bluuebunny/crossref_metadata_2025 huggingface dataset:
Design and Performance of a Miniaturized, Low-Energy, Large Beam Spot Electron Flood Gun
Authors: Deng Chenhui , Han Li , Wang Yan
Published: March 2021

Charge accumulation often occurs in electron optics equipment and interferes with their operation. The trouble can be handled by using the electron flood gun. However, there are still some scenarios where neutralization is not as desired. To achieve a better charge neutralization effect and to facilitate work in confined spaces, a small size, low-energy electron flood gun providing a large area and uniform electron beam has been required. This article employs Munro’s Electron Beam Software (MEBS) to simulate the effect of the structure parameters on the performance of the beam. Based on the simulation results, the electron flood gun is processed and assembled. To verify the performance of the electron flood gun, this paper proposes a new “pinhole scanning method”. By using the method, we have achieved in-situ measurements of beam current and beam spot. The experimental results generally match the simulation results, which demonstrates that the electron flood gun has good performance and is likely to have many applications.
State Estimation for Cooperative Lateral Vehicle Following Using Vehicle-to-Vehicle Communication
Authors: Schinkel Wouter , van der Sande Tom , Nijmeijer Henk
Published: March 2021

A cooperative state estimation framework for automated vehicle applications is presented and demonstrated via simulations, the estimation framework is used to estimate the state of a lead and following vehicle simultaneously. Recent developments in the field of cooperative driving require novel techniques to ensure accurate and stable vehicle following behavior. Control schemes for the cooperative control of longitudinal and lateral vehicle dynamics generally require vehicle state information about the lead vehicle, which in some cases cannot be accurately measured. Including vehicle-to-vehicle communication in the state estimation process can provide the required input signals for the practical implementation of cooperative control schemes. This study is focused on demonstrating the benefits of using vehicle-to-vehicle communication in the state estimation of a lead and following vehicle via simulations. The state estimator, which uses a cascaded Kalman filtering process, takes the operating frequencies of different sensors into account in the estimation process. Simulation results of three different driving scenarios demonstrate the benefits of using vehicle-to-vehicle communication as well as the attenuation of measurement noise. Furthermore, in contrast to relying on low frequency measurement data for the input signals of cooperative control schemes, the state estimator provides a state estimate at every sample.
Carry-Propagation-Adder-Factored Gemmini Systolic Array for Machine Learning Acceleration
Authors: Inayat Kashif , Chung Jaeyong
Published: March 2021

Systolic arrays are the primary part of modern deep learning accelerators and are being used widely in real-life applications such as self-driving cars. This paper presents a novel factored systolic array, where the carry propagation adder for accumulation and the rounding logic are extracted out from each processing element, which reduces the area, power and delay of the processing elements substantially. The factoring is performed in the column-wise manner and the cost of the factored logic, placed at each column output, is amortized by the processing elements in a column. We demonstrate the proposed factoring in an open source systolic array, Gemmini. The factoring technique does not change the functionality of the base design and is transparent to applications. We show that the proposed technique leads to substantial reduction in area and delay up to 45.3% and 23.7%, respectively, compared to the Gemmini baseline.
Modelling and Control of a Grid-Connected RES-Hydrogen Hybrid Microgrid
Authors: Villa Londono Jonny Esteban , Mazza Andrea , Pons Enrico , Lok Harm , Bompard Ettore
Published: March 2021

This paper proposes a Hybrid Microgrid (HμG) model including distributed generation (DG) and a hydrogen-based storage system, controlled through a tailored control strategy. The HμG is composed of three DG units, two of them supplied by solar and wind sources, and the latter one based on the exploitation of theProton Exchange Membrane (PEM) technology. Furthermore, the system includes an alkaline electrolyser, which is used as a responsive load to balance the excess of Variable Renewable Energy Sources (VRES) production, and to produce the hydrogen that will be stored into the hydrogen tank and that will be used to supply the fuel cell in case of lack of generation. The main objectives of this work are to present a validated dynamic model for every component of the HμG and to provide a strategy to reduce as much as possible the power absorption from the grid by exploiting the VRES production. The alkaline electrolyser and PEM fuel cell models are validated through real measurements. The State of Charge (SoC) of the hydrogen tank is adjusted through an adaptive scheme. Furthermore, the designed supervisor power control allows reducing the power exchange and improving the system stability. Finally, a case, considering a summer load profile measured in an electrical substation of Politecnico di Torino, is presented. The results demonstrates the advantages of a hydrogen-based micro-grid, where the hydrogen is used as medium to store the energy produced by photovoltaic and wind systems, with the aim to improve the self-sufficiency of the system.
Study of the EU-DEMO WCLL Breeding Blanket Primary Cooling Circuits Thermal-Hydraulic Performances during Transients Belonging to LOFA Category
Authors: Ciurluini Cristiano , Giannetti Fabio , Del Nevo Alessandro , Caruso Gianfranco
Published: March 2021

The Breeding Blanket (BB) is one of the key components of the European Demonstration (EU-DEMO) fusion reactor. Its main subsystems, the Breeder Zone (BZ) and the First Wall (FW), are cooled by two independent cooling circuits, called Primary Heat Transfer Systems (PHTS). Evaluating the BB PHTS performances in anticipated transient and accident conditions is a relevant issue for the design of these cooling systems. Within the framework of the EUROfusion Work Package Breeding Blanket, it was performed a thermal-hydraulic analysis of the PHTS during transient conditions belonging to the category of “Decrease in Coolant System Flow Rate”, by using Reactor Excursion Leak Analysis Program (RELAP5) Mod3.3. The BB, the PHTS circuits, the BZ Once Through Steam Generators and the FW Heat Exchangers were included in the study. Selected transients consist in partial and complete Loss of Flow Accident (LOFA) involving either the BZ or the FW PHTS Main Coolant Pumps (MCPs). The influence of the loss of off-site power, combined with the accident occurrence, was also investigated. The transient analysis was performed with the aim of design improvement. The current practice of a standard Pressurized Water Reactor (PWR) was adopted to propose and study actuation logics related to each accidental scenario. The appropriateness of the current PHTS design was demonstrated by simulation outcomes.
Operation Regimes: A Comparison Based on Nannochloropsis oceanica Biomass and Lipid Productivity
Authors: Guerra Inês , Pereira Hugo , Costa Margarida , Silva Joana T. , Santos Tamára , Varela João , Mateus Marília , Silva Joana
Published: March 2021

Microalgae are currently considered to be a promising feedstock for biodiesel production. However, significant research efforts are crucial to improve the current biomass and lipid productivities under real outdoor production conditions. In this context, batch, continuous and semi-continuous operation regimes were compared during the Spring/Summer seasons in 2.6 m3 tubular photobioreactors to select the most suitable one for the production of the oleaginous microalga Nannochloropsis oceanica. Results obtained revealed that N. oceanica grown using the semi-continuous and continuous operation regimes enabled a 1.5-fold increase in biomass volumetric productivity compared to that cultivated in batch. The lipid productivity was 1.7-fold higher under semi-continuous cultivation than that under a batch operation regime. On the other hand, the semi-continuous and continuous operation regimes spent nearly the double amount of water compared to that of the batch regime. Interestingly, the biochemical profile of produced biomass using the different operation regimes was not affected regarding the contents of proteins, lipids and fatty acids. Overall, these results show that the semi-continuous operation regime is more suitable for the outdoor production of N. oceanica, significantly improving the biomass and lipid productivities at large-scale, which is a crucial factor for biodiesel production.
Anti-Apoptotic and Anti-Inflammatory Role of Trans ε-Viniferin in a Neuron–Glia Co-Culture Cellular Model of Parkinson’s Disease
Authors: Sergi Domenico , Gélinas Alex , Beaulieu Jimmy , Renaud Justine , Tardif-Pellerin Emilie , Guillard Jérôme , Martinoli Maria-Grazia
Published: March 2021

The polyphenol trans-ε-viniferin (viniferin) is a dimer of resveratrol, reported to hold antioxidant and anti-inflammatory properties. The aims of our study were to evaluate the neuroprotective potential of viniferin in the nerve growth factor (NGF)-differentiated PC12 cells, a dopaminergic cellular model of Parkinson’s disease (PD) and assess its anti-inflammatory properties in a N9 microglia–neuronal PC12 cell co-culture system. The neuronal cells were pre-treated with viniferin, resveratrol or their mixture before the administration of 6-hydroxydopamine (6-OHDA), recognized to induce parkinsonism in rats. Furthermore, N9 microglia cells, in a co-culture system with neuronal PC12, were pre-treated with viniferin, resveratrol or their mixture to investigate whether these polyphenols could reduce lipopolysaccharide (LPS)-induced inflammation. Our results show that viniferin as well as a mixture of viniferin and resveratrol protects neuronal dopaminergic cells from 6-OHDA-induced cytotoxicity and apoptosis. Furthermore, when viniferin, resveratrol or their mixture was used to pre-treat microglia cells in our co-culture system, they reduced neuronal cytotoxicity induced by glial activation. Altogether, our data highlight a novel role for viniferin as a neuroprotective and anti-inflammatory molecule in a dopaminergic cellular model, paving the way for nutraceutical therapeutic avenues in the complementary treatments of PD.
Analysis of MCM Proteins’ Role as a Potential Target of Statins in Patients with Acute Type A Aortic Dissection through Bioinformatics
Authors: Liang Zheyong , Zhang Yongjian , Chen Qiang , Hao Junjun , Wang Haichen , Li Yongxin , Yan Yang
Published: March 2021

Acute aortic dissection is one of the most severe vascular diseases. The molecular mechanisms of aortic expansion and dissection are unclear. Clinical studies have found that statins play a protective role in aortic dissection development and therapy; however, the mechanism of statins’ effects on the aorta is unknown. The Gene Expression Omnibus (GEO) dataset GSE52093, GSE2450and GSE8686 were analyzed, and genes expressed differentially between aortic dissection samples and normal samples were determined using the Networkanalyst and iDEP tools. Weight gene correlation network analysis (WGCNA), functional annotation, pathway enrichment analysis, and the analysis of the regional variations of genomic features were then performed. We found that the minichromosome maintenance proteins (MCMs), a family of proteins targeted by statins, were upregulated in dissected aortic wall tissues and play a central role in cell-cycle and mitosis regulation in aortic dissection patients. Our results indicate a potential molecular target and mechanism for statins’ effects in patients with acute type A aortic dissection.
Expansion and Diversification of Fluorescent Protein Genes in Fifteen Acropora Species during the Evolution of Acroporid Corals
Authors: Kashimoto Rio , Hisata Kanako , Shinzato Chuya , Satoh Noriyuki , Shoguchi Eiichi
Published: March 2021

In addition to a purple, non-fluorescent chromoprotein (ChrP), fluorescent proteins (FPs) account for the vivid colors of corals, which occur in green (GFP), cyan (CFP), and red (RFP) FPs. To understand the evolution of the coral FP gene family, we examined the genomes of 15 Acropora species and three confamilial taxa. This genome-wide survey identified 219 FP genes. Molecular phylogeny revealed that the 15 Acropora species each have 9–18 FP genes, whereas the other acroporids examined have only two, suggesting a pronounced expansion of the FP genes in the genus Acropora. The data estimates of FP gene duplication suggest that the last common ancestor of the Acropora species that survived in the period of high sea surface temperature (Paleogene period) has already gained 16 FP genes. Different evolutionary histories of lineage-specific duplication and loss were discovered among GFP/CFPs, RFPs, and ChrPs. Synteny analysis revealed core GFP/CFP, RFP, and ChrP gene clusters, in which a tandem duplication of the FP genes was evident. The expansion and diversification of Acropora FPs may have contributed to the present-day richness of this genus.
Evaluation of Social Values for Ecosystem Services in Urban Riverfront Space Based on the SolVES Model: A Case Study of the Fenghe River, Xi’an, China
Authors: Zhang Zhicheng , Zhang Hongjuan , Feng Juan , Wang Yirong , Liu Kang
Published: March 2021

Urban riverfront space has diversified ecosystem services, but due to excessive changes in the geographical environment, such as drastic changes in land use, people gain social value at a great ecological cost. Obtaining benefits from the ecosystem in this way is not sustainable. Therefore, this paper uses the SolVES model to evaluate the social value of ecosystem services on the east bank of the Fenghe River, while also studying the contribution of different environmental variables to social value. The main results are as follows. (1) Environmental variables affect the spatial distribution characteristics of social value. The distance to water (DTW) means the social value was distributed in strips, and the distance to road (DTR) concentrated the social value along the road. The landscape type (LT) means the social value was concentrated in the landscape space. (2) When DTW, DTR, and LT were collectively used as environmental variables, the distribution characteristics of various social values were similar to when LT was used as the only environmental variable. (3) The results of MaxEnt show that LT made a greater contribution to the aesthetic, recreation, therapeutic, and historic values, and was the largest contribution factor to the aesthetic, therapeutic, and historic values, with contribution rates of 47.6, 50.5, and 80.0%, respectively. DTW is the factor that contributed the most to recreation, with a contribution rate of 43.1%. Improving social value based on the influence and contribution of environmental variables can reduce the damage to the ecological environment caused by changes in geographic factors. This is sustainable for both the ecosystem and the services it provides to mankind.
Lean Six Sigma Approach for Reducing Length of Hospital Stay for Patients with Femur Fracture in a University Hospital
Authors: Scala Arianna , Ponsiglione Alfonso Maria , Loperto Ilaria , Della Vecchia Antonio , Borrelli Anna , Russo Giuseppe , Triassi Maria , Improta Giovanni
Published: March 2021

Surgical intervention within 48 h of hospital admission is the gold standard procedure for the management of elderly patients with femur fractures, since the increase in preoperative waiting time is correlated with the onset of complications and longer overall length of stay (LOS) in the hospital. However, national evidence demonstrates that there is still the need to provide timely intervention for this type of patient, especially in some regions of central southern Italy. Here we discuss the introduction of a diagnostic–therapeutic assistance pathway (DTAP) to reduce the preoperative LOS for patients undergoing femur fracture surgery in a university hospital. A Lean Six Sigma methodology, based on the DMAIC cycle (Define, Measure, Analyze, Improve, Control), is implemented to evaluate the effectiveness of the DTAP. Data were retrospectively collected and analyzed from two groups of patients before and after the implementation of DTAP over a period of 10 years. The statistics of the process measured before the DTAP showed an average preoperative LOS of 5.6 days (standard deviation of 3.2), thus confirming the need for corrective actions to reduce the LOS in compliance with the national guidelines. The influence of demographic and anamnestic variables on the LOS was evaluated, and the impact of the DTAP was measured and discussed, demonstrating the effectiveness of the improvement actions implemented over the years and leading to a significant reduction in the preoperative LOS, which decreased to an average of 3.5 days (standard deviation of 3.60). The obtained reduction of 39% in the average LOS proved to be in good agreement with previously developed DTAPs for femur fracture available in the literature.
Accentuating the Interrelation between Consumer Intention and Healthy Packaged Food Selection during COVID-19: A Case Study of Pakistan
Authors: Zafar Muhammad Zeeshan , Maqbool Adnan , Cioca Lucian-Ionel , Shah Syed Ghulam Meran , Masud Shahjahan
Published: March 2021

This study contemplates the factors that influence consumer intention, before and during the eruption of COVID-19, for the selection of healthy packaged food in Pakistan. The extant studies have identified two distinct attitudes of consumers about food label information: one is its usefulness and the second elucidates the avoidance. Hence forth, the current study contributes to the extant literature while signifying both reasons which motivate consumers to read food labels and reasons which discourage consumers from consult food labels at the point of purchase. Moreover, the impact of subjective norms and self-efficacy for healthy packaged food intentions has also been examined for both before the emergence of COVID-19 and during the spread of COVID-19. The underpinning of the proposed model has been justified by the behavioral reasoning theory. The cross-sectional data of 14,455 students has was collected from 10 universities through Microsoft Teams and Zoom. AMOS 21 was employed for the final analysis. The results indicate that before COVID-19 the subjective norms and self-efficacy were not the stimulating factors for the selection of healthy packaged food. On the contrary, during the COVID-19 outbreak, the subjective norms and self-efficacy divulged a significant effect. Moreover, the reasons to consult food labels are positively significant whereas the reasons to avoid food labels have negatively affected the consumer, both before COVID-19 and during COVID-19 outbreak, while endorsing healthy packaged food. Conclusively, COVID-19 has been proved to be a deterrent for unhealthy packaged food lovers while being a blessing for healthy packaged food.
The Effects of Cold Atmospheric Pressure Plasma on Germination Parameters, Enzyme Activities and Induction of DNA Damage in Barley
Authors: Peťková Mária , Švubová Renáta , Kyzek Stanislav , Medvecká Veronika , Slováková Ľudmila , Ševčovičová Andrea , Gálová Eliška
Published: March 2021

Climate change, environmental pollution and pathogen resistance to available chemical agents are part of the problems that the food industry has to face in order to ensure healthy food for people and livestock. One of the promising solutions to these problems is the use of cold atmospheric pressure plasma (CAPP). Plasma is suitable for efficient surface decontamination of seeds and food products, germination enhancement and obtaining higher yields in agricultural production. However, the plasma effects vary due to plasma source, treatment conditions and seed type. In our study, we tried to find the proper conditions for treatment of barley grains by diffuse coplanar surface barrier discharge, in which positive effects of CAPP, such as enhanced germination or decontamination effects, would be maximized and harmful effects, such as oxidation and genotoxic potential, minimized. Besides germination parameters, we evaluated DNA damage and activities of various germination and antioxidant enzymes in barley seedlings. Plasma exposure resulted in changes in germination parameters and enzyme activities. Longer exposures had also genotoxic effects. As such, our findings indicate that appropriate plasma exposure conditions need to be carefully optimized in order to preserve germination, oxidation balance and genome stability, should CAPP be used in agricultural practice.
Coagulation and Fibrinolysis in Obstructive Sleep Apnoea
Authors: Bikov Andras , Meszaros Martina , Schwarz Esther Irene
Published: March 2021

Obstructive sleep apnoea (OSA) is a common disease which is characterised by repetitive collapse of the upper airways during sleep resulting in chronic intermittent hypoxaemia and frequent microarousals, consequently leading to sympathetic overflow, enhanced oxidative stress, systemic inflammation, and metabolic disturbances. OSA is associated with increased risk for cardiovascular morbidity and mortality, and accelerated coagulation, platelet activation, and impaired fibrinolysis serve the link between OSA and cardiovascular disease. In this article we briefly describe physiological coagulation and fibrinolysis focusing on processes which could be altered in OSA. Then, we discuss how OSA-associated disturbances, such as hypoxaemia, sympathetic system activation, and systemic inflammation, affect these processes. Finally, we critically review the literature on OSA-related changes in markers of coagulation and fibrinolysis, discuss potential reasons for discrepancies, and comment on the clinical implications and future research needs.
Transcriptional Regulatory Networks Associate with Early Stages of Potato Virus X Infection of Solanum tuberosum
Authors: Herath Venura , Verchot Jeanmarie
Published: March 2021

Potato virus X (PVX) belongs to genus Potexvirus. This study characterizes the cellular transcriptome responses to PVX infection in Russet potato at 2 and 3 days post infection (dpi). Among the 1242 differentially expressed genes (DEGs), 268 genes were upregulated, and 37 genes were downregulated at 2 dpi while 677 genes were upregulated, and 265 genes were downregulated at 3 dpi. DEGs related to signal transduction, stress response, and redox processes. Key stress related transcription factors were identified. Twenty-five pathogen resistance gene analogs linked to effector triggered immunity or pathogen-associated molecular pattern (PAMP)-triggered immunity were identified. Comparative analysis with Arabidopsis unfolded protein response (UPR) induced DEGs revealed genes associated with UPR and plasmodesmata transport that are likely needed to establish infection. In conclusion, this study provides an insight on major transcriptional regulatory networked involved in early response to PVX infection and establishment.
Characteristics of MSCs in Synovial Fluid and Mode of Action of Intra-Articular Injections of Synovial MSCs in Knee Osteoarthritis
Authors: Sekiya Ichiro , Katano Hisako , Ozeki Nobutake
Published: March 2021

We have been studying mesenchymal stem cells (MSCs) in synovial fluid and the intra-articular injection of synovial MSCs in osteoarthritis (OA) knees. Here, mainly based on our own findings, we overview the characteristics of endogenous MSCs in the synovial fluid of OA knees and their mode of action when injected exogenously into OA knees. Many MSCs similar to synovial MSCs were detected in the synovial fluid of human OA knees, and their number correlated with the radiological OA grade. Our suspended synovium culture model demonstrated the release of MSCs from the synovium through a medium into a non-contacting culture dish. In OA knees, endogenous MSCs possibly mobilize in a similar manner from the synovium through the synovial fluid and act protectively. However, the number of mobilized MSCs is limited; therefore, OA progresses in its natural course. Synovial MSC injections inhibited the progression of cartilage degeneration in a rat OA model. Injected synovial MSCs migrated into the synovium, maintained their MSC properties, and increased the gene expressions of TSG-6, PRG-4, and BMP-2. Exogenous synovial MSCs can promote anti-inflammation, lubrication, and cartilage matrix synthesis in OA knees. Based on our findings, we have initiated a human clinical study of synovial MSC injections in OA knees.
Bioluminescence Measurement of Time-Dependent Dynamic Changes of CYP-Mediated Cytotoxicity in CYP-Expressing Luminescent HepG2 Cells
Authors: Iwado Satoru , Abe Satoshi , Oshimura Mitsuo , Kazuki Yasuhiro , Nakajima Yoshihiro
Published: March 2021

We sought to develop a cell-based cytotoxicity assay using human hepatocytes, which reflect the effects of drug-metabolizing enzymes on cytotoxicity. In this study, we generated luminescent human hepatoblastoma HepG2 cells using the mouse artificial chromosome vector, in which click beetle luciferase alone or luciferase and major drug-metabolizing enzymes (CYP2C9, CYP2C19, CYP2D6, and CYP3A4) are expressed, and monitored the time-dependent changes of CYP-mediated cytotoxicity expression by bioluminescence measurement. Real-time bioluminescence measurement revealed that compared with CYP-non-expressing cells, the luminescence intensity of CYP-expressing cells rapidly decreased when the cells were treated with low concentrations of aflatoxin B1 or primaquine, which exhibits cytotoxicity in the presence of CYP3A4 or CYP2D6, respectively. Using kinetics data obtained by the real-time bioluminescence measurement, we estimated the time-dependent changes of 50% inhibitory concentration (IC50) values in the aflatoxin B1- and primaquine-treated cell lines. The first IC50 value was detected much earlier and at a lower concentration in primaquine-treated CYP-expressing HepG2 cells than in primaquine-treated CYP-non-expressing cells, and the decrease of IC50 values was much faster in the former than the latter. Thus, we successfully monitored time- and concentration-dependent dynamic changes of CYP-mediated cytotoxicity expression in CYP-expressing luminescent HepG2 cells by means of real-time bioluminescence measurement.
Skp2 and Slug Are Coexpressed in Aggressive Prostate Cancer and Inhibited by Neddylation Blockade
Authors: Mickova Alena , Kharaishvili Gvantsa , Kurfurstova Daniela , Gachechiladze Mariam , Kral Milan , Vacek Ondrej , Pokryvkova Barbora , Mistrik Martin , Soucek Karel , Bouchal Jan
Published: March 2021

Prostate cancer (PCa) is the second leading cause of cancer-related deaths in men in Western countries, and there is still an urgent need for a better understanding of PCa progression to inspire new treatment strategies. Skp2 is a substrate-recruiting component of the E3 ubiquitin ligase complex, whose activity is regulated through neddylation. Slug is a transcriptional repressor involved in the epithelial-to-mesenchymal transition, which may contribute to therapy resistance. Although Skp2 has previously been associated with a mesenchymal phenotype and prostate cancer progression, the relationship with Slug deserves further elucidation. We have previously shown that a high Gleason score (≥8) is associated with higher Skp2 and lower E-cadherin expression. In this study, significantly increased expression of Skp2, AR, and Slug, along with E-cadherin downregulation, was observed in primary prostate cancer in patients who already had lymph node metastases. Skp2 was slightly correlated with Slug and AR in the whole cohort (Rs 0.32 and 0.37, respectively), which was enhanced for both proteins in patients with high Gleason scores (Rs 0.56 and 0.53, respectively) and, in the case of Slug, also in patients with metastasis to lymph nodes (Rs 0.56). Coexpression of Skp2 and Slug was confirmed in prostate cancer tissues by multiplex immunohistochemistry and confocal microscopy. The same relationship between these two proteins was observed in three sets of prostate epithelial cell lines (PC3, DU145, and E2) and their mesenchymal counterparts. Chemical inhibition of Skp2, but not RNA interference, modestly decreased Slug protein in PC3 and its docetaxel-resistant subline PC3 DR12. Importantly, chemical inhibition of Skp2 by MLN4924 upregulated p27 and decreased Slug expression in PC3, PC3 DR12, and LAPC4 cells. Novel treatment strategies targeting Skp2 and Slug by the neddylation blockade may be promising in advanced prostate cancer, as recently documented for other aggressive solid tumors.
An Inverse QSAR Method Based on a Two-Layered Model and Integer Programming
Authors: Shi Yu , Zhu Jianshen , Azam Naveed Ahmed , Haraguchi Kazuya , Zhao Liang , Nagamochi Hiroshi , Akutsu Tatsuya
Published: March 2021

A novel framework for inverse quantitative structure–activity relationships (inverse QSAR) has recently been proposed and developed using both artificial neural networks and mixed integer linear programming. However, classes of chemical graphs treated by the framework are limited. In order to deal with an arbitrary graph in the framework, we introduce a new model, called a two-layered model, and develop a corresponding method. In this model, each chemical graph is regarded as two parts: the exterior and the interior. The exterior consists of maximal acyclic induced subgraphs with bounded height, the interior is the connected subgraph obtained by ignoring the exterior, and the feature vector consists of the frequency of adjacent atom pairs in the interior and the frequency of chemical acyclic graphs in the exterior. Our method is more flexible than the existing method in the sense that any type of graphs can be inferred. We compared the proposed method with an existing method using several data sets obtained from PubChem database. The new method could infer more general chemical graphs with up to 50 non-hydrogen atoms. The proposed inverse QSAR method can be applied to the inference of more general chemical graphs than before.
A Tryptophan ‘Gate’ in the CRISPR-Cas3 Nuclease Controls ssDNA Entry into the Nuclease Site, That When Removed Results in Nuclease Hyperactivity
Authors: He Liu , Matošević Zoe Jelić , Mitić Damjan , Markulin Dora , Killelea Tom , Matković Marija , Bertoša Branimir , Ivančić-Baće Ivana , Bolt Edward L.
Published: March 2021

Cas3 is a ssDNA-targeting nuclease-helicase essential for class 1 prokaryotic CRISPR immunity systems, which has been utilized for genome editing in human cells. Cas3-DNA crystal structures show that ssDNA follows a pathway from helicase domains into a HD-nuclease active site, requiring protein conformational flexibility during DNA translocation. In genetic studies, we had noted that the efficacy of Cas3 in CRISPR immunity was drastically reduced when temperature was increased from 30 °C to 37 °C, caused by an unknown mechanism. Here, using E. coli Cas3 proteins, we show that reduced nuclease activity at higher temperature corresponds with measurable changes in protein structure. This effect of temperature on Cas3 was alleviated by changing a single highly conserved tryptophan residue (Trp-406) into an alanine. This Cas3W406A protein is a hyperactive nuclease that functions independently from temperature and from the interference effector module Cascade. Trp-406 is situated at the interface of Cas3 HD and RecA1 domains that is important for maneuvering DNA into the nuclease active site. Molecular dynamics simulations based on the experimental data showed temperature-induced changes in positioning of Trp-406 that either blocked or cleared the ssDNA pathway. We propose that Trp-406 forms a ‘gate’ for controlling Cas3 nuclease activity via access of ssDNA to the nuclease active site. The effect of temperature in these experiments may indicate allosteric control of Cas3 nuclease activity caused by changes in protein conformations. The hyperactive Cas3W406A protein may offer improved Cas3-based genetic editing in human cells.
Regulation of Osteoblast Differentiation by Cytokine Networks
Authors: Amarasekara Dulshara Sachini , Kim Sumi , Rho Jaerang
Published: March 2021

Osteoblasts, which are bone-forming cells, play pivotal roles in bone modeling and remodeling. Osteoblast differentiation, also known as osteoblastogenesis, is orchestrated by transcription factors, such as runt-related transcription factor 1/2, osterix, activating transcription factor 4, special AT-rich sequence-binding protein 2 and activator protein-1. Osteoblastogenesis is regulated by a network of cytokines under physiological and pathophysiological conditions. Osteoblastogenic cytokines, such as interleukin-10 (IL-10), IL-11, IL-18, interferon-γ (IFN-γ), cardiotrophin-1 and oncostatin M, promote osteoblastogenesis, whereas anti-osteoblastogenic cytokines, such as tumor necrosis factor-α (TNF-α), TNF-β, IL-1α, IL-4, IL-7, IL-12, IL-13, IL-23, IFN-α, IFN-β, leukemia inhibitory factor, cardiotrophin-like cytokine, and ciliary neurotrophic factor, downregulate osteoblastogenesis. Although there are gaps in the body of knowledge regarding the interplay of cytokine networks in osteoblastogenesis, cytokines appear to be potential therapeutic targets in bone-related diseases. Thus, in this study, we review and discuss our osteoblast, osteoblast differentiation, osteoblastogenesis, cytokines, signaling pathway of cytokine networks in osteoblastogenesis.
Strategic Challenges of Human Resources Allocation in Industry 4.0
Authors: Ziaei Nafchi Majid , Mohelská Hana
Published: March 2021

The emergence of the fourth industrial revolution (Industry 4.0, hereinafter I 4.0) has led to an entirely fresh approach to production, helping to enhance the key industrial processes and therefore increase the growth of labor productivity and competitiveness. Simultaneously, I 4.0 compels changes in the organization of work and influences the lives of employees. The paper intends to construct a model for predicting the allocation of human resources in the sectors of the national economy of the Czech Republic in connection with I 4.0. The model used in this research visualizes the shift of labor in the economic sectors of the Czech Republic from the year 2013 to the following years in the near future. The main contribution of this article is to show the growth of employment in the high-tech services sector, which will have an ascending trend.
Combined Transcutaneous Spinal Stimulation and Locomotor Training to Improve Walking Function and Reduce Spasticity in Subacute Spinal Cord Injury: A Randomized Study of Clinical Feasibility and Efficacy
Authors: Estes Stephen , Zarkou Anastasia , Hope Jasmine M. , Suri Cazmon , Field-Fote Edelle C.
Published: March 2021

Locomotor training (LT) is intended to improve walking function and can also reduce spasticity in motor-incomplete spinal cord injury (MISCI). Transcutaneous spinal stimulation (TSS) also influences these outcomes. We assessed feasibility and preliminary efficacy of combined LT + TSS during inpatient rehabilitation in a randomized, sham-controlled, pragmatic study. Eighteen individuals with subacute MISCI (2–6 months post-SCI) were enrolled and randomly assigned to the LT + TSS or the LT + TSSsham intervention group. Participants completed a 4-week program consisting of a 2-week wash-in period (LT only) then a 2-week intervention period (LT + TSS or LT + TSSsham). Before and after each 2-week period, walking (10 m walk test, 2-min walk test, step length asymmetry) and spasticity (pendulum test, clonus drop test, modified spinal cord injury—spasticity evaluation tool) were assessed. Sixteen participants completed the study. Both groups improved in walking speed and distance. While there were no significant between-groups differences, the LT + TSS group had significant improvements in walking outcomes following the intervention period; conversely, improvements in the LT + TSSsham group were not significant. Neither group had significant changes in spasticity, and the large amount of variability in spasticity may have obscured ability to observe change in these measures. TSS is a feasible adjunct to LT in the subacute stage of SCI and may have potential to augment training-related improvements in walking outcomes.
Elderly Patients in a Large Nephrology Unit: Who Are Our Old, Old-Old and Oldest-Old Patients?
Authors: Torreggiani Massimo , Chatrenet Antoine , Fois Antioco , Moio Maria Rita , Mazé Béatrice , Coindre Jean Philippe , Crochette Romain , Sigogne Mickael , Wacrenier Samuel , Lecointre Léna , Breuer Conrad , Fessi Hafedh , Piccoli Giorgina Barbara
Published: March 2021

The world population is aging, and the prevalence of chronic kidney disease (CKD) is increasing. Whether this increase is also due to the methods currently being used to assess kidney function in the elderly is still a matter of discussion. We aimed to describe the actual referral pattern of CKD patients in a large nephrology unit and test whether the use of different formulae to estimate kidney function could affect the staging and the need for specialist care in the older subset of our population. In 2019, 1992 patients were referred to our center. Almost 28% of the patients were aged ≥80 and about 6% were ≥90 years old. Among the causes of kidney disease, glomerulonephritis displayed a higher prevalence in younger patients whereas hypertensive or diabetic kidney disease were more prevalent in older patients. The prevalence of referred patients in advanced CKD stages increased with age; estimated glomerular filtration rate (eGFR) decreased with age regardless of which equation was used (chronic kidney disease epidemiology collaboration (CKD-EPI), Lund–Malmö Revised (LMR), modification of diet in renal disease (MDRD), Full Age Spectrum (FAS), or Berlin Initiative Study 1 (BIS)). With CKD-EPI as a reference, MDRD and FAS underestimated the CKD stage while LMR overestimated it. The BIS showed the highest heterogeneity. Considering an eGFR threshold limit of 45 mL/min for defining “significant” CKD in patients over 65 years of age, the variability in CKD staging was 10% no matter which equation was used. Our study quantified the weight of “old” and “old-old” patients on follow-up in a large nephrology outpatient unit and suggested that with the current referral pattern, the type of formula used does not affect the need for CKD care within the context of a relatively late referral, particularly in elderly patients.
Challenges and Advances in Managing Thrombocytopenic Cancer Patients
Authors: Leader Avi , Hofstetter Liron , Spectre Galia
Published: March 2021

Cancer patients have varying incidence, depth and duration of thrombocytopenia. The mainstay of managing severe chemotherapy-induced thrombocytopenia (CIT) in cancer is the use of platelet transfusions. While prophylactic platelet transfusions reduce the bleeding rate, multiple unmet needs remain, such as high residual rates of bleeding, and anticancer treatment dose reductions/delays. Accordingly, the following promising results in other settings, antifibrinolytic drugs have been evaluated for prevention and treatment of bleeding in patients with hematological malignancies and solid tumors. In addition, Thrombopoeitin receptor agonists have been studied for two major implications in cancer: treatment of severe thrombocytopenia associated with myelodysplastic syndrome and acute myeloid leukemia; primary and secondary prevention of CIT in solid tumors in order to maintain dose density and intensity of anti-cancer treatment. Furthermore, thrombocytopenic cancer patients are often prescribed antithrombotic medication for indications arising prior or post cancer diagnosis. Balancing the bleeding and thrombotic risks in such patients represents a unique clinical challenge. This review focuses upon non-transfusion-based approaches to managing thrombocytopenia and the associated bleeding risk in cancer, and also addresses the management of antithrombotic therapy in thrombocytopenic cancer patients.
Epidemiological Studies of Brown Rot in Spanish Cherry Orchards in the Jerte Valley
Authors: Larena Inmaculada , Villarino Maria , Melgarejo Paloma , Cal Antonieta De
Published: March 2021

Cherry brown rot caused by Monilinialaxa was observed and estimated in organic cherry orchard located in the Jerte Valley between 2013 and 2018 (Cáceres, Spain). Climatic variables were collected from this orchard and also from a nearby weather station. The primary inoculum of the pathogen recorded in March was detected in overwintered mummified fruits, ground mummies, and necrotic twigs and was a function of the average temperature of the previous three months (December, January, and February). The first symptoms of brown rot could be observed on flowers until fruit set in April. The months of March and April were identified as the critical period for cherry brown-rot development. A significant positive correlation was identified between brown rot observed at harvest and the mean number of consecutive days in each fortnight of March and April when the percent relative humidity was above 80%. Brown-rot incidence observed over the 6 years ranged from 0 to 38%. More than 11 days with relative humidity >80% in each fortnight of critical period would mean 100% of cherry brown rot at harvest. A forecasting model could be used to predict brown rot infection in Jerte Valley cherries.
Novel Cyclic Peptides from Lethal Amanita Mushrooms through a Genome-Guided Approach
Authors: Zhou Shengwen , Li Xincan , Lüli Yunjiao , Li Xuan , Chen Zuo H. , Yuan Pengcheng , Yang Zhu L. , Li Guohong , Luo Hong
Published: March 2021

Most species in the genus Amanita are ectomycorrhizal fungi comprising both edible and poisonous mushrooms. Some species produce potent cyclic peptide toxins, such as α-amanitin, which places them among the deadliest organisms known to mankind. These toxins and related cyclic peptides are encoded by genes of the “MSDIN” family (named after the first five amino acid residues of the precursor peptides), and it is largely unknown to what extent these genes are expressed in the basidiocarps. In the present study, Amanita rimosa and Amanita exitialis were sequenced through the PacBio and Illumina techniques. Together with our two previously sequenced genomes, Amanita subjunquillea and Amanita pallidorosea, in total, 46 previously unknown MSDIN genes were discovered. The expression of over 80% of the MSDIN genes was demonstrated in A. subjunquillea. Through a combination of genomics and mass spectrometry, 12 MSDIN genes were shown to produce novel cyclic peptides. To further confirm the results, three of the cyclic peptides were chemically synthesized. The tandem mass spectrometry (MS/MS) spectra of the natural and the synthetic peptides shared a majority of the fragment ions, demonstrating an identical structure between each peptide pair. Collectively, the results suggested that the genome-guided approach is reliable for identifying novel cyclic peptides in Amanita species and that there is a large peptide reservoir in these mushrooms.
Dissection of the Activity of Agricultural Fungicides against Clinical Aspergillus Isolates with and without Environmentally and Medically Induced Azole Resistance
Authors: Jørgensen Karin Meinike , Helleberg Marie , Hare Rasmus Krøger , Jørgensen Lise Nistrup , Arendrup Maiken Cavling
Published: March 2021

Azole resistance is an emerging problem in patients with aspergillosis. The role of fungicides for resistance development and occurrence is not fully elucidated. EUCAST reference MICs of 17 fungicides (11 azoles and 6 others), five azole fungicide metabolites and four medical triazoles were examined against two reference and 28 clinical isolates of A. fumigatus, A. flavus and A. terreus with (n = 12) and without (n = 16) resistance mutations. Eight/11 azole fungicides were active against wild-type A. fumigatus, A. flavus and A. terreus, including four (metconazole, prothioconazole-desthio, prochloraz and imazalil) with low MIC50 (≤2 mg/L) against all three species and epoxiconazole, propiconazole, tebuconazole and difenoconazole also against wild-type A. terreus. Mefentrifluconazole, azole metabolites and non-azole fungicides MICs were >16 mg/L against A. fumigatus although partial growth inhibition was found with mefentrifluconazole. Moreover, mefentrifluconazole and axozystrobin were active against wild-type A. terreus. Increased MICs (≥3 dilutions) were found for TR34/L98H, TR34(3)/L98H, TR46/Y121F/T289A and G432S compared to wild-type A. fumigatus for epoxiconazole, propiconazole, tebuconazole, difenoconazole, prochloraz, imazalil and metconazole (except G432S), and for prothioconazole-desthio against TR46/Y121F/T289A, specifically. Increased MICs were found in A. fumigatus harbouring G54R, M220K and M220R alterations for five, one and one azole fungicides, respectively, compared to MICs against wild-type A. fumigatus. Similarly, increased MICs wer found for A. terreus with G51A, M217I and Y491H alterations for five, six and two azole fungicides, respectively. Azole fungicides showed activity against wild-type A. fumigatus, A. terreus and A. flavus, but not against all mutant isolates, suggesting the environmental route of azole resistance may have a role for all three species.
Mechanical Characteristics of Cement Paste in the Presence of Carbon Nanotubes and Silica Oxide Nanoparticles: An Experimental Study
Authors: Karakouzian Moses , Farhangi Visar , Farani Marzieh Ramezani , Joshaghani Alireza , Zadehmohamad Mehdi , Ahmadzadeh Mohammad
Published: March 2021

Considering the remarkable characteristics of nanomaterials, previous research studies investigated the effects of incorporating different types of these materials on improving the concrete properties. However, further studies are required to evaluate the complementary hybridization and synergistic influence of nanomaterials. In this research, the combined effect of adding nano silica particles (NS) and multi-walled carbon nanotubes (MWCNT) on enhancing both the compressive and flexural strengths of the cement paste was investigated. Moreover, the morphology of the interface between cement paste and aggregates was studied by scanning electron microscopy (SEM). The mixtures were prepared using three different portions of MWCNT and NS. Electron microscopy images indicated a uniform distribution of nanoparticles in the cement matrix, enhanced hydration reactions, and increased density. Based on the experiments’ outcomes, the combined utilization of silica and carbon nanomaterials in the cement paste did not necessarily result in the maximum compressive and flexural strengths. Furthermore, it was observed that the use of higher percentages of pristine NS in the absence of MWCNT can lead to further enhancement of strength properties of the cement paste.