bluuebunny/crossref_metadata_2025 huggingface dataset:
A Trp574Leu Target-Site Mutation Confers Imazamox Resistance in Multiple Herbicide-Resistant Wild Poinsettia Populations from Brazil
Authors: Mendes Rafael R. , Takano Hudson K. , Oliveira Rubem S. , Adegas Fernando S. , Gaines Todd A. , Dayan Franck E.
Published: July 2020

Wild poinsettia (Euphorbia heterophylla L.) is an important weed species in southern Brazil, especially due to the evolution of multiple herbicide resistance (e.g., acetolactate synthase (ALS)- inhibitors, protoporphyrinogen oxidase inhibitors, and glyphosate). The mechanism of resistance to imazamox was investigated in two wild poinsettia populations (R1 and R2) from southern Brazil and compared to a known susceptible (S) population. Imazamox dose-response experiments revealed high levels of resistance: 45-fold and 224.5-fold based on dry biomass reduction, for R1 and R2, respectively. Extremely high concentrations of imazamox (20,000 µM) were not sufficient to provide 50% inhibition of ALS enzyme activity (I50) for R1 or R2. Hence, resistance levels were estimated to be greater than 123-fold for both populations based on in vitro ALS assays. The ALS gene from all R1 and R2 plants had a Trp574Leu mutation. A genotyping assay was developed to discriminate resistant and susceptible alleles based on the Trp574Leu mutation.
Environmental Dissemination of Selected Antibiotics from Hospital Wastewater to the Aquatic Environment
Authors: Ekwanzala Mutshiene , Lehutso Raisibe , Kasonga Teddy , Dewar John , Momba Maggy
Published: July 2020

The environmental dissemination of selected antibiotics from hospital wastewater into municipal wastewater and lastly to a receiving water body was investigated. Selected antibiotics (azithromycin (AZM), ciprofloxacin (CIP), clindamycin (CDM), doxycycline (DXC) and sulfamethoxazole (SMZ)) present in effluents of academic hospital wastewater, influents, sewage sludge, and effluents of municipal wastewater, receiving water, and its benthic sediment samples were quantified using the Acquity® Waters Ultra-Performance Liquid Chromatography System hyphenated with a Waters Synapt G2 coupled to a quadrupole time-of-flight mass spectrometer. The overall results showed that all assessed antibiotics were found in all matrices. For solid matrices, river sediment samples had elevated concentrations with mean concentrations of 34,834, 35,623, 50,913, 55,263, and 41,781 ng/g for AZM, CIP, CDM, DXC, and SMZ, respectively, whereas for liquid samples, hospital wastewater and influent of wastewater had the highest concentrations. The lowest concentrations were observed in river water, with mean concentrations of 11, 97, 15, and 123 ng/L, except for CDM, which was 18 ng/L in the effluent of wastewater. The results showed that the highest percentages of antibiotics removed was SMZ with 90%, followed by DXC, AZM and CIP with a removal efficiency of 85%, 83%, and 83%, respectively. The antibiotic that showed the lowest removal percentage was CDM with 66%. However, the calculated environmental dissemination analysis through the use of mass load calculations revealed daily release of 15,486, 14,934, 1526, 922, and 680 mg/d for SMZ, CIP, AZM, DXC, and CDM, respectively, indicating a substantial release of selected antibiotics from wastewater to the river system, where they are possibly adsorbed in the river sediment. Further research into the efficient removal of antibiotics from wastewater and the identification of antibiotic sources in river sediment is needed.
Brazilian Red Propolis Is as Effective as Amoxicillin in Controlling Red-Complex of Multispecies Subgingival Mature Biofilm In Vitro
Authors: de Figueiredo Kadmo Azevedo , da Silva Helio Doyle Pereira , Miranda Stela Lima Farias , Gonçalves Francisco Jerfeson dos Santos , de Sousa Arlene Pereira , de Figueiredo Luciene Cristina , Feres Magda , Bueno-Silva Bruno
Published: July 2020

This study investigated the effects of Brazilian Red Propolis (BRP) extract on seven-day-old multispecies subgingival biofilms. Mixed biofilm cultures containing 31 species associated with periodontal health or disease were grown for six days on a Calgary device. Then, mature biofilms were treated for 24 h with BRP extract at different concentrations (200–1600 µg/mL), amoxicillin (AMOXI) at 54 µg/mL (positive control) or vehicle (negative control). Biofilm metabolic activity was determined by colorimetry, and bacterial counts/proportions were determined by DNA–DNA hybridization. Data were analyzed by Kruskal–Wallis and Dunn’s tests. Treatment with BRP at 1600, 800 and 400 μg/mL reduced biofilm metabolic activity by 56%, 56% and 57%, respectively, as compared to 65% reduction obtained with AMOXI. Mean total cell counts were significantly reduced in all test groups (~50–55%). Lower proportions of red, green and yellow complex species were observed upon treatment with BRP (400 µg/mL) and AMOXI, but only AMOXI reduced the proportions of Actinomyces species. In conclusion, BRP extract was as effective as AMOXI in killing seven-day-old multispecies biofilm pathogens and did not affect the levels of the host-compatible Actinomyces species. These data suggest that BRP may be an alternative to AMOXI as an adjunct in periodontal therapy. In vivo studies are needed to validate these results.
Co-Administration of Iron and a Bioavailable Curcumin Supplement Increases Serum BDNF Levels in Healthy Adults
Authors: Tiekou Lorinczova Helena , Fitzsimons Owen , Mursaleen Leah , Renshaw Derek , Begum Gulshanara , Zariwala Mohammed Gulrez
Published: July 2020

Brain-derived neurotrophic factor (BDNF) is key for the maintenance of normal neuronal function and energy homeostasis and has been suggested to improve cognitive function, including learning and memory. Iron and the antioxidant curcumin have been shown to influence BDNF homeostasis. This 6-week, double blind, randomized, placebo-controlled study examined the effects of oral iron supplementation at low (18 mg) and high (65 mg) ferrous (FS) iron dosages, compared to a combination of these iron doses with a bioavailable formulated form of curcumin (HydroCurcTM; 500 mg) on BDNF levels in a healthy adult cohort of 155 male (26.42 years ± 0.55) and female (25.82 years ± 0.54) participants. Participants were randomly allocated to five different treatment groups: both iron and curcumin placebo (FS0+Plac), low dose iron and curcumin placebo (FS18+Plac), low dose iron and curcumin (FS18+Curc), high dose iron and curcumin placebo (FS65+Plac) and high dose iron and curcumin (FS65+Curc). Results showed a significant increase in BDNF over time (26%) in the FS18+Curc group (p = 0.024), and at end-point between FS18+Curc and FS18+Plac groups (35%, p = 0.042), demonstrating for the first time that the combination with curcumin, rather than iron supplementation alone, results in increased serum BDNF. The addition of curcumin to iron supplementation may therefore provide a novel approach to further enhance the benefits associated with increased BDNF levels.
Effect of Strain Hardening on Increase in Collapse Pressure during the Manufacture of ERW Pipe
Authors: Han Seong-Wook , Park Yeun Chul , Kim Ho-Kyung , Kang Soo-Chang
Published: July 2020

Plastic deformation during the manufacture process of electric resistance welded (ERW) pipe determines the stress–strain relationship of the steel pipe, which affects the collapse pressure of offshore pipelines. To track the deformation history of the pipe, the entire process was simulated via finite element analysis using a solid element. A material model that considered both the Bauschinger effect and strain hardening was adopted. Various sizes of pipe cross-sections were simulated. As greater compression was applied during the sizing process, the strain hardening effect became more significant, so that the compressive yield strength was increased in the circumferential direction. The strain hardening effect was most prominent for a smaller diameter-to-thickness ratio (D/t), so that an increase in the collapse pressure could be obtained with a larger sizing ratio. Therefore, current design criteria for the collapse pressure recommended by Det Norske Verita (DNV) and API could be enhanced for a smaller D/t to consider the strain hardening effect during the sizing process.
Layer-by-Layer Deposition of Copper and Phosphorus Compounds to Develop Flame-Retardant Polyamide 6/Montmorillonite Hybrid Composites
Authors: Majka Tomasz M. , Witek Monika , Radzik Paulina , Komisarz Karolina , Mitoraj Agnieszka , Pielichowski Krzysztof
Published: July 2020

Nowadays, increasing attention is devoted to the search for polymeric composite materials that are characterized by reduced flammability. In this work, the layer-by-layer (LbL) technique was applied to form multilayered protective coatings for polyamide 6/montmorillonite (PA6/MMT) hybrid nanocomposites. This time, the double layers LbL deposition was used in order to improve the thermal properties or flammability of PA6 materials. Our goal was to check how five, 10, and 15 triple-layer deposition onto the surface of PA6 and PA6/MMT composites influenced these relevant properties. For this reason, disodium H-phosphonate, sodium montmorillonite, and iodo-bis(triphenylphosphino)copper were used for polyelectrolyte solution preparation. It was found that the LbL method could be successfully used to improve the flammability characteristics of polyamide 6-based composites. Nevertheless, the deposition of the copper complex should be combined with other flame retardants—preferentially containing phosphorus—which enable synergistic effects to occur. Moreover, microscopic observations confirmed that the surfaces on which the formation of interwoven fibrous crystal structures was observed had a tendency to protect the entire material against the destructive effects of heat, contributing, among other things, to reduce the maximum point of heat release rate (PHRR).
Micro-Structured Patches for Dermal Regeneration Obtained via Electrophoretic Replica Deposition
Authors: Ghalayani Esfahani Arash , Altomare Lina , Bonetti Lorenzo , Nejaddehbashi Fereshteh , Boccafoschi Francesca , Chiesa Roberto , Boschetti Federica , Bayati Vahid , De Nardo Luigi
Published: July 2020

Artificial substrates supporting the healing of skin wounds require specific structural and chemical architectures that promote a recapitulation of the complexity of the native organ. Bottom-up fabrication technologies are emerging as effective strategies to fine tune biochemical, morphological, and structural features intended for regenerative applications. Here, we proposed an electrophoretic replica deposition (EPrD) approach to realize chitosan three-dimensional structures specifically designed to treat patients with serious cutaneous damages or losses. The EPrD process has been optimized to consistently obtain random porosity vs. hierarchical lattice structures, showing mechanical properties in the range of skin tissue (E = 0.2–20 MPa). The obtained patches were tested in vivo via a one-stage grafting procedure in a full thickness skin wound rat model. Chitosan patches showed no adverse reactions throughout the experimental period (14 days). Hair follicles and sebaceous glands were observed in histological sections, indicating the regeneration of a thin epidermal layer with more skin appendages. Immunohistochemistry results demonstrated that keratin 10 was mostly expressed in basal and suprabasal layers, like normal skin, in structures with random porosity and with smaller lattice structures. The obtained results show the potential of EPrD to innovate the design of artificial substrates in skin healing therapies.
Food Chains and Food Webs in Aquatic Ecosystems
Authors: Kwak Ihn-Sil , Park Young-Seuk
Published: July 2020

Food chains and food webs describe the structure of communities and their energy flows, and they present interactions between species. Recently, diverse methods have been developed for both experimental studies and theoretical/computational studies on food webs as well as species interactions. They are effectively used for various applications, including the monitoring and assessment of ecosystems. This Special Issue includes six empirical studies on food chains and food webs as well as effects of environmental factors on organisms in aquatic ecosystems. They confirmed the usefulness of their methods including isotope, DNA-barcoding with gut contents, and environmental DNA for biological monitoring and ecosystem assessment.
Development Cycle Modeling: Resource Estimation
Authors: Denard Samuel , Ertas Atila , Mengel Susan , Ekwaro-Osire Stephen
Published: July 2020

This paper presents results produced by a domain-independent system development model that enables objective and quantitative calculation of certain development cycle characteristics. The presentation recounts the model’s motivation and includes an outline of the model’s structure. The outline shows that the model is constructive. As such, it provides an explanatory mechanism for the results that it produces, not just a representation of qualitative observations or measured data. The model is a Statistical Agent-based Model of Development and Evaluation (SAbMDE); and it appears to be novel with respect to previous design theory and methodology work. This paper focuses on one development cycle characteristic: resource utilization. The model’s resource estimation capability is compared to Boehm’s long-used software development estimation techniques. His Cone of Uncertainty (COU) captures project estimation accuracy empirically at project start but intuitively over a project’s duration. SAbMDE calculates estimation accuracy at start up and over project duration; and SAbMDE duplicates the COU’s empirical values. Additionally, SAbMDE produces results very similar to the Constructive Cost Model (COCOMO) effort estimation for a wide range of input values.
Measurement of Head-Related Transfer Functions: A Review
Authors: Li Song , Peissig Jürgen
Published: July 2020

A head-related transfer function (HRTF) describes an acoustic transfer function between a point sound source in the free-field and a defined position in the listener’s ear canal, and plays an essential role in creating immersive virtual acoustic environments (VAEs) reproduced over headphones or loudspeakers. HRTFs are highly individual, and depend on directions and distances (near-field HRTFs). However, the measurement of high-density HRTF datasets is usually time-consuming, especially for human subjects. Over the years, various novel measurement setups and methods have been proposed for the fast acquisition of individual HRTFs while maintaining high measurement accuracy. This review paper provides an overview of various HRTF measurement systems and some insights into trends in individual HRTF measurements.
Modeling the Long-Term Deformation of a Geodesic Spherical Frame Structure Made from Wood Plastic Composite Lumber
Authors: Alrubaie Murtada Abass A. , Gardner Douglas J. , Lopez-Anido Roberto A.
Published: July 2020

The long-term deformation of a geodesic spherical frame structure with a diameter of 20 m made from wood plastic composite (WPC) lumber (struts) is described using the Norton-Bailey power law model to predict the service life creep behavior (the creep strain ( ε c r )) of the WPC. The Norton-Bailey power law model parameters, A the power law multiplier, n the stress order, and m the time order, were obtained from experimental four-point bending flexural creep measurements of WPC lumber subjected to three levels of flexural stress: 7, 14, and 29% of the ultimate flexural strength for 200 days. The parameters obtained from the experiments showed good agreement to the model of the WPC lumber in flexure. The Norton-Bailey power law parameters were then implemented to describe the long-term deformation of the spherical frame structure. The limit of failure was considered when the WPC creep strain reaches the value of 1%. However, the FEA predicted the maximum creep strain to be 20% of the failure strain. This modeling approach is considered useful to describe and predict the long-term deformation of aquacultural structures made from viscoelastic materials during the envisioned service life (10 years) based on experimental creep data for the members that form the structure.
Applications of the Open-Source Hardware Arduino Platform in the Mining Industry: A Review
Authors: Kim Sung-Min , Choi Yosoon , Suh Jangwon
Published: July 2020

In this study, applications of the Arduino platform in the mining industry were reviewed. Arduino, a representative and popular open-source hardware, can acquire information from various sensors, transmit data using communication technology, and control devices through actuators. The review was conducted by classifying previous studies into three types of Arduino applications: field monitoring systems, wearable systems, and autonomous systems. With regard to field monitoring systems, most studies in mines were classified as atmospheric or geotechnical monitoring. In wearable systems, the health status of the miner was an important consideration, in addition to the environmental conditions of the mine. Arduino can be a useful tool as an initial prototype for autonomous mine systems. Arduino has advantages in that it can be combined with various electronic products and is cost-effective. Therefore, although many studies have been conducted in the laboratory (as opposed to field tests), Arduino applications can be further expanded in the mining field in the future.
The Hyaluronan Pericellular Coat and Cold Atmospheric Plasma Treatment of Cells
Authors: Bergemann Claudia , Waldner Anna-Christin , Emmert Steffen , Nebe J. Barbara
Published: July 2020

In different tumors, high amounts of hyaluronan (HA) are correlated with tumor progression. Therefore, new tumor therapy strategies are targeting HA production and degradation. In plasma medicine research, antiproliferative and apoptosis-inducing effects on tumor cells were observed using cold atmospheric plasma (CAP) or plasma-activated media (PAM). Until now, the influence of PAM on the HA pericellular coat has not been the focus of research. PAM was generated by argon-plasma treatment of Dulbecco’s modified Eagle’s Medium via the kINPen®09 plasma jet. The HA expression on PAM-treated HaCaT cells was determined by flow cytometry and confocal laser scanning microscopy. Changes in the adhesion behavior of vital cells in PAM were observed by impedance measurement using the xCELLigence system. We found that PAM treatment impaired the HA pericellular coat of HaCaT cells. The time-dependent adhesion was impressively diminished. However, a disturbed HA coat alone was not the reason for the inhibition of cell adhesion because cells enzymatically treated with HAdase did not lose their adhesion capacity completely. Here, we showed for the first time that the plasma-activated medium (PAM) was able to influence the HA pericellular coat.
Generalized Single Stage Class C Amplifier: Analysis from the Viewpoint of Chaotic Behavior
Authors: Petrzela Jiri
Published: July 2020

This paper briefly describes a recent discovery that occurred during the study of the simplest mathematical model of a class C amplifier with a bipolar transistor. It is proved both numerically and experimentally that chaos can be observed in this simple network structure under three conditions: (1) the transistor is considered non-unilateral, (2) bias point provides cubic polynomial feedforward and feedback transconductance, and (3) the LC tank has very high resonant frequency. Moreover, chaos is generated by an autonomous class C amplifier; i.e., an isolated system without a driving force is analyzed. By the connection of a harmonic input signal, much more complex behavior can be observed. Additionally, due to the high degree of generalization of the amplifier cell, similar fundamental circuits can be ordinarily found as subparts of typical building blocks of a radio frequency signal path.
Auditory Device Voice Activity Detection Based on Statistical Likelihood-Ratio Order Statistics
Authors: Kim Seon Man
Published: July 2020

This paper proposes a technique for improving statistical-model-based voice activity detection (VAD) in noisy environments to be applied in an auditory hearing aid. The proposed method is implemented for a uniform polyphase discrete Fourier transform filter bank satisfying an auditory device time latency of 8 ms. The proposed VAD technique provides an online unified framework to overcome the frequent false rejection of the statistical-model-based likelihood-ratio test (LRT) in noisy environments. The method is based on the observation that the sparseness of speech and background noise cause high false-rejection error rates in statistical LRT-based VAD—the false rejection rate increases as the sparseness increases. We demonstrate that the false-rejection error rate can be reduced by incorporating likelihood-ratio order statistics into a conventional LRT VAD. We confirm experimentally that the proposed method relatively reduces the average detection error rate by 15.8% compared to a conventional VAD with only minimal change in the false acceptance probability for three different noise conditions whose signal-to-noise ratio ranges from 0 to 20 dB.
Analysis of Fluid Velocity inside an Agricultural Sprayer Using Generalized Linear Mixed Models
Authors: Aguirre Ángel Javier , Guevara-Viera Guillermo E. , Torres-Inga Carlos S. , Guevara-Viera Raúl V. , Boné Antonio , Vidal Mariano , García-Ramos Francisco Javier
Published: July 2020

The fluid velocity inside the tank of agricultural sprayers is an indicator of the quality of the mixture. This study aims to formulate the best generalized linear mixed model to infer the fluid velocity inside a tank under specific operational parameters of the agitation system, such as liquid level, circuit pressures, and number of active nozzles. A complex model was developed that included operational parameters as fixed effects (FE) and the section of the tank as the random effect. The goodness of fit of the model was evaluated by considering the lowest values of Akaike’s information criteria and Bayesian information criterion, and by estimating the residual variance. The gamma distribution and log-link function enhanced the goodness of fit of the best model. The Toeplitz structure was chosen as the structure of the covariance matrix. SPSS and SAS software were used to compute the model. The analysis showed that the greatest influence on the fluid velocity was exerted by the liquid level in the tank, followed by the circuit pressure and, finally, the number of active nozzles. The development presented here could serve as a guide for formulating models to evaluate the efficiency of the agitation system of agricultural sprayers.
A Note on the Opacity of the Sun’s Atmosphere
Authors: Bhatia Anand. K. , Pesnell William. D.
Published: July 2020

The opacity of the atmosphere of the Sun is due to processes such as Thomson scattering, bound–bound transitions and photodetachment (bound–free) of hydrogen and positronium ions. The well-studied free–free transitions involving photons, electrons, and hydrogen atoms are re-examined, while free–free transitions involving positrons are considered for the first time. Cross sections, averaged over a Maxwellian velocity distribution, involving positrons are comparable to those involving electrons. This indicates that positrons do contribute to the opacity of the atmosphere of the Sun. Accurate results are obtained because definitive phase shifts are known for electron–hydrogen and positron–hydrogen scattering.
Radial and Angular Electron Ejection Patterns of Two-Electron Quantum Dots in THz Fields
Authors: Prior Adam , Bachau Henri , Nikolopoulos Lampros A. A.
Published: July 2020

In this work, we develop and apply an ab-initio method to calculate the joint radial- and- angular electron distributions following the interaction of two-electron spherical quantum dots (QD) with intense terahertz pulses of subpicosecond duration. By applying the method to two QDs of different size, we could investigate two particular ionization mechanisms: the direct and the sequential two-photon double ionization. According to our results, the two ionization mechanisms show some similarity in the angular distribution patterns, whereas the corresponding radial distributions are distinctly different, associated with their joint kinetic energy spectrum. We also discuss the time-evolution of the ionization process in the context of the different nature of the interaction of the QD with the external radiation and the electron–electron correlation interactions.
SARS-CoV-2 Positive Hospitalized Cancer Patients during the Italian Outbreak: The Cohort Study in Reggio Emilia
Authors: Pinto Carmine , Berselli Annalisa , Mangone Lucia , Damato Angela , Iachetta Francesco , Foracchia Marco , Zanelli Francesca , Gervasi Erika , Romagnani Alessandra , Prati Giuseppe , Lui Stefania , Venturelli Francesco , Vicentini Massimo , Besutti Giulia , De Palma Rossana , Giorgi Rossi Paolo
Published: July 2020

In the coronavirus disease (COVID-19) pandemic, cancer patients could be a high-risk group due to their immunosuppressed status; therefore, data on cancer patients must be available in order to consider the most adequate strategy of care. We carried out a cohort study on the risk of hospitalization for COVID-19, oncological history, and outcomes on COVID-19 infected cancer patients admitted to the Hospital of Reggio Emilia. Between 1 February and 3 April 2020, a total of 1226 COVID-19 infected patients were hospitalized. The number of cancer patients hospitalized with COVID-19 infection was 138 (11.3%). The median age was slightly higher in patients with cancers than in those without (76.5 vs. 73.0). The risk of intensive care unit (ICU) admission (10.1% vs. 6.7%; RR 1.23, 95% Confidence Interval (CI) 0.63–2.41) and risk of death (34.1% vs. 26.0%; RR 1.07, 95% CI 0.61–1.71) were similar in cancer and non-cancer patients. In the cancer patients group, 89/138 (64.5%) patients had a time interval >5 years between the diagnosis of the tumor and hospitalization. Male gender, age > 74 years, metastatic disease, bladder cancer, and cardiovascular disease were associated with mortality risk in cancer patients. In the Reggio Emilia Study, the incidence of hospitalization for COVID-19 in people with previous diagnosis of cancer is similar to that in the general population (standardized incidence ratio 98; 95% CI 73–131), and it does not appear to have a more severe course or a higher mortality rate than patients without cancer. The phase II of the COVID-19 epidemic in cancer patients needs a strategy to reduce the likelihood of infection and identify the vulnerable population, both in patients with active antineoplastic treatment and in survivors with frequently different coexisting medical conditions.
Compound Prunetin Induces Cell Death in Gastric Cancer Cell with Potent Anti-Proliferative Properties: In Vitro Assay, Molecular Docking, Dynamics, and ADMET Studies
Authors: Vetrivel Preethi , Kim Seong Min , Ha Sang Eun , Kim Hun Hwan , Bhosale Pritam Bhagwan , Senthil Kalaiselvi , Kim Gon Sup
Published: July 2020

Gastric cancer is the common type of malignancy positioned at second in mortality rate causing burden worldwide with increasing treatment options. Prunetin (PRU) is an O-methylated flavonoid that belongs to the group of isoflavone executing beneficial activities. In the present study, we investigated the anti-proliferative and cell death effect of the compound PRU in AGS gastric cancer cell line. The in vitro cytotoxic potential of PRU was evaluated and significant proliferation was observed. We identified that the mechanism of cell death was due to necroptosis through double staining and was confirmed by co-treatment with inhibitor necrostatin (Nec-1). We further elucidated the mechanism of action of necroptosis via receptor interacting protein kinase 3 (RIPK3) protein expression and it has been attributed by ROS generation through JNK activation. Furthermore, through computational analysis by molecular docking and dynamics simulation, the efficiency of compound prunetin against RIPK3 binding was validated. In addition, we also briefed the pharmacokinetic properties of the compound by in silico ADMET analysis.
Significance of Immunosuppressive Cells as a Target for Immunotherapies in Melanoma and Non-Melanoma Skin Cancers
Authors: Fujimura Taku , Aiba Setsuya
Published: July 2020

Tumor-associated macrophages (TAMs) have been detected in most skin cancers. TAMs produce various chemokines and angiogenic factors that promote tumor development, along with other immunosuppressive cells such as myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs) and tumor-associated neutrophils. TAMs generated from monocytes develop into functional, fully activated macrophages, and TAMs obtain various immunosuppressive functions to maintain the tumor microenvironment. Since TAMs express PD1 to maintain the immunosuppressive M2 phenotype by PD1/PD-L1 signaling from tumor cells, and the blockade of PD1/PD-L1 signaling by anti-PD1 antibodies (Abs) activate and re-polarize TAMs into immunoreactive M1 phenotypes, TAMs represent a potential target for anti-PD1 Abs. The main population of TAMs comprises CD163+ M2 macrophages, and CD163+ TAMs release soluble (s)CD163 and several proinflammatory chemokines (CXCL5, CXCL10, CCL19, etc.) as a result of TAM activation to induce an immunosuppressive tumor microenvironment together with other immunosuppressive cells. Since direct blockade of PD1/PD-L1 signaling between tumor cells and tumor-infiltrating T cells (both effector T cells and Tregs) is mandatory for inducing an anti-immune response by anti-PD1 Abs, anti-PD1 Abs need to reach the tumor microenvironment to induce anti-immune responses in the tumor-bearing host. Taken together, TAM-related factors could offer a biomarker for anti-PD1 Ab-based immunotherapy. Understanding the crosstalk between TAMs and immunosuppressive cells is important for optimizing PD1 Ab-based immunotherapy.
In-Vitro Identification and In-Vivo Confirmation of DNA Methylation Biomarkers for Urothelial Cancer
Authors: Köhler Christina U. , Walter Michael , Lang Kerstin , Plöttner Sabine , Roghmann Florian , Noldus Joachim , Tannapfel Andrea , Tam Yu Chun , Käfferlein Heiko U. , Brüning Thomas
Published: July 2020

We identified DNA methylation targets specific for urothelial cancer (UC) by genome-wide methylation difference analysis of human urothelial (RT4, J82, 5637), prostate (LNCAP, DU-145, PC3) and renal (RCC-KP, CAKI-2, CAL-54) cancer cell lines with their respective primary epithelial cells. A large overlap of differentially methylated targets between all organs was observed and 40 Cytosine-phosphate-Guanine motifs (CpGs) were only specific for UC cells. Of those sites, two also showed high methylation differences (≥47%) in vivo when we further compared our data to those previously obtained in our array-based analyses of urine samples in 12 UC patients and 12 controls. Using mass spectrometry, we finally assessed seven CpG sites in this “bladder-specific” region of interest in urine samples of patients with urothelial (n = 293), prostate (n = 75) and renal (n = 23) cancer, and 143 controls. DNA methylation was significantly increased in UC compared to non-UC individuals. The differences were more pronounced for males rather than females. Male UC cases could be distinguished from non-UC individuals with >30% sensitivity at 95% specificity (Area under the curve (AUC) 0.85). In summary, methylation sites highly specific in UC cell lines were also specific in urine samples of UC patients showing that in-vitro data can be successfully used to identify biomarker candidates of in-vivo relevance.
Immunotherapy for Early Stage Colorectal Cancer: A Glance into the Future
Authors: Cohen Romain , Shi Qian , André Thierry
Published: July 2020

Immune checkpoint inhibitors (ICI) have reshaped therapeutic strategies for cancer patients. The development of ICI for early stage colorectal cancer is accompanied by specific challenges: (i) the selection of patients who are likely to benefit from these treatments, i.e., patients with tumors harboring predictive factors of efficacy of ICI, such as microsatellite instability and/or mismatch repair deficiency (MSI/dMMR), or other potential parameters (increased T cell infiltration using Immunoscore® or others, high tumor mutational burden, POLE mutation), (ii) the selection of patients at risk of disease recurrence (poor prognostic features), and (iii) the choice of an accurate clinical trial methodological framework. In this review, we will discuss the ins and outs of clinical research of ICI for early stage MSI/dMMR CC patients in adjuvant and neoadjuvant settings. We will then summarize data that might support the development of ICI in localized colorectal cancer beyond MSI/dMMR.
Clinico-Biological Features and Clonal Hematopoiesis in Patients with Severe COVID-19
Authors: Duployez Nicolas , Demonchy Jordane , Berthon Céline , Goutay Julien , Caplan Morgan , Moreau Anne-Sophie , Bignon Anne , Marceau-Renaut Alice , Garrigue Delphine , Raczkiewicz Imelda , Geffroy Sandrine , Bucci Maxime , Alidjinou Kazali , Demaret Julie , Labalette Myriam , Brousseau Thierry , Dupont Annabelle , Rauch Antoine , Poissy Julien , Susen Sophie , Preudhomme Claude , Quesnel Bruno
Published: July 2020

Advanced age or preexisting comorbidities have been characterized as risk factors for severe coronavirus disease 2019 (COVID-19) cases requiring hospitalization and intensive care. In recent years, clonal hematopoiesis (CH) of indeterminate potential (CHIP) has emerged as a risk factor for chronic inflammatory background and subsequent aging-associated diseases. The purpose of this study was to identify biological factors (particularly leukocyte subtypes and inflammatory markers) associated with a risk of clinical deterioration (i.e., orotracheal intubation (OTI)) and to determine whether CH was likely to influence clinical and biological behavior in patients with severe COVID-19 requiring hospitalization. Here, we describe clinical and biological features, including the screening of CHIP mutants in a well-annotated cohort of 122 hospitalized patients with a laboratory-confirmed diagnosis of COVID-19 (55% requiring OTI). We showed that elevated white blood cell counts, especially neutrophils and high C-reactive protein (CRP) levels at admission, were associated with an increased requirement of OTI. We noticed a high prevalence of CH (25%, 38%, 56%, and 82% of patients aged <60 years, 60–70 years, 70–80 years, and >80 years) compared to a retrospective cohort of patients free of hematological malignancy explored with the same pipelines (10%, 21%, 37%, and 44%). However, the existence of CH did not significantly impact clinical outcome, including OTI or death, and did not correlate with other laboratory findings.
Plasma Treatment Limits Cutaneous Squamous Cell Carcinoma Development In Vitro and In Vivo
Authors: Pasqual-Melo Gabriella , Nascimento Thiago , Sanches Larissa Juliani , Blegniski Fernanda Paschoal , Bianchi Julya Karen , Sagwal Sanjeev Kumar , Berner Julia , Schmidt Anke , Emmert Steffen , Weltmann Klaus-Dieter , von Woedtke Thomas , Gandhirajan Rajesh Kumar , Cecchini Alessandra Lourenço , Bekeschus Sander
Published: July 2020

Cutaneous squamous cell carcinoma (SCC) is the most prevalent cancer worldwide, increasing the cost of healthcare services and with a high rate of morbidity. Its etiology is linked to chronic ultraviolet (UV) exposure that leads to malignant transformation of keratinocytes. Invasive growth and metastasis are severe consequences of this process. Therapy-resistant and highly aggressive SCC is frequently fatal, exemplifying the need for novel treatment strategies. Cold physical plasma is a partially ionized gas, expelling therapeutic doses of reactive oxygen and nitrogen species that were investigated for their anticancer capacity against SCC in vitro and SCC-like lesions in vivo. Using the kINPen argon plasma jet, a selective growth-reducing action of plasma treatment was identified in two SCC cell lines in 2D and 3D cultures. In vivo, plasma treatment limited the progression of UVB-induced SSC-like skin lesions and dermal degeneration without compromising lesional or non-lesional skin. In lesional tissue, this was associated with a decrease in cell proliferation and the antioxidant transcription factor Nrf2 following plasma treatment, while catalase expression was increased. Analysis of skin adjacent to the lesions and determination of global antioxidant parameters confirmed the local but not systemic action of the plasma anticancer therapy in vivo.
Adjuvant Therapy for Melanoma: Past, Current, and Future Developments
Authors: Testori Alessandro A. E. , Chiellino Silvia , van Akkooi Alexander C.J.
Published: July 2020

This review describes the progress that the concept of adjuvant therapies has undergone in the last 50 years and focuses on the most recent development where an adjuvant approach has been scientifically evaluated in melanoma clinical trials. Over the past decade the development of immunotherapies and targeted therapies has drastically changed the treatment of stage IV melanoma patients. These successes led to trials studying the same therapies in the adjuvant setting, in high risk resected stage III and IV melanoma patients. Adjuvant immune checkpoint blockade with anti-CTLA-4 antibody ipilimumab was the first drug to show an improvement in recurrence-free and overall survival but this was accompanied by high severe toxicity rates. Therefore, these results were bypassed by adjuvant treatment with anti-PD-1 agents nivolumab and pembrolizumab and BRAF-directed target therapy, which showed even better recurrence-free survival rates with more favorable toxicity rates. The whole concept of adjuvant therapy may be integrated with the new neoadjuvant approaches that are under investigation through several clinical trials. However, there is still no data available on whether the effective adjuvant therapy that patients finally have at their disposal could be offered to them while waiting for recurrence, sparing at least 50% of them a potentially long-term toxic side effect but with the same rate of overall survival (OS). Adjuvant therapy for melanoma has radically changed over the past few years—anti-PD-1 or BRAF-directed therapy is the new standard of care.
Discovery of Prognostic Markers for Early-Stage High-Grade Serous Ovarian Cancer by Maldi-Imaging
Authors: Kulbe Hagen , Klein Oliver , Wu Zhiyang , Taube Eliane T. , Kassuhn Wanja , Horst David , Darb-Esfahani Silvia , Jank Paul , Abobaker Salem , Ringel Frauke , du Bois Andreas , Heitz Florian , Sehouli Jalid , Braicu Elena I.
Published: July 2020

With regard to relapse and survival, early-stage high-grade serous ovarian (HGSOC) patients comprise a heterogeneous group and there is no clear consensus on first-line treatment. Currently, no prognostic markers are available for risk assessment by standard targeted immunohistochemistry and novel approaches are urgently required. Here, we applied MALDI-imaging mass spectrometry (MALDI-IMS), a new method to identify distinct mass profiles including protein signatures on paraffin-embedded tissue sections. In search of prognostic biomarker candidates, we compared proteomic profiles of primary tumor sections from early-stage HGSOC patients with either recurrent (RD) or non-recurrent disease (N = 4; each group) as a proof of concept study. In total, MALDI-IMS analysis resulted in 7537 spectra from the malignant tumor areas. Using receiver operating characteristic (ROC) analysis, 151 peptides were able to discriminate between patients with RD and non-RD (AUC > 0.6 or < 0.4; p < 0.01), and 13 of them could be annotated to proteins. Strongest expression levels of specific peptides linked to Keratin type1 and Collagen alpha-2(I) were observed and associated with poor prognosis (AUC > 0.7). These results confirm that in using IMS, we could identify new candidates to predict clinical outcome and treatment extent for patients with early-stage HGSOC.
Genomic Alterations and Their Implications on Survival in Nonmetastatic Colorectal Cancer: Status Quo and Future Perspectives
Authors: Mukherji Reetu , Marshall John , Seeber Andreas
Published: July 2020

The selection of treatment according to genomic alterations is a standard approach in metastatic colorectal cancer but is only starting to have an impact in the earlier stages of the disease. The status of genes like KRAS, BRAF, and MMR has substantial survival implications, and concerted research efforts have revolutionized treatment towards precision oncology. In contrast, a genomic-based approach has not changed the adjuvant setting after curative tumor-resection in the daily routine so far. This review focuses on the current knowledge regarding prognostic and predictive genomic biomarkers in patients with locally advanced nonmetastasized colorectal cancer. Furthermore, we provide an outlook on future challenges for a personalized adjuvant treatment approach in patients with colorectal cancer.
Composites of Crosslinked Aggregates of Eversa® Transform and Magnetic Nanoparticles. Performance in the Ethanolysis of Soybean Oil
Authors: Miranda Letícia Passos , Guimarães José Renato , Giordano Roberto Campos , Fernandez-Lafuente Roberto , Tardioli Paulo Waldir
Published: July 2020

Eversa® Transform 2.0 has been launched to be used in free form, but its immobilization may improve its performance. This work aimed to optimize the immobilization of Eversa® Transform 2.0 by the crosslinked enzyme aggregates (CLEAs) technique, using almost all the available tools to improve its performance. Several variables in the CLEA preparation were optimized to improve the recovered activity, such as precipitant nature and crosslinker concentration. Moreover, some feeders were co-precipitated to improve the crosslinking step, such as bovine serum albumin, soy protein, or polyethyleneimine. Starch (later enzymatically degraded) was utilized as a porogenic agent to decrease the substrate diffusion limitations. Silica magnetic nanoparticles were also utilized to simplify the CLEA handling, but it was found that a large percentage of the Eversa activity could be immobilized on these nanoparticles before aggregation. The best CLEA protocol gave a 98.9% immobilization yield and 30.1% recovered activity, exhibited a porous structure, and an excellent performance in the transesterification of soybean oil with ethanol: 89.8 wt% of fatty acid ethyl esters (FAEEs) yield after 12 h of reaction, while the free enzyme required a 48 h reaction to give the same yield. A caustic polishing step of the product yielded a biodiesel containing 98.9 wt% of FAEEs and a free fatty acids content lower than 0.25%, thus the final product met the international standards for biodiesel. The immobilized biocatalyst could be reused for at least five 12 h-batches maintaining 89.6% of the first-batch yield, showing the efficient catalyst recovery by applying an external magnetic field.
The Lipid Receptor G2A (GPR132) Mediates Macrophage Migration in Nerve Injury-Induced Neuropathic Pain
Authors: Osthues Tabea , Zimmer Béla , Rimola Vittoria , Klann Kevin , Schilling Karin , Mathoor Praveen , Angioni Carlo , Weigert Andreas , Geisslinger Gerd , Münch Christian , Scholich Klaus , Sisignano Marco
Published: July 2020

Nerve injury-induced neuropathic pain is difficult to treat and mechanistically characterized by strong neuroimmune interactions, involving signaling lipids that act via specific G-protein coupled receptors. Here, we investigated the role of the signaling lipid receptor G2A (GPR132) in nerve injury-induced neuropathic pain using the robust spared nerve injury (SNI) mouse model. We found that the concentrations of the G2A agonist 9-HODE (9-Hydroxyoctadecadienoic acid) are strongly increased at the site of nerve injury during neuropathic pain. Moreover, G2A-deficient mice show a strong reduction of mechanical hypersensitivity after nerve injury. This phenotype is accompanied by a massive reduction of invading macrophages and neutrophils in G2A-deficient mice and a strongly reduced release of the proalgesic mediators TNFα, IL-6 and VEGF at the site of injury. Using a global proteome analysis to identify the underlying signaling pathways, we found that G2A activation in macrophages initiates MyD88-PI3K-AKT signaling and transient MMP9 release to trigger cytoskeleton remodeling and migration. We conclude that G2A-deficiency reduces inflammatory responses by decreasing the number of immune cells and the release of proinflammatory cytokines and growth factors at the site of nerve injury. Inhibiting the G2A receptor after nerve injury may reduce immune cell-mediated peripheral sensitization and may thus ameliorate neuropathic pain.
Highly Efficient Conversion of Motor Neuron-Like NSC-34 Cells into Functional Motor Neurons by Prostaglandin E2
Authors: Nango Hiroshi , Kosuge Yasuhiro , Sato Masaki , Shibukawa Yoshiyuki , Aono Yuri , Saigusa Tadashi , Ito Yoshihisa , Ishige Kumiko
Published: July 2020

Motor neuron diseases are a group of progressive neurological disorders that degenerate motor neurons. The neuroblastoma × spinal cord hybrid cell line NSC-34 is widely used as an experimental model in studies of motor neuron diseases. However, the differentiation efficiency of NSC-34 cells to neurons is not always sufficient. We have found that prostaglandin E2 (PGE2) induces morphological differentiation in NSC-34 cells. The present study investigated the functional properties of PGE2-differentiated NSC-34 cells. Retinoic acid (RA), a widely-used agent inducing cell differentiation, facilitated neuritogenesis, which peaked on day 7, whereas PGE2-induced neuritogenesis took only 2 days to reach the same level. Whole-cell patch-clamp recordings showed that the current threshold of PGE2-treated cell action potentials was lower than that of RA-treated cells. PGE2 and RA increased the protein expression levels of neuronal differentiation markers, microtubule-associated protein 2c and synaptophysin, and to the same extent, motor neuron-specific markers HB9 and Islet-1. On the other hand, protein levels of choline acetyltransferase and basal release of acetylcholine in PGE2-treated cells were higher than in RA-treated cells. These results suggest that PGE2 is a rapid and efficient differentiation-inducing factor for the preparation of functionally mature motor neurons from NSC-34 cells.
Hyaluronic Acid: Redefining Its Role
Authors: Abatangelo G. , Vindigni V. , Avruscio G. , Pandis L. , Brun P.
Published: July 2020

The discovery of several unexpected complex biological roles of hyaluronic acid (HA) has promoted new research impetus for biologists and, the clinical interest in several fields of medicine, such as ophthalmology, articular pathologies, cutaneous repair, skin remodeling, vascular prosthesis, adipose tissue engineering, nerve reconstruction and cancer therapy. In addition, the great potential of HA in medicine has stimulated the interest of pharmaceutical companies which, by means of new technologies can produce HA and several new derivatives in order to increase both the residence time in a variety of human tissues and the anti-inflammatory properties. Minor chemical modifications of the molecule, such as the esterification with benzyl alcohol (Hyaff-11® biomaterials), have made possible the production of water-insoluble polymers that have been manufactured in various forms: membranes, gauzes, nonwoven meshes, gels, tubes. All these biomaterials are used as wound-covering, anti-adhesive devices and as scaffolds for tissue engineering, such as epidermis, dermis, micro-vascularized skin, cartilage and bone. In this review, the essential biological functions of HA and the applications of its derivatives for pharmaceutical and tissue regeneration purposes are reviewed.