bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantitative Super-Resolution Microscopy to Assess Adhesion of Neuronal Cells on Single-Layer Graphene Substrates
Authors: Scalisi Silvia , Pennacchietti Francesca , Keshavan Sandeep , Derr Nathan D. , Diaspro Alberto , Pisignano Dario , Pierzynska-Mach Agnieszka , Dante Silvia , Cella Zanacchi Francesca
Published: November 2021

Single Layer Graphene (SLG) has emerged as a critically important nanomaterial due to its unique optical and electrical properties and has become a potential candidate for biomedical applications, biosensors, and tissue engineering. Due to its intrinsic 2D nature, SLG is an ideal surface for the development of large-area biosensors and, due to its biocompatibility, can be easily exploited as a substrate for cell growth. The cellular response to SLG has been addressed in different studies with high cellular affinity for graphene often detected. Still, little is known about the molecular mechanism that drives/regulates the cellular adhesion and migration on SLG and SLG-coated interfaces with respect to other substrates. Within this scenario, we used quantitative super-resolution microscopy based on single-molecule localization to study the molecular distribution of adhesion proteins at the nanoscale level in cells growing on SLG and glass. In order to reveal the molecular mechanisms underlying the higher affinity of biological samples on SLG, we exploited stochastic optical reconstruction microscopy (STORM) imaging and cluster analysis, quantifying the super-resolution localization of the adhesion protein vinculin in neurons and clearly highlighting substrate-related correlations. Additionally, a comparison with an epithelial cell line (Chinese Hamster Ovary) revealed a cell dependent mechanism of interaction with SLG.
Investigation on the Correlation between Inclusions and High Temperature Urea Corrosion Behavior in Ferritic Stainless Steel
Authors: Wang Xuelin , Lu Qingsong , Zhang Wei , Xie Zhenjia , Shang Chengjia
Published: November 2021

The influence of inclusion size and number density on high-temperature urea corrosion (HTUC) behavior of ferritic stainless steels was investigated in a simulated working environment of selective catalytic reduction (SCR) system in commercial vehicles. There is a positive correlation between the control level of inclusions and the resistance of HTUC. By slightly increasing the content of Nb in ferritic stainless steels, the inclusions, especially TiN, were significantly refined, and thus displayed an improvement in HTUC resistance. The interface between inclusions and the matrix becomes a fast channel for chromium precipitation during high-temperature nitriding induced by the decomposition of urea. Chromium nitrides will precipitate around the inclusions and wrap the inclusions, which will decrease the chromium equivalent of the matrix and reduce the resistance of ferritic stainless steels to HTUC. In addition, the high-temperature oxidation accompanied with thermal fatigue also makes the inclusions more likely to become the crack nucleation source, which can accelerate the material thinning and reduce its service life.
Practical Aspects of the Design and Use of the Artificial Neural Networks in Materials Engineering
Authors: Sitek Wojciech , Trzaska Jacek
Published: November 2021

Artificial neural networks are an effective and frequently used modelling method in regression and classification tasks in the area of steels and metal alloys. New publications show examples of the use of artificial neural networks in this area, which appear regularly. The paper presents an overview of these publications. Attention was paid to critical issues related to the design of artificial neural networks. There have been presented our suggestions regarding the individual stages of creating and evaluating neural models. Among other things, attention was paid to the vital role of the dataset, which is used to train and test the neural network and its relationship to the artificial neural network topology. Examples of approaches to designing neural networks by other researchers in this area are presented.
Reduction of Die Wear and Structural Defects of Railway Screw Spike Heads Estimated by FEM
Authors: Alcázar Jackeline , Abate Germán , Antunez Nazareno , Simoncelli Alejandro , Egea Antonio J. Sánchez , Krahmer Daniel Martinez , López de Lacalle Norberto
Published: November 2021

Railway spike screws are manufactured by hot forging on a massive scale, due to each kilometer of railway track needing 8600 spike screws. These components have a low market value, so the head must be formed in a single die stroke. The service life of the dies is directly related to the amount of energy required to form a single screw. The existing standard for spike screws specifies only the required tolerances for the head dimensions, particularly the angle of the hub faces and the radius of agreement of the hub with the cap. Both geometrical variables of the head and process conditions (as-received material diameter and flash thickness) are critical parameters in spike production. This work focuses on minimizing the energy required for forming the head of a railway spike screw by computational simulation. The variables with the highest degree of incidence on the energy, forging load, and filling of the die are ordered statistically. The results show that flash thickness is the variable with the most significant influence on forming energy and forming load, as well as on die filling. Specifically, the minimum forming energy was obtained for combining of a hub wall angle of 1.3° an as-received material diameter of 23.54 mm and a flash thickness of 2.25 mm. Flash thickness generates a lack of filling at the top vertices of the hub, although this defect does not affect the functionality of the part or its serviceability. Finally, the wear is mainly concentrated on the die splice radii, where the highest contact pressure is concentrated according to the computational simulation results.
BRAF-Inhibitor-Induced Metabolic Alterations in A375 Melanoma Cells
Authors: Karki Prashant , Sensenbach Shayne , Angardi Vahideh , Orman Mehmet A.
Published: November 2021

Acquired drug tolerance has been a major challenge in cancer therapy. Recent evidence has revealed the existence of slow-cycling persister cells that survive drug treatments and give rise to multi-drug-tolerant mutants in cancer. Cells in this dynamic persister state can escape drug treatment by undergoing various epigenetic changes, which may result in a transient metabolic rewiring. In this study, with the use of untargeted metabolomics and phenotype microarrays, we characterize the metabolic profiles of melanoma persister cells mediated by treatment with vemurafenib, a BRAF inhibitor. Our findings demonstrate that metabolites associated with phospholipid synthesis, pyrimidine, and one-carbon metabolism and branched-chain amino acid metabolism are significantly altered in vemurafenib persister cells when compared to the bulk cancer population. Our data also show that vemurafenib persisters have higher lactic acid consumption rates than control cells, further validating the existence of a unique metabolic reprogramming in these drug-tolerant cells. Determining the metabolic mechanisms underlying persister cell survival and maintenance will facilitate the development of novel treatment strategies that target persisters and enhance cancer therapy.
Development of HIV Drug Resistance in a Cohort of Adults on First-Line Antiretroviral Therapy in Tanzania during the Stavudine Era
Authors: Sangeda Raphael Z. , Gómes Perpétua , Rhee Soo-Yon , Mosha Fausta , Camacho Ricardo J. , Van Wijngaerden Eric , Lyamuya Eligius F. , Vandamme Anne-Mieke
Published: November 2021

As more HIV patients start combination antiretroviral therapy (cART), the emergence of HIV drug resistance (HIVDR) is inevitable. This will have consequences for the transmission of HIVDR, the success of ART, and the nature and trend of the epidemic. We recruited a cohort of 223 patients starting or continuing their first-line cART in Tanzania towards the end of the stavudine era in 2010. Patients were then followed for one year. Of those with a viral load test at baseline and follow-up time, 34% had a detectable viral load at the one-year endpoint. For 41 patients, protease and reverse transcriptase genotyping were successful. Eighteen samples were from cART-naïve patients, and 23 samples were taken under therapy either at baseline for cART-experienced patients or from follow-up samples for both cART–naïve and cART–experienced patients. The isolates were subtype A, followed by C and D in 41.5%, 22%, and 12.2% of the patients, respectively. No transmitted HIVDR was detected, as scored using the surveillance drug resistance mutations (DRMs) list. However, in 3 of the 18 samples from cART-naïve patients, the clinical Rega interpretation algorithm scored 44D or 138A as non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance-associated polymorphisms. The most observed nucleoside reverse transcriptase inhibitor (NRTI) mutation was 184V. The mutation was found in 16 patients, causing resistance to lamivudine and emtricitabine. Nineteen patients had NNRTI resistance mutations, the most common of which was 103N, observed in eight patients. These high levels of resistance call for regular drug resistance surveillance in Tanzania to inform the control of the emergence and transmission of HIVDR.
Consequences of Both Coxsackievirus B4 and Type 1 Diabetes on Female Non-Obese Diabetic Mouse Kidneys
Authors: Walter Debra L. , Thuma Jean R. , Malgor Ramiro , Schwartz Frank L. , McCall Kelly D. , Coschigano Karen T.
Published: November 2021

Despite the 2019 Executive Order on Advancing American Kidney Health Initiative, kidney disease has moved up in rank from the 9th to the 8th leading cause of death in the United States. A recent push in the field of nephrology has been to identify molecular markers and/or molecular profiles involved in kidney disease process or injury that can help identify the cause of injury and predict patient outcomes. While these studies have had moderate success, they have not yet considered that many of the health conditions that cause kidney disease (diabetes, hypertension, etc.) can also be caused by environmental factors (such as viruses), which in and of themselves can cause kidney disease. Thus, the goal of this study was to identify molecular and phenotypic profiles that can differentiate kidney injury caused by diabetes (a health condition resulting in kidney disease) and coxsackievirus B4 (CVB4) exposure (which can cause diabetes and/or kidney disease), both alone and together. Non-obese diabetic (NOD) mice were used for this study due to their susceptibility to both type 1 diabetes (T1D)- and CVB4-mediated kidney injury, in order to glean a better understanding of how hyperglycemia and viral exposure, when occurring on their own and in combination, may alter the kidneys’ molecular and phenotypic profiles. While no changes in kidney function were observed, molecular biomarkers of kidney injury were significantly up- and downregulated based on T1D and CVB4 exposure, both alone and together, but not in a predictable pattern. By combining individual biomarkers with function and phenotypic measurements (i.e., urinary albumin creatinine ratio, serum creatinine, kidney weight, and body weight), we were able to perform an unbiased separation of injury group based on the type of injury. This study provides evidence that unique kidney injury profiles within a kidney disease health condition are identifiable, and will help us to identify the causes of kidney injury in the future.
Origin of the Pd-Rich Pentlandite in the Massive Sulfide Ores of the Talnakh Deposit, Norilsk Region, Russia
Authors: Kalugin Valery , Gusev Viktor , Tolstykh Nadezhda , Lavrenchuk Andrey , Nigmatulina Elena
Published: November 2021

Pd-rich pentlandite (PdPn) along with ore-forming pentlandite (Pn) occurs in the cubanite and chalcopyrite massive sulfide ores in the EM-7 well of the Southern-2 ore body of the Talnakh deposit. PdPn forms groups of small grains and comprises marginal areas in large crystals of Pn. The palladium content in PdPn reaches up to 11.26 wt.%. EDS elemental mapping and a contour map of palladium concentrations indicate distinct variations in the palladium content within and between individual grains. Palladium distribution in the large grains is uneven and non-zoned. PdPn was formed as the result of a superimposed process, which is not associated with either the sulfide liquid crystallization or the subsolidus transformations of sulfides. Deming regression calculations demonstrated the isomorphic substitution character of Ni by 0.71 Pd and 0.30 Fe (apfu), leading to PdPn occurrence. The replacement of Ni by Fe may also indicate a change in sulfur fugacity, compared to that taking place during the crystallization of the primary Pn. The transformation of Pn into PdPn could have occurred under the influence of a Pd-bearing fluid, which separated from the crystallizing body of the massive sulfide ores.
Advances on the Fragmentation-Energy Fan Concept and the Swebrec Function in Modeling Drop Weight Testing
Authors: Ouchterlony Finn , Sanchidrián José A. , Genç Ömürden
Published: November 2021

The breakage index equation (BIE), or t10 model from drop weight testing (DWT) data for rocks and ores is used in the design of crushers and mills. Such models are becoming increasingly difficult to visualize as the number of variables increases. The so-called double fan BIE, combined with the Swebrec distribution’s accurate description of the sieving curves, is applied to the modelling of drop-weight test fragmentation. The key parameters are geometric properties visible in the fan plot; slopes of straight lines and their point of convergence. The ability of the double fan BIE to reproduce DWT data had been previously established for 8 rocks with 480 DWT data sets. Here the fidelity of the double fan BIE is further evaluated for 18 new materials, based on 281 data sets. The fidelity of the double fan BIE with three fan lines is on par with the fidelity of the current state-of-the-art models for the new materials. Besides the breakage index equation, the new double fan BIE’s t10 equation produces, without additional parameters or fitted constants, the general breakage surface equation tn for an arbitrary n value as a bonus. The specific sieving curve for any combination of particle size and impact energy is also contained in the same formula. The result is an accurate, compact and transparent model.
Anthracene and Pyrene Biodegradation Performance of Marine Sponge Symbiont Bacteria Consortium
Authors: Marzuki Ismail , Asaf Ruzkiah , Paena Mudian , Athirah Admi , Nisaa Khairun , Ahmad Rasheed , Kamaruddin Mudyawati
Published: November 2021

Every petroleum-processing plant produces sewage sludge containing several types of polycyclic aromatic hydrocarbons (PAHs). The degradation of PAHs via physical, biological, and chemical methods is not yet efficient. Among biological methods, the use of marine sponge symbiont bacteria is considered an alternative and promising approach in the degradation of and reduction in PAHs. This study aimed to explore the potential performance of a consortium of sponge symbiont bacteria in degrading anthracene and pyrene. Three bacterial species (Bacillus pumilus strain GLB197, Pseudomonas stutzeri strain SLG510A3-8, and Acinetobacter calcoaceticus strain SLCDA 976) were mixed to form the consortium. The interaction between the bacterial consortium suspension and PAH components was measured at 5 day intervals for 25 days. The biodegradation performance of bacteria on PAH samples was determined on the basis of five biodegradation parameters. The analysis results showed a decrease in the concentration of anthracene (21.89%) and pyrene (7.71%), equivalent to a ratio of 3:1, followed by a decrease in the abundance of anthracene (60.30%) and pyrene (27.52%), equivalent to a ratio of 2:1. The level of pyrene degradation was lower than that of the anthracene due to fact that pyrene is more toxic and has a more stable molecular structure, which hinders its metabolism by bacterial cells. The products from the biodegradation of the two PAHs are alcohols, aldehydes, carboxylic acids, and a small proportion of aromatic hydrocarbon components.
Preparation and Application of a Hydrochar-Based Palladium Nanocatalyst for the Reduction of Nitroarenes
Authors: Çalışkan Melike , Akay Sema , Kayan Berkant , Baran Talat , Kalderis Dimitrios
Published: November 2021

In the present study, a novel heterogeneous catalyst was successfully fabricated through the decoration of palladium nanoparticles on the surface of designed Fe3O4-coffee waste composite (Pd-Fe3O4-CWH) for the catalytic reduction of nitroarenes. Various characterization techniques such as XRD, FE-SEM and EDS were used to establish its nano-sized chemical structure. It was determined that Pd-Fe3O4-CWH is a useful nanocatalyst, which can efficiently reduce various nitroarenes, including 4-nitrobenzoic acid (4-NBA), 4-nitroaniline (4-NA), 4-nitro-o-phenylenediamine (4-NPD), 2-nitroaniline (2-NA) and 3-nitroanisole (3-NAS), using NaBH4 in aqueous media and ambient conditions. Catalytic reactions were monitored with the help of high-performance liquid chromatography. Additionally, Pd-Fe3O4-CWH was proved to be a reusable catalyst by maintaining its catalytic activity through six successive runs. Moreover, the nanocatalyst displayed a superior catalytic performance compared to other catalysts by providing a shorter reaction time to complete the reduction in nitroarenes.
T-2 Toxin—The Most Toxic Trichothecene Mycotoxin: Metabolism, Toxicity, and Decontamination Strategies
Authors: Janik Edyta , Niemcewicz Marcin , Podogrocki Marcin , Ceremuga Michal , Stela Maksymilian , Bijak Michal
Published: November 2021

Among trichothecenes, T-2 toxin is the most toxic fungal secondary metabolite produced by different Fusarium species. Moreover, T-2 is the most common cause of poisoning that results from the consumption of contaminated cereal-based food and feed reported among humans and animals. The food and feed most contaminated with T-2 toxin is made from wheat, barley, rye, oats, and maize. After exposition or ingestion, T-2 is immediately absorbed from the alimentary tract or through the respiratory mucosal membranes and transported to the liver as a primary organ responsible for toxin's metabolism. Depending on the age, way of exposure, and dosage, intoxication manifests by vomiting, feed refusal, stomach necrosis, and skin irritation, which is rarely observed in case of mycotoxins intoxication. In order to eliminate T-2 toxin, various decontamination techniques have been found to mitigate the concentration of T-2 toxin in agricultural commodities. However, it is believed that 100% degradation of this toxin could be not possible. In this review, T-2 toxin toxicity, metabolism, and decontamination strategies are presented and discussed.
Antioxidant and Starch-Hydrolyzing Enzymes Inhibitory Properties of Striga-Resistant Yellow-Orange Maize Hybrids
Authors: Elemosho Abdulazeez Olamilekan , Irondi Emmanuel Anyachukwu , Alamu Emmanuel Oladeji , Ajani Emmanuel Oladipo , Menkir Abebe , Maziya-Dixon Busie
Published: November 2021

Most of the health benefits derived from cereals are attributed to their bioactive compounds. This study evaluated the levels of the bioactive compounds, and the antioxidant and starch-hydrolyzing enzymes inhibitory properties of six pipeline Striga-resistant yellow-orange maize hybrids (coded AS1828-1, 4, 6, 8, 9, 11) in vitro. The maize hybrids were grown at the International Institute of Tropical Agriculture (IITA), Nigeria. The bioactive compounds (total phenolics, tannins, flavonoids, and phytate) levels, antioxidant (DPPH• and ABTS•+ scavenging capacity and reducing power) and starch-hydrolyzing enzymes (α-amylase and α-glucosidase) inhibitory activities of the maize hybrids were determined by spectrophotometry. At the same time, carotenoids were quantified using a reverse-phase HPLC system. The ranges of the bioactive compounds were: 11.25–14.14 mg GAE/g (total phenolics), 3.62–4.67 mg QE/g (total flavonoids), 3.63–6.29 mg/g (tannins), 3.66–4.31% (phytate), 8.92–12.11 µg/g (total xanthophylls), 2.42–2.89 µg/g (total β-carotene), and 3.17–3.77 µg/g (total provitamin A carotenoids). Extracts of the maize hybrids scavenged DPPH• (SC50: 9.07–26.35 mg/mL) and ABTS•+ (2.65–7.68 TEAC mmol/g), reduced Fe3+ to Fe2+ (0.25 ± 0.64–0.43 ± 0.01 mg GAE/g), and inhibited α-amylase and α-glucosidase, with IC50 ranges of 26.28–52.55 mg/mL and 47.72–63.98 mg/mL, respectively. Among the six clones of the maize hybrids, AS1828-9 had the highest (p < 0.05) levels of tannins and phytate and the strongest antioxidant and starch-hydrolyzing enzymes inhibitory activities. Significant correlations were observed between total phenolics and the following: ABTS•+ (p < 0.01, r = 0.757), DPPH• SC50 (p < 0.01, r = −0.867), reducing power (p < 0.05, r = 0.633), α-amylase IC50 (p < 0.01, r = −0.836) and α-glucosidase IC50 (p < 0.05, r = −0.582). Hence, the Striga-resistant yellow-orange maize hybrids (especially AS1828-9) may be beneficial for alleviating oxidative stress and postprandial hyperglycemia.
32 × 32 Pixelated High-Power Flip-Chip Blue Micro-LED-on-HFET Arrays for Submarine Optical Communication
Authors: Kim Tae Kyoung , Islam Abu Bashar Mohammad Hamidul , Cha Yu-Jung , Kwak Joon Seop
Published: November 2021

This work proposes the use of integrated high-power InGaN/GaN multiple-quantum-well flip-chip blue micro light-emitting diode (μ-LED) arrays on an AlGaN/GaN-based heterojunction field-effect transistor (HFET), also known as a high electron mobility transistor (HEMT), for various applications: underwater wireless optical communication (UWOC) and smart lighting. Therefore, we demonstrate high-power μ-LED-on-HEMT arrays that consist of 32 × 32 pixelated μ-LED arrays and 32 × 32 pixelated HEMT arrays and that are interconnected by a solder bump bonding technique. Each pixel of the μ-LED arrays emits light in the HEMT on-state. The threshold voltage, the off-state leakage current, and the drain current of the HEMT arrays are −4.6 V, <~1.1 × 10−9 A at gate-to-source voltage (VGS) = −10 V, and 21 mA at VGS = 4 V, respectively. At 12 mA, the forward voltage and the light output power (LOP) of μ-LED arrays are ~4.05 V and ~3.5 mW, respectively. The LOP of the integrated μ-LED-on-HEMT arrays increases from 0 to ~4 mW as the VGS increases from −6 to 4 V at VDD = 10 V. Each pixel of the integrated μ-LEDs exhibits a modulated high LOP at a peak wavelength of ~450 nm, showing their potential as candidates for use in UWOC.
Surfactant-Mediated Co-Existence of Single-Walled Carbon Nanotube Networks and Cellulose Nanocrystal Mesophases
Authors: Attia David , Yekymov Evgenee , Shmidov Yulia , Levi-Kalisman Yael , Mendelson Orit , Bitton Ronit , Yerushalmi-Rozen Rachel
Published: November 2021

Hybrids comprising cellulose nanocrystals (CNCs) and percolated networks of single-walled carbon nanotubes (SWNTs) may serve for the casting of hybrid materials with improved optical, mechanical, electrical, and thermal properties. However, CNC-dispersed SWNTs are depleted from the chiral nematic (N*) phase and enrich the isotropic phase. Herein, we report that SWNTs dispersed by non-ionic surfactant or triblock copolymers are incorporated within the surfactant-mediated CNC mesophases. Small-angle X-ray measurements indicate that the nanostructure of the hybrid phases is only slightly modified by the presence of the surfactants, and the chiral nature of the N* phase is preserved. Cryo-TEM and Raman spectroscopy show that SWNTs networks with typical mesh size from hundreds of nanometers to microns are distributed equally between the two phases. We suggest that the adsorption of the surfactants or polymers mediates the interfacial interaction between the CNCs and SWNTs, enhancing the formation of co-existing meso-structures in the hybrid phases.
Ultrasonic-Assisted Diels–Alder Reaction Exfoliation of Graphite into Graphene with High Resveratrol Adsorption Capacity
Authors: Zhang Jinxing , Ouyang Qi , Gui Qilin , Chen Xiaonong
Published: November 2021

Scalable preparation of graphene with high adsorption capacity is an important prerequisite for fully realizing its commercial application. Herein, we propose an environmentally friendly route for exfoliation of graphene, which is established based on the Diels–Alder reaction. In our route, N-(4-hydroxyl phenyl) maleimide enters between the flakes as an intercalating agent and participates in the Diels–Alder reaction as a dienophile to increase the interlayer spacing of graphite. Then, graphite is exfoliated into graphene with the aid of ultrasound. The exfoliated product is hydroxyl phenyl functionalized graphene with a thickness of 0.5–1.5 nm and an average lateral size of about 500–800 nm. The exfoliation process does not involve any acid or catalyst and would be a safe and environmentally friendly approach. In addition, the exfoliated graphite shows high resveratrol adsorption capacity, which is ten times that of macroporous resins reported in the literature. Thus, the method proposed herein yields exfoliated graphite with high resveratrol adsorption capacity and is of great significance for the mass production of graphene for practical applications.
Kramers–Kronig Relation for Attenuated Total Reflection from a Metal–Dielectric Interface Where Surface Plasmon Polaritons Are Excited
Authors: Ju Heongkyu
Published: November 2021

The applicability of the Kramers–Kronig relation for attenuated total reflection (ATR) from a metal–dielectric interface that can excite surface plasmon polaritons (SPP) is theoretically investigated. The plasmon-induced attenuation of reflected light can be taken as the resonant absorption of light through a virtual absorptive medium. The optical phase shift of light reflected from the SPP-generating interface is calculated using the KK relation, for which the spectral dependence of ATR is used at around the plasmonic resonance. The KK relation-calculated phase shift shows good agreement with that directly obtained from the reflection coefficient, calculated by a field transfer matrix formula at around the resonance. This indicates that physical causality also produces the spectral dependence of the phase of the leakage field radiated by surface plasmons that would interfere with the reflected part of light incident to the interface. This is analogous with optical dispersion in an absorptive medium where the phase of the secondary field induced by a medium polarization, which interferes with a polarization-stimulating incident field, has a spectral dependence that stems from physical causality.
Identification of CTX-M Type ESBL E. coli from Sheep and Their Abattoir Environment Using Whole-Genome Sequencing
Authors: Atlaw Nigatu Aklilu , Keelara Shivaramu , Correa Maria , Foster Derek , Gebreyes Wondwossen , Aidara-Kane Awa , Harden Lyndy , Thakur Siddhartha , Cray Paula J. Fedorka
Published: November 2021

Widespread dissemination of extended-spectrum beta-lactamase (ESBL) Escherichia coli (E. coli) in animals, retail meats, and patients has been reported worldwide except for limited information on small ruminants. Our study focused on the genotypic characterization of ESBL E. coli from healthy sheep and their abattoir environment in North Carolina, USA. A total of 113 ESBL E. coli isolates from sheep (n = 65) and their abattoir environment (n = 48) were subjected to whole-genome sequencing (WGS). Bioinformatics tools were used to analyze the WGS data. Multiple CTX-M-type beta-lactamase genes were detected, namely blaCTX-M-1, blaCTX-M-14, blaCTX-M-15, blaCTX-M-27, blaCTX-M-32, blaCTX-M-55, and blaCTX-M-65. Other beta-lactamase genes detected included blaCMY-2, blaTEM-1A/B/C, and blaCARB-2. In addition, antimicrobial resistance (AMR) genes and/or point mutations that confer resistance to quinolones, aminoglycosides, phenicols, tetracyclines, macrolides, lincosamides, and folate-pathway antagonists were identified. The majority of the detected plasmids were shared between isolates from sheep and the abattoir environment. Sequence types were more clustered around seasonal sampling but dispersed across sample types. In conclusion, our study reported wide dissemination of ESBL E. coli in sheep and the abattoir environment and associated AMR genes, point mutations, and plasmids. This is the first comprehensive AMR and WGS report on ESBL E. coli from sheep and abattoir environments in the United States.
Fusaricidins, Polymyxins and Volatiles Produced by Paenibacillus polymyxa Strains DSM 32871 and M1
Authors: Mülner Pascal , Schwarz Elisa , Dietel Kristin , Herfort Stefanie , Jähne Jennifer , Lasch Peter , Cernava Tomislav , Berg Gabriele , Vater Joachim
Published: November 2021

Paenibacilli are efficient producers of potent agents against bacterial and fungal pathogens, which are of great interest both for therapeutic applications in medicine as well as in agrobiotechnology. Lipopeptides produced by such organisms play a major role in their potential to inactivate pathogens. In this work we investigated two lipopeptide complexes, the fusaricidins and the polymyxins, produced by Paenibacillus polymyxa strains DSM 32871 and M1 by MALDI-TOF mass spectrometry. The fusaricidins show potent antifungal activities and are distinguished by an unusual variability. For strain DSM 32871 we identified numerous yet unknown variants mass spectrometrically. DSM 32871 produces polymyxins of type E (colistins), while M1 forms polymyxins P. For both strains, novel but not yet completely characterized polymyxin species were detected, which possibly are glycosylated. These compounds may be of interest therapeutically, because polymyxins have gained increasing attention as last-resort antibiotics against multiresistant pathogenic Gram-negative bacteria. In addition, the volatilomes of DSM 32781 and M1 were investigated with a GC–MS approach using different cultivation media. Production of volatile organic compounds (VOCs) was strain and medium dependent. In particular, strain M1 manifested as an efficient VOC-producer that exhibited formation of 25 volatiles in total. A characteristic feature of Paenibacilli is the formation of volatile pyrazine derivatives.
Combating Biofilm by Targeting Its Formation and Dispersal Using Gallic Acid against Single and Multispecies Bacteria Causing Dental Plaque
Authors: Albutti Aqel , Gul Muhammad Shoaib , Siddiqui Muhammad Faisal , Maqbool Farhana , Adnan Fazal , Ullah Ihsan , Rahman Ziaur , Qayyum Sadia , Shah Muhammad Ajmal , Salman Muhammad
Published: November 2021

Exploring biological agents to control biofilm is a vital alternative in combating pathogenic bacteria that cause dental plaque. This study was focused on antimicrobial, biofilm formation and biofilm dispersal efficacy of Gallic acid (GA) against bacteria, including Proteus spp., Escherichia coli, Pseudomonas spp., Salmonella spp., Streptococcus mutans, and Staphylococcus aureus and multispecies bacteria. Biofilm was qualitatively and quantitatively assessed by crystal violet assay, florescence microscopy (bacterial biomass (µm2), surface coverage (%)) and extracellular polymeric substances (EPS). It was exhibited that GA (1–200 mg/L) can reduce bacterial growth. However, higher concentrations (100–200 mg/L) markedly reduced (86%) bacterial growth and biofilm formation (85.5%), while GA did not exhibit any substantial dispersal effects on pre-formed biofilm. Further, GA (20–200 mg/L) exhibited 93.43% biomass reduction and 88.6% (p < 0.05) EPS (polysaccharide) reduction. Microscopic images were processed with BioImageL software. It was revealed that biomass surface coverage was reduced to 2% at 200 mg/L of GA and that 13,612 (µm2) biomass was present for control, while it was reduced to 894 (µm2) at 200 mg/L of GA. Thus, this data suggest that GA have antimicrobial and biofilm control potential against single and multispecies bacteria causing dental plaque.
The Response of Iranian Melon (Cucumis melo L.) Accessions to 2,4-D Drift
Authors: Zangoueinejad Rouzbeh , Sirooeinejad Behnaz , Alebrahim Mohammad Taghi , Bajwa Ali Ahsan
Published: November 2021

One of the most widely used auxinic herbicides in southern Iran’s cereal crop fields is 2,4-D; however, the concurrent growing season of off-season melons in this region potentially leads to herbicide drift from cereal fields to the melon fields. To study the response of some Iranian wild melon accessions to three simulated drift rates of 2,4-D, including 112.1, 11.2, and 3.7 g ae ha−1, a field experiment was conducted during 2019 and 2020 growing seasons. It was found that by increasing the herbicide rate from 3.7 to 112.1 g ae ha−1, the level of visual injury increased in all accessions. However, significant variation in herbicide tolerance was observed among different melon accessions. The MEL-R1 was the most tolerant accession with only 20% injury, while MEL-D8 displayed very high injury rate (ca. 90%) as assessed at 6 weeks after treatment during 2019. The accession MEL-S3 was the most tolerant to 2,4-D drift rates (20% injury) at 6 weeks after treatment during 2020. There was no significant difference between the accessions MEL-R1 and MEL-S3 in terms of their response to 2,4-D treatment during both years of the study, as these accessions fully recovered from injury over 6 weeks after herbicide treatment. In addition, only these two accessions were able to produce yield after the application of 2,4-D at the highest rate tested (112.1 g ae ha−1). Therefore, the melon accessions MEL-R1 and MEL-S3 could be recommended for cultivation and even for breeding programs in order to develop 2,4-D-tolerant commercial cultivars in regions where this herbicide is commonly used in cereal crop production adjacent to the melon fields.
Agronomic Strategies to Improve N Efficiency Indices in Organic Durum Wheat Grown in Mediterranean Area
Authors: Carucci Federica , Gatta Giuseppe , Gagliardi Anna , De Vita Pasquale , Bregaglio Simone , Giuliani Marcella Michela
Published: November 2021

Organic farming systems are often constrained by limited soil nitrogen (N) availability. Here we evaluated the effect of foliar organic N and sulphur (S), and selenium (Se) application on durum wheat, considering N uptake, utilization efficiency (NUtE), grain yield, and protein concentration as target variables. Field trials were conducted in 2018 and 2019 on two old (Cappelli and old Saragolla) and two modern (Marco Aurelio and Nadif) Italian durum wheat varieties. Four organic fertilization strategies were evaluated, i.e., the control (CTR, dry blood meal at sowing), the application of foliar N (CTR + N) and S (CTR + S), and their joint use (CTR + NS). Furthermore, a foliar application of sodium selenate was evaluated. Three factors—variety, fertilization strategies and selenium application—were arranged in a split-split-plot design and tested in two growing seasons. The modern variety Marco Aurelio led to the highest NUtE and grain yield in both seasons. S and N applications had a positive synergic effect, especially under drought conditions, on pre-anthesis N uptake, N translocation, NUtE, and grain yield. Se treatment improved post-anthesis N uptake and NUtE, leading to 17% yield increase in the old variety Cappelli, and to 13% and 14% yield increase in Marco Aurelio and Nadif, mainly attributed to NUtE increase. This study demonstrated that the synergistic effect of foliar applications could improve organic durum wheat yields in Mediterranean environments, especially on modern varieties.
Trichoderma hamatum Increases Productivity, Glucosinolate Content and Antioxidant Potential of Different Leafy Brassica Vegetables
Authors: Velasco Pablo , Rodríguez Víctor Manuel , Soengas Pilar , Poveda Jorge
Published: November 2021

Brassica crops include important vegetables known as “superfoods” due to the content of phytochemicals of great interest to human health, such as glucosinolates (GSLs) and antioxidant compounds. On the other hand, Trichoderma is a genus of filamentous fungi that includes several species described as biostimulants and/or biological control agents in agriculture. In a previous work, an endophytic strain of Trichoderma hamatum was isolated from kale roots (Brassica oleracea var. acephala), describing its ability to induce systemic resistance in its host plant. In the present work, some of the main leafy Brassica crops (kale, cabbage, leaf rape and turnip greens) have been root-inoculated with T. hamatum, having the aim to verify the possible capacity of the fungus as a biostimulant in productivity as well as the foliar content of GSLs and its antioxidant potential, in order to improve these “superfoods”. The results reported, for the first time, an increase in the productivity of kale (55%), cabbage (36%) and turnip greens (46%) by T. hamatum root inoculation. Furthermore, fungal inoculation reported a significant increase in the content of total GSLs in cabbage and turnip greens, mainly of the GSLs sinigrin and gluconapin, respectively, along with an increase in their antioxidant capacity. Therefore, T. hamatum could be a good agricultural biostimulant in leafy Brassica crops, increasing the content of GSLs and antioxidant potential of great food and health interest.
Assessing the Weed-Suppressing Potential of Cotton Chromosome Substitution Lines Using the Stair-Step Assay
Authors: Fuller Mary Gracen , Saha Sukumar , Stelly David M. , Jenkins Johnie N. , Tseng Te Ming
Published: November 2021

Palmer amaranth is a problematic common weed species, especially in cotton. With the wide use of chemical herbicide and herbicide-tolerant transgenic cotton lines, Palmer amaranth populations have developed tolerance to commonly used herbicides. It is imperative to develop alternative weed control methods to slow the evolution of herbicide-resistant weed populations and provide new strategies for weed management. Eleven chromosome substitution (CS) cotton lines (CS-B26lo, CS-T17, CS-B16-15, CS-B17-11, CS-B12, CS-T05sh, CS-T26lo, CS-T11sh, CS-M11sh, CS-B22sh, and CS-B22lo) were screened for weed-suppressing abilities in this study. The cotton lines were tested using the established stair-step assay. Height (cm) and chlorophyll concentration (cci) were measured for each plant in the system. The most significant variation in Palmer amaranth height reduction among the CS lines was observed 21 days after establishment. CS-B22sh (76.82%) and T26lo (68.32%) were most effective in reducing Palmer amaranth height. The cluster analysis revealed that CS-B22sh, and CS-T26lo were clustered in one group, suggesting similar genetic potential with reference to Palmer amaranth growth and development. CS-B22sh showed novel genetic potential to control the growth and development of Palmer amaranth, a problematic weed in cotton fields. Future experimentation should implement more parameters and chemical testing to explore allelopathic interactions among CS lines and Palmer amaranth.
Contrasting Metabolic Fingerprints and Seed Protein Profiles of Cucurbita foetidissima and C. radicans Fruits from Feral Plants Sampled in Central Mexico
Authors: Mejía-Morales Claudia , Rodríguez-Macías Ramón , Salcedo-Pérez Eduardo , Zamora-Natera Juan Francisco , Rodríguez-Zaragoza Fabián Alejandro , Molina-Torres Jorge , Délano-Frier John Paul , Zañudo-Hernández Julia
Published: November 2021

Cucurbita foetidissima and C. radicans are scarcely studied wild pumpkin species that grow in arid and semi-arid areas of Mexico and the United States. This study describes the morphological, proximal composition, metabolic finger-prints and seed protein profiles of C. foetidissima and C. radicans fruits collected in the wild during a one-year period in different locations of central-western Mexico. The results obtained complement the limited information concerning the fruit composition of C. foetidissima and greatly expand information in this respect regarding C. radicans. Morphology and proximal composition of their fruits varied significantly. Different metabolic fingerprints and seed protein profiles were detected between them and also with the chemical composition of domesticated Cucurbita fruits. The neutral lipids in seed, pulp and peels were rich in wax content and in unsaturated compounds, probably carotenoids and tocopherols, in addition to tri-, di- and mono-acylglycerols. The tri- and diacylglycerol profiles of their seed oils were different from commercial seed oils and between each other. They also showed unusual fatty acid compositions. Evidence of a possible alkaloid in the pulp and peel of both species was obtained in addition to several putative cucurbitacins. An abundance of phenolic acids was found in all fruit parts, whereas flavonoids were only detected in the peels. Unlike most cucurbits, globulins were not the main protein fraction in the seeds of C. radicans, whereas the non-structural carbohydrate and raffinose oligosaccharide content in their fruit parts was lower than in other wild cucurbit species. These results emphasize the significantly different chemical composition of these two marginally studied Cucurbita species, which was more discrepant in C. radicans, despite the notion regarding C. foetidissima as an aberrant species with no affinity to any other Cucurbita species.
Inhibitory Activity of Shrimp Waste Extracts on Fungal and Oomycete Plant Pathogens
Authors: El boumlasy Soumia , La Spada Federico , Tuccitto Nunzio , Marletta Giovanni , Mínguez Carlos Luz , Meca Giuseppe , Rovetto Ermes Ivan , Pane Antonella , Debdoubi Abderrahmane , Cacciola Santa Olga
Published: November 2021

(1) Background: This study was aimed at determining the in vitro inhibitory effect of new natural substances obtained by minimal processing from shrimp wastes on fungi and oomycetes in the genera Alternaria, Colletotrichum, Fusarium, Penicillium, Plenodomus and Phytophthora; the effectiveness of the substance with the highest in vitro activity in preventing citrus and apple fruit rot incited by P. digitatum and P. expansum, respectively, was also evaluated. (2) Methods: The four tested substances, water-extract, EtOAc-extract, MetOH-extract and nitric-extract, were analyzed by HPLC-ESI-MS-TOF; in vitro preliminary tests were carried out to determine the minimal inhibitory/fungicidal concentrations (MIC and MFC, respectively) of the raw dry powder, EtOAc-extract, MetOH-extract and nitric-extract for each pathogen. (3) Results: in the agar-diffusion-assay, nitric-extract showed an inhibitory effect on all pathogens, at all concentrations tested (100, 75, 50 and 25%); the maximum activity was on Plenodomus tracheiphilus, C. gloeosporioides and Ph. nicotianae; the diameters of inhibition halos were directly proportional to the extract concentration; values of MIC and MFC of this extract for all pathogens ranged from 2 to 3.5%; the highest concentrations (50 to 100%) tested in vivo were effective in preventing citrus and apple fruit molds. (4) Conclusions: This study contributes to the search for natural and ecofriendly substances for the control of pre- and post-harvest plant pathogens.
A Novel Distinct Genetic Variant of Tomato Torrado Virus with Substantially Shorter RNA1-Specific 3’Untranslated Region (3’UTR)
Authors: Budziszewska Marta , Wieczorek Przemysław
Published: November 2021

Tomato torrado virus (ToTV) induces severe systemic necrosis in Solanum lycopersicum. This work aimed at describing the genetic variability of necrosis-inducing ToTV-Wal’17 collected in 2017, derived from the ToTV-Wal’03 after long-term passages in plants. Sequence analyses of the ToTV-Wal’17 indicated twenty-eight single nucleotide substitutions in coding sequence of both RNAs, twelve of which resulted in amino acid changes in viral polyproteins. Moreover the sequencing data revealed that the 3’UTR of ToTV-Wal’17 RNA1 was 394 nts shorter in comparison to Wal’03. The performed sequence analyses revealed that 3’UTR of RNA1 of ToTV-Wal’17 is the most divergent across all previously described European isolates.
The Relationships between Plant Developmental Traits and Winter Field Survival in Rye (Secale cereale L.)
Authors: Bahrani Hirbod , Båga Monica , Larsen Jamie , Graf Robert J. , Laroche Andre , Chibbar Ravindra N.
Published: November 2021

Overwintering cereals accumulate low temperature tolerance (LTT) during cold acclimation in the autumn. Simultaneously, the plants adjust to the colder season by making developmental changes at the shoot apical meristem. These processes lead to higher winter hardiness in winter rye varieties (Secale cereale L.) adapted to Northern latitudes as compared to other cereal crops. To dissect the winter-hardiness trait in rye, a panel of 96 genotypes of different origins and growth habits was assessed for winter field survival (WFS), LTT, and six developmental traits. Best Linear Unbiased Estimates for WFS determined from five field trials correlated strongly with LTT (r = 0.90, p < 0.001); thus, cold acclimation efficiency was the major contributor to WFS. WFS also correlated strongly (p < 0.001) with final leaf number (r = 0.80), prostrate growth habit (r = 0.61), plant height (r = 0.34), but showed weaker associations with top internode length (r = 0.30, p < 0.01) and days to anthesis (r = 0.25, p < 0.05). The heritability estimates (h2) for WFS-associated traits ranged from 0.45 (prostrate growth habit) to 0.81 (final leaf number) and were overall higher than for WFS (h2 = 0.48). All developmental traits associated with WFS and LTT are postulated to be regulated by phytohormone levels at shoot apical meristem.
Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis)
Authors: Rizwan Hafiz Muhammad , Yang Qiang , Yousef Ahmed Fathy , Zhang Xiaoxue , Sharif Yasir , Kaijie Jia , Shi Meng , Li Han , Munir Nigarish , Yang Xuelian , Wei Xiaoxia , Oelmüller Ralf , Cheng Chunzhen , Chen Faxing
Published: November 2021

Passion fruit (Passiflora edulis) is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient Agrobacterium-mediated in planta transformation system for passion fruit was established, using Agrobacterium virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.