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    Conductive Gas Plasma Treatment Augments Tumor Toxicity of Ringer’s Lactate Solutions in a Model of Peritoneal Carcinomatosis
    (Basel : MDPI, 2022) Miebach, Lea; Freund, Eric; Cecchini, Alessandra Lourenço; Bekeschus, Sander
    Reactive species generated by medical gas plasma technology can be enriched in liquids for use in oncology targeting disseminated malignancies, such as metastatic colorectal cancer. Notwithstanding, reactive species quantities depend on the treatment mode, and we recently showed gas plasma exposure in conductive modes to be superior for cancer tissue treatment. However, evidence is lacking that such a conductive mode also equips gas plasma-treated liquids to confer augmented intraperitoneal anticancer activity. To this end, employing atmospheric pressure argon plasma jet kINPen-treated Ringer’s lactate (oxRilac) in a CT26-model of colorectal peritoneal carcinomatosis, we tested repeated intraabdominal injection of such remotely or conductively oxidized liquid for antitumor control and immunomodulation. Enhanced reactive species formation in conductive mode correlated with reduced tumor burden in vivo, emphasizing the advantage of conduction over the free mode for plasma-conditioned liquids. Interestingly, the infiltration of lymphocytes into the tumors was equally enhanced by both treatments. However, significantly lower levels of interleukin (IL)4 and IL13 and increased levels of IL2 argue for a shift in intratumoral T-helper cell subpopulations correlating with disease control. In conclusion, our data argue for using conductively over remotely prepared plasma-treated liquids for anticancer treatment.
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    Characterisation of Methicillin-Resistant Staphylococcus aureus from Alexandria, Egypt
    (Basel : MDPI, 2023) Monecke, Stefan; Bedewy, Amira K.; Müller, Elke; Braun, Sascha D.; Diezel, Celia; Elsheredy, Amel; Kader, Ola; Reinicke, Martin; Ghazal, Abeer; Rezk, Shahinda; Ehricht, Ralf
    The present study aims to characterise clinical MRSA isolates from a tertiary care centre in Egypt’s second-largest city, Alexandria. Thirty isolates collected in 2020 were genotypically characterised by microarray to detect their resistance and virulence genes and assign them to clonal complexes (CC) and strains. Isolates belonged to 11 different CCs and 14 different strains. CC15-MRSA-[V+fus] (n = 6), CC1-MRSA-[V+fus+tir+ccrA/B-1] (PVL+) (n = 5) as well as CC1-MRSA-[V+fus+tir+ccrA/B-1] and CC1153-MRSA-[V+fus] (PVL+) (both with n = 3) were the most common strains. Most isolates (83%) harboured variant or composite SCCmec V or VI elements that included the fusidic acid resistance gene fusC. The SCCmec [V+fus+tir+ccrA/B-1] element of one of the CC1 isolates was sequenced, revealing a presence not only of fusC but also of blaZ, aacA-aphD and other resistance genes. PVL genes were also common (40%). The hospital-acquired MRSA CC239-III strain was only found twice. A comparison to data from a study on strains collected in 2015 (Montelongo et al., 2022) showed an increase in fusC and PVL carriage and a decreasing prevalence of the CC239 strain. These observations indicate a diffusion of community-acquired strains into hospital settings. The beta-lactam use in hospitals and the widespread fusidic acid consumption in the community might pose a selective pressure that favours MRSA strains with composite SCCmec elements comprising mecA and fusC. This is an unsettling trend, but more MRSA typing data from Egypt are required.
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    Spatial Distribution Patterns for Identifying Risk Areas Associated with False Smut Disease of Rice in Southern India
    (Basel : MDPI, 2022) Huded, Sharanabasav; Pramesh, Devanna; Chittaragi, Amoghavarsha; Sridhara, Shankarappa; Chidanandappa, Eranna; Prasannakumar, Muthukapalli K.; Manjunatha, Channappa; Patil, Balanagouda; Shil, Sandip; Pushpa, Hanumanthappa Deeshappa; Raghunandana, Adke; Usha, Indrajeet; Balasundram, Siva K.; Shamshiri, Redmond R.
    False smut disease (FSD) of rice incited by Ustilaginoidea virens is an emerging threat to paddy cultivation worldwide. We investigated the spatial distribution of FSD in different paddy ecosystems of South Indian states, viz., Andhra Pradesh, Karnataka, Tamil Nadu, and Telangana, by considering the exploratory data from 111 sampling sites. Point pattern and surface interpolation analyses were carried out to identify the spatial patterns of FSD across the studied areas. The spatial clusters of FSD were confirmed by employing spatial autocorrelation and Ripley’s K function. Further, ordinary kriging (OK), indicator kriging (IK), and inverse distance weighting (IDW) were used to create spatial maps by predicting the values at unvisited locations. The agglomerative hierarchical cluster analysis using the average linkage method identified four main clusters of FSD. From the Local Moran’s I statistic, most of the areas of Andhra Pradesh and Tamil Nadu were clustered together (at I > 0), except the coastal and interior districts of Karnataka (at I < 0). Spatial patterns of FSD severity were determined by semi-variogram experimental models, and the spherical model was the best fit. Results from the interpolation technique, the potential FSD hot spots/risk areas were majorly identified in Tamil Nadu and a few traditional rice-growing ecosystems of Northern Karnataka. This is the first intensive study that attempted to understand the spatial patterns of FSD using geostatistical approaches in India. The findings from this study would help in setting up ecosystem-specific management strategies to reduce the spread of FSD in India.
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    Optical Spectrometry to Determine Nutrient Concentrations and other Physicochemical Parameters in Liquid Organic Manures: A Review
    (Basel : MDPI, 2022) Horf, Michael; Vogel, Sebastian; Drücker, Harm; Gebbers, Robin; Olfs, Hans-Werner
    Nutrient concentrations in livestock manures and biogas digestates show a huge variability due to disparities in animal husbandry systems concerning animal species, feed composition, etc. Therefore, a nutrient estimation based on recommendation tables is not reliable when the exact chemical composition is needed. The alternative, to analyse representative fertilizer samples in a standard laboratory, is too time-and cost-intensive to be an accepted routine method for farmers. However, precise knowledge about the actual nutrient concentrations in liquid organic fertilizers is a prerequisite to ensure optimal nutrient supply for growing crops and on the other hand to avoid environmental problems caused by overfertilization. Therefore, spectrometric methods receive increasing attention as fast and low-cost alternatives. This review summarizes the present state of research based on optical spectrometry used at laboratory and field scale for predicting several parameters of liquid organic manures. It emphasizes three categories: (1) physicochemical parameters, e.g., dry matter, pH, and electrical conductivity; (2) main plant nutrients, i.e., total nitrogen, ammonium nitrogen, phosphorus, potassium, magnesium, calcium, and sulfur; and (3) micronutrients, i.e., manganese, iron, copper, and zinc. Furthermore, the commonly used sample preparation techniques, spectrometer types, measuring modes, and chemometric methods are presented. The primarily promising scientific results of the last 30 years contributed to the fact that near-infrared spectrometry (NIRS) was established in commercial laboratories as an alternative method to wet chemical standard methods. Furthermore, companies developed technical setups using NIRS for on-line applications of liquid organic manures. Thus, NIRS seems to have evolved to a competitive measurement procedure, although parts of this technique still need to be improved to ensure sufficient accuracy, especially in quality management.
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    A Flashback on Control Logic Injection Attacks against Programmable Logic Controllers
    (Basel : MDPI, 2022) Alsabbagh, Wael; Langendörfer, Peter
    Programmable logic controllers (PLCs) make up a substantial part of critical infrastructures (CIs) and industrial control systems (ICSs). They are programmed with a control logic that defines how to drive and operate critical processes such as nuclear power plants, petrochemical factories, water treatment systems, and other facilities. Unfortunately, these devices are not fully secure and are prone to malicious threats, especially those exploiting vulnerabilities in the control logic of PLCs. Such threats are known as control logic injection attacks. They mainly aim at sabotaging physical processes controlled by exposed PLCs, causing catastrophic damage to target systems as shown by Stuxnet. Looking back over the last decade, many research endeavors exploring and discussing these threats have been published. In this article, we present a flashback on the recent works related to control logic injection attacks against PLCs. To this end, we provide the security research community with a new systematization based on the attacker techniques under three main attack scenarios. For each study presented in this work, we overview the attack strategies, tools, security goals, infected devices, and underlying vulnerabilities. Based on our analysis, we highlight the current security challenges in protecting PLCs from such severe attacks and suggest security recommendations for future research directions.
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    Dielectrophoresis: An Approach to Increase Sensitivity, Reduce Response Time and to Suppress Nonspecific Binding in Biosensors?
    (Basel : MDPI, 2022) Henriksson, Anders; Neubauer, Peter; Birkholz, Mario
    The performance of receptor-based biosensors is often limited by either diffusion of the analyte causing unreasonable long assay times or a lack of specificity limiting the sensitivity due to the noise of nonspecific binding. Alternating current (AC) electrokinetics and its effect on biosensing is an increasing field of research dedicated to address this issue and can improve mass transfer of the analyte by electrothermal effects, electroosmosis, or dielectrophoresis (DEP). Accordingly, several works have shown improved sensitivity and lowered assay times by order of magnitude thanks to the improved mass transfer with these techniques. To realize high sensitivity in real samples with realistic sample matrix avoiding nonspecific binding is critical and the improved mass transfer should ideally be specific to the target analyte. In this paper we cover recent approaches to combine biosensors with DEP, which is the AC kinetic approach with the highest selectivity. We conclude that while associated with many challenges, for several applications the approach could be beneficial, especially if more work is dedicated to minimizing nonspecific bindings, for which DEP offers interesting perspectives.
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    Combined In Vitro Toxicity and Immunogenicity of Cold Plasma and Pulsed Electric Fields
    (Basel : MDPI, 2022) Wolff, Christina M.; Kolb, Juergen F.; Bekeschus, Sander
    In modern oncology, therapies are based on combining monotherapies to overcome treatment resistance and increase therapy precision. The application of microsecond-pulsed electric fields (PEF) is approved to enhance local chemotherapeutic drug uptake within combination electrochemotherapy regimens. Reactive oxygen species (ROS) have been implicated in anticancer effects, and cold physical plasma produces vast amounts of ROS, which have recently been shown to benefit head and neck cancer patients. PEF and cold plasma technology have been linked to immunogenic cell death (ICD) induction, a regulated cell death accompanied by sterile inflammation that promotes antitumor immunity. To this end, we investigated the combined effect of both treatments regarding their intracellular ROS accumulation, toxicity, ICD-related marker expression, and optimal exposure sequence in a leukemia model cell line. The combination treatment substantially increased ROS and intracellular glutathione levels, leading to additive cytotoxic effects accompanied by a significantly increased expression of ICD markers, such as the eat-me signal calreticulin (CRT). Preconditioned treatment with cold plasma followed by PEF exposure was the most potent treatment sequence. The results indicate additive effects of cold plasma and PEF, motivating further studies in skin and breast tumor models for the future improvement of ECT in such patients.
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    Surface Plasmon Resonance Sensitivity Enhancement Based on Protonated Polyaniline Films Doped by Aluminum Nitrate
    (Basel : MDPI, 2022) Al-Bataineh, Qais M.; Shpacovitch, Victoria; Sadiq, Diyar; Telfah, Ahmad; Hergenröder, Roland
    Complex composite films based on polyaniline (PANI) doped hydrochloric acid (HCl) incorporated with aluminum nitrate (Al(NO3)3) on Au-layer were designed and synthesized as a surface plasmon resonance (SPR) sensing device. The physicochemical properties of (PANI-HCl)/Al(NO3)3 complex composite films were studied for various Al(NO3)3 concentrations (0, 2, 4, 8, 16, and 32 wt.%). The refractive index of the (PANI-HCl)/Al(NO3)3 complex composite films increased continuously as Al(NO3)3 concentrations increased. The electrical conductivity values increased from 5.10 µS/cm to 10.00 µS/cm as Al(NO3)3 concentration increased to 32 wt.%. The sensitivity of the SPR sensing device was investigated using a theoretical approach and experimental measurements. The theoretical system of SPR measurement confirmed that increasing Al(NO3)3 in (PANI-HCl)/Al(NO3)3 complex composite films enhanced the sensitivity from about 114.5 [Deg/RIU] for Au-layer to 159.0 [Deg/RIU] for Au-((PANI-HCl)/Al(NO3)3 (32 wt.%)). In addition, the signal-to-noise ratio for Au-layer was 3.95, which increased after coating by (PANI-HCl)/Al(NO3)3 (32 wt.%) complex composite layer to 8.82. Finally, we conclude that coating Au-layer by (PANI-HCl)/Al(NO3)3 complex composite films enhances the sensitivity of the SPR sensing device.
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    Influence of redox stress on crosstalk between fibroblasts and keratinocytes
    (Basel : MDPI, 2021) Bhartiya, Pradeep; Masur, Kai; Shome, Debarati; Kaushik, Neha; Nguyen, Linh N.; Kaushik, Nagendra Kumar; Choi, Eun Ha
    Although the skin is constantly subjected to endogenous and exogenous stress, it maintains a homeostatic state through wound repair and regeneration pathways. Treatment for skin diseases and injury requires a significant understanding of the various mechanisms and interactions that occur within skin cells. Keratinocytes and fibroblasts interact with each other and act as key players in the repair process. Although fibroblasts and keratinocytes are widely studied in wound healing and skin remodeling under different conditions, the influence of redox stress on keratinocyte-fibroblast crosstalk has not been thoroughly investigated. In this study, we used cold atmospheric plasma (CAP) to generate and deliver oxidative stress to keratinocytes and fibroblasts and to assess its impact on their interactions. To this end, we used a well-established in vitro 3D co-culture model imitating a realistic scenario. Our study shows that low CAP exposure is biocompatible and does not affect the viability or energetics of fibroblasts and keratinocytes. Exposure to low doses of CAP enhanced the proliferation rate of cells and stimulated the expression of key genes (KGF, MMP2, GMCSF, IL-6, and IL-8) in fibroblasts, indicating the activation and initiation of the skin repair process. Additionally, enhanced migration was observed under co-culture conditions under the given redox stress conditions, and expression of the upstream regulator and the effectors of the Hippo pathway (YAP and CYR61, respectively), which are associated with enhanced migration, were elevated. Overall, this study reinforces the application of CAP and redox stress in skin repair physiology.
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    Particulate Matter Dispersion Modeling in Agricultural Applications: Investigation of a Transient Open Source Solver
    (Basel : MDPI, 2021) Janke, David; Swaminathan, Senthilathiban; Hempel, Sabrina; Kasper, Robert; Amon, Thomas
    Agriculture is a major emitter of particulate matter (PM), which causes health problems and can act as a carrier of the pathogen material that spreads diseases. The aim of this study was to investigate an open-source solver that simulates the transport and dispersion of PM for typical agricultural applications. We investigated a coupled Eulerian–Lagrangian solver within the open source software package OpenFOAM. The continuous phase was solved using transient large eddy simulations, where four different subgrid-scale turbulence models and an inflow turbulence generator were tested. The discrete phase was simulated using two different Lagrangian solvers. For the validation case of a turbulent flow of a street canyon, the flowfield could be recaptured very well, with errors of around 5% for the non-equilibrium turbulence models (WALE and dynamicKeq) in the main regions. The inflow turbulence generator could create a stable and accurate boundary layer for the mean vertical velocity and vertical profile of the turbulent Reynolds stresses R11. The validation of the Lagrangian solver showed mixed results, with partly good agreements (simulation results within the measurement uncertainty), and partly high deviations of up to 80% for the concentration of particles. The higher deviations were attributed to an insufficient turbulence regime of the used validation case, which was an experimental chamber. For the simulation case of PM dispersion from manure application on a field, the solver could capture the influence of features such as size and density on the dispersion. The investigated solver is especially useful for further investigations into time-dependent processes in the near-source area of PM sources.