bluuebunny/crossref_metadata_2025 huggingface dataset:
A Retrospective Analysis of Dog–Dog and Dog–Human Cases of Aggression in Northern Italy
Authors: Notari Lorella , Cannas Simona , Di Sotto Ylenia Agata , Palestrini Clara
Published: September 2020

In this survey, a caseload of aggressive dogs (n = 170) was analysed to increase the available information about biting dog characteristics, contribute to risk evaluation and improve bite prevention tools. All dog data were collected from questionnaires completed by veterinary behaviourists in Northern Italy. All dogs were referred to them by public authorities to be evaluated and treated due to the incidence of one or more episodes of aggression. Between the two groups of human-directed and dog-directed aggressive dogs, significant associations were found: the dog-directed aggressive group inflicted significantly more severe bites (p < 0.01) and offensive aggression (p < 0.01), whereas defensive aggression was significantly more numerous in the human-directed aggression group (p < 0.01) and more significantly located in private homes (p < 0.01). No significant associations were found between the severity of bites and one or more specific breeds in either group. The prevalence of defensive bites in private homes in human-directed aggression indicate that owners’ understanding of their dogs’ behaviour and communication is fundamental to preventing aggression, and that owner education programmes are fundamental tools to reduce aggression risk factors and prevent aggression.
Effects of Pre-Cooling on Thermophysiological Responses in Elite Eventing Horses
Authors: Klous Lisa , Siegers Esther , van den Broek Jan , Folkerts Mireille , Gerrett Nicola , van Oldruitenborgh-Oosterbaan Marianne Sloet , Munsters Carolien
Published: September 2020

In this study, we examined the effects of pre-cooling on thermophysiological responses in horses exercising in moderate environmental conditions (average wet bulb globe temperature: 18.5 ± 3.8 °C). Ten international eventing horses performed moderate intensity canter training on two separate days, and were either pre-cooled with cold-water rinsing (5–9 °C for 8 ± 3 min; cooling) or were not pre-cooled (control). We determined velocity (V), heart rate (HR), rectal temperature (Tre,), shoulder and rump skin temperature (Tshoulder and Trump), plasma lactate concentration (LA), gross sweat loss (GSL), and local sweat rate (LSR), as well as sweat sodium, chloride and potassium concentrations. The effect of pre-cooling on Tre was dependent on time; after 20 min of exercise the effect was the largest (estimate: 0.990, 95% likelihood confidence intervals (95% CI): 0.987, 0.993) compared to the control condition, resulting in a lower median Tre of 0.3 °C. Skin temperature was also affected by pre-cooling compared to the control condition (Tshoulder: −3.30 °C, 95% CI: −3.739, −2.867; Trump: −2.31 °C, 95% CI: −2.661, −1.967). V, HR, LA, GSL, LSR and sweat composition were not affected by pre-cooling. In conclusion, pre-cooling by cold-water rinsing could increase the margin for heat storage, allowing a longer exercise time before a critical Tre is reached and, therefore, could potentially improve equine welfare during competition.
Pork Production with Entire Males: Directions for Control of Boar Taint
Authors: Squires E. James , Bone Christine , Cameron Jocelyn
Published: September 2020

Boar taint is caused by the accumulation of androstenone and skatole and other indoles in the fat; this is regulated by the balance between synthesis and degradation of these compounds and can be affected by a number of factors, including environment and management practices, sexual maturity, nutrition, and genetics. Boar taint can be controlled by immunocastration, but this practice has not been accepted in some countries. Genetics offers a long-term solution to the boar taint problem via selective breeding or genome editing. A number of short-term strategies to control boar taint have been proposed, but these can have inconsistent effects and there is too much variability between breeds and individuals to implement a blanket solution for boar taint. Therefore, we propose a precision livestock management approach to developing solutions for controlling taint. This involves determining the differences in metabolic processes and the genetic variations that cause boar taint in specific groups of pigs and using this information to design custom treatments based on the cause of boar taint. Genetic, proteomic or metabolomic profiling can then be used to identify and implement effective solutions for boar taint for specific populations of animals.
Effects of Growing-Finishing Pig Stocking Rates on Bermudagrass Ground Cover and Soil Properties
Authors: Pietrosemoli Silvana , Raczkowski Charles , Green James T. , Villamide Maria Jesús
Published: September 2020

This study compares four stocking rates (37, 74, 111 and 148 pigs ha−1) for growing to finishing pigs (18.4 ± 0.5 kg and 118.5 ± 2.0 kg and 35.7 ± 2.1 kg and 125.7 ± 2.3 kg initial and final BW for grazing periods 1 and 2, respectively) and their effect on ground cover and soil traits in bermudagrass (Cynodon dactylon [L.] Pers) pastures, over two 14-week grazing periods (July–September and May–August). The study was conducted at the Center for Environmental Farming systems at the Cherry Research Station, Goldsboro North Carolina. A continuous stocking method was implemented to manage the pasture. The percent ground cover was estimated with a modified step point technique. Soil samples were collected in three sampling positions (center, inner and outer areas of the paddocks) and two soil sampling depths (0–30 and 30–90 cm). The experimental design was a completely randomized block with three field replicates. Data were analyzed using the PROC GLIMMIX procedure of SAS/STAT ® Version 9.4. Greater ground cover and lesser soil nutrient concentrations were registered in bermudagrass paddocks managed with 37 pigs ha−1. The results of this study also validated the existence of a spatial pattern of soil properties, which differed among sampling positions and depths.
Delayed Antibiotic Prescription by General Practitioners in the UK: A Stated-Choice Study
Authors: Morrell Liz , Buchanan James , Roope Laurence , Pouwels Koen , Butler Christopher , Hayhoe Benedict , Moore Michael , Tonkin-Crine Sarah , McLeod Monsey , Robotham Julie , Walker A. , Wordsworth Sarah ,   on behalf of the STEPUP team
Published: September 2020

Delayed antibiotic prescription in primary care has been shown to reduce antibiotic consumption, without increasing risk of complications, yet is not widely used in the UK. We sought to quantify the relative importance of factors affecting the decision to give a delayed prescription, using a stated-choice survey among UK general practitioners. Respondents were asked whether they would provide a delayed or immediate prescription in fifteen hypothetical consultations, described by eight attributes. They were also asked if they would prefer not to prescribe antibiotics. The most important determinants of choice between immediate and delayed prescription were symptoms, duration of illness, and the presence of multiple comorbidities. Respondents were more likely to choose a delayed prescription if the patient preferred not to have antibiotics, but consultation length had little effect. When given the option, respondents chose not to prescribe antibiotics in 51% of cases, with delayed prescription chosen in 21%. Clinical features remained important. Patient preference did not affect the decision to give no antibiotics. We suggest that broader dissemination of the clinical evidence supporting use of delayed prescription for specific presentations may help increase appropriate use. Establishing patient preferences regarding antibiotics may help to overcome concerns about patient acceptance. Increasing consultation length appears unlikely to affect the use of delayed prescription.
Estimating the Heat Capacity of Non-Newtonian Ionanofluid Systems Using ANN, ANFIS, and SGB Tree Algorithms
Authors: Daneshfar Reza , Bemani Amin , Hadipoor Masoud , Sharifpur Mohsen , Ali Hafiz Muhammad , Mahariq Ibrahim , Abdeljawad Thabet
Published: September 2020

This work investigated the capability of multilayer perceptron artificial neural network (MLP–ANN), stochastic gradient boosting (SGB) tree, radial basis function artificial neural network (RBF–ANN), and adaptive neuro-fuzzy inference system (ANFIS) models to determine the heat capacity (Cp) of ionanofluids in terms of the nanoparticle concentration (x) and the critical temperature (Tc), operational temperature (T), acentric factor (ω), and molecular weight (Mw) of pure ionic liquids (ILs). To this end, a comprehensive database of literature reviews was searched. The results of the SGB model were more satisfactory than the other models. Furthermore, an analysis was done to determine the outlying bad data points. It showed that most of the experimental data points were located in a reliable zone for the development of the model. The mean squared error and R2 were 0.00249 and 0.987, 0.0132 and 0.9434, 0.0320 and 0.8754, and 0.0201 and 0.9204 for the SGB, MLP–ANN, ANFIS, and RBF–ANN, respectively. According to this study, the ability of SGB for estimating the Cp of ionanofluids was shown to be greater than other models. By eliminating the need for conducting costly and time-consuming experiments, the SGB strategy showed its superiority compared with experimental measurements. Furthermore, the SGB displayed great generalizability because of the stochastic element. Therefore, it can be highly applicable to unseen conditions. Furthermore, it can help chemical engineers and chemists by providing a model with low parameters that yields satisfactory results for estimating the Cp of ionanofluids. Additionally, the sensitivity analysis showed that Cp is directly related to T, Mw, and Tc, and has an inverse relation with ω and x. Mw and Tc had the highest impact and ω had the lowest impact on Cp.
Correction: Han et al. Detection of Spray-Dried Porcine Plasma (SDPP) Based on Electronic Nose and Near-Infrared Spectroscopy Data. Appl. Sci. 2020, 10, 2967
Authors: Han Xiaoteng , Lü Enli , Lu Huazhong , Zeng Fanguo , Qiu Guangjun , Yu Qiaodong , Zhang Min
Published: September 2020

We, the authors, wish to make the following corrections to our paper [...]
Rheology and Curability Characterization of Photosensitive Slurries for 3D Printing of Si3N4 Ceramics
Authors: Li Xingbang , Zhang Jingxian , Duan Yusen , Liu Ning , Jiang Jinhua , Ma Ruixin , Xi Hongan , Li Xiaoguang
Published: September 2020

Among a series of 3D printing techniques, stereolithography provides a new route to produce ceramic architectures with the advantages of high-precision and short cycle time. However, up to now the stereolithography of non-oxide ceramics still face complex and difficult problems. This work focused on the analysis of rheological and curing ability of Si3N4 photocurable slurries. The effects of monomer type, coarse silicon powder, solid loading and ambient temperature on the rheological behavior were intensively studied. The relationships between powder characteristic (involving refractive index, absorbance and the introduce of coarse silicon powder), monomer type and curing ability were discussed in detail. It is expected that this study may benefit the development of Si3N4 or other non-oxide ceramic slurries for stereolithography.
Biomedical Application of a Herbal Product Based on Two Asteraceae Species
Authors: Grigore Alice , Neagu Georgeta , Nita Sultana , Ionita Carmen , Ionita Lucian , Goanta Ana-Maria , Fernoaga Cristina , Danacu Valerica , Albulescu Radu
Published: September 2020

Background: Species belonging to the Asteraceae family have been widely studied for their healing properties. The paper introduces STOMAPET (patent pending A2016/00888/23.11.2016) based on Eupatorium cannabinum (hemp-agrimony) and Inula helenium (elecampagne) and its potential application in diseases of the oral system. Methods: STOMAPET was prepared according to the patent application and HPLC analysis was performed for chemical analysis. In vitro studies were performed using human monocytes and the cells’ viability was determined by MTS assay. In vivo studies involved lipopolysaccharide (LPS)-induced murine endotoxemia and veterinary clinical case reports. Results: HPLC analysis revealed various phenolic compounds and alantolactone. In in vitro studies, monocytes viability remained stable, around and above 100% at all concentrations (7.8 to 250 µg/mL), and a high stimulatory effect on cell proliferation was noted. In LPS-induced murine endotoxemia, STOMAPET prevents mortality, improves the decrease in body mass, and prevents the drop in temperature. Case reports of companion animals with oral diseases ameliorated after STOMAPET treatment are presented. Conclusions: Preclinical studies confirmed the safety of STOMAPET and its pharmacological potential. The results of the veterinary case reports are also encouraging. More studies are needed to find the precise mechanism of action in correlation to chemical composition.
Copernicus Sentinel-1 MT-InSAR, GNSS and Seismic Monitoring of Deformation Patterns and Trends at the Methana Volcano, Greece
Authors: Gatsios Theodoros , Cigna Francesca , Tapete Deodato , Sakkas Vassilis , Pavlou Kyriaki , Parcharidis Issaak
Published: September 2020

The Methana volcano in Greece belongs to the western part of the Hellenic Volcanic Arc, where the African and Eurasian tectonic plates converge at a rate of approximately 3 cm/year. While volcanic hazard in Methana is considered low, the neotectonic basin constituting the Saronic Gulf area is seismically active and there is evidence of local geothermal activity. Monitoring is therefore crucial to characterize any activity at the volcano that could impact the local population. This study aims to detect surface deformation in the whole Methana peninsula based on a long stack of 99 Sentinel-1 C-band Synthetic Aperture Radar (SAR) images in interferometric wide swath mode acquired in March 2015–August 2019. A Multi-Temporal Interferometric SAR (MT-InSAR) processing approach is exploited using the Interferometric Point Target Analysis (IPTA) method, involving the extraction of a network of targets including both Persistent Scatterers (PS) and Distributed Scatterers (DS) to augment the monitoring capability across the varied land cover of the peninsula. Satellite geodetic data from 2006–2019 Global Positioning System (GPS) benchmark surveying are used to calibrate and validate the MT-InSAR results. Deformation monitoring records from permanent Global Navigation Satellite System (GNSS) stations, two of which were installed within the peninsula in 2004 (METH) and 2019 (MTNA), are also exploited for interpretation of the regional deformation scenario. Geological, topographic, and 2006–2019 seismological data enable better understanding of the ground deformation observed. Line-of-sight displacement velocities of the over 4700 PS and 6200 DS within the peninsula are from −18.1 to +7.5 mm/year. The MT-InSAR data suggest a complex displacement pattern across the volcano edifice, including local-scale land surface processes. In Methana town, ground stability is found on volcanoclasts and limestone for the majority of the urban area footprint while some deformation is observed in the suburban zones. At the Mavri Petra andesitic dome, time series of the exceptionally dense PS/DS network across blocks of agglomerate and cinder reveal seasonal fluctuation (5 mm amplitude) overlapping the long-term stable trend. Given the steepness of the slopes along the eastern flank of the volcano, displacement patterns may indicate mass movements. The GNSS, seismological and MT-InSAR analyses lead to a first account of deformation processes and their temporal evolution over the last years for Methana, thus providing initial information to feed into the volcano baseline hazard assessment and monitoring system.
Tonic and Phasic Amperometric Monitoring of Dopamine Using Microelectrode Arrays in Rat Striatum
Authors: Lundblad Martin , Price David A. , Burmeister Jason J. , Quintero Jorge E. , Huettl Peter , Pomerleau Francois , Zahniser Nancy R. , Gerhardt Greg A.
Published: September 2020

Here we report a novel microelectrode array recording approach to measure tonic (resting) and phasic release of dopamine (DA) in DA-rich areas such as the rat striatum and nucleus accumbens. The resulting method is tested in intact central nervous system (CNS) and in animals with extensive loss of the DA pathway using the neurotoxin, 6-hydroxyDA (6-OHDA). The self-referencing amperometric recording method employs Nafion-coated with and without m-phenylenediamine recording sites that through real-time subtraction allow for simultaneous measures of tonic DA levels and transient changes due to depolarization and amphetamine-induced release. The recording method achieves low-level measures of both tonic and phasic DA with decreased recording drift allowing for enhanced sensitivity normally not achieved with electrochemical sensors in vivo.
XOR Binary Gravitational Search Algorithm with Repository: Industry 4.0 Applications
Authors: Ahmadieh Khanesar Mojtaba , Bansal Ridhi , Martínez-Arellano Giovanna , Branson David T.
Published: September 2020

Industry 4.0 is the fourth generation of industry which will theoretically revolutionize manufacturing methods through the integration of machine learning and artificial intelligence approaches on the factory floor to obtain robustness and speed-up process changes. In particular, the use of the digital twin in a manufacturing environment makes it possible to test such approaches in a timely manner using a realistic 3D environment that limits incurring safety issues and danger of damage to resources. To obtain superior performance in an Industry 4.0 setup, a modified version of a binary gravitational search algorithm is introduced which benefits from an exclusive or (XOR) operator and a repository to improve the exploration property of the algorithm. Mathematical analysis of the proposed optimization approach is performed which resulted in two theorems which show that the proposed modification to the velocity vector can direct particles to the best particles. The use of repository in this algorithm provides a guideline to direct the particles to the best solutions more rapidly. The proposed algorithm is evaluated on some benchmark optimization problems covering a diverse range of functions including unimodal and multimodal as well as those which suffer from multiple local minima. The proposed algorithm is compared against several existing binary optimization algorithms including existing versions of a binary gravitational search algorithm, improved binary optimization, binary particle swarm optimization, binary grey wolf optimization and binary dragonfly optimization. To show that the proposed approach is an effective method to deal with real world binary optimization problems raised in an Industry 4.0 environment, it is then applied to optimize the assembly task of an industrial robot assembling an industrial calculator. The optimal movements obtained are then implemented on a real robot. Furthermore, the digital twin of a universal robot is developed, and its path planning is done in the presence of obstacles using the proposed optimization algorithm. The obtained path is then inspected by human expert and validated. It is shown that the proposed approach can effectively solve such optimization problems which arises in Industry 4.0 environment.
Detection of Precipitation and Fog Using Machine Learning on Backscatter Data from Lidar Ceilometer
Authors: Kim Yong-Hyuk , Moon Seung-Hyun , Yoon Yourim
Published: September 2020

The lidar ceilometer estimates cloud height by analyzing backscatter data. This study examines weather detectability using a lidar ceilometer by making an unprecedented attempt at detecting weather phenomena through the application of machine learning techniques to the backscatter data obtained from a lidar ceilometer. This study investigates the weather phenomena of precipitation and fog, which are expected to greatly affect backscatter data. In this experiment, the backscatter data obtained from the lidar ceilometer, CL51, installed in Boseong, South Korea, were used. For validation, the data from the automatic weather station for precipitation and visibility sensor PWD20 for fog, installed at the same location, were used. The experimental results showed potential for precipitation detection, which yielded an F1 score of 0.34. However, fog detection was found to be very difficult and yielded an F1 score of 0.10.
Modeling of Woodworkers’ Exposure to Occupational Noises by Integrating Frequency Spectra Generated by Power Tools: A Pilot Study
Authors: Zheng Yu-Ping , Juang Yow-Jer , Yiin Lih-Ming
Published: September 2020

Woodwork is one of the occupations with high levels of noises. This pilot study attempted to simulate woodworkers’ occupational noise exposure by integrating frequency spectra measured from individual power tools with the respective time of tool use. Five volunteering woodworkers participated in the study, and each wore a noise dosimeter with 1/1 octave-band analysis during the work for exposure assessment. The information of use of tools and time of tool use was recorded by an on-site technician. Frequency spectra of common power tools, including circular saw, electric curve saw, orbit sander, router trimmer, drill machine, pillar drill machine, nail gun, and air compressor, were also individually measured by the dosimeter. Monte Carlo simulation was used to simulate the distances between tools and workers, which were used to determine noise levels in the modeling. The personal measurements of noise exposure were around 80 dBA with peaks locating between 1 and 4 kHz and were fairly matched by the simulated results. This pilot modeling is seemingly feasible and promising, and noise exposure could be assessed by multiple times of tool use with known noise characteristics of the tools. The convenient dose estimates could be useful for the hearing protection of woodworkers.
Durability Assessment of Recycled Aggregate HVFA Concrete
Authors: Corinaldesi Valeria , Donnini Jacopo , Giosué Chiara , Mobili Alessandra , Tittarelli Francesca
Published: September 2020

The possibility of producing high-volume fly ash (HVFA) recycled aggregate concrete represents an important step towards the development of sustainable building materials. In fact, there is a growing need to reduce the use of non-renewable natural resources and, at the same time, to valorize industrial by-products, such as fly ash, that would otherwise be sent to the landfill. The present experimental work investigates the physical and mechanical properties of concrete by replacing natural aggregates and cement with recycled aggregates and fly ash, respectively. First, the mechanical properties of four different mixtures have been analyzed and compared. Then, the effectiveness of recycled aggregate and fly ash on reducing carbonation and chloride penetration depth has been also evaluated. Finally, the corrosion behavior of the different concrete mixtures, reinforced with either bare or galvanized steel plates, has been evaluated. The results obtained show that high-volume fly ash (HVFA) recycled aggregate concrete can be produced without significative reduction in mechanical properties. Furthermore, the addition of high-volume fly ash and the total replacement of natural aggregates with recycled ones did not modify the corrosion behavior of embedded bare and galvanized steel reinforcement.
Thermal Action on Normal and High Strength Cement Mortars
Authors: Ripani Marianela , Xargay Hernán , Iriarte Ignacio , Bernardo Kevin , Caggiano Antonio , Folino Paula
Published: September 2020

High temperature effect on cement-based composites, such as concrete or mortars, represents one of the most important damaging process that may drastically affect the mechanical and durability characteristics of structures. In this paper, the results of an experimental campaign on cement mortars submitted to high temperatures are reported and discussed. Particularly, two mixtures (i.e., Normal (MNS) and High Strength Mortar (MHS)) having different water-to-binder ratios were designed and evaluated in order to investigate the incidence of both the mortar composition and the effects of thermal treatments on their physical and mechanical properties. Mortar specimens were thermally treated in an electrical furnace, being submitted to the action of temperatures ranging from 100 to 600 °C. After that and for each mortar quality and considered temperature, including the room temperature case of 20 °C, water absorption was measured by following a capillary water absorption test. Furthermore, uniaxial compression, splitting tensile and three-points bending tests were performed under residual conditions. A comparative analysis of the progressive damage caused by temperature on physical and mechanical properties of the considered mortars types is presented. On one hand, increasing temperatures produced increasing water absorption coefficients, evidencing the effect of thermal damages which may cause an increase in the mortars accessible porosity. However, under these circumstances, the internal porosity structure of lower w/b ratio mixtures results much more thermally-damaged than those of MNS. On the other hand, strengths suffered a progressive degradation due to temperature rises. While at low to medium temperatures, strength loss resulted similar for both mortar types, at higher temperature, MNS presented a relatively greater strength loss than that of MHS. The action of temperature also caused in all cases a decrease of Young’s Modulus and an increase in the strain corresponding to peak load. However, MHS showed a much more brittle behavior in comparison with that of MNS, for all temperature cases. Finally, the obtained results demonstrated that mortar quality cannot be neglected when the action of temperature is considered, being the final material performance dependent on the physical properties which, in turn, mainly depend on the mixture proportioning.
A Model-Based Analysis of Spatio-Temporal Changes of the Urban Expansion in Arid Area of Western China: A Case Study in North Xinjiang Economic Zone
Authors: Qian Jing , Zhou Qiming , Chen Xi , Sun Bo
Published: September 2020

Investigation of urban expansion can provide a better understanding of the urbanization process and its driving forces, which is critical for environmental management and land use planning. Total of 514 sampling points from the aerial photos and field sampling were applied to assess the image accuracy. A Conversion of Land Use and its Effect at Small Region Extent (CLUE-S) model was established to simulate the urbanization process at the township level in the North Xinjiang Economic Zone (NXEZ) of western China. Historical land use and land cover changes with multi-temporal remote sensing data were retrieved, and the underlying driving forces were explored by training the CLUE-S model. Moreover, future changes in urban development were simulated under different scenarios. Results showed that the overall accuracy reaches larger than 80% for the years of 2002, 2005, and 2007, and the corresponding kappa coefficient is bigger than 0.8. The NXEZ is at a premature development stage compared with urban clusters in eastern China. Before 1999, the driving force in this region was primary industry development. In recent years, secondary industries started to show significance in urbanization. These findings indicate that the industrial base and economic development in the NXEZ are still relatively weak and have not taken a strong leading role. When industry and population become the main driving factors, the regional economy will enter a new stage of leap-forward development, which in turn will stimulate a new round of rapid urbanization.
Temporal Variability of Drought in Nine Agricultural Regions of China and the Influence of Atmospheric Circulation
Authors: Sun Haowei , Hu Haiying , Wang Zhaoli , Lai Chengguang
Published: September 2020

In recent decades, the severe drought across agricultural regions of China has had significant impact on agriculture. The standardized precipitation evapotranspiration index (SPEI) has been widely used for drought analyses; however, SPEI is prone to be affected by potential evapotranspiration (PET). We thus examined the correlations between soil moisture anomalies and the SPEI calculated by the Thornthwaite, Hargreaves, and Penman–Monteith (PM) equations to select the most suitable for drought research. Additionally, the Mann–Kendall and wavelet analysis were used to investigate drought trends and to analyze and the impact of atmospheric circulation on drought in China from 1961 to 2018. The results showed that (1) PET obtained from the PM equation is the most suitable for SPEI calculation; (2) there were significant wetting trends in Northern China and the whole Chinese mainland and most of the wetting mutation points occurred in the 1970s and 1980s and the significant inter-annual oscillations period in the Chinese mainland was 2–4 years; (3) the Chinese mainland and Northern China are strongly influenced by West Pacific Trade Wind, while Western Pacific Subtropical High Intensity and Pacific Subtropical High Area have primary impact on Southern China.
Regulation of Poly(ADP-Ribose) Polymerase 1 Activity by Y-Box-Binding Protein 1
Authors: Naumenko Konstantin N. , Sukhanova Mariya V. , Hamon Loic , Kurgina Tatyana A. , Alemasova Elizaveta E. , Kutuzov Mikhail M. , Pastré David , Lavrik Olga I.
Published: September 2020

Y-box-binding protein 1 (YB-1) is a multifunctional positively charged protein that interacts with DNA or RNA and poly(ADP-ribose) (PAR). YB-1 is poly(ADP-ribosyl)ated and stimulates poly(ADP-ribose) polymerase 1 (PARP1) activity. Here, we studied the mechanism of YB-1-dependent PAR synthesis by PARP1 in vitro using biochemical and atomic force microscopy assays. PAR synthesis activity of PARP1 is known to be facilitated by co-factors such as Mg2+. However, in contrast to an Mg2+-dependent reaction, the activation of PARP1 by YB-1 is accompanied by overall up-regulation of protein PARylation and shortening of the PAR polymer. Therefore, YB-1 and cation co-factors stimulated PAR synthesis in divergent ways. PARP1 autoPARylation in the presence of YB-1 as well as trans-PARylation of YB-1 are greatly affected by the type of damaged DNA, suggesting that PARP1 activation depends on the formation of a PARP1–YB-1–DNA ternary complex. An unstructured C-terminal part of YB-1 involved in an interaction with PAR behaves similarly to full-length YB-1, indicating that both DNA and PAR binding are involved in the stimulation of PARP1 activity by YB-1. Thus, YB-1 is likely linked to the regulation of PARylation events in cells via an interaction with PAR and damaged DNA.
Hyperhomocysteinemia and Endothelial Dysfunction in Multiple Sclerosis
Authors: Dubchenko Ekaterina , Ivanov Alexander , Spirina Natalia , Smirnova Nina , Melnikov Mikhail , Boyko Alexey , Gusev Evgeniy , Kubatiev Aslan
Published: September 2020

Endothelial dysfunction is recognized as one of the leading factors in the pathogenesis of diseases of the central nervous system of various etiologies. Numerous studies have shown the role of hyperhomocysteinemia in the development of endothelial dysfunction and the prothrombogenic state. The most important condition in the development of multiple sclerosis (MS) is a dysregulation of the blood-brain barrier (BBB) and transendothelial leukocyte migration. It has been proven that homocysteine also contributes to the damage of neurons by the mechanism of excitotoxicity and the induction of the apoptosis of neurons. These processes can be one of the factors of neurodegenerative brain damage, which plays a leading role in the progression of MS. This review describes the pleiotropic effect of homocysteine on these processes and its role in MS pathogenesis.
Differentiating High-Grade Gliomas from Brain Metastases at Magnetic Resonance: The Role of Texture Analysis of the Peritumoral Zone
Authors: Csutak Csaba , Ștefan Paul-Andrei , Lenghel Lavinia Manuela , Moroșanu Cezar Octavian , Lupean Roxana-Adelina , Șimonca Larisa , Mihu Carmen Mihaela , Lebovici Andrei
Published: September 2020

High-grade gliomas (HGGs) and solitary brain metastases (BMs) have similar imaging appearances, which often leads to misclassification. In HGGs, the surrounding tissues show malignant invasion, while BMs tend to displace the adjacent area. The surrounding edema produced by the two cannot be differentiated by conventional magnetic resonance (MRI) examinations. Forty-two patients with pathology-proven brain tumors who underwent conventional pretreatment MRIs were retrospectively included (HGGs, n = 16; BMs, n = 26). Texture analysis of the peritumoral zone was performed on the T2-weighted sequence using dedicated software. The most discriminative texture features were selected using the Fisher and the probability of classification error and average correlation coefficients. The ability of texture parameters to distinguish between HGGs and BMs was evaluated through univariate, receiver operating, and multivariate analyses. The first percentile and wavelet energy texture parameters were independent predictors of HGGs (75–87.5% sensitivity, 53.85–88.46% specificity). The prediction model consisting of all parameters that showed statistically significant results at the univariate analysis was able to identify HGGs with 100% sensitivity and 66.7% specificity. Texture analysis can provide a quantitative description of the peritumoral zone encountered in solitary brain tumors, that can provide adequate differentiation between HGGs and BMs.
Impact of New Systemic Treatment and Radiotherapy in Melanoma Patients with Leptomeningeal Metastases
Authors: Tétu Pauline , Sirven-Villaros Lila , Cuzzubbo Stefania , Ursu Renata , Baroudjian Barouyr , Delyon Julie , Nataf François , De Margerie-Mellon Constance , Allayous Clara , Lefevre Wendy , Carpentier Antoine F. , Lebbé Céleste
Published: September 2020

Importance: Few data are available on patients with leptomeningeal disease (LM) from melanoma treated with new systemic therapies. Objective: To gain a better understanding of patients, disease characteristics, and therapeutic interventions in melanoma patients with LM in the era of new systemic treatment. Design: Clinical characteristics, treatments, and survival of melanoma patients diagnosed with LM, isolated or associated with brain metastases, were collected. The Cox regression model assessed the influence of patient and melanoma characteristics on survival. Setting: Monocentric, retrospective, real-life cohort of patients with LM from melanoma. Participants: All patients followed up at Saint-Louis University Hospital and diagnosed with LM between December 2013 and February 2020 were included. For each patient identified, a central review by dermato-oncologist and neuro-oncologist experts was performed to confirm the diagnosis of LM. Exposure: Impact of new systemic therapies and radiotherapy. Results: Among the 452 advanced melanoma patients followed at St Louis Hospital between 2013 and 2020, 41 patients with LM from melanoma were identified. Among them, 29 patients with a diagnosis of LM “confirmed” or “probable” after central neuro-oncologists reviewing were included. Nineteen patients had known melanoma brain metastases at LM diagnosis. Among the 27 patients treated with systemic therapy, 17 patients were treated with immunotherapy, 5 patients received targeted therapy, 1 was treated with chemotherapy, and 4 patients were treated with anti-PD-1 in combination with BRAF inhibitor. The median overall survival (OS) from LM diagnosis was 5.1 months. Median OS was 7.1 months for the 9 patients receiving systemic therapy combined with radiotherapy, and 3.2 months for the 20 patients not receiving combined radiotherapy. Elevated serum lactate dehydrogenase (LDH) (HR 1.44, 95% CI 1.09–1.90, p < 0.01) and presence of neurological symptoms at LM diagnosis (HR 2.96, 95% CI 1.25–6.99, p = 0.01) were associated with poor survival. At the time of data analysis, five patients were still alive with a median follow-up of 47.4 months and had persistent complete response. Conclusion: Targeted therapy and immunotherapy are promising new treatment options in LM from melanoma that can increase overall survival, and may induce long lasting remission in some patients.
Trial Sponsorship and Time to Reporting for Phase 3 Randomized Cancer Clinical Trials
Authors: Lin Timothy A. , Fuller Clifton David , Verma Vivek , Mainwaring Walker , Espinoza Andres F. , Miller Austin B. , Jethanandani Amit , Pasalic Dario , Das Prajnan , Minsky Bruce D. , Thomas Charles R. , Fogelman David R. , Subbiah Vivek , Subbiah Ishwaria M. , Ludmir Ethan B.
Published: September 2020

The pace of clinical trial data generation and publication is an area of interest within clinical oncology; however, little is known about the dynamics and covariates of time to reporting (TTR) of trial results. To assess these, ClinicalTrials.gov was queried for phase three clinical trials for patients with metastatic solid tumors, and the factors associated with TTR from enrollment completion to publication were analyzed. Based on the 319 included trials, cooperative-group-sponsored trials were reported at a slower rate than non-cooperative-group trials (median 37.5 vs. 31.0 months; p < 0.001), while industry-funded studies were reported at a faster rate than non-industry-supported trials (31.0 vs. 40.0 months; p = 0.005). Furthermore, successful trials (those meeting their primary endpoint) were reported at a faster rate than unsuccessful studies (27.5 vs. 36.0 months; p < 0.001). Multivariable analysis confirmed that industry funding was independently associated with a shorter TTR (p = 0.006), while cooperative group sponsorship was not associated with a statistically significant difference in TTR (p = 0.18). These data underscore an opportunity to improve cooperative group trial efficiency by reducing TTR.
Expression of AR-V7 (Androgen Receptor Variant 7) Protein in Granular Cytoplasmic Structures Is an Independent Prognostic Factor in Prostate Cancer Patients
Authors: König Paul , Eckstein Markus , Jung Rudolf , Abdulrahman Amer , Guzman Juan , Weigelt Katrin , Serrero Ginette , Hayashi Jun , Geppert Carol , Stöhr Robert , Hartmann Arndt , Wullich Bernd , Wach Sven , Taubert Helge , Lieb Verena
Published: September 2020

Prostate cancer (PCa) is the second most common cancer, causing morbidity and mortality among men world-wide. The expression of the androgen receptor (AR) and its splice variants is a crucial factor of prostate cancer biology that has not been comprehensively studied in PCa tumors. The aim of this study was to characterize the protein expression of the AR and its splice variant, AR-V7, and their subcellular distributions in PCa by immunohistochemistry and to correlate the results to the clinicopathological data and prognosis. Immunohistochemical staining for AR and AR-V7 was performed on a tissue microarray (TMA) with specimens from 410 PCa patients using an immunoreactive score (IRS) or only the percentage of AR-V7 staining in cytoplasmic granules. Nuclear or cytoplasmic AR staining was not associated with prognosis. AR-V7 staining was only occasionally observed in the nucleus. However, AR-V7 staining in the cytoplasm or in cytoplasmic granules was associated with relapse-free survival (RFS). AR-V7 staining of the cytoplasm was associated with a shorter RFS, whereas AR-V7 staining of cytoplasmic granules was associated with a longer RFS. In a multivariate Cox’s regression analysis, only negative (<5%) AR-V7 staining of cytoplasmic granules remained an independent prognostic factor for RFS (HR = 5.3; p = 0.006). In a further subgroup analysis by multivariate Cox’s regression analysis, AR-V7 was an independent prognostic factor in the following groups: age ≤ 65 (HR = 9.7; p = 0.029), negative CK20 staining (HR = 7.0; p = 0.008), and positive perineural invasion (HR = 3.7; p = 0.034). Altogether, AR-V7 protein in granular cytoplasmic structures is an independent prognostic factor for RFS in PCa patients.
Isolation and Enumeration of CTC in Colorectal Cancer Patients: Introduction of a Novel Cell Imaging Approach and Comparison to Cellular and Molecular Detection Techniques
Authors: Hendricks Alexander , Brandt Burkhard , Geisen Reinhild , Dall Katharina , Röder Christian , Schafmayer Clemens , Becker Thomas , Hinz Sebastian , Sebens Susanne
Published: September 2020

Circulating tumour cells (CTC) were proven to be prognostically relevant in cancer treatment, e.g., in colorectal cancer (CRC). This study validates a molecular detection technique through using a novel cell imaging approach for CTC detection and enumeration, in comparison to a size-based cellular and correlated the data to clinico-pathological characteristics. Overall, 57 CRC patients were recruited for this prospective study. Blood samples were analysed for CTCs by three methods: (1) Epithelial marker immunofluorescence staining combined with automated microscopy using the NYONE® cell imager; (2) isolation by size using membrane filtration with the ScreenCell® Cyto IS device and immunofluorescence staining; (3) detection by semi-quantitative Cytokeratin-20 RT-qPCR. Enumeration data were compared and correlated with clinic-pathological parameters. CTC were detected by either approach; however, with varying positivity rates: NYONE® 36.4%, ScreenCell® 100%, and PCR 80.5%. All methods revealed a positive correlation of CTC presence and higher tumour burden, which was most striking using the ScreenCell® device. Generally, no intercorrelation of CTC presence emerged amongst the applied techniques. Overall, enumeration of CTC after isolation by size demonstrated to be the most reliable strategy for the detection of CTC in CRC patients. Ongoing studies will have to unravel the prognostic value of this finding, and validate this approach in a larger cohort.
B Cells and Tertiary Lymphoid Structures Influence Survival in Lung Cancer Patients with Resectable Tumors
Authors: Tang Jun , Ramis-Cabrer Daniel , Curull Víctor , Wang Xuejie , Mateu-Jiménez Mercé , Pijuan Lara , Duran Xavier , Qin Liyun , Rodríguez-Fuster Alberto , Aguiló Rafael , Barreiro Esther
Published: September 2020

Immune profile of B and T cells and tertiary lymphoid structures (TLSs) may differ in tumors of lung cancer (LC) patients with/without chronic obstructive pulmonary disease (COPD), and may also influence patient survival. We sought to analyze: (1) TLSs, germinal centers (GCs), B and T cells, and (2) associations of the immune biomarkers with the patients’ 10-year overall survival (OS). TLSs (numbers and area), B [cluster of differentiation (CD) 20], and T (CD3), and GCs cells were identified in both tumor and non-tumor specimens (thoracotomy) from 90 LC-COPD patients and 43 LC-only patients. Ten-year OS was analyzed in the patients. Immune profile in tumors of LC-COPD versus LC: TLS numbers and areas significantly decreased in tumors of LC-COPD compared to LC patients. No significant differences were observed in tumors between LC-COPD and LC patients for B or T cells. Immune profile in tumors versus non-tumor specimens: TLS areas and B cells significantly increased, T cells significantly decreased in tumors of both LC and LC-COPD patients. Survival: in LC-COPD patients: greater area of TLSs and proportion of B cells were associated with longer survival rates. The immune tumor microenvironment differs in patients with underlying COPD and these different phenotypes may eventually impact the response to immunotherapy in patients with LC.
Inhibition of DNA Repair Pathways and Induction of ROS Are Potential Mechanisms of Action of the Small Molecule Inhibitor BOLD-100 in Breast Cancer
Authors: Bakewell Suzanne , Conde Isabel , Fallah Yassi , McCoy Mathew , Jin Lu , Shajahan-Haq Ayesha N.
Published: September 2020

BOLD-100, a ruthenium-based complex, sodium trans-[tetrachloridobis (1H-indazole) ruthenate (III)] (also known as IT-139, NKP1339 or KP1339), is a novel small molecule drug that demonstrated a manageable safety profile at the maximum tolerated dose and modest antitumor activity in a phase I clinical trial. BOLD-100 has been reported to inhibit the upregulation of the endoplasmic reticulum stress sensing protein GRP78. However, response to BOLD-100 varies in different cancer models and the precise mechanism of action in high-response versus low-response cancer cells remains unclear. In vitro studies have indicated that BOLD-100 induces cytostatic rather than cytotoxic effects as a monotherapy. To understand BOLD-100-mediated signaling mechanism in breast cancer cells, we used estrogen receptor positive (ER+) MCF7 breast cancer cells to obtain gene-metabolite integrated models. At 100 μM, BOLD-100 significantly reduced cell proliferation and expression of genes involved in the DNA repair pathway. BOLD-100 also induced reactive oxygen species (ROS) and phosphorylation of histone H2AX, gamma-H2AX (Ser139), suggesting disruption of proper DNA surveillance. In estrogen receptor negative (ER−) breast cancer cells, combination of BOLD-100 with a PARP inhibitor, olaparib, induced significant inhibition of cell growth and xenografts and increased gamma-H2AX. Thus, BOLD-100 is a novel DNA repair pathway targeting agent and can be used with other chemotherapies in ER− breast cancer.
Screening of Biocatalysts for Synthesis of the Wieland–Miescher Ketone
Authors: Patel Mitul P. , Green Nathaneal T. , Burch Jacob K. , Kew Kimberly A. , Hughes Robert M.
Published: September 2020

Lipases, a versatile class of biocatalysts, have been shown to function in non-aqueous media/organic solvents and to possess “promiscuous” catalytic activity for a wide range of organic transformations. In this study, we explored the biocatalytic properties of a library of commercially available lipases by screening them for catalysis of a one-pot synthesis of Wieland–Miescher ketone, an important intermediate in the synthesis of biologically active compounds such as steroids and terpenoids, from methyl vinyl ketone and 2-methyl-1,3-cyclohexanedione. As a direct outgrowth of this screen, we created an optimized procedure for Wieland–Miescher ketone (WMK) synthesis using crude lipase preparations, characterizing both reaction yield and enantiomeric excess. We also identified principal components of the crude lipase mixture through proteomics and present evidence for a non-lipolytic origin of the observed catalysis. Finally, using the optimized conditions developed in this study, we propose a general absorbance-based screening methodology for assessing biocatalytic potential of crude enzyme preparations for synthesis of WMK.
Aspergillus: A Powerful Protein Production Platform
Authors: Ntana Fani , Mortensen Uffe Hasbro , Sarazin Catherine , Figge Rainer
Published: September 2020

Aspergilli have been widely used in the production of organic acids, enzymes, and secondary metabolites for almost a century. Today, several GRAS (generally recognized as safe) Aspergillus species hold a central role in the field of industrial biotechnology with multiple profitable applications. Since the 1990s, research has focused on the use of Aspergillus species in the development of cell factories for the production of recombinant proteins mainly due to their natively high secretion capacity. Advances in the Aspergillus-specific molecular toolkit and combination of several engineering strategies (e.g., protease-deficient strains and fusions to carrier proteins) resulted in strains able to generate high titers of recombinant fungal proteins. However, the production of non-fungal proteins appears to still be inefficient due to bottlenecks in fungal expression and secretion machinery. After a brief overview of the different heterologous expression systems currently available, this review focuses on the filamentous fungi belonging to the genus Aspergillus and their use in recombinant protein production. We describe key steps in protein synthesis and secretion that may limit production efficiency in Aspergillus systems and present genetic engineering approaches and bioprocessing strategies that have been adopted in order to improve recombinant protein titers and expand the potential of Aspergilli as competitive production platforms.