bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation, Identification and Biotechnological Applications of a Novel, Robust, Free-living Chlorococcum (Oophila) amblystomatis Strain Isolated from a Local Pond
Authors: Correia Nádia , Pereira Hugo , Silva Joana T. , Santos Tamára , Soares Maria , Sousa Carolina B. , Schüler Lisa M. , Costa Margarida , Varela João , Pereira Leonel , Silva Joana
Published: April 2020

Bioprospection of novel autochthonous strains is key to the successful industrial-scale production of microalgal biomass. A novel Chlorococcum strain was recently isolated from a pond inside the industrial production facility of Allmicroalgae (Leiria, Portugal). Phylogenetic analysis based on 18S ribosomal ribonucleic acid (rRNA) gene sequences suggests that this isolate is a novel, free-living Oophila amblystomatis strain. However, as our phylogenetic data strongly suggests that the aforementioned taxon belongs to the genus Chlorococcum, it is here proposed to rename this species as Chlorococcum amblystomatis. In order to characterize the biotechnological potential of this novel isolate, growth performance and biochemical composition were evaluated from the pilot (2.5-m3) to industrial (10-m3) scale. The highest maximum areal productivity (36.56 g·m−2·day−1) was reached in a 10-m3 tubular photobioreactor (PBR), as compared to that obtained in a 2.5-m3 PBR (26.75 g·m−2·day−1). Chlorococcum amblystomatis displayed high protein content (48%–56% dry weight (DW)) and moderate levels of total lipids (18%–31% DW), carbohydrates (6%–18% DW) and ashes (9%–16% DW). Furthermore, the lipid profile was dominated by polyunsaturated fatty acids (PUFAs). The highest pigment contents were obtained in the 2.5-m3 PBR, where total chlorophylls accounted for 40.24 mg·g−1 DW, followed by lutein with 5.37 mg·g−1 DW. Overall, this free-living Chlorococcum amblystomatis strain shows great potential for nutritional applications, coupling a promising growth performance with a high protein content as well as relevant amounts of PUFAs, chlorophyll, and carotenoids.
Effort Estimation Approach through Extracting Use Cases via Informal Requirement Specifications
Authors: Park Bo Kyung , Kim R. Young Chul
Published: April 2020

Sometimes unclearly describing the requirement specifications of satisfied customer’s needs, means it may be difficult to develop the production of high-quality software systems. A persistent issue of requirement engineering is how to clearly understand the requirements of the large and complex software project, and also how to analyze them exactly. To solve this problem, we propose a linguistic analysis method based on the semantic analysis of the Fillmore’s textual approach. This method extracts use-cases from informal requirement specifications. For applied requirement engineering with this method, we suggest extracting a use-case diagram, as well as calculating the software effort estimation with the original use-case point (UCP). To simply explanations of our use-case extraction method, we use one example of a simple postal information system.
Towards an Ergonomic Assessment Framework for Industrial Assembly Workstations—A Case Study
Authors: Colim Ana , Faria Carlos , Braga Ana Cristina , Sousa Nuno , Rocha Luís , Carneiro Paula , Costa Nélson , Arezes Pedro
Published: April 2020

Work-related musculoskeletal disorders (WMSD) are one of the main occupational health problems. The best strategy to prevent them lies on ergonomic interventions. The variety of industrial processes and environments, however, makes it difficult to define an all-purpose framework to guide these ergonomic interventions. This undefinition is exacerbated by recurrent introduction of new technologies, e.g., collaborative robots. In this paper, we propose a framework to guide ergonomics and human factors practitioners through all stages of assessment and redesign of workstations. This framework was applied in a case study at an assembly workstation of a large furniture enterprise. Direct observation of work activity and questionnaires were applied to characterize the workstations, the process, and the workers’ profiles and perceptions. An ergonomic multi-method approach, based on well-known and validated methods (such as the Finnish Institute of Occupational Health and Rapid Upper Limb Assessment), was applied to identify the most critical risk factors. We concluded that this approach supports the process redesign and tasks’ allocation of the future workstation. From these conclusions, we distill a list of requirements for the creation of a collaborative robot cell, specifying which tasks are performed by whom, as well as the scheduling of the human-robot collaboration (HRC).
Comparison of Wear Performance of Austempered and Quench-Tempered Gray Cast Irons Enhanced by Laser Hardening Treatment
Authors: Wang Bingxu , Barber Gary C. , Wang Rui , Pan Yuming
Published: April 2020

The current research studied the effects of laser surface hardening treatment on the phase transformation and wear properties of gray cast irons heat treated by austempering or quench-tempering, respectively. Three austempering temperatures of 232 °C, 288 °C, and 343 °C with a constant holding duration of 120 min and three tempering temperatures of 316 °C, 399 °C, and 482 °C with a constant holding duration of 60 min were utilized to prepare austempered and quench-tempered gray cast iron specimens with equivalent macro-hardness values. A ball-on-flat reciprocating wear test configuration was used to investigate the wear resistance of austempered and quench-tempered gray cast iron specimens before and after applying laser surface-hardening treatment. The phase transformation, hardness, mass loss, and worn surfaces were evaluated. There were four zones in the matrix of the laser-hardened austempered gray cast iron. Zone 1 contained ledeburite without the presence of graphite flakes. Zone 2 contained martensite and had a high hardness, which was greater than 67 HRC. Zone 4 was the substrate containing the acicular ferrite and carbon-saturated austenite with a hardness of 41–27 HRC. In Zone 3, the substrate was tempered by the low thermal radiation. For the laser-hardened quench-tempered gray cast iron specimens, three zones were observed beneath the laser-hardened surface. Zone 1 also contained ledeburite, and Zone 2 was full martensite. Zone 3 was the substrate containing the tempered martensite. The tempered martensite became coarse with increasing tempering temperature due to the decomposition of the as-quenched martensite and precipitation of cementite particles. In the wear tests, the gray cast iron specimens without heat treatment had the highest wear loss. The wear performance was improved by applying quench-tempering heat treatment and further enhanced by applying austempering heat treatment. Austempered gray cast iron specimens had lower mass loss than the quench-tempered gray cast iron specimens, which was attributed to the high fracture toughness of acicular ferrite and stable austenite. After utilizing the laser surface hardening treatment, both austempered and quench-tempered gray cast iron specimens had decreased wear loss due to the high surface protection provided by the ledeburitic and martensitic structures with high hardness. In the worn surfaces, it was found that cracks were the dominant wear mechanism. The results of this work have significant value in the future applications of gray cast iron engineering components and provide valuable references for future studies on laser-hardened gray cast iron.
Semiempirical Model for Assessing Dewatering Process by Flocculation of Dredged Sludge in an Artificial Reservoir
Authors: Molino Bruno , Bufalo Gennaro , De Vincenzo Annamaria , Ambrosone Luigi
Published: April 2020

Understanding sedimentation behaviour of clay material is crucial in planning project for sediment removal from bottom of a reservoir. The sedimentation of samples taken from Occhito reservoir ( Italy) is investigated. Samples containing and not containing polyacrylamide have been monitored. Results reveal that polymer induces bridging flocculation and the particle-size distribution tends to become uniform. The sedimentation profiles follow a mater curve. Such experimental observation is used to develop a semi-empirical model for assessment of dewatering process by flocculation of dredged sludge in artificial reservoir. A two-step stage model for assessing the volume of solids in a geotextile tube is suggested. Such model is based on the idea that for very long dewatering times solids reach the configuration of free sedimentation.
Study on Pressure Characteristics and Its Evolution Law at the Ellipsoidal End Cover Pole of Cylindrical Explosion Containment Vessels
Authors: Hu Yunhao , Gu Wenbin , Wang Zhen , Han Yangming
Published: April 2020

To explore the postposition of the maximum pressure at the pole of the ellipsoidal end cover of cylindrical explosion containment vessels and to reveal the mechanism of the load evolution, the experimental method was used to measure the pressure curve at the pole under different charges, and the numerical simulation method was used to analyze the evolution law of the explosion flow field within the end cover. The results show that the end cover pole was subjected to three types of pressure: the primary explosion wave, the secondary shock wave and the convergence wave. In addition, the pressure peaks increased in sequence. The evolution of the flow field in the end cover was affected by the amount of charge and the aspect ratio of the vessel. When the scaled distance due to a small charge increased, or when the aspect ratio of the vessel was reduced, the time interval between the convergence wave and the secondary shock wave at the end cover pole decreased gradually. When the scaled distance increased to 4.05 m·kg−1/3, the convergence wave at the pole superimposed on the secondary shock wave. As the aspect ratio of the vessel ranged from 1.75 to 2.50, the time interval between the two peaks was about 150 μs. However, if the aspect ratio was less than 1.40, the convergence wave and the secondary shock wave were fused through complex interaction.
Functional Roles of Saccades for a Hand Movement
Authors: Sakazume Yuki , Furubayashi Sho , Miyashita Eizo
Published: April 2020

An eye saccade provides appropriate visual information for motor control. The present study was aimed to reveal the role of saccades in hand movements. Two types of movements, i.e., hitting and circle-drawing movements, were adopted, and saccades during the movements were classified as either a leading saccade (LS) or catching saccade (CS) depending on the relative gaze position of the saccade to the hand position. The ratio of types of the saccades during the movements was heavily dependent on the skillfulness of the subjects. In the late phase of the movements in a less skillful subject, CS tended to occur in less precise movements, and precision of the movement tended to be improved in the subsequent movement in the hitting. While LS directing gaze to a target point was observed in both types of the movements regardless of skillfulness of the subjects, LS in between a start point and a target point, which led gaze to a local minimum variance point on a hand movement trajectory, was exclusively found in the drawing in a less skillful subject. These results suggest that LS and some types of CS may provide positional information of via-points in addition to a target point and visual information to improve precision of a feedforward controller in the brain, respectively.
Influence on Bone-to-Implant Contact of Non-Thermal Low-Pressure Argon Plasma: An Experimental Study in Rats
Authors: Sanchez-Perez Arturo , Nicolas-Silvente Ana I. , Sanchez-Matas Carmen , Muñoz-Guzon Fernando , Navarro-Cuellar Carlos , Romanos Georgios E.
Published: April 2020

Roughness characteristics play an essential role in osseointegration. However, there is a concern about the susceptibility of those surfaces to bacterial colonization. New techniques for cleaning and surface treatment have appeared that could favor osseointegration without the need to create surfaces as rough. Such is the case of non-thermal low-pressure argon plasma (NTLP-ArP). One hundred and forty-four implants were placed in the tibiae of 36 Sprague Dawley rats, distributed in four experimental groups: I: mechanized surface; II: mechanized surface treated with NTLP-ArP, III: resorbable blast media (RBM) surface; and IV: RBM surface treated with (NTLP-ArP). Bone-to-implant contact (BIC) percentages were calculated by microtomographic evaluation and histological analysis at one, two, and four weeks after implant placement. ANOVA and Mann–Whitney tests were used for statistical analysis, establishing p < 0.05. No significant differences were found at one-week comparisons. The groups treated with NTLP-ArP obtained higher BIC% than those not treated at two and four weeks. Mechanized surfaces treated with NTLP-ArP obtained BIC values similar to RBM surfaces.
Understanding of Feedback Field-Effect Transistor and Its Applications
Authors: Lee Changhoon , Sung Juho , Shin Changhwan
Published: April 2020

Feedback field-effect transistors (FBFETs) are devices based on a positive feedback loop in which the electrons and holes in the channel region act on the energy states of the potential barrier and wall. Owing to the positive feedback phenomenon, FBFETs have an excellent subthreshold swing (~0 mV/decade at 300 K), a high on-/off current ratio (~1010), and a clear saturation region. The power consumption of both the turn-on state and turn-off state is significantly low until operation commences. In addition, the hysteresis caused by the carriers accumulated in the potential wall allows the FBFET to act as a memory device. Moreover, the power consumption of neuromorphic devices can be suppressed by ~100 times with the use of FBFETs. In this work, we analyze the device structure and operating principle of the FBFET and summarize its applications.
A Novel Way to Automatically Plan Cellular Networks Supported by Linear Programming and Cloud Computing
Authors: Godinho André , Fernandes Daniel , Soares Gabriela , Pina Paulo , Sebastião Pedro , Correia Américo , Ferreira Lucio S.
Published: April 2020

With the increasing number of mobile subscribers worldwide, there is a need for fast and reliable algorithms for planning/optimization of mobile networks, especially because, in order to maintain a network’s quality of service, an operator might need to deploy more equipment. This paper presents a quick and reliable way to automatically plan a set of frequencies in a cellular network, using both cloud technologies and linear programming. We evaluate our pattern in a realistic scenario of a Global System for Mobile communications protocol (GSM) network and compare the results to another already implemented commercial tool. Results show that even though network quality was similar, our algorithm was twelve times faster and used four times less memory. It was also able to frequency plan seventy cells simultaneously in less than three minutes. This mechanism was successfully integrated in the professional tool Metric, and is currently being used for cellular planning. Its extension for application to 3/4/5G networks is under study.
Nonlinear Control Design of a Half-Car Model Using Feedback Linearization and an LQR Controller
Authors: Khan Muhammad Aseer , Abid Muhammad , Ahmed Nisar , Wadood Abdul , Park Herie
Published: April 2020

Effective control of ride quality and handling performance are challenges for active vehicle suspension systems, particularly for off-road applications. The nonlinearities tend to degrade the performance of active suspension systems; these introduce harshness to the ride quality and reduce off-road mobility. Typical control strategies rely on linear models of the suspension dynamics. While these models are convenient, nominally accurate, and controllable due to the abundance of linear control techniques, they neglect the nonlinearities present in real suspension systems. The techniques already implemented and methods used to cope with problem of Half-Car model were studied. Every method and technique had some drawbacks in terms of complexity, cost-effectiveness, and ease of real time implementation. In this paper, an improved control method for Half-Car model was proposed. First, input/output feedback linearization was performed to convert the nonlinear system of Half-Car model into an equivalent linear system. This was followed by a Linear Quadratic Regulator (LQR) controller. This controller had minimized the effects of road disturbances by designing a gain matrix with optimal robustness properties. The proposed control technique was applied in the presence of the deterministic road disturbance. The results were verified using the Matlab/Simulink toolbox.
Flexural Behavior of a 30-Meter Full-Scale Simply Supported Prestressed Concrete Box Girder
Authors: Wang Jianqun , Tang Shenghua , Zheng Hui , Zhou Cong , Zhu Mingqiao
Published: April 2020

Compared with scaled-model testing, full-scale destructive testing is more reliable since the test has no size effect and can truly record the mechanical performance of the structure. However, due to the high cost, only very few full-scale destructive tests have been conducted on the flexural behavior of prestressed concrete (PC) box girders with girders removed from decommissioned bridges. Moreover, related destructive testing on the flexural behavior of a new precast box girder has been rarely reported. To investigate the flexural behavior and optimize the design, destructive testing of a 30-meter full-scale simply supported prestressed box girder was conducted at the construction site. It is illustrated that the failure mode of the tested girder was fracture of the prestressing tendon, and the corresponding maximum compressive strain in the top flange was only 1456 μ ε , which is far less than the ultimate compressive strain (3300 μ ε ). Therefore, the concrete in the top flange was not fully utilized. A nonlinear analysis procedure was performed using the finite strip method (FSM). The validity of the analysis was demonstrated by comparing the analytical results with those of the full-scale test in the field and a scaled model test in a laboratory. Using the developed numerical method, parametric analyses of the ratio of reinforcement were carried out. The prestressing tendon of the tested girder was increased from four strands to six strands in each duct. After the optimization of the prestressed reinforcement, the girder was ductile and the bearing capacity could be increased by 44.3%.
Performance and Mechanism of Asphalt Modified by Buton-Rock Asphalt and Different Types of Styrene-Butadiene-Rubber
Authors: Qu Fangting , Lv Songtao , Gao Junfeng , Liu Chaochao
Published: April 2020

In this paper, two types of Styrene-Butadiene-Rubber (SBR) were adopted to modify the Buton-rock asphalt (BRA) modified asphalt, aiming to select a binder with excellent comprehensive performances. Powder SBR and latex SBR (0%, 2%, 4%, 6%, and 8%), were mixed with the 15 w% BRA modified asphalt. The characterization of rheological properties included dynamic shear rheometer, rotational viscometer, and bending beam rheometer test. The short-term aging performance was characterized by the ratio of the complex shear modulus from the un-aged and rolling thin film oven (RTFOT) -aged asphalt. Besides, Fourier transform infrared spectroscopy and scanning electron microscopy were conducted to reveal the modification mechanism. It was observed that the two kinds of BRA-SBR modified asphalt had preferable anti-crack capacity at low temperatures than the BRA modified asphalt. Compared with latex SBR, the powder SBR significantly improved the high-temperature performance, and the anti-aging capacity was stable. However, some negative influence occurred by the addition of latex SBR on the anti-rutting and short-term aging property. According to the micro-mechanism analysis, adding powder SBR and latex SBR into BRA modified asphalt was a physical blending process, and they improved the dispersion state of BRA in asphalt. Based on the comprehensive performance, the recommended combination was BRA and powder SBR.
A Simplified Method for Assessing the Response of RC Frame Structures to Sudden Column Removal
Authors: Biagi Valerio De , Kiakojouri Foad , Chiaia Bernardino , Sheidaii Mohammad Reza
Published: April 2020

Column loss is a type of damage that can occur in frame structures subjected to explosions or impacts. The response of such structures largely depends on the capacity of the assembly of elements and on the inertia effects due to the sudden nature of the phenomenon. Frame structures are able to develop various resisting mechanisms that prevent the collapse to progress. The assessment of the robustness often requires complex and detailed numerical modelling. For the preliminary design of a robust frame, simplified methods to assess the effectiveness of the redistribution of the loads after the removal of a member are welcome. In the present paper, an approach based on the idealisation of the damaged structure into a single degree-of-freedom system with an elastic-plastic compliance law is proposed. The output of the method is the dynamic response of a target point, which can serve for assessing the residual safety of the structure. Comparing the obtained results with the outputs of a more sophisticated FE (Finite Elements) analysis, a satisfying accuracy is found.
Support Vector Regression for the Relationships between Ground Motion Parameters and Macroseismic Intensity in the Sichuan–Yunnan Region
Authors: Tao Dongwang , Ma Qiang , Li Shuilong , Xie Zhinan , Lin Dexin , Li Shanyou
Published: April 2020

In this paper, a nonlinear regression method called a support vector regression (SVR) is presented to establish the relationship between engineering ground motion parameters and macroseismic intensity (MSI). Sixteen ground motion parameters, including peak ground acceleration (PGA), peak ground velocity (PGV), Arias intensity, Housner intensity, acceleration spectrum intensity, velocity spectrum intensity, and others, are considered as candidates for feature selection to generate optimal SVR models. The datasets with both useable strong ground motion records and corresponding investigated MSIs in the Sichuan–Yunnan region, China, are all collected, and these 125 pairs of datasets are used for selecting features and comparing regression results. Nine ground motion parameters are selected as the most relevant features: PGA is the first fundamental one and PGV is the fifth relevant feature. Based on performance measures on the testing dataset, the best SVR model is given when the number of features is one all the way up to nine. According to predicted accuracy, SVR models with Gaussian kernel give much better MSI prediction than linear kernel SVR models and linear regression models. In particular, the Gaussian kernel SVR of PGA gives much higher MSI prediction accuracy than the linear regression model of PGV and PGA. The proposed SVR models are valid for MSI values from VI to IX, and they can be used for rapid mapping damage potential and reporting seismic intensity for this high-seismic-activity region.
Special Issue Editorial: Green Buildings and Indoor Air Quality
Authors: Licina Dusan
Published: April 2020

Since the inception of the first green certification scheme several decades ago, expectations for high-performance buildings have been evolving [...]
Lyapunov Type Theorems for Exponential Stability of Linear Skew-Product Three-Parameter Semiflows with Discrete Time
Authors: Dragičević Davor , Preda Ciprian
Published: April 2020

For linear skew-product three-parameter semiflows with discrete time acting on an arbitrary Hilbert space, we obtain a complete characterization of exponential stability in terms of the existence of appropriate Lyapunov functions. As a nontrivial application of our work, we prove that the notion of an exponential stability persists under sufficiently small linear perturbations.
Next-Generation Sequencing and MALDI Mass Spectrometry in the Study of Multiresistant Processed Meat Vancomycin-Resistant Enterococci (VRE)
Authors: Sabença Carolina , de Sousa Telma , Oliveira Soraia , Viala Didier , Théron Laetitia , Chambon Christophe , Hébraud Michel , Beyrouthy Racha , Bonnet Richard , Caniça Manuela , Poeta Patrícia , Igrejas Gilberto
Published: April 2020

Vancomycin-resistant enterococci (VRE), due to their intrinsic resistance to various commonly used antibiotics and their malleable genome, make the treatment of infections caused by these bacteria less effective. The aims of this work were to characterize isolates of Enterococcus spp. that originated from processed meat, through phenotypic and genotypic techniques, as well as to detect putative antibiotic resistance biomarkers. The 19 VRE identified had high resistance to teicoplanin (89%), tetracycline (94%), and erythromycin (84%) and a low resistance to kanamycin (11%), gentamicin (11%), and streptomycin (5%). Based on a Next-Generation Sequencing NGS technique, most isolates were vanA-positive. The most prevalent resistance genes detected were erm(B) and aac(6’)-Ii, conferring resistance to the classes of macrolides and aminoglycosides, respectively. MALDI-TOF mass spectrometry (MS) analysis detected an exclusive peak of the Enterococcus genus at m/z (mass-to-charge-ratio) 4428 ± 3, and a peak at m/z 6048 ± 1 allowed us to distinguish Enterococcus faecium from the other species. Several statistically significant protein masses associated with resistance were detected, such as peaks at m/z 6358.27 and m/z 13237.3 in ciprofloxacin resistance isolates. These results reinforce the relevance of the combined and complementary NGS and MALDI-TOF MS techniques for bacterial characterization.
The Water Content Drives the Susceptibility of the Lichen Evernia prunastri and the Moss Brachythecium sp. to High Ozone Concentrations
Authors: Vannini Andrea , Canali Giulia , Pica Mario , Nali Cristina , Loppi Stefano
Published: April 2020

The aim of this study was to evaluate the tolerance of lichens (Evernia prunastri) and mosses (Brachythecium sp.) to short-term (1 h), acute (1 ppm) O3 fumigation under different hydration states (dry, <10% water content, metabolism almost inactive; wet, >200% water content, metabolism fully active). We hypothesized that stronger damage would occur following exposure under wet conditions. In addition, we checked for the effect of recovery (1 week) after the exposure. Ozone fumigation negatively affected the content of chlorophyll only in wet samples, but in the moss, such a difference was no longer evident after one week of recovery. Photosynthetic efficiency was always impaired by O3 exposure, irrespective of the dry or wet state, and also after one week of recovery, but the effect was much stronger in wet samples. The antioxidant power was increased in wet moss and in dry lichen, while a decrease was found for wet lichens after 1 week. Our results confirm that the tolerance to O3 of lichens and mosses may be determined by their low water content, which is the case during the peaks of O3 occurring during the Mediterranean summer. The role of antioxidant power as a mechanism of resistance to high O3 concentrations needs to be further investigated.
Epigenetic Changes in Host Ribosomal DNA Promoter Induced by an Asymptomatic Plant Virus Infection
Authors: Pérez-Cañamás Miryam , Hevia Elizabeth , Hernández Carmen
Published: April 2020

DNA cytosine methylation is one of the main epigenetic mechanisms in higher eukaryotes and is considered to play a key role in transcriptional gene silencing. In plants, cytosine methylation can occur in all sequence contexts (CG, CHG, and CHH), and its levels are controlled by multiple pathways, including de novo methylation, maintenance methylation, and demethylation. Modulation of DNA methylation represents a potentially robust mechanism to adjust gene expression following exposure to different stresses. However, the potential involvement of epigenetics in plant-virus interactions has been scarcely explored, especially with regard to RNA viruses. Here, we studied the impact of a symptomless viral infection on the epigenetic status of the host genome. We focused our attention on the interaction between Nicotiana benthamiana and Pelargonium line pattern virus (PLPV, family Tombusviridae), and analyzed cytosine methylation in the repetitive genomic element corresponding to ribosomal DNA (rDNA). Through a combination of bisulfite sequencing and RT-qPCR, we obtained data showing that PLPV infection gives rise to a reduction in methylation at CG sites of the rDNA promoter. Such a reduction correlated with an increase and decrease, respectively, in the expression levels of some key demethylases and of MET1, the DNA methyltransferase responsible for the maintenance of CG methylation. Hypomethylation of rDNA promoter was associated with a five-fold augmentation of rRNA precursor levels. The PLPV protein p37, reported as a suppressor of post-transcriptional gene silencing, did not lead to the same effects when expressed alone and, thus, it is unlikely to act as suppressor of transcriptional gene silencing. Collectively, the results suggest that PLPV infection as a whole is able to modulate host transcriptional activity through changes in the cytosine methylation pattern arising from misregulation of methyltransferases/demethylases balance.
Clinically Feasible and Accurate View Classification of Echocardiographic Images Using Deep Learning
Authors: Kusunose Kenya , Haga Akihiro , Inoue Mizuki , Fukuda Daiju , Yamada Hirotsugu , Sata Masataka
Published: April 2020

A proper echocardiographic study requires several video clips recorded from different acquisition angles for observation of the complex cardiac anatomy. However, these video clips are not necessarily labeled in a database. Identification of the acquired view becomes the first step of analyzing an echocardiogram. Currently, there is no consensus whether the mislabeled samples can be used to create a feasible clinical prediction model of ejection fraction (EF). The aim of this study was to test two types of input methods for the classification of images, and to test the accuracy of the prediction model for EF in a learning database containing mislabeled images that were not checked by observers. We enrolled 340 patients with five standard views (long axis, short axis, 3-chamber view, 4-chamber view and 2-chamber view) and 10 images in a cycle, used for training a convolutional neural network to classify views (total 17,000 labeled images). All DICOM images were rigidly registered and rescaled into a reference image to fit the size of echocardiographic images. We employed 5-fold cross validation to examine model performance. We tested models trained by two types of data, averaged images and 10 selected images. Our best model (from 10 selected images) classified video views with 98.1% overall test accuracy in the independent cohort. In our view classification model, 1.9% of the images were mislabeled. To determine if this 98.1% accuracy was acceptable for creating the clinical prediction model using echocardiographic data, we tested the prediction model for EF using learning data with a 1.9% error rate. The accuracy of the prediction model for EF was warranted, even with training data containing 1.9% mislabeled images. The CNN algorithm can classify images into five standard views in a clinical setting. Our results suggest that this approach may provide a clinically feasible accuracy level of view classification for the analysis of echocardiographic data.
Local Translation in Growth Cones and Presynapses, Two Axonal Compartments for Local Neuronal Functions
Authors: Sasaki Yukio
Published: April 2020

During neural development, growth cones, very motile compartments of tips of axons, lead axonal extension to the correct targets. Subsequently, presynapses, another axonal compartment with vigorous trafficking of synaptic vesicles, emerge to form functional synapses with postsynapses. In response to extracellular stimuli, the immediate supply of proteins by local translation within these two axonal compartments far from cell bodies confers high motility of growth cones and active vesicle trafficking in presynapses. Although local translation in growth cones and presynapses occurs at a very low level compared with cell bodies and even dendrites, recent progress in omics and visualization techniques with subcellular fractionation of these compartments has revealed the actual situation of local translation within these two axonal compartments. Here, the increasing evidence for local protein synthesis in growth cones and presynapses for axonal and synaptic functions has been reviewed. Furthermore, the mechanisms regulating local translation in these two compartments and pathophysiological conditions caused by dysregulated local translation are highlighted.
Structures of FOX-4 Cephamycinase in Complex with Transition-State Analog Inhibitors
Authors: Lefurgy Scott T. , Caselli Emilia , Taracila Magdalena A. , Malashkevich Vladimir N. , Biju Beena , Papp-Wallace Krisztina M. , Bonanno Jeffrey B. , Prati Fabio , Almo Steven C. , Bonomo Robert A.
Published: April 2020

Boronic acid transition-state analog inhibitors (BATSIs) are partners with β-lactam antibiotics for the treatment of complex bacterial infections. Herein, microbiological, biochemical, and structural findings on four BATSIs with the FOX-4 cephamycinase, a class C β-lactamase that rapidly hydrolyzes cefoxitin, are revealed. FOX-4 is an extended-spectrum class C cephalosporinase that demonstrates conformational flexibility when complexed with certain ligands. Like other β-lactamases of this class, studies on FOX-4 reveal important insights into structure–activity relationships. We show that SM23, a BATSI, shows both remarkable flexibility and affinity, binding similarly to other β-lactamases, yet retaining an IC50 value < 0.1 μM. Our analyses open up new opportunities for the design of novel transition-state analogs of class C enzymes.
Molecularly Imprinted Polymers and Magnetic Molecularly Imprinted Polymers for Selective Determination of Estrogens in Water by ESI-MS/FAPA-MS
Authors: Guć Maria , Schroeder Grzegorz
Published: April 2020

Qualitative and quantitative analysis of estrogens content in natural water is a difficult task. An important problem in the analysis of hormones in water is the quantitative determination of their individual species. Low detection limits and instability of estrogen derivatives are the main challenges. Magnetic molecularly imprinted polymers (mag-MIPs) in combination with Flowing Atmospheric-Pressure Afterglow Mass Spectrometry (FAPA-MS) were successfully used for analysis of estrogen hormones in water samples. The aim of the study was to obtain mag-MIPs selective to estrone (E1) and β-estradiol (E2) for solid phase extraction and pre-concentration of estrogens. Due to their superior analyte binding properties at low concentrations (0.03 g in 1 g of polymer structure) and possibility of magnetic separation, mag-MIPs were proven to be very convenient and efficient adsorbent materials. In addition, MS analyses were performed using two ionization sources: ESI- and FAPA-MS. For both estrogens, LOD was significantly lower for FAPA-MS analysis (0.135 μg L−1 for E1 and E2) than for ESI-MS analysis (27 μg L−1 for E1 and 13.6 μg L−1 for E2). The total estrogen concentration in the environmental water sample was determined as: cE1 = 0.271 μg L−1 and cE2 = 0.275 μg L−1.
Genomic and Metabolomic Analysis of Antarctic Bacteria Revealed Culture and Elicitation Conditions for the Production of Antimicrobial Compounds
Authors: Núñez-Montero Kattia , Quezada-Solís Damián , Khalil Zeinab , Capon Robert , Andreote Fernando , Barrientos Leticia
Published: April 2020

Concern about finding new antibiotics against drug-resistant pathogens is increasing every year. Antarctic bacteria have been proposed as an unexplored source of bioactive metabolites; however, most biosynthetic gene clusters (BGCs) producing secondary metabolites remain silent under common culture conditions. Our work aimed to characterize elicitation conditions for the production of antibacterial secondary metabolites from 34 Antarctic bacterial strains based on MS/MS metabolomics and genome mining approaches. Bacterial strains were cultivated under different nutrient and elicitation conditions, including the addition of lipopolysaccharide (LPS), sodium nitroprusside (SNP), and coculture. Metabolomes were obtained by HPLC-QTOF-MS/MS and analyzed through molecular networking. Antibacterial activity was determined, and seven strains were selected for genome sequencing and analysis. Biosynthesis pathways were activated by all the elicitation treatments, which varies among strains and dependents of culture media. Increased antibacterial activity was observed for a few strains and addition of LPS was related with inhibition of Gram-negative pathogens. Antibiotic BGCs were found for all selected strains and the expressions of putative actinomycin, carotenoids, and bacillibactin were characterized by comparison of genomic and metabolomic data. This work established the use of promising new elicitors for bioprospection of Antarctic bacteria and highlights the importance of new “-omics” comparative approaches for drug discovery.
Connexin-46 Contained in Extracellular Vesicles Enhance Malignancy Features in Breast Cancer Cells
Authors: Acuña Rodrigo A. , Varas-Godoy Manuel , Berthoud Viviana M. , Alfaro Ivan E. , Retamal Mauricio A.
Published: April 2020

Under normal conditions, almost all cell types communicate with their neighboring cells through gap junction channels (GJC), facilitating cellular and tissue homeostasis. A GJC is formed by the interaction of two hemichannels; each one of these hemichannels in turn is formed by six subunits of transmembrane proteins called connexins (Cx). For many years, it was believed that the loss of GJC-mediated intercellular communication was a hallmark in cancer development. However, nowadays this paradigm is changing. The connexin 46 (Cx46), which is almost exclusively expressed in the eye lens, is upregulated in human breast cancer, and is correlated with tumor growth in a Xenograft mouse model. On the other hand, extracellular vesicles (EVs) have an important role in long-distance communication under physiological conditions. In the last decade, EVs also have been recognized as key players in cancer aggressiveness. The aim of this work was to explore the involvement of Cx46 in EV-mediated intercellular communication. Here, we demonstrated for the first time, that Cx46 is contained in EVs released from breast cancer cells overexpressing Cx46 (EVs-Cx46). This EV-Cx46 facilitates the interaction between EVs and the recipient cell resulting in an increase in their migration and invasion properties. Our results suggest that EV-Cx46 could be a marker of cancer malignancy and open the possibility to consider Cx46 as a new therapeutic target in cancer treatment.
D-Aspartate Upregulates DAAM1 Protein Levels in the Rat Testis and Induces Its Localization in Spermatogonia Nucleus
Authors: Venditti Massimo , Santillo Alessandra , Falvo Sara , Di Fiore Maria Maddalena , Chieffi Baccari Gabriella , Minucci Sergio
Published: April 2020

Cell differentiation during spermatogenesis requires a proper actin dynamic, regulated by several proteins, including formins. Disheveled-Associated-Activator of Morphogenesis1 (DAAM1) belongs to the formins and promotes actin polymerization. Our results showed that oral D-Aspartate (D-Asp) administration, an excitatory amino acid, increased DAAM1 protein levels in germ cells cytoplasm of rat testis. Interestingly, after the treatment, DAAM1 also localized in rat spermatogonia (SPG) and mouse GC-1 cells nuclei. We provided bioinformatic evidence that DAAM1 sequence has two predicted NLS, supporting its nuclear localization. The data also suggested a role of D-Asp in promoting DAAM1 shuttling to the nuclear compartment of those proliferative cells. In addition, the proliferative action induced by D-Asp is confirmed by the increased levels of PCNA, a protein expressed in the nucleus of cells in the S phase and p-H3, a histone crucial for chromatin condensation during mitosis and meiosis. In conclusion, we demonstrated, for the first time, an increased DAAM1 protein levels following D-Asp treatment in rat testis and also its localization in the nucleus of rat SPG and in mouse GC-1 cells. Our results suggest an assumed role for this formin as a regulator of actin dynamics in both cytoplasm and nuclei of the germ cells.
Investigating the Effects of Chiropractic Spinal Manipulation on EEG in Stroke Patients
Authors: Navid Muhammad Samran , Niazi Imran Khan , Lelic Dina , Nedergaard Rasmus Bach , Holt Kelly , Amjad Imran , Drewes Asbjørn Mohr , Haavik Heidi
Published: April 2020

Objective: The purpose of this study was to evaluate the impact of chiropractic spinal manipulation on the early somatosensory evoked potentials (SEPs) and resting-state electroencephalography (EEG) recorded from chronic stroke patients. Methods: Seventeen male patients (53 ± 12 years old) participated in this randomized cross-over study. The patients received chiropractic spinal manipulation and control intervention, in random order, separated by at least 24 hours. EEG was recorded before and after each intervention during rest and stimulation of the non-paretic median nerve. For resting-state EEG, the delta-alpha ratio, brain-symmetry index, and power-spectra were calculated. For SEPs, the amplitudes and latencies of N20 and N30 peaks were assessed. Source localization was performed on the power-spectra of resting-state EEG and the N30 SEP peak. Results: Following spinal manipulation, the N30 amplitude increased by 39%, which was a significant increase compared to the control intervention (p < 0.01). The latency and changes to the strength of the cortical sources underlying the N30 peak were not significant. The N20 peak, the resting-state power-spectra, delta-alpha ratio, brain-symmetry index, and resting-state source localization showed no significant changes after either intervention. Conclusion: A single session of chiropractic spinal manipulation increased the amplitude of the N30 SEP peak in a group of chronic stroke patients, which may reflect changes to early sensorimotor function. More research is required to investigate the long-term effects of chiropractic spinal manipulation, to better understand what impact it may have on the neurological function of stroke survivors.
Site-Specific Effects of Online rTMS during a Working Memory Task in Healthy Older Adults
Authors: Beynel Lysianne , Davis Simon W. , Crowell Courtney A. , Dannhauer Moritz , Lim Wesley , Palmer Hannah , Hilbig Susan A. , Brito Alexandra , Hile Connor , Luber Bruce , Lisanby Sarah H. , Peterchev Angel V. , Cabeza Roberto , Appelbaum Lawrence G.
Published: April 2020

The process of manipulating information within working memory is central to many cognitive functions, but also declines rapidly in old age. Improving this process could markedly enhance the health-span in older adults. The current pre-registered, randomized and placebo-controlled study tested the potential of online repetitive transcranial magnetic stimulation (rTMS) applied at 5 Hz over the left lateral parietal cortex to enhance working memory manipulation in healthy elderly adults. rTMS was applied, while participants performed a delayed-response alphabetization task with two individually titrated levels of difficulty. Coil placement and stimulation amplitude were calculated from fMRI activation maps combined with electric field modeling on an individual-subject basis in order to standardize dosing at the targeted cortical location. Contrary to the a priori hypothesis, active rTMS significantly decreased accuracy relative to sham, and only in the hardest difficulty level. When compared to the results from our previous study, in which rTMS was applied over the left prefrontal cortex, we found equivalent effect sizes but opposite directionality suggesting a site-specific effect of rTMS. These results demonstrate engagement of cortical working memory processing using a novel TMS targeting approach, while also providing prescriptions for future studies seeking to enhance memory through rTMS.
Therapy of Dysphagia by Prolonged Pharyngeal Electrical Stimulation (Phagenyx) in a Patient with Brainstem Infarction
Authors: Florea Cristina , Bräumann Christine , Mussger Christine , Leis Stefan , Hauer Larissa , Sellner Johann , Golaszewski Stefan M.
Published: April 2020

Dysphagia after stroke impacts quality of life and is a risk factor for respiratory infections. Patients frequently require prophylactic measures including nasogastric tube or percutaneous endoscopic gastrostomy. Until recently, therapy for dysphagia was limited to training with a speech and language specialist. Intraluminal pharyngeal electrical stimulation (PES) is a new technique that stimulates the pharyngeal sensory afferents to the higher swallowing center in cortex. The clinical trials published to date involved stimulation for 10 minutes over three days. We present a case of brainstem infarction with severe dysphagia in a 53-year-old woman with preserved cognitive functions. For airway protection, she had a surgical tracheotomy. The initial swallowing training achieved slight improvements, but stagnated after three months so PES was tried. Under good PES tube tolerance, a prolonged and repeated stimulation protocol was administered, with the main purpose of relieving her of the tracheal tube. Although the swallowing improved, she stayed tube-dependent with minimal attempts with puréed food during therapy, and could not be decannulated. Further studies are required to assess the value of this promising approach for the treatment of dysphagia.
Examining the Effects of Anabolic–Androgenic Steroids on Repetitive Mild Traumatic Brain Injury (RmTBI) Outcomes in Adolescent Rats
Authors: Tabor Jason , Wright David. K. , Christensen Jennaya , Zamani Akram , Collins Reid , Shultz Sandy R. , Mychasiuk Richelle
Published: April 2020

Background: Repetitive mild traumatic brain injury (RmTBI) is increasingly common in adolescents. Anabolic–androgenic steroid (AAS) consumption among younger professional athletes is a significant risk factor for impaired neurodevelopment. Given the increased rates and overlapping symptomology of RmTBI and AAS use, we sought to investigate the behavioural and neuropathological outcomes associated with the AAS Metandienone (Met) and RmTBI on rats. Methods: Rats received either Met or placebo and were then administered RmTBIs or sham injuries, followed by a behavioural test battery. Post-mortem MRI was conducted to examine markers of brain integrity and qRT-PCR assessed mRNA expression of markers for neurodevelopment, neuroinflammation, stress responses, and repair processes. Results: Although AAS and RmTBI did not produce cumulative deficits, AAS use was associated with detrimental outcomes including changes to depression, aggression, and memory; prefrontal cortex (PFC) atrophy and amygdala (AMYG) enlargement; damaged white matter integrity in the corpus callosum; and altered mRNA expression in the PFC and AMYG. RmTBI affected general activity and contributed to PFC atrophy. Conclusions: Findings corroborate previous results indicating that RmTBI negatively impacts neurodevelopment but also demonstrates that AAS results in significant neuropathological insult to the developing brain.