bluuebunny/crossref_metadata_2025 huggingface dataset:
Acute Effects of Whole-Body Vibration Exercise on Pain Level, Functionality, and Rating of Exertion of Elderly Obese Knee Osteoarthritis Individuals: A Randomized Study
Authors: Moura-Fernandes Marcia Cristina , Moreira-Marconi Eloá , de Meirelles Alexandre Gonçalves , Reis-Silva Aline , de Souza Luiz Felipe Ferreira , Lírio Pereira da Silva Adriana , de Oliveira Bruno Bessa Monteiro , de Souza Gama Marco Antonio , de Oliveira Ana Carolina Coelho , Batouli-Santos Daniel , da Silva Ygor Teixeira , Torres-Nunes Luiza , da Silva Caiado Vanessa , de Souza Melo-Oliveira Maria Eduarda , de Aguiar Eliane de Oliveira Guedes , de Oliveira Liszt Palmeira , de Paula Mozella Alan , Paineiras-Domingos Laisa Liane , dos Santos Pereira Mario José , Xavier Vinicius Layter , Lacerda Ana Cristina Rodrigues , Mendonça Vanessa Amaral , Sañudo Borja , Seixas Adérito , Taiar Redha , da Cunha de Sá-Caputo Danúbia , Bernardo-Filho Mario
Published: August 2020

Introduction: Among chronic diseases, knee osteoarthritis (KOA) is a joint disease that causes important progressive alterations in the articular and periarticular structures, including synovial inflammation. Exercise has been suggested as an intervention to KOA individuals, and studies suggest that whole-body vibration (WBV) exercise decreases pain levels and favours the functionality of KOA individuals. Objective: The aim of the present study is to analyze the acute effects of WBV exercise on pain levels, functionality (Timed Up and Go (TUG test), anterior trunk flexion (ATF)), and rating of exertion of elderly obese KOA individuals. Methods: Thirty-seven individuals with KOA were allocated to a WBV exercise group (WBVEG), n = 19 (15 females/4 males), and a control group (CG), n = 18 (15 females/3 males). WBVEG performed one session of WBV exercise (11 min, using 5 Hz, 2.5, 5.0, and 7.5 mm, 0.12, 0.25, and 0.37 g). Three bouts were performed (working time of 3 min and rest time of 1 min) using a side-alternating vibrating platform (VP). The same position was used in CG; however, the VP was turned off and there was equipment coupled to the VP that emitted a sound like the vibrations. The pain level was evaluated using a visual analog scale (VAS). Functionality was evaluated with a TUG test and ATF. The rating of subjectively perceived exertion was measured with the category ratio CR-10 (BORG Scale CR-10), Results: A reduction of pain levels in WBVEG after the intervention (p = 0.001) and intergroups (p = 0.041) was found. A decrease of TUG test time in both groups (p = 0.001) and intergroups (p = 0.045) was found, while no statistical changes were observed in the Borg Scale score. Significant improvements of flexibility in both groups (p = 0.001) and intergroups (p = 0.043) were found. Conclusion: One session of WBV exercise can lead to important improvements in individuals with KOA, possibly triggered by physiological responses. However, more studies are needed, in this clinical context, to confirm these results.
On the Differential Analysis of Enterprise Valuation Methods as a Guideline for Unlisted Companies Assessment (I): Empowering Discounted Cash Flow Valuation
Authors: Vayas-Ortega Germania , Soguero-Ruiz Cristina , Rojo-Álvarez José-Luis , Gimeno-Blanes Francisco-Javier
Published: August 2020

The Discounted Cash Flow (DCF) method is probably the most extended approach used in company valuation, its main drawbacks being probably the known extreme sensitivity to key variables such as Weighted Average Cost of Capital (WACC) and Free Cash Flow (FCF) estimations not unquestionably obtained. In this paper we propose an unbiased and systematic DCF method which allows us to value private equity by leveraging on stock markets evidences, based on a twofold approach: First, the use of the inverse method assesses the existence of a coherent WACC that positively compares with market observations; second, different FCF forecasting methods are benchmarked and shown to correspond with actual valuations. We use financial historical data including 42 companies in five sectors, extracted from Eikon-Reuters. Our results show that WACC and FCF forecasting are not coherent with market expectations along time, with sectors, or with market regions, when only historical and endogenous variables are taken into account. The best estimates are found when exogenous variables, operational normalization of input space, and data-driven linear techniques are considered (Root Mean Square Error of 6.51). Our method suggests that FCFs and their positive alignment with Market Capitalization and the subordinate enterprise value are the most influencing variables. The fine-tuning of the methods presented here, along with an exhaustive analysis using nonlinear machine-learning techniques, are developed and discussed in the companion paper.
Prediction of the Energy Consumption of School Buildings
Authors: Alshibani Adel
Published: August 2020

The energy consumption of a constructed facility is a primary concern as a result of its impact on the total energy expenditure. It has been found that up to 70% of the power consumption in Saudi Arabia are caused by building structures and air conditioning (AC). Energy consumption in government-constructed buildings constitutes a considerable ≈13% of the consumption of the total energy in Saudi Arabia. Therefore, the government of Saudi Arabia initiated the Saudi Energy Efficiency Program (SEEP) that goals to lower the domestic energy severity by roughly 30% by 2030. This paper introduces a study carried out in Eastern Province in Saudi Arabia to identify factors influencing the consumption of energy in school facilities (which are built of concrete in hot and humid climate zones), investigate the correlation between those factors and their impacts on the consumption of energy in school facilities, and finally, develop a prediction model for the energy consumption of school facilities. The study was based on the utilization of 352 real-world datasets of energy consumption of operating schools across Eastern Province in Saudi Arabia. The developed energy prediction model considers eleven identified factors that influence the consumption of energy of constructed schools. The identified factors were utilized as input variables to build the model. A systematic search among different neural network (NN) design architectures was conducted to identify the optimal network model. Validation of the developed model on eight real-world cases demonstrated that the accuracy of the developed model was about 87.5%. Moreover, the findings of this study indicate that the weakest correlation between the input variables was recorded as −0.015 between “type of school” and “AC capacity,” while the strongest correlation was recorded as 0.95 between the variables of “number of classrooms” and “total air-conditioned area (sqm),” followed by “total air-conditioned area (sqm)” and “number of students,” which was recorded as 0.90. It is worth noting that “AC capacity” was the most significant predictor, which increased exponentially for high values of energy consumption, followed by “total school roof area.” The study also found that the age of the schools had a very small impact on energy consumption, although the age of the schools varied from 11 to 51 years. This was probably due to a good maintenance system applied by the Ministry of Education. The implication of the developed prediction model was that the model can be used by the Ministry of Education to predict the energy consumption and its associated cost for public school buildings for the purpose of budget allocation. The model may be utilized as a stand-alone application, or it can be integrated with an existing building information module (BIM)-based system.
Automatic Tomato and Peduncle Location System Based on Computer Vision for Use in Robotized Harvesting
Authors: Benavides M. , Cantón-Garbín M. , Sánchez-Molina J. A. , Rodríguez F.
Published: August 2020

Protected agriculture is a field in which the use of automatic systems is a key factor. In fact, the automatic harvesting of delicate fruit has not yet been perfected. This issue has received a great deal of attention over the last forty years, although no commercial harvesting robots are available at present, mainly due to the complexity and variability of the working environments. In this work we developed a computer vision system (CVS) to automate the detection and localization of fruit in a tomato crop in a typical Mediterranean greenhouse. The tasks to be performed by the system are: (1) the detection of the ripe tomatoes, (2) the location of the ripe tomatoes in the XY coordinates of the image, and (3) the location of the ripe tomatoes’ peduncles in the XY coordinates of the image. Tasks 1 and 2 were performed using a large set of digital image processing tools (enhancement, edge detection, segmentation, and the feature’s description of the tomatoes). Task 3 was carried out using basic trigonometry and numerical and geometrical descriptors. The results are very promising for beef and cluster tomatoes, with the system being able to classify 80.8% and 87.5%, respectively, of fruit with visible peduncles as “collectible”. The average processing time per image for visible ripe and harvested tomatoes was less than 30 ms.
Recent Development of Morphology Controlled Conducting Polymer Nanomaterial-Based Biosensor
Authors: Cho Sunghun , Lee Jun Seop
Published: August 2020

Biosensors are of particular importance for the detection of biological analytes at low concentrations. Conducting polymer nanomaterials, which often serve as sensing transducers, are renowned for their small dimensions, high surface-to-volume ratio, and amplified sensitivity. Despite these traits, the widespread implementation of conventional conducting polymer nanomaterials is hampered by their scarcity and lack of structural uniformity. Herein, a brief overview of the latest developments in the synthesis of morphologically tunable conducting polymer-based biosensors is discussed. Research related to the dimensional (0, 1, 2, and 3D) hetero-nanostructures of conducting polymers are highlighted in this paper, and how these structures affect traits such as the speed of charge transfer processes, low-working temperature, high sensitivity and cycle stability are discussed.
Numerical Simulation and Design of a High-Temperature, High-Pressure Fluid Transport Pipe
Authors: Yoon Jiyoung , Park Junkyu , Park Jinhyoung
Published: August 2020

When designing a hand caliber with a high-temperature, high-pressure internal fluid transport pipe, reliability, safe use, and performance must be considered. Reliability refers to the stress caused by thermo-mechanical load; safe use refers to the low-temperature burns that might occur upon contact, and high-temperature burns caused by gas leakage occurring in the cylinder gap; and performance refers to projectile velocity. In this study, numerical simulation methods for heat transfer, structure analysis, and gas leakage are proposed so that solutions can be designed to account for the above three criteria. Furthermore, a hand-caliber design guide is presented. For heat transfer and structural analysis, mesh size, the transient convective heat transfer coefficient, and boundary conditions are described. Regarding gas leakage, methods reflecting projectile motion and determination of the molecular weight of the propellant are described. As a result, a designed hand caliber will have a high reliability, because the thermo-mechanical stress is lower than the yield stress. There will be little risk of low-temperature burns, but there will be a high temperature-burn risk, owing to gas leakage in the cylinder gap. The larger the cylinder-gap size, the greater the gas leakage and the smaller projectile velocity. The presented numerical simulation method can be applied to evaluate various aspects of other structures that require high-temperature, high-pressure fluid-transport pipes.
Effects of Adding Batteries in Household Installations: Savings, Efficiency and Emissions
Authors: Villanueva Daniel , Cordeiro Moisés , Feijóo Andrés , Míguez Edelmiro , Fernández Antonio
Published: August 2020

Nowadays, common electrical household appliances are mostly being powered by means of alternate current (AC), although there are cases where direct current (DC) is used instead. In all cases, internal devices are supplied with DC, and this fact involves there are losses due to the need for AC/DC converters. At the same time, most electrical home consumption takes place during peak hours when electricity is more expensive in many electricity markets. The addition of a battery in these installations permits storing electrical energy during certain periods of the day with the aim of supplying it during other ones—when this operation is more efficient or convenient—simultaneously reducing costs and greenhouse gas emissions. In this paper, a comparison is proposed between three possible home consumption scenarios, i.e., one consisting of a current AC system, one consisting of an AC system with a battery, and a third consisting of a hybrid AC/DC system with a battery.
A Review of Climate-Smart Agriculture Applications in Cyprus
Authors: Adamides George
Published: August 2020

Climate-smart agriculture is an approach for developing agricultural strategies to modernize agricultural systems using digital techniques, aiming for sustainable agriculture and ensuring food security under climate change. This article provides a systematic literature review of smart agriculture technologies towards climate-smart agriculture in Cyprus, including robotics, Internet of Things, and remote sensing. The paper starts with a review of climate-smart agriculture, presenting its importance in terms of optimizing agricultural production processes in order to address the interlinked challenges of food security and climate change. An extensive literature review of works published in the areas of robotics, Internet of Things, and remote sensing is undertaken, with particular attention paid to works carried out in relation to agriculture in Cyprus. The paper analyzes aspects of the climate-smart agriculture research situation in Cypriot agriculture, identifies gaps, and concludes with new directions.
Flow Regimes and Föhn Types Characterize the Local Climate of Southern Patagonia
Authors: Temme Franziska , Turton Jenny V. , Mölg Thomas , Sauter Tobias
Published: August 2020

The local climate in Southern Patagonia is strongly influenced by the interaction between the topography and persistent westerlies, which can generate föhn events, dry and warm downslope winds. The upstream flow regime influences different föhn types which dictate the lee-side atmospheric response regarding the strength, spatial extent and phenomenology. We use a combination of observations from four automatic weather stations (AWSs) and high-resolution numerical modeling with the Weather Research and Forecasting (WRF) model for a region in Southern Patagonia (48° S–52° S, 72° W–76.5° W) including the Southern Patagonian Icefield (SPI). The application of a föhn identification algorithm to a 10-month study period (June 2018–March 2019) reveals 81 föhn events in total. A simulation of three events of differing flow regimes (supercritical, subcritical, transition) suggests that a supercritical flow regime leads to a linear föhn event with a large spatial extent but moderate intensity. In contrast, a spatially limited but locally strong föhn response is induced by a subcritical regime with upstream blocking and by a transition regime with a hydraulic jump present. Our results imply that the hydraulic jump-type föhn event (transition case) is the most critical for glacier mass balances since it shows the strongest warming, drying, wind velocities and solar radiation over the SPI. The consideration of flow regimes over the last 40 years shows that subcritical flow occurs most frequently (78%), however transitional flow occurs 14% of the time, implying the potential impact on Patagonian glaciers.
H″IT″ting the Barriers for Exercising during Social Isolation
Authors: Souza Daniel , Coswig Victor , de Lira Claudio Andre Barbosa , Gentil Paulo
Published: August 2020

Aerobic exercise is traditionally recommended to improve general health and prevent many non-communicable diseases. However, the measures adopted to control the novel Coronavirus (COVID-19) outbreak culminated with closing of exercise facilities and fitness centers and, as a primary consequence, impaired aerobic exercise practice. This contributed to an increase in risk factors associated with physical inactivity such as insulin resistance, high blood pressure, low-grade inflammation, weight gain, and mental health problems. The scenario is worrisome, and it is important to propose alternatives for exercise practice during the COVID-19 pandemic. Interval training (IT) emerges as an exercise mode that might be feasible, low-cost, and potentially safe to be performed in many different places. IT consists of interspersing relative brief bouts of high-intensity exercise with recovery periods and promotes similar or greater health benefits when compared to moderate-intensity continuous exercise. Among the different types of IT, sprint interval training and “Tabata protocols” might be particularly useful during social isolation. These protocols can be controlled and performed without the need of complex equipment and can be adapted to different places, including domestic environments. In this article, we present variations of IT as possible alternatives to cope physical inactivity during COVID-19 pandemics with a focus on its practical applications. The protocols suggested can be performed without the need of specialized equipment or facilities, in a time-efficient manner, and aiming to prevent detraining or even improve physical fitness and general health.
Current Trends in Proteomic Advances for Food Allergen Analysis
Authors: López-Pedrouso María , Lorenzo José , Gagaoua Mohammed , Franco Daniel
Published: August 2020

Food allergies are a global food challenge. For correct food labelling, the detection and quantification of allergens are necessary. However, novel product formulations and industrial processes produce new scenarios, which require much more technological developments. For this purpose, OMICS technologies, especially proteomics, seemed to be relevant in this context. This review summarises the current knowledge and studies that used proteomics to study food allergens. In the case of the allergenic proteins, a wide variety of isoforms, post-translational modifications and other structural changes during food processing can increase or decrease the allergenicity. Most of the plant-based food allergens are proteins with biological functions involved in storage, structure, and plant defence. The allergenicity of these proteins could be increased by the presence of heavy metals, air pollution, and pesticides. Targeted proteomics like selected/multiple reaction monitoring (SRM/MRM) have been very useful, especially in the case of gluten from wheat, rye and barley, and allergens from lentil, soy, and fruit. Conventional 1D and 2-DE immunoblotting have been further widely used. For animal-based food allergens, the widely used technologies are 1D and 2-DE immunoblotting followed by MALDI-TOF/TOF, and more recently LC-MS/MS, which is becoming useful to assess egg, fish, or milk allergens. The detection and quantification of allergenic proteins using mass spectrometry-based proteomics are promising and would contribute to greater accuracy, therefore improving consumer information.
The Transcriptomic Evidence on the Role of Abdominal Visceral vs. Subcutaneous Adipose Tissue in the Pathophysiology of Diabetes in Asian Indians Indicates the Involvement of Both
Authors: Kumar Anshul , Tiwari Pradeep , Saxena Aditya , Purwar Naincy , Wahi Nitin , Sharma Balram , Mathur Sandeep Kumar
Published: August 2020

The roles of abdominal visceral (VAT) and subcutaneous adipose tissue (SAT) in the molecular pathogenesis type-2 diabetics (T2D) among Asian Indians showing a “thin fat” phenotype largely remains obscure. In this study, we generated transcription profiles in biopsies of these adipose depots obtained during surgery in 19 diabetics (M: F ratio, 8:11) and 16 (M: F ratio 5:11) age- and BMI-matched non-diabetics. Gene set enrichment analysis (GSEA) was used for comparing transcription profile and showed that 19 gene sets, enriching inflammation and immune system-related pathways, were upregulated in diabetics with F.D.R. <25% and >25%, respectively, in VAT and SAT. Moreover, 13 out of the 19 significantly enriched pathways in VAT were among the top 20 pathways in SAT. On comparison of VAT vs. SAT among diabetics, none of the gene sets were found significant at F.D.R. <25%. The Weighted Gene Correlation Analysis (WGCNA) analysis of the correlation between measures of average gene expression and overall connectivity between VAT and SAT was significantly positive. Several modules of co-expressed genes in both the depots showed a bidirectional correlation with various diabetes-related intermediate phenotypic traits. They enriched several diabetes pathogenicity marker pathways, such as inflammation, adipogenesis, etc. It is concluded that, in Asian Indians, diabetes pathology inflicts similar molecular alternations in VAT and SAT, which are more intense in the former. Both adipose depots possibly play a role in the pathophysiology of T2D, and whether it is protective or pathogenic also depends on the nature of modules of co-expressed genes contained in them.
Rhizobacteria Inoculation Effects on Phytohormone Status of Potato Microclones Cultivated In Vitro under Osmotic Stress
Authors: Arkhipova Tatiana N. , Evseeva Nina V. , Tkachenko Oksana V. , Burygin Gennady L. , Vysotskaya Lidiya B. , Akhtyamova Zarina A. , Kudoyarova Guzel R.
Published: August 2020

Water deficits inhibit plant growth and decrease crop productivity. Remedies are needed to counter this increasingly urgent problem in practical farming. One possible approach is to utilize rhizobacteria known to increase plant resistance to abiotic and other stresses. We therefore studied the effects of inoculating the culture medium of potato microplants grown in vitro with Azospirillum brasilense Sp245 or Ochrobactrum cytisi IPA7.2. Growth and hormone content of the plants were evaluated under stress-free conditions and under a water deficit imposed with polyethylene glycol (PEG 6000). Inoculation with either bacterium promoted the growth in terms of leaf mass accumulation. The effects were associated with increased concentrations of auxin and cytokinin hormones in the leaves and stems and with suppression of an increase in the leaf abscisic acid that PEG treatment otherwise promoted in the potato microplants. O. cytisi IPA7.2 had a greater growth-stimulating effect than A. brasilense Sp245 on stressed plants, while A. brasilense Sp245 was more effective in unstressed plants. The effects were likely to be the result of changes to the plant’s hormonal balance brought about by the bacteria.
Galectins in Intra- and Extracellular Vesicles
Authors: Bänfer Sebastian , Jacob Ralf
Published: August 2020

Carbohydrate-binding galectins are expressed in various tissues of multicellular organisms. They are involved in autophagy, cell migration, immune response, inflammation, intracellular transport, and signaling. In recent years, novel roles of galectin-interaction with membrane components have been characterized, which lead to the formation of vesicles with diverse functions. These vesicles are part of intracellular transport pathways, belong to the cellular degradation machinery, or can be released for cell-to-cell communication. Several characteristics of galectins in the lumen or at the membrane of newly formed vesicular structures are discussed in this review and illustrate the need to fully elucidate their contributions at the molecular and structural level.
Green Synthesis of Antileishmanial and Antifungal Silver Nanoparticles Using Corn Cob Xylan as a Reducing and Stabilizing Agent
Authors: Silva Viana Rony Lucas , Pereira Fidelis Gabriel , Jane Campos Medeiros Mayara , Antonio Morgano Marcelo , Gabriela Chagas Faustino Alves Monique , Domingues Passero Luiz Felipe , Lima Pontes Daniel , Cordeiro Theodoro Raquel , Domingos Arantes Thales , Araujo Sabry Diego , Lanzi Sassaki Guilherme , Fagundes Melo-Silveira Raniere , Rocha Hugo Alexandre Oliveira
Published: August 2020

Corn cob is an agricultural byproduct that produces an estimated waste burden in the thousands of tons annually, but it is also a good source of xylan, an important bioactive polysaccharide. Silver nanoparticles containing xylan (nanoxylan) were produced using an environmentally friendly synthesis method. To do this, we extracted xylan from corn cobs using an ultrasound technique, which was confirmed by both chemical and NMR analyses. This xylan contained xylose, glucose, arabinose, galactose, mannose, and glucuronic acid in a molar ratio of 50:21:14:9:2.5:2.5, respectively. Nanoxylan synthesis was analyzed using UV–vis spectroscopy at kmax = 469 nm and Fourier transform infrared spectroscopy (FT-IR), which confirmed the presence of both silver and xylan in the nanoxylan product. Dynamic light scattering (DLS) and atomic force microscopy (AFM) revealed that the nanoxylan particles were ~102.0 nm in size and spherical in shape, respectively. DLS also demonstrated that nanoxylan was stable for 12 months and coupled plasma optical emission spectrometry (ICP-OES) showed that the nanoxylan particles were 19% silver. Nanoxylan reduced Leishmania amazonensis promastigote viability with a half maximal inhibitory concentration (IC50) value of 25 μg/mL, while xylan alone showed no effective. Additionally, nanoxylan exhibited antifungal activity against Candida albicans (MIC = 7.5 μg/mL), C. parapsilosis (MIC = 7.5 μg/mL), and Cryptococcus neoformans (MIC = 7.5 μg/mL). Taken together, these data suggest that it is possible to synthesize silver nanoparticles using xylan and that these nanoxylan exert improved antileishmanial and antifungal activities when compared to the untreated polysaccharide or silver nitrate used for their synthesis. Thus, nanoxylan may represent a promising new class of antiparasitic agents for use in the treatment of these microorganisms.
A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors
Authors: Galkin Simon , Rozina Anna , Zalevsky Arthur , Gottikh Marina , Anisenko Andrey
Published: August 2020

The search for compounds that can inhibit the interaction of certain viral proteins with their cellular partners is a promising trend in the development of antiviral drugs. We have previously shown that binding of HIV-1 integrase with human Ku70 protein is essential for viral replication. Here, we present a novel, cheap, and fast assay to search for inhibitors of these proteins’ binding based on the usage of genetically encoded fluorescent tags linked to both integrase and Ku70. Using this approach, we have elucidated structure-activity relationships for a set of oligonucleotide conjugates with eosin and shown that their inhibitory activity is primarily achieved through interactions between the conjugate nucleic bases and integrase. Molecular modeling of HIV-1 integrase in complex with the conjugates suggests that they can shield E212/L213 residues in integrase, which are crucial for its efficient binding to Ku70, in a length-dependent manner. Using the developed system, we have found the 11-mer phosphorothioate bearing 3’-end eosin-Y to be the most efficient inhibitor among the tested conjugates.
Integrin-Targeting Peptides for the Design of Functional Cell-Responsive Biomaterials
Authors: Zhao Junwei , Santino Federica , Giacomini Daria , Gentilucci Luca
Published: August 2020

Integrins are a family of cell surface receptors crucial to fundamental cellular functions such as adhesion, signaling, and viability, deeply involved in a variety of diseases, including the initiation and progression of cancer, of coronary, inflammatory, or autoimmune diseases. The natural ligands of integrins are glycoproteins expressed on the cell surface or proteins of the extracellular matrix. For this reason, short peptides or peptidomimetic sequences that reproduce the integrin-binding motives have attracted much attention as potential drugs. When challenged in clinical trials, these peptides/peptidomimetics let to contrasting and disappointing results. In the search for alternative utilizations, the integrin peptide ligands have been conjugated onto nanoparticles, materials, or drugs and drug carrier systems, for specific recognition or delivery of drugs to cells overexpressing the targeted integrins. Recent research in peptidic integrin ligands is exploring new opportunities, in particular for the design of nanostructured, micro-fabricated, cell-responsive, stimuli-responsive, smart materials.
Targeting HER3, a Catalytically Defective Receptor Tyrosine Kinase, Prevents Resistance of Lung Cancer to a Third-Generation EGFR Kinase Inhibitor
Authors: Romaniello Donatella , Marrocco Ilaria , Belugali Nataraj Nishanth , Ferrer Irene , Drago-Garcia Diana , Vaknin Itay , Oren Roni , Lindzen Moshit , Ghosh Soma , Kreitman Matthew , Kittel Jeanette Clarissa , Gaborit Nadege , Bergado Baez Gretchen , Sanchez Belinda , Eilam Raya , Pikarsky Eli , Paz-Ares Luis , Yarden Yosef
Published: August 2020

Although two growth factor receptors, EGFR and HER2, are amongst the best targets for cancer treatment, no agents targeting HER3, their kinase-defective family member, have so far been approved. Because emergence of resistance of lung tumors to EGFR kinase inhibitors (EGFRi) associates with compensatory up-regulation of HER3 and several secreted forms, we anticipated that blocking HER3 would prevent resistance. As demonstrated herein, a neutralizing anti-HER3 antibody we generated can clear HER3 from the cell surface, as well as reduce HER3 cleavage by ADAM10, a surface metalloproteinase. When combined with a kinase inhibitor and an anti-EGFR antibody, the antibody completely blocked patient-derived xenograft models that acquired resistance to EGFRi. We found that the underlying mechanism involves posttranslational downregulation of HER3, suppression of MET and AXL upregulation, as well as concomitant inhibition of AKT signaling and upregulation of BIM, which mediates apoptosis. Thus, although HER3 is nearly devoid of kinase activity, it can still serve as an effective drug target in the context of acquired resistance. Because this study simulated in animals the situation of patients who develop resistance to EGFRi and remain with no obvious treatment options, the observations presented herein may warrant clinical testing.
Molecular Characterisation of Canine Osteosarcoma in High Risk Breeds
Authors: Simpson Siobhan , Dunning Mark , de Brot Simone , Alibhai Aziza , Bailey Clara , Woodcock Corinne L. , Mestas Madeline , Akhtar Shareen , Jeyapalan Jennie N. , Lothion-Roy Jennifer , Emes Richard D. , Allegrucci Cinzia , Rizvanov Albert A. , Mongan Nigel P. , Rutland Catrin S.
Published: August 2020

Dogs develop osteosarcoma (OSA) and the disease process closely resembles that of human OSA. OSA has a poor prognosis in both species and disease-free intervals and cure rates have not improved in recent years. Gene expression in canine OSAs was compared with non-tumor tissue utilising RNA sequencing, validated by qRT-PCR and immunohistochemistry (n = 16). Polymorphic polyglutamine (polyQ) tracts in the androgen receptor (AR/NR3C4) and nuclear receptor coactivator 3 (NCOA3) genes were investigated in control and OSA patients using polymerase chain reaction (PCR), Sanger sequencing and fragment analysis (n = 1019 Rottweilers, 379 Irish Wolfhounds). Our analysis identified 1281 significantly differentially expressed genes (>2 fold change, p < 0.05), specifically 839 lower and 442 elevated gene expression in osteosarcoma (n = 3) samples relative to non-malignant (n = 4) bone. Enriched pathways and gene ontologies were identified, which provide insight into the molecular pathways implicated in canine OSA. Expression of a subset of these genes (SLC2A1, DKK3, MMP3, POSTN, RBP4, ASPN) was validated by qRTPCR and immunohistochemistry (MMP3, DKK3, SLC2A1) respectively. While little variation was found in the NCOA3 polyQ tract, greater variation was present in both polyQ tracts in the AR, but no significant associations in length were made with OSA. The data provides novel insights into the molecular mechanisms of OSA in high risk breeds. This knowledge may inform development of new prevention strategies and treatments for OSA in dogs and supports utilising spontaneous OSA in dogs to improve understanding of the disease in people.
Detection of Ovarian Cancer through Exhaled Breath by Electronic Nose: A Prospective Study
Authors: Raspagliesi Francesco , Bogani Giorgio , Benedetti Simona , Grassi Silvia , Ferla Stefano , Buratti Susanna
Published: August 2020

Background: Diagnostic methods for the early identification of ovarian cancer (OC) represent an unmet clinical need, as no reliable diagnostic tools are available. Here, we tested the feasibility of electronic nose (e-nose), composed of ten metal oxide semiconductor (MOS) sensors, as a diagnostic tool for OC detection. Methods: Women with suspected ovarian masses and healthy subjects had volatile organic compounds analysis of the exhaled breath using e-nose. Results: E-nose analysis was performed on breath samples collected from 251 women divided into three groups: 86 OC cases, 51 benign masses, and 114 controls. Data collected were analyzed by Principal Component Analysis (PCA) and K-Nearest Neighbors’ algorithm (K-NN). A first 1-K-NN (cases vs. controls) model has been developed to discriminate between OC cases and controls; the model performance tested in the prediction gave 98% of sensitivity and 95% of specificity, when the strict class prediction was applied; a second 1-K-NN (cases vs. controls + benign) model was built by grouping the non-cancer groups (controls + benign), thus considering two classes, cases and controls + benign; the model performance in the prediction was of 89% for sensitivity and 86% for specificity when the strict class prediction was applied. Conclusions: Our preliminary results suggested the potential role of e-nose for the detection of OC. Further studies aiming to test the potential adoption of e-nose in the early diagnosis of OC are needed.
Kv10.1 Regulates Microtubule Dynamics during Mitosis
Authors: Movsisyan Naira , Pardo Luis A.
Published: August 2020

Kv10.1 (potassium voltage-gated channel subfamily H member 1, known as EAG1 or Ether-à-go-go 1), is a voltage-gated potassium channel, prevailingly expressed in the central nervous system. The aberrant expression of Kv10.1 is detected in over 70% of all human tumor tissues and correlates with poorer prognosis. In peripheral tissues, Kv10.1 is expressed almost exclusively during the G2/M phase of the cell cycle and regulates its progression—downregulation of Kv10.1 extends the duration of the G2/M phase both in cancer and healthy cells. Here, using biochemical and imaging techniques, such as live-cell measurements of microtubule growth and of cytosolic calcium, we elucidate the mechanisms of Kv10.1-mediated regulation at the G2/M phase. We show that Kv10.1 has a dual effect on mitotic microtubule dynamics. Through the functional interaction with ORAI1 (calcium release-activated calcium channel protein 1), it modulates cytosolic calcium oscillations, thereby changing microtubule behavior. The inhibition of either Kv10.1 or ORAI1 stabilizes the microtubules. In contrast, the knockdown of Kv10.1 increases the dynamicity of mitotic microtubules, resulting in a stronger spindle assembly checkpoint, greater mitotic spindle angle, and a decrease in lagging chromosomes. Understanding of Kv10.1-mediated modulation of the microtubule architecture will help to comprehend how cancer tissue benefits from the presence of Kv10.1, and thereby increase the efficacy and safety of Kv10.1-directed therapeutic strategies.
A Distinctive microRNA (miRNA) Signature in the Blood of Colorectal Cancer (CRC) Patients at Surgery
Authors: Gasparello Jessica , Papi Chiara , Allegretti Matteo , Giordani Elena , Carboni Fabio , Zazza Settimio , Pescarmona Edoardo , Romania Paolo , Giacomini Patrizio , Scapoli Chiara , Gambari Roberto , Finotti Alessia
Published: August 2020

Background: Liquid biopsy (LB) provides an examination of the peripheral blood of cancer patients for circulating tumor cells, cell-free nucleic acids and microRNAs (miRNAs) and is an established tool of precision medicine. Unlike most previous LB studies that focused on advanced metastatic colorectal cancer (CRC), we assessed miRNA dysregulation in blood samples obtained on the day of surgery from patients with primary CRC lesions but no clinical evidence of extra-colonic diffusion. In this study, plasma preparation included miRNAs associated to exosomes, but excluded large macrovesicles from the preparation. Methods: The miRNA profile in plasma isolated from a cohort of 35 CRC patients at the day of surgery was analyzed by Next Generation Sequencing (NGS) and further confirmed by droplet digital RT-PCR (dd-RT-PCR). Results: A miR-141-3p/miR-221-3p/miR-222-3p upregulation signature previously described in advanced CRC did not discriminate the analyzed early-CRC cohort from six tumor-free donors (Tf-D). In contrast, NGS-based miRNome analysis of a training cohort of five CRC and three tumor-free donors identified a novel, distinct nine miRNA signature comprising five up-regulated and four down-regulated miRNAs, six of which could be confirmed in the full CRC and tumor-free donor validation dataset by dd-RT-PCR. Additionally, a KRAS (Kirsten Rat Sarcoma Viral Oncogene Homolog) mutant status was correlated with the plasma content of three identified miRNAs. Conclusions: When the data obtained were comparatively evaluated, at least one of the miRNAs belonging to the signature list was found to be dysregulated in 34/35 (97.1%) of our early-CRC plasma samples. The miRNA list provides diagnostic markers as well as possible molecular targets for protocols focusing on “microRNA therapeutics”.
Colorectal Cancer Survival in 50- to 69-Year-Olds after Introducing the Faecal Immunochemical Test
Authors: Gutierrez-Stampa María Angeles , Aguilar Vanessa , Sarasqueta Cristina , Cubiella Joaquín , Portillo Isabel , Bujanda Luis
Published: August 2020

Population screening has improved early diagnosis of colorectal cancer (CRC). Nonetheless, most cases are diagnosed in symptomatic patients. Faecal immunochemical testing has been recommended for assessing patients with lower gastrointestinal symptoms, but whether it improves patient survival is unknown. Our objective was to compare CRC survival in 50- to 69-year-olds between asymptomatic screen-detected patients and symptomatic patients by route to diagnosis. Methods: We identified all cases of CRC diagnosed in 50-to 69-year-olds between 2009 and 2016, in Donostialdea (Gipuzkoa, Spain). Three groups were created: 1-screen-detected CRC; 2-CRC detected in symptomatic patients after a positive faecal immunochemical test(FIT); and 3-CRC detected in symptomatic patients without a FIT or after a negative result. We analysed survival using the Kaplan-Meier method and log-rank tests. Results: Of 930 patients diagnosed with CRC, 433 cases were detected through screening and 497 in symptomatic patients, 7.9% after a positive FIT and 45.5% by other means. The 3-year CRC survival was significantly lower in group 3 (69.5%) than groups 1 (93%; p = 0.007) or 2 (87.5%; p = 0.02). The risk of death was lower in groups 1 (HR 0.42, 95% CI 0.30–0.58) and 2 (HR 0.51; 95% CI 0.29–0.87). Conclusion: Half of CRC cases in 50- to 69-year-olds are diagnosed outside screening. Use of the FIT as a diagnostic strategy in symptomatic patients may improve survival.
Aza-BODIPY: A New Vector for Enhanced Theranostic Boron Neutron Capture Therapy Applications
Authors: Kalot Ghadir , Godard Amélie , Busser Benoît , Pliquett Jacques , Broekgaarden Mans , Motto-Ros Vincent , Wegner Karl David , Resch-Genger Ute , Köster Ulli , Denat Franck , Coll Jean-Luc , Bodio Ewen , Goze Christine , Sancey Lucie
Published: August 2020

Boron neutron capture therapy (BNCT) is a radiotherapeutic modality based on the nuclear capture of slow neutrons by stable 10B atoms followed by charged particle emission that inducing extensive damage on a very localized level (<10 μm). To be efficient, a sufficient amount of 10B should accumulate in the tumor area while being almost cleared from the normal surroundings. A water-soluble aza-boron-dipyrromethene dyes (BODIPY) fluorophore was reported to strongly accumulate in the tumor area with high and BNCT compatible Tumor/Healthy Tissue ratios. The clinically used 10B-BSH (sodium borocaptate) was coupled to the water-soluble aza-BODIPY platform for enhanced 10B-BSH tumor vectorization. We demonstrated a strong uptake of the compound in tumor cells and determined its biodistribution in mice-bearing tumors. A model of chorioallantoic membrane-bearing glioblastoma xenograft was developed to evidence the BNCT potential of such compound, by subjecting it to slow neutrons. We demonstrated the tumor accumulation of the compound in real-time using optical imaging and ex vivo using elemental imaging based on laser-induced breakdown spectroscopy. The tumor growth was significantly reduced as compared to BNCT with 10B-BSH. Altogether, the fluorescent aza-BODIPY/10B-BSH compound is able to vectorize and image the 10B-BSH in the tumor area, increasing its theranostic potential for efficient approach of BNCT.
Role of EPAC1 Signalosomes in Cell Fate: Friends or Foes?
Authors: Formoso Karina , Lezoualc’h Frank , Mialet-Perez Jeanne
Published: August 2020

The compartmentation of signaling processes is accomplished by the assembly of protein complexes called signalosomes. These signaling platforms colocalize enzymes, substrates, and anchoring proteins into specific subcellular compartments. Exchange protein directly activated by cAMP 1 (EPAC1) is an effector of the second messenger, 3′,5′-cyclic adenosine monophosphate (cAMP) that is associated with multiple roles in several pathologies including cardiac diseases. Both EPAC1 intracellular localization and molecular partners are key players in the regulation of cell fate, which may have important therapeutic potential. In this review, we summarize the recent findings on EPAC1 structure, regulation, and pharmacology. We describe the importance of EPAC1 subcellular distribution in its biological action, paying special attention to its nuclear localization and mechanism of action leading to cardiomyocyte hypertrophy. In addition, we discuss the role of mitochondrial EPAC1 in the regulation of cell death. Depending on the cell type and stress condition, we present evidence that supports either a protective or detrimental role of EPAC1 activation.
Clinical Importance of the Human Umbilical Artery Potassium Channels
Authors: Lorigo Margarida , Oliveira Nelson , Cairrao Elisa
Published: August 2020

Potassium (K+) channels are usually predominant in the membranes of vascular smooth muscle cells (SMCs). These channels play an important role in regulating the membrane potential and vessel contractility—a role that depends on the vascular bed. Thus, the activity of K+ channels represents one of the main mechanisms regulating the vascular tone in physiological and pathophysiological conditions. Briefly, the activation of K+ channels in SMC leads to hyperpolarization and vasorelaxation, while its inhibition induces depolarization and consequent vascular contraction. Currently, there are four different types of K+ channels described in SMCs: voltage-dependent K+ (KV) channels, calcium-activated K+ (KCa) channels, inward rectifier K+ (Kir) channels, and 2-pore domain K+ (K2P) channels. Due to the fundamental role of K+ channels in excitable cells, these channels are promising therapeutic targets in clinical practice. Therefore, this review discusses the basic properties of the various types of K+ channels, including structure, cellular mechanisms that regulate their activity, and new advances in the development of activators and blockers of these channels. The vascular functions of these channels will be discussed with a focus on vascular SMCs of the human umbilical artery. Then, the clinical importance of K+ channels in the treatment and prevention of cardiovascular diseases during pregnancy, such as gestational hypertension and preeclampsia, will be explored.
Proteomic Profiling of Extracellular Vesicles Derived from Cerebrospinal Fluid of Alzheimer’s Disease Patients: A Pilot Study
Authors: Muraoka Satoshi , Jedrychowski Mark P. , Yanamandra Kiran , Ikezu Seiko , Gygi Steven P. , Ikezu Tsuneya
Published: August 2020

Pathological hallmarks of Alzheimer’s disease (AD) are deposits of amyloid beta (Aβ) and hyper-phosphorylated tau aggregates in brain plaques. Recent studies have highlighted the importance of Aβ and tau-containing extracellular vesicles (EVs) in AD. We therefore examined EVs separated from cerebrospinal fluid (CSF) of AD, mild cognitive impairment (MCI), and control (CTRL) patient samples to profile the protein composition of CSF EV. EV fractions were separated from AD (n = 13), MCI (n = 10), and CTRL (n = 10) CSF samples using MagCapture Exosome Isolation kit. The CSF-derived EV proteins were identified and quantified by label-free and tandem mass tag (TMT)-labeled mass spectrometry. Label-free proteomics analysis identified 2546 proteins that were significantly enriched for extracellular exosome ontology by Gene Ontology analysis. Canonical Pathway Analysis revealed glia-related signaling. Quantitative proteomics analysis, moreover, showed that EVs expressed 1284 unique proteins in AD, MCI and CTRL groups. Statistical analysis identified three proteins—HSPA1A, NPEPPS, and PTGFRN—involved in AD progression. In addition, the PTGFRN showed a moderate correlation with amyloid plaque (rho = 0.404, p = 0.027) and tangle scores (rho = 0.500, p = 0.005) in AD, MCI and CTRL. Based on the CSF EV proteomics, these data indicate that three proteins, HSPA1A, NPEPPS and PTGFRN, may be used to monitor the progression of MCI to AD.
Targeting the sAC-Dependent cAMP Pool to Prevent SARS-Cov-2 Infection
Authors: Aslam Muhammad , Ladilov Yury
Published: August 2020

An outbreak of the novel coronavirus (CoV) SARS-CoV-2, the causative agent of COVID-19 respiratory disease, infected millions of people since the end of 2019, led to high-level morbidity and mortality and caused worldwide social and economic disruption. There are currently no antiviral drugs available with proven efficacy or vaccines for its prevention. An understanding of the underlying cellular mechanisms involved in virus replication is essential for repurposing the existing drugs and/or the discovery of new ones. Endocytosis is the important mechanism of entry of CoVs into host cells. Endosomal maturation followed by the fusion with lysosomes are crucial events in endocytosis. Late endosomes and lysosomes are characterized by their acidic pH, which is generated by a proton transporter V-ATPase and required for virus entry via endocytic pathway. The cytoplasmic cAMP pool produced by soluble adenylyl cyclase (sAC) promotes V-ATPase recruitment to endosomes/lysosomes and thus their acidification. In this review, we discuss targeting the sAC-specific cAMP pool as a potential strategy to impair the endocytic entry of the SARS-CoV-2 into the host cell. Furthermore, we consider the potential impact of sAC inhibition on CoV-induced disease via modulation of autophagy and apoptosis.