicochemical processes occurring upon receipt of ceramic materials using production waste at the firing stage are studied. It has been established that an increase in the duration of grinding of calcium-containing raw materials at the stage of preparation of the starting materials determines a change in its particle size distribution, an improvement in the plasticity of the molding materials and an increase in the coherence of the raw product and provides an increase in the mechanical strength of the raw product in comparison with the initial indicators.
Heat engineering problems of phosphogypsum utilization during its processing into construction gypsum
Authors: Nedoseko I V , Vinnichenko V I , Ryazanov A N , Ryazanova V A
Published: April 2020

The shortcomings of common equipment for utilization of phosphogypsum when processing it into semi-aqueous gypsum have been analyzed. Specific energy consumption per ton of produced construction gypsum in different equipment is shown. Calcination duration is compared. The reasons for the uncompetitivity of the process of production of construction gypsum compared to the production of natural gypsum stone are analyzed. Two problems are considered. The first - common equipment for burning construction gypsum is energy-intensive. The second is the established practice of washing impurities in phosphogypsum with water. At the same time, energy consumption increases several times due to the need for energy consumption for moisture evaporation. It is theoretically reasonable to carry out the firing in a suspended state. The firing time is a few seconds. The experimental studies show the possibility of producing construction gypsum by firing in a suspended state.
Experimental investigation of reinforced concrete beams using post- tension
Authors: Pischulev A , Panfilov D , Zhiltsov Yu
Published: April 2020

The paper presents the results of an experimental investigation of statically determinable pre-stressed concrete beams with tension on concrete (using post-tension). The authors consider the existing practice of applying post-tension in reinforced concrete structures, the current regulatory documents of the Russian Federation containing information on the strength and deformation properties of post-tension. The paper contains information about the loading pattern, beam-fixing conditions, geometric characteristics of a reinforced concrete beam, physical characteristics of concrete and reinforcement of an experimental sample, technical characteristics of pre-stressed reinforcement. The authors present the research results using graphs of structural deflection changes under load, the distribution of relative strain along the depth of a concrete cross-section, as well as cracking and failure schemes of experimental samples. The research presented in the paper showed that the destruction of the samples occurred along a standard cross-section and was brittle. As a result of processing the results of experimental data, loads of crack formation and critical crack width were determined. The authors confirmed the effect of pre-stressing on crack resistance and the magnitude of the deflection of bent reinforced concrete elements.
Optimization of fractional composition of Volga River sand for improvement of characteristics of mortars and concrete
Authors: Zhigulina A Y , Mizuryaev S A
Published: April 2020

The article is devoted to the use of fine sands to produce high-quality mortars and concrete. The problem of improving the quality (rheological and strength characteristics) of mortars and concrete without increasing cement consumption is being solved. The results of the study of the fractional composition of typical fine river Volga sand are presented. The results of the studies show that the method of fractional selection can significantly improve the rheology and strength of mortars and concrete with the effective use of screenings. Optimization of fine sands should not only consist in their enlargement due to the removal of fine fractions, but also in the selection of the optimal fractional composition.
Modeling of stress-strain state of external reinforcement of single-span reinforced concrete beams
Authors: Merkulov S I , Esipov S M
Published: April 2020

The paper touches upon the issues and features of computer modeling of external reinforcement of reinforced concrete beams with the use of computer complex ANSYS. The method of modeling of bent reinforced concrete beams reinforced in the stretched zone by external reinforcement based on carbon fiber composite materials, approved by experimental studies, is presented. The explanations on application of a technique at various approaches to strengthening of a beam depending on the current stress-strain state are given. Qualitative and quantitative dependences between experimental and numerical studies are established, confirming the initial prerequisites of the study.
Numerical study of stress-strain state of reinforced concrete slab in punching zone
Authors: Filatov V B , Galyautdinov Z Sh , Blinkova E V , Zhiltsov Yu V
Published: April 2020

The article presents the analysis of the stress state of a flat reinforced concrete slab under punching through a numerical study on finite element models. Three models with different cross – section shape of the column-square, rectangular and L-shaped-are considered. The calculation of the punching according to the standard method is carried out under the assumption of a uniform distribution of shear stresses on the control perimeter of the punching. The results of the study allow drawing a conclusion about a significant uneven distribution of shear stresses on the control perimeter, depending on the shape of the cross-section of the column. The conclusion corresponds to the results of previously performed experimental studies on test samples. According to the results of the study, the effective length of the control perimeter of the plate punching for the considered forms of the column cross-section is determined.
Study of non-formwork hollow-core slabs with support zone restraint in brick walls
Authors: Prokopovich A A , Repetto V V , Aleshin A N
Published: April 2020

The purpose of the research is to develop a method for calculating the crack formation in the support zones of hollow core slabs manufactured by the non-formwork concreting technology, clamped into brick walls. The testing results of six samples were obtained by a longitudinal cut. On the basis of the experimental data and finite-element modeling using the software complex Lira SAPR-2016 a method for calculating cracks formation in the support slab zones pitched in masonry has been developed and presented. The developed method of calculation allows taking into account the ductility of the slab joints with bricklaying, the length of the sealing of the support zones, the value of the average compressive stresses in the masonry, as well as spans, stiffness characteristics of slabs and load application schemes. The recommendations concerning support points reduction at designing support slab assemblies clamped into a brick wall are provided.
Discrete-analytical nonlinear analysis with improved computation accuracy for steel frame lateral response evaluation
Authors: Rutman Yu L , Ivanov A Yu , Meleshko V A
Published: April 2020

The structural performance evaluation under the monotonic increase of a lateral load due the whole loading process in modern earthquake engineering practice can be performed with the help of different methods and software and can be dramatically represented by «Base reaction – Roof displacement» relationship, which is also known as capacity curve. This kind of relationship is used by nonlinear static Pushover-analysis for seismic performance evaluation of buildings considering any possible damage, which is acceptable by serviceability requirements of a building. However, there are many problems arising due to physically nonlinear analysis of structure, primarily associated with features of the employing finite elements analysis software. Main problems are rather long analysis time among the necessity of high performance computing, or shortcomings of nonlinear analysis methods inherent to the program used. One of the most important shortcomings of structural analysis software is the necessity of single-handed assignment of a failure mechanism for the structure by nonlinear hinges formation zones emplacement to elements of analytical model. This circumstance may lead to results substantially different from reality. These problems can be resolved with the aid of discrete-analytical methods.
Welded joints of offshore fixed platforms for the extraction of oil, gas and other minerals on shelf
Authors: Starokon I V
Published: April 2020

The world is actively developing offshore mineral deposits (oil, gas, coal, steel and other minerals). Offshore fixed platforms are actively used for offshore production. During operation, these platforms are subject to various impacts (wave, wind, etc.), which leads to the occurrence of alternating stresses. Current AC voltages activate fatigue processes in welded joints. Therefore, the knowledge of the peculiarities of the stress state of the welded joints of offshore platforms is the key to understanding the fatigue processes taking place in them. Nowadays various theories offer different understanding of the stresses acting in a welded joint. For example, the strength theory does not take into account the presence of “hot spots” in the welded joint, assuming a uniform distribution of stresses on the welding seam. The influence of static load on the general picture of the stress state in the welded joint was also investigated. As a result of the carried out researches the location of points with the greatest concentration of pressure, influence of thicknesses, diameters and an angle of interface of connected elements is established. The calculations showed that the sharper the angle of connection of the branch to the main pipe, the lower the value of stress concentration coefficients. Stresses acting in the welded joint also increase in case of increase in the diameter and wall thickness of the offshore platform columns. The analysis of the results of the computer model showed that the greatest contribution to the stress state of the platform is the tensile force and bending moment outside the plane, which indicates that the points with the highest stress concentration will be the points located in the direction of the wave beam.
Study of deformations of building structures thin-walled elements by the method of moire strips
Authors: Yemets V N , Kalmova M A , Akhmedov A D
Published: April 2020

The article deals with the application of one of the experimental optical-geometric methods which is the method of moire strips with a reflected grid (according to the Ligtenberg scheme) to the study of I-beam corrugated beams with different wall configurations of constant thickness, rigidly fixed on one side and free on the other (console). The bending deformation process was observed from the action of a concentrated force at the free end with the gradual loading at different intervals of the applied force action. A model of a beam made of low-modulus material (plexiglass) with two types of wall profile: wavy and triangular was tested. The modeling process of the studied beams was developed. Virtual technique of moire patterns formation with the subsequent computer processing by standard programs was applied. The analysis of the deflections along the beam length depending on the load is given and deflections graphs are constructed in comparison with the theoretical calculation for models with corrugated and flat walls (standard I-beam).
Esterified Hyaluronic Acid Matrix in Lower Extremity Reconstruction With Exposed Tendon and Bone: A Retrospective Review
Authors: Kozusko Steven D , Hassouba Mahmoud , Hill David M , Liu Xiangxia , Dadireddy Kalyan , Velamuri Sai R
Published: April 2020

Lower extremity wounds with exposed bone and tendon often need coverage to allow the underlying tissue to regenerate prior to skin graft. The surgeon is limited in his or her choices to augment tissue regeneration in these types of complicated cases; for instance, autologous skin should not be placed on exposed bone or tendon and is at risk for contracture when placed over the joints. Therefore, novel technologies are necessary to provide a scaffolding for tissue to regenerate and allow for a successful graft. One such technology is an esterified hyaluronic acid matrix (eHAM), which can provide a proper scaffold for endothelial cell migration and aid in angiogenesis. The eHAM is made of two layers: a layer of hyaluronic acid covered with a silicone layer. In this retrospective chart review, we describe our usage of eHAM to provide scaffolding for tissue regeneration prior to grafting in 15 cases of complicated lower extremity wounds with exposed bone and tendon. The average patient age was 45.8 years, and all patients had multiple medical comorbidities, such as poorly controlled diabetes mellitus, hypertension, and nicotine addiction. Patient wound types were diverse, including traumatic wounds, chronic diabetic foot ulcers, and thermal or electric burns. Thirteen of the 15 cases were treated successfully with eHAM. In these cases, definitive coverage with split-thickness skin grafting was effective and limb salvage was successful. In the 13 successful cases, the mean time to split-thickness skin graft was 22.9 ± 7.0 days. All patients continue to do well at follow-up (ranging from 6 to 48 weeks), with minimal complications reported. Given the success rate with eHAM in this challenging population, we conclude that eHAM can be a treatment option for similar cases.
When brain stimulation backfires: the effects of prefrontal cortex stimulation on impulsivity
Authors: Bell Sarah Beth , Turner Brian , Sawaki Lumy , DeWall Nathan
Published: April 2020

Transcranial direct current stimulation (tDCS) can sometimes cause the opposite of its intended effect. These reverse effects may be related in part to individual differences in personality and neurochemistry. Previous studies have demonstrated that dopamine levels can impact the effects of tDCS. In the present study, 124 healthy participants took the UPPS impulsive behavior scale. Participants then underwent a single, randomized anodal or sham tDCS session on the prefrontal cortex. While the effects of tDCS were still active, they performed the Stop Signal Task, a measure of state impulsivity. tDCS was associated with increased errors on this task in people who had higher scores on the UPPS in two facets of impulsivity that correlate with dopamine levels. tDCS had no effects on people who are low in trait impulsivity. These results suggest that the reverse effects of tDCS could be associated with inter-individual differences in personality and neurochemistry.
Multi‐omics approach identifies differences between cranial and spinal motor neuron – A step towards understanding ALS
Authors: Maity Shuvadeep , An Disi , Randleman Justin , Iannitelli Dylan , Mazzoni Esteban , Vogel Christine
Published: April 2020

Amyotrophic Lateral Sclerosis (ALS) is an adult‐onset fatal neurodegenerative disease with a selective, highly progressive loss of both spinal and upper motor neurons. However, cranial motor neurons are much less sensitive and survive until the late stages of the disease. The inability of the cells to respond appropriately to Endoplasmic Reticulum (ER) stress is thought to be one of the major reasons for the ALS progression. ER stress is a condition that arises after the accumulation of misfolded proteins inside the ER and activates a repertoire of transcriptional and translational events collectively called unfolded protein response (UPR). In the context of ALS progression, the dynamics of transcriptomics and proteomic profile over time in the presence of ER stress conditions are poorly understood. Here, we established a condition where ER stress shows differential vulnerability of stem cell derived cranial (CrMN) and spinal motor (SpMN) neurons. Using high throughput quantitative omics approaches, we quantified >8,200 genes over time and found that subtle expression changes influence several pathways that are essential for cranial and spinal motor neuron function. We demonstrate that pathways related to overall protein turnover differ in these two cell lines. Specifically, we identified expression differences in Rab proteins that have not been recognized to‐date. These Rab proteins affect synaptic recycling, endocytic sorting, and autophagic flux. Further, we discovered a novel link between the core proteasome and endocytic pathways that potentially help CrMN to turn over proteins more efficiently than SpMN. Combined, these results illustrate new insights into the complex behavior and subtle differences between two motor neuron cell lines and provide new avenues towards therapeutic intervention.

Support or Funding Information

National Institute of General Medical Sciences (R01 GM113237), National Institute of General Medical Sciences (1R35GM127089‐01), Project ALS (A13‐0416)
NEIL2 plays a critical role in limiting inflammation and preserving genomic integrity in H. pylori‐infected gastric epithelial cells
Authors: Sayed Ibrahim M. , Sahan Ayse Z. , Venkova Tatiana , Chakraborty Anirban , Reyes Victor , Mukhopadhyay Dibyabrata , Pinchuk Irina , Beswick Eileen , Ghosh Pradipta , Hazra Tapas K. , Das Soumita
Published: April 2020

The accumulation of Helicobacter pylori infection‐ induced DNA damage in gastric epithelial cells is a risk factor for developing gastric cancer; however, the underlying mechanisms remain poorly understood. Here we report that suppression of the DNA repair enzyme NEIL2 is one such mechanism via which H. pylori infection may fuel the accumulation of DNA damage during the initiation and progression of gastric cancers (GCs). We found that NEIL2 was significantly downregulated upon H. pylori infection both in vitro and in human gastric biopsies. The H. pylori infection‐induced downregulation of NEIL2 is specific, as Campylobacter jejuni has no such effect. Using gastric organoids isolated from the murine and human stomach in co‐culture studies with live microbes, mimicking the infected stomach lining, we found that H. pylori infection was associated with proinflammatory cytokines release; this response was more pronounced in Neil2 knock out (KO) mouse cells compared to wild type (WT) cells suggesting that NEIL2 suppresses inflammation under physiological conditions. In vivo studies in mice showed that despite a lower bacterial load, as determined by UreB expression, DNA damage was significantly higher in Neil2 KO mice compared to WT mice. Interestingly, after infection with H. pylori, Neil2 KO mice has higher inflammation and epithelial cell damage compared to WT cells. Taken together, our data suggest that downregulation of NEIL2 is a plausible way of how H. pylori infection derails DNA damage repair mechanisms and initiates GCs.

Support or Funding Information

This work was supported, in whole or in part, by National Institute of Health Grants: DK107585 and DK099275 (to SD); R01 NS073976 (to TH).
Inhibition of P2X4 purinoceptor reduces growth of the cerebral aneurysm
Authors: Fukuda Miyuki , Fukuda Shunichi , Suzuki Takashi , Niwa Youko , Hasegawa Koji , Shimatsu Akira , Tsukahara Tetsuya
Published: April 2020

Background

Rupture of a cerebral aneurysm is a major cause of life‐threatening subarachnoid hemorrhage. However, there are no effective therapeutic drugs for the treatment of aneurysms, partly because the pathogenesis of the aneurysm remains unresolved. Although chronic inflammation of the cerebral arterial wall plays an important role in aneurysm formation, it is not clear what triggers the inflammation. We previously reported that inhibition of the endothelial shear stress sensor, P2X4 purinoceptor, significantly reduces mRNA expressions of known inflammatory contributors to aneurysm formation, leading to suppression of aneurysm induction. We examined here whether inhibition of P2X4 has any influence on cerebral aneurysm ‘growth’ or not.

Methods

Sprague‐Dawley rats were subjected to cerebral aneurysm‐generating surgery with ligation of unilateral common carotid artery and renal hypertension. P2X4 purinoceptor inhibitor, paroxetine (8 mg/kg/day) was administered to rats. Three weeks after the surgery, the aneurysm size was examined with light microscopy, and protein expressions of contributors to aneurysm formation using Western blotting.

Results

The cerebral aneurysm‐inducing surgery induced an aneurysm in 100% of rats 2 weeks after the surgery. Application of paroxetine from 2 weeks after surgery led to a significant reduction in aneurysm size in the rats sacrificed 3 weeks after aneurysm‐inducing surgery (p=0.0145), indicating that paroxetine suppressed enlargement of formed aneurysms. The protein expression levels of known inflammatory contributors to aneurysm formation (MCP‐1, IL1ß, TNFα, iNOS, and COX‐2) were all significantly elevated in the rats that underwent the aneurysm‐inducing surgery compared to the non‐surgical group, and the values in the surgical group were all significantly decreased by paroxetine administration.

Conclusion

The data suggest that P2X4 is required for the cerebral aneurysm growth. Enhanced shear stress‐associated hemodynamic stress on the vascular endothelium may trigger the cerebral aneurysm enlargement. Since paroxetine has been used for human as an antidepressant, clinical use of this P2X4 purinoceptor inhibitor is expected as a novel therapy for cerebral aneurysms.

Support or Funding Information

This work was supported by the Japan Agency for Medical Research and Development (AMED) under grant no. JP15gm0810006h0301, a JSPS KAKENHI grant (no. 15K10323), and The Shimizu Foundation for Immunology and Neuroscience.
ZEBRAFISH AS MODELS OF TYROSINE KINASE‐DRIVEN CANCER
Authors: Medishetty Raghavender , Challa Anil , Rao C. Mallukarjuna , Chatti Kiranam
Published: April 2020

The protein kinase gene family encodes proteins involved in several essential functions, with cell growth and migration being essential outcomes. Mutations in kinases underlie a broad range of diseases, primarily inflammation and cancer. Tyrosine kinase inhibitors (TKIs) in particular, are successfully used as a targeted therapy for cancer and inflammation‐driven disorders. Current unmet needs include addressing the adverse effects of TKIs, the emergence of resistance, and the necessity of newer TKIs. Experiments using accessible animal models such as zebrafish can help address these needs. Zebrafish studies can help in efficient characterization of toxicological and pharmacological effects of inhibition using known or potential TKIs, via the combination of phenotypic and molecular analysis. The recent application of CRISPR‐Cas9 further enables the development of genetically engineered cancer models in zebrafish. Following our previous work showing conservation of zebrafish tyrosine kinases compared to humans, we are now studying the effects of chemical or genetic tyrosine kinase modulation in zebrafish.

To identify the phenotypic endpoints of tyrosine kinase inhibition in zebrafish, we selected a few drugs approved by the USFDA for treating cancer and inflammation‐related disorders in humans. We evaluated phenotypic effects including adverse effects to identify NOELs and detectable and consistent phenotypic endpoints of such compounds in zebrafish. These effects provide a basis to evaluate novel TKI compounds. Further, tyrosine kinase genes involved in inflammation and cancer were selected for targeted genetic modification in zebrafish using CRISPR Cas9. Our initial data demonstrate the development of MET (HGFR) homozygous mutant zebrafish and the phenotypes associated with this mutant. Detailed characterization of this mutant including molecular analysis is under way. The characteristics of this mutant provide the basis to identify MET‐specific inhibitors. Development of zebrafish models carrying oncogenic TK mutants, and detailed characterization of the effects of TKIs in zebrafish are ongoing.

In summary, our work is focussed on identifying TKI treatment‐related phenotypes in zebrafish, and generation of TK‐crispant zebrafish as cancer models.

Support or Funding Information

Department of Biotechnology and the Indian Council of Medical Research, Govt. of India
Transcription hub dynamics revealed by nascent RNA‐chromatin interactomes
Authors: Li Xiao
Published: April 2020

In mammalian nucleus, many types of RNA are physically associated with chromatin. Specific and diverse functions have been increasingly ascribed to these chromatin‐associated RNAs, particularly in transcription and organization of 3‐dimensional chromatin architecture. Yet the full scope of molecular mechanisms that chromatin‐associated RNAs employ is far from understood. These mechanisms may be key to delineating regulatory redundancy and plasticity of transcription factors and enhancers. There is thus a compelling need to integrate chromatin‐associated RNA into our understanding of gene expression regulation. Among these chromatin‐associated RNA species, over 85% are nascent mRNAs being tethered to their sites of transcription. These nascent transcripts form individual ‘clouds’ over regulatory elements in cis. Such clouds enable faithful depiction of the landscape of transcription hubs. While decoration by these clouds of nascent mRNA on chromatin seems passive, evidence has indicated that it may dictate the expression of neighboring genes within the hubs. We therefore hypothesize that, before traveling to the cytoplasm as the messengers for protein synthesis, nascent mRNAs may modulate gene expression in the nucleus via interacting with chromatin. Two distinct types of interplay exist between nascent mRNA and the local transcription activity have been identified. In the RNA‐dependent model, nascent transcripts reinforce the local transcription microenvironment to promote expression of itself and neighboring gene(s). In the transcription‐dependent model, neighboring genes within a hub compete for enhancer activation and thereby repress the expression of one another. These findings underscore the importance of nascent mRNAs in their regulatory roles outside of the central dogma, adding a new layer to the paradigm in gene expression regulation.
Impaired Tricellular Tight Junctions Open the Barrier for Lipopolysaccharide Uptake
Authors: Krug Susanne M. , Lee In-Fah M. , Weiß Franziska , Hu Jia-Chen E. , Fromm Michael , Schulzke Jörg D.
Published: April 2020

Lipopolysaccharides (LPS) are essential membrane components of most Gram‐negative bacteria and several pathologies have been linked to LPS‐induced inflammation. However, understanding the mechanisms of intestinal uptake of LPS is still at its beginning. Uptake is thought to happen either via toll‐like‐receptor‐4‐induced phagocytosis or via paracellular passage. In ulcerative colitis (UC), the paracellular barrier formed by tight junctions (TJ) is impaired and tricellulin, a protein of the tricellular TJ of importance for regulation of paracellular macromolecule passage, is downregulated.

We hypothesize that a sole reduction in tricellulin expression leads to enhanced antigen uptake with the consequence that LPS is passing the intestinal epithelium via the tricellular TJ.

For analyzing this, we generated clones of intestinal epithelial cell monolayers HT‐29/B6 at different tricellulin knockdown levels and measured the permeability for LPS. The passage of LPS through the tricellular TJ was visualized using a dedicated fluorescence microscopical technique. We then analyzed the capability of passaged LPS to stimulate immune cells and by that to induce inflammation. For this, we added LPS apically to the monolayers containing different levels of tricellulin and co‐cultured them with basolaterally located macrophages. Passing LPS should stimulate these after passage. The basolateral medium thus containing secreted cytokines was added to otherwise uninvolved HT‐29/B6 cells and the effect on transepithelial resistance (TER) was monitored.

Using concentrations of LPS which were below to affect the epithelial barrier of HT‐29/B6 cells, at a decreased level of tricellulin of 50% the permeability for LPS was three times higher than in controls (shTric50: 2.09±0.56 × 10−9 cm/s; control: 0.70±0.13 × 10−9 cm/s; n=7, p=0.03). Although the permeability was quite low in absolute terms, LPS passage in shTric50 was sufficient to stimulate macrophages to secrete cytokines. This was not the case for cells with high tricellulin expression. TER of HT‐29/B6 cells treated with the supernatants of the macrophages of the co‐culture with LPS‐incubated controls did not change.

We conclude that reduced tricellulin levels allow for paracellular passage of LPS via tricellular TJs and thereby may stimulate immune cells supporting inflammatory processes as happening in UC. This signifies a new pathomechanistic pathway for LPS‐induced inflammation.

Support or Funding Information

Supported by DFG GRK 2318 and DFG TRR 241