Final Step Research Articles

Structure, mechanism, and evolution of the last step in vitamin C biosynthesis.

Photosynthetic organisms, fungi, and animals comprise distinct pathways for vitamin C biosynthesis. Besides this diversity, the final biosynthetic step consistently involves an oxidation reaction carried out by the aldonolactone oxidoreductases. Here, we study the origin and evolution of the diversified activities and substrate preferences featured by these flavoenzymes using molecular phylogeny, kinetics, mutagenesis, and crystallographic experiments. We find clear evidence that they share a common ancestor. A flavin-interacting amino acid modulates the reactivity with the electron acceptors, including oxygen, and determines whether an enzyme functions as an oxidase or a dehydrogenase. We show that a few side chains in the catalytic cavity impart the reaction stereoselectivity. Ancestral sequence reconstruction outlines how these critical positions were affixed to specific amino acids along the evolution of the major eukaryotic clades. During Eukarya evolution, the aldonolactone oxidoreductases adapted to the varying metabolic demands while retaining their overarching vitamin C-generating function.

Troubling and diffracting Winnicott’s pioneering approach to playing through Deleuze’s ontology for early childhood education

ABSTRACT This paper diffracts Deleuze and quantum physics through Winnicott’s work to argue for an enrichment to playing. The roots of the play-cognitive hierarchy in Freudian psychoanalysis makes visible that progression and the stages that a child must pass en route to rationality continue to feed educational assumptions that a child must leave playing behind in order to learn. Addressing critiques of Freud’s psychosexual theory of child development, I introduce Winnicott’s work on playing as creative activity and transitional phenomena which cast playing in a positive light. I then turn to Deleuze’s critique of Freud’s foundational concepts of child development influenced by Newtonian physics to argue that a Deleuzian concept of energy aligns with quantum physics, which unsettles the binary play-cognition hierarchy. I introduce a vignette to illustrate a rich array of concepts involved in playing that are read through Winnicott. In the final step, I argue that Winnicott’s transitional object provides a juncture with Deleuze’s concept of the partial/virtual object paving the way to graft an alternative and more processual ontology onto Winnicott’s work on playing that acts beside cognitive connections in ways that bring learning alive and liberate teachers to enter less constrained relationships with children.

Novel Tetrazolium-Based Colorimetric Assay for Helicase nsp13 in SARS-CoV-2

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a human pathogenic virus that encodes for a helicase (SC2Hel) that is essential for viral replication. SC2Hel has the ability to unravel dsRNA or dsDNA in an NTP-dependent manner from the 5′ to 3′ directionality. The standard helicase assay from studies involving SARS-CoV and SARS-CoV-2 have relied on the concept of fluorescence resonance energy transfer. Adding to the collection of helicase assays, herein, we have developed a novel tetrazolium-based colorimetric assay system for the detection of ADP that is produced via SC2Hel activity. This SC2Hel assay combines three enzyme-coupled steps involving the ADP-dependent Thermococcus litoralis glucokinase (TlGlcK), Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase (LmG6PDH), and Clostridium kluyveri diaphorase (CkDIA). Iodonitrotetrazolium chloride (INT), a colorimetric tetrazolium reagent, was used in the final step of the assay that converted into INT-formazan during reduction. INT-formazan in the assay’s buffered solution at pH 7.6 exhibited an intense colorimetric response at a wavelength maximum of 505 nm. The assay exhibited excellent performance characteristics as it revealed a Z’ factor of 0.87 and it has the potential to be further adopted into high-throughput screening studies for therapeutic drug discovery research.

Image Robust Watermarking Method Based on DWT-SVD Transform and Chaotic Map

The existing watermarking algorithms make it difficult to balance the invisibility and robustness of the watermark. This paper proposes a robust image watermarking method based on discrete wavelet transform (DWT), singular value decomposition (SVD), and chaotic maps. This method is a semiblind watermarking method. First, a chaotic logistic-tent map is introduced, employing an extensive chaotic parameter domain. This map is amalgamated with Arnold’s transformation to encrypt the watermark image, thereby bolstering the security of the watermark information. Subsequently, the frequency domain is obtained by applying DWT to the carrier image. Embedding watermarks in the frequency domain ensures the invisibility of the watermark, with a preference for a high-frequency subband after the DWT of the carrier image for enhanced watermark robustness. SVD is then applied to both the high-frequency subband of the carrier image after DWT and the encrypted watermark image. The final step involves embedding the singular values of the encrypted watermark image into the carrier image’s singular values, thereby completing the watermark information embedding process. In simulation experiments, an invisibility test was conducted on various carrier images, yielding peak signal-to-noise ratio (PSNR) values consistently exceeding 43, and structural similarity (SSIM) close to 1. Robustness testing against various types of attacks resulted in normalized correlation (NC) values consistently surpassing 0.9, with bit error rate (BER) values approaching 0. In conclusion, the proposed algorithm satisfies imperceptibility requirements while demonstrating formidable robustness.

Development of a clinical pathway for bariatric surgery as an integral part of a comprehensive treatment for adolescents with severe obesity in the Netherlands.

Introduction In the Netherlands, bariatric surgery in adolescents is currently only allowed in the context of scientific research. Besides this, there was no clinical pathway for bariatric surgery in adolescents. In this paper, the development of a comprehensive clinical pathway for bariatric surgery in adolescents with severe obesity in the is described. Methods The clinical pathway for bariatric surgery in adolescents consists of an eligibility assessment as well as comprehensive peri- and postoperative care. Regarding the eligibility assessment, the adolescents need to be identified by their attending pediatricians and afterwards be evaluated by specialized pediatric obesity units. If the provided treatment is considered to be insufficiently effective, the adolescent will anonymously be evaluated by a national board. This is an additional diligence procedure specifically established for bariatric surgery in adolescents. The national board consists of independent experts regarding adolescent bariatric surgery, and evaluates whether the adolescents meet the criteria defined by the national professional associations. The final step is an assessment by a multidisciplinary team for adolescent bariatric surgery. The various disciplines (pediatrician, bariatric surgeon, psychologist, dietician) evaluate whether an adolescent is eligible for bariatric surgery. In this decision-making process, it is crucial to assess whether the adolescent is expected to adhere to postoperative behavioral changes and follow-up. When an adolescent is deemed eligible for bariatric surgery, he or she will receive preoperative counselling by a bariatric surgeon to decide on the type of bariatric procedure (Roux-en-Y gastric bypass or sleeve gastrectomy). Postoperative care consists of intensive guidance by the multidisciplinary team for adolescent bariatric surgery. In this guidance, several regular appointments are included and additional care will be provided based on the needs of the adolescent and his or her family. Furthermore, the multidisciplinary lifestyle intervention, in which the adolescents participated before bariatric surgery, continues in coordination with the multidisciplinary team for adolescent bariatric surgery, and this ensures long-term counselling and follow-up. Conclusion The implementation of bariatric surgery as an integral part of a comprehensive treatment for adolescents with severe obesity requires the development of a clinical pathway with a variety of disciplines.

Impact of DNA ligase inhibition on the nick sealing of polβ nucleotide insertion products at the downstream steps of base excision repair pathway.

DNA ligase (LIG) I and IIIα finalize base excision repair (BER) by sealing a nick product after nucleotide insertion by DNA polymerase (pol) β at the downstream steps. We previously demonstrated that a functional interplay between polβ and BER ligases is critical for efficient repair, and polβ mismatch or oxidized nucleotide insertions confound final ligation step. Yet, how targeting downstream enzymes with small molecule inhibitors could affect this coordination remains unknown. Here, we report that DNA ligase inhibitors, L67 and L82-G17, slightly enhance hypersensitivity to oxidative stress-inducing agent, KBrO3, in polβ+/+ cells more than polβ-/- null cells. We showed less efficient ligation after polβ nucleotide insertions in the presence of the DNA ligase inhibitors. Furthermore, the mutations at the ligase inhibitor binding sites (G448, R451, A455) of LIG1 significantly affect nick DNA binding affinity and nick sealing efficiency. Finally, our results demonstrated that the BER ligases seal a gap repair intermediate by the effect of polβ inhibitor that diminishes gap filling activity. Overall, our results contribute to understand how the BER inhibitors against downstream enzymes, polβ, LIG1, and LIGIIIα, could impact the efficiency of gap filling and subsequent nick sealing at the final steps leading to the formation of deleterious repair intermediates.

Thermogravimetry as a research tool for the development of an ammonium sulphate roasting process for selective metal extraction from minerals

AbstractThe ammonium sulphate roasting process involves reacting mineral-bearing materials with ammonium sulphate via a solid–solid roasting process and subjecting the resulting roast residue to aqueous leaching. This process enables the simultaneous, non-selective co-extraction of strategic metals from the starting materials. However, effective separation of the extracted metals is often mandatory to produce quality products of high purity. In this study, the combined application of thermogravimetric analysis, X-ray powder diffraction and inductively coupled plasma optical emission spectrometry confirmed the non-selectivity of the process when applied to a South African diamond mine residue residue roasted with ammonium sulphate in a 1:2 mass ratio (m/m) at 450 °C for 2 h, with magnesium, iron and aluminium being co-extracted into water-soluble metal sulphates. Thermogravimetry was then applied to develop a multi-step, multi-temperature selective roasting process using mixtures of pure commercial metal sulphate salts. The first step of the modified process successfully separated iron and aluminium sulphates from magnesium-sulphates in the roast residues by thermally decomposing soluble iron and aluminium sulphates into insoluble oxides via calcination at 750 °C for 2 h. This temperature was lower than the one at which magnesium sulphates convert into magnesium oxide. In the second and final step, iron and aluminium were recovered from the oxide minerals via solid–solid re-roasting with ammonium sulphate at 450 °C for 1 h, causing the oxides to revert back to their water-soluble sulphate forms. The effectiveness of the modified process was subsequently verified using a diamond mine residue, showing that the soluble iron and aluminium contents in the magnesium-bearing leachate could be reduced by over 90%.

Development and Calibration of a Phenomenological Material Model for Steel-Fiber-Reinforced High-Performance Concrete Based on Unit Cell Calculations

A phenomenological material model has been developed to facilitate the efficient numerical analysis of fiber-reinforced high-performance concrete (HPC). The formulation integrates an elasto-plastic phase-field model for simulating fractures within the HPC matrix, along with a superimposed one-dimensional elasto-plasticity model that represents the behavior of the embedded fibers. The Drucker–Prager plasticity and one-dimensional von-Mises plasticity formulations are incorporated to describe the nonlinear material behavior of both the HPC matrix and the fibers, respectively. Specific steps are undertaken during the development and calibration of the phenomenological material model. In the initial step, an experimental and numerical analysis of the pullout test of steel fibers embedded in an HPC matrix is conducted. This process is used to calibrate the micro-mechanical model based on the elasto-plastic phase-field formulation for fracture. In the subsequent step, virtual experiments based on an ellipsoidal unit cell, also with the resolution of fibers (used as a representative volume element, RVE), are simulated to analyze the impact of fiber–matrix interactions and their physical properties on the effective material behavior of fiber-reinforced HPC. In the final step, macroscopic boundary value problems (BVPs) based on a cuboid are simulated on a single scale using the developed phenomenological material model. The resulting macroscopic stress–strain characteristics obtained from both types of simulations, namely simulations of virtual experiments and macroscopic BVPs, are compared. This comparison is utilized for the calibration of material parameters to obtain a regularized solution and to assess the effectiveness of the presented phenomenological material model.

Solving a Mystery with Classical and Dual Photoredox Catalysis: Application of Nickel in the Synthesis of Ergot Alkaloids.

A short synthesis of the ergot alkaloid lysergene and a formal total synthesis of lysergic acid diethylamide (LSD) under the avoidance of palladium and including two nickel-catalyzed steps instead have been developed. A key intermediate of this approach has already been reported by Hendrickson et al. in 2004 (Hendrickson, J.B. et al. Org. Lett. 2004, 6, 3-5), yet the spectral data do not match, adding to doubts about the course of their route. While the final steps of the Hendrickson synthesis could not be reproduced, we were able to leverage the elusive intermediate.

Engineering a coenzyme-independent dioxygenase for one-step production of vanillin from ferulic acid.

The final step of vanillin biosynthesis in plants is reportedly catalyzed by the enzyme VpVAN. Prior to our study, VpVAN was the only reported enzyme that directly converts ferulic acid to vanillin. However, as many characteristics of VpVAN remain unknown, this enzyme is not yet suitable for biocatalytic applications. We show that an enzyme that converts ferulic acid to vanillin in one step could be constructed by modifying a microbial dioxygenase-type enzyme. The engineered enzyme is of biotechnological importance as a tool for the production of vanillin and related compounds via biocatalytic processes and metabolic engineering. The results of this study may also provide useful insights for understanding vanillin biosynthesis in plants.

Reversible Capture Mechanism of CO2 as a Zn(II)-Methylcarbonate.

In this study, we elucidate the reaction mechanism for capturing CO2 with the ZnL1(MeOH) complex (L1 = diacetyl-2-(4-methyl-3-thiosemicarbazone)-3-(2-hydrazinatopyridine)) in a methanol solution, using density functional theory calculations. One pathway involves the protonation of ZnL1(MeOH) by methylcarbonic acid, followed by ligand exchange of MeOH with MeOCO2-. An alternative mechanism suggests a tautomerization between ZnL1(MeOH) and Zn(HL1)(OMe), followed by CO2 insertion. This latter pathway is energetically more favorable than the former and more complex than initially proposed. In fact, we unveiled that the solvent catalyzes tautomerization, as one explicit methanol molecule acts as a proton transfer agent. Then, Zn(HL1)(OMe) captures CO2, yielding a methylcarbonate bound to the metal center. The final step involves a rearrangement that leads to the cleavage of the Zn-O(Me)(COO) bond and the formation of a new Zn-O(COOMe) bond, along with the rotation of the methylcarbonate group. Furthermore, we evaluated the ligand basicity through the pKa calculated values of the Zn(II) complexes, the effects of varying the ligand from 4-methyl-thiosemicarbazone to 4-ethyl (L2), 4-phenethyl (L3), and 4-benzyl (L4) derivatives, and reversibility of the reaction in an argon environment.

Preparation and investigation of green and cheap deep eutectic solvents for determination of essential and toxic metal ions at trace levels in different coffee products

In this research, a vortex-assisted hydrophobic deep eutectic solvent-based dispersive liquid phase microextraction (VA-HDES-DLPME) method was developed to extract some essential (Fe, Mn and Zn) and toxic (Cd and Pb) metal ions from coffee products. After the VA-HDES-DLPME method, the amounts of the metal ions in the HDES-phase were determined by flame atomic absorption spectrophotometry (FAAS). For selective extraction of metal ions, five HDESs (as extraction solvent) were prepared and tested according to the ultrasonic-assisted procedure. Some dispersive solvents were investigated to ensure the dispersion of HDES in the sample solution. In the VA-HDES-DLPME method, heating and centrifugation steps were not required to separate the HDES-phase from the sample solution. Key factors were investigated and optimized to achieve selective and rapid extraction of metal ions. Under optimized conditions, the working range of the VA-HDES-DLPME method for metal ions ranged from 0.03 to 280 μg kg−1. The detection limits of Fe(II), Mn(II), Zn(II), Cd(II), and Pb(II) ions were 0.01, 0.27, 0.02, 0.015, and 0.18 μg kg−1, respectively. Relative standard deviation and extraction recovery for five replicate runs of 5 μg kg−1 of metal ions were in the range of 1.3%–1.9% and 94 ± 4%-98 ± 2%, respectively. Validation of the VA-HDES-DLPME method was tested by studies on three reference materials. In the final step, the VA-HDES-DLPME method was successfully applied to microwave digestion of coffee samples using a standard addition method. The obtained quantitative recoveries showed the good accuracy of the method.

Finite-time resilient control for uncertain periodic piecewise polynomial time-varying systems

This study explores the problem of finite-time resilient control for periodic piecewise polynomial time-varying systems in the face of parameter uncertainties, time-varying state delays and external disturbances. Particularly, the considered system is characterized by dividing the fundamental period of periodic systems into numerous subintervals, each of which can be expressed by using matrix polynomial functions. The foremost intention of this work is to lay out a resilient controller such that the resulting closed-loop system is finite-time bounded and satisfies a mixed [Formula: see text] and passivity performance index. Furthermore, by constructing a periodic piecewise time-varying Lyapunov–Krasovskii functional, a delay-dependent sufficient condition is established in line with Wiritinger’s inequality and matrix polynomial lemma to guarantee the needed outcomes of the system under study. Following this, the gain matrix of the devised controller can be calculated by solving the established constraints. As a final step, we conclude with a numerical example that validates the potential and importance of the theoretical discoveries and the developed control scheme.

Notch signaling without the APH-2/Nicastrin subunit of Gamma Secretase in C. elegans germline stem cells.

The final step in Notch signaling activation is the transmembrane cleavage of Notch receptor by γ secretase. Thus far, genetic and biochemical evidence indicate that four subunits are essential for γ secretase activity in vivo: presenilin (the catalytic core), APH-1, PEN-2, and APH-2/Nicastrin. Although some γ secretase activity has been detected in APH-2/Nicastrin-deficient mammalian cell lines, the lack of biological relevance for this activity has left the quaternary γ secretase model unchallenged. Here we provide the first example of in vivo Notch signal transduction without APH-2/Nicastrin. The surprising dispensability of APH-2/Nicastrin is observed in C. elegans germline stem cells (GSCs), and contrasts with its essential role in previously described C. elegans Notch signaling events. Depletion of GLP-1/Notch, presenilin, APH-1, or PEN-2 causes a striking loss of GSCs. In contrast, aph-2/Nicastrin mutants maintain GSCs, and exhibit robust and localized expression of the downstream Notch target sygl-1. Interestingly, APH-2/Nicastrin is present in GSCs and becomes essential under conditions of compromised Notch function. Further insight is provided by reconstituting the C. elegans γ secretase complex in yeast, where we find that APH-2/Nicastrin increases, but is not essential for γ secretase activity. Together, our results are most consistent with a revised model of γ secretase in which the APH-2/Nicastrin subunit has a modulatory, rather than obligatory role. We propose that a trimeric presenilin-APH-1-PEN-2 γ secretase complex can provide a low level of γ secretase activity, and that cellular context determines whether or not APH-2/Nicastrin is essential for effective Notch signal transduction.

A physics and data co-driven surrogate modeling method for high-dimensional rare event simulation

This paper presents a physics and data co-driven surrogate modeling method for efficient rare event simulation of civil and mechanical systems with high-dimensional input uncertainties. The method fuses interpretable low-fidelity physical models with data-driven error corrections. The hypothesis is that a well-designed and well-trained simplified physical model can preserve salient features of the original model, while data-fitting techniques can fill the remaining gaps between the surrogate and original model predictions. The coupled physics-data-driven surrogate model is adaptively trained using active learning, aiming to achieve a high correlation and small bias between the surrogate and original model responses in the critical parametric region of a rare event. A final importance sampling step is introduced to correct the surrogate model-based probability estimations. Static and dynamic problems with input uncertainties modeled by random field and stochastic process are studied to demonstrate the proposed method.

Zn Oxide Nanoparticles and Fine Particles: Synthesis, Characterization and Evaluation of the Toxic Effect on Germination and Vigour of Solanum licopersicum L.

The micronutrient Zinc (Zn) is essential for the proper growth and development of crops. Zinc oxide nanoparticles (NPs) and fine particles are presented as an emerging alternative to more traditional fertilizers. In this study, the possible toxic effect of four laboratory-synthesized ZnO NPs and fine particles with different characteristics on tomato seed germination and vigor parameters was examined in comparison to bulk ZnO. Different metal precursors were used for the chemical synthesis of the particles: Zn(NO3)2 and ZnSO4, for particles named NIT-. and SUL-., respectively. In addition, the synthesis process was modified to obtain coated particles (denoted as UW-, NIT-UW and SUL-UW) and washed particles (denoted as W-, NIT-W and SUL-W). These particles were applied at different toxic doses (0, 1.4, 2.8, 5.6 and 11.2 g L−1). The results indicated that although the constant contact time between the ZnO particles did not affect the germination capacity of the seeds, it affected the growth of roots and hypocotyls, with a higher impact on the roots’ development. This toxicity was more evident from the lowest particle dose used, although it did not prevent radicle and hypocotyl elongation during the development period studied (14 days). The synthesized coated particles (NIT-UW, SUL-UW) generated high toxicity on radicle and hypocotyl development, and this effect was observed from the first days of contact with the particles. The observed toxic effects on radicle length were minimized by the application of bulk ZnO particles. In the case of hypocotyl growth, these minor toxic effects were observed by using NIT-W particles and bulk ZnO. The possibility of positive effects on seed germination and development (radicle and hypocotyl length) when in continuous contact with ZnO, whether in fine particles, nanoparticles, or bulk sizes, was excluded. Furthermore, no benefits on germination parameters were observed by suppressing the final washing step in the particle’s synthesis process, suggesting that particle coating did not provide any advantage for seed germination under these continuous contact conditions.

Physical activity moderates the association between school start time and sleep duration in a cross-sectional national sample of adolescents

BackgroundAdolescent insufficient sleep is an endemic issue that may result in compromised functioning throughout the course of the day and is associated with increased risk for a variety of adverse outcomes. Early school start time (SST) has been consistently found to be detrimental to adolescents’ sleep achievement on school nights. However, there are logistical barriers to changing SST. Evidence supports daily engagement in moderate-to-vigorous intensity physical activity (MVPA) to enhance adolescents’ nightly sleep achievement. However, the role of MVPA in the association between SST and sleep duration is unknown. This study examines the potential moderating effect of MVPA in the association between SST and sleep duration on a typical school day among adolescents.MethodsThis study examined data (collected in April and October 2014) from a national sample of 1132 adolescents (mage = 14.5 years) living in the United States from the Family Life, Activity, Sun, Health, and Eating study, a cross-sectional, internet-based survey. First, three linear regressions were computed to examine bivariate associations between SST, MVPA, and sleep duration while controlling for participant sex, race and ethnicity, household income, school level, and the presence of a TV in the bedroom. Next, a three-step multiple regression was computed with sleep duration as the dependent variable, and the final step included an interaction term between SST and MVPA.ResultsLater SST (b1 = 0.41, p < 0.001) and increased MVPA (b1 = 0.39, p < 0.001) were both associated with increased sleep duration, while SST and MVPA were not significantly associated. In the final multiple regression model, which included the interaction term, school day MVPA moderated the positive association between SST and school night sleep duration (b1 = − 3.7, p < 0.05), such that the greater the MVPA on a typical school day, the weaker the positive association between early SST and sleep duration. In post-hoc analysis, the interaction effect was only significant for females and not males.ConclusionsThe significant buffering effect of MVPA on the association between SST and sleep duration suggests that in the absence of SST changes, promoting MVPA among adolescents may be a promising strategy to mitigate insufficient sleep among US adolescents.

A simple and unified protocol to purify all seven Escherichia coli RNA polymerase sigma factors.

RNA polymerase sigma factors are indispensable in the process of bacterial transcription. They are responsible for a given gene's promoter region recognition on template DNA and hence determine specificity of RNA polymerase and play a significant role in gene expression regulation. Here, we present a simple and unified protocol for purification of all seven Escherichia coli RNA polymerase sigma factors. In our approach, we took advantage of the His8-SUMO tag, known to increase protein solubilization. Sigma factors were first purified in N-terminal fusions with this tag, which was followed by tag removal with Ulp1 protease. This allowed to obtain proteins in their native form. In addition, the procedure is simple and requires only one resin type. With the general protocol we employed, we were able to successfully purify σD, σE, σS, and σN. Final step modification was required for σF, while for σH and σFecI, denaturing conditions had to be applied. All seven sigma factors were fully functional in forming an active holoenzyme with core RNA polymerase which we demonstrated with EMSA studies.

67 Decontamination of a feed manufacturing environment following inoculation of porcine epidemic diarrhea virus, porcine reproductive and respiratory virus, and Seneca Valley virus 1

Abstract Feed mill decontamination is difficult because equipment is not designed to be cleaned with water. Alternate strategies may improve the ability of a mill to decontaminate in the event of viral contamination. The objective was to evaluate different decontamination strategies within a mill following the inoculation of porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV), and Seneca Valley virus 1 (SVV1) contaminated feed. A batch of feed was inoculated with PEDV, PRRSV, and SVV1 and ran through a mixer, bucket elevator, corn cleaner, screw conveyor, distributor head, and spout. Following mill inoculation, decontamination strategies were implemented with environmental samples collected after each decontamination step. Decontamination strategies included: 1) complete facility decontamination including organic matter removal with hot water power washing, disinfection with 1% peroxygen (Virkon S, Lanxess, Germany) followed by a rinse, and a second disinfection with 5% sodium hypochlorite (Clorox, Oakland, CA) followed by a heating period for 48 h once the facility reached 60°C; 2) chlorine dioxide application (ProOxine AH, Bio-Cide International, Inc., Norman, OK); 3) organic matter removal using vacuums (Ridge Tool Company, Elyria, OH) and chlorine dioxide application; 4) heat up with portable electric heaters for exactly 48 h; and 5) organic matter removal and heat up with portable heaters for exactly 48 h. Samples were analyzed via triplex PCR. Cycle threshold and proportion PCR positive were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC). A swine bioassay was completed to determine the infectivity of samples following the final decontamination step of each treatment. A treatment × decontamination step × location interaction was observed (P &amp;lt; 0.05) for SVV1, where less RNA was detected post-treatment compared with pre-treatment following complete facility decontamination on surfaces including the mixer, corn cleaner, screw conveyor, and floor around the feed discharge (P &amp;lt; 0.05). There was no statistical difference between pre- and post-treatment samples for other combinations of decontamination treatment and location (P &amp;gt; 0.05), although numerical increases in Ct were generally observed following treatment. Across all treatments, the act of decontamination reduced detectable PEDV (P &amp;lt; 0.05) and PRRSV (P &amp;lt; 0.05) RNA when compared with samples immediately following inoculation, but complete facility decontamination was the only treatment where PEDV and PRRSV was non-detectable in all locations. Pigs given samples post-treatment showed no evidence of SVV1 or PEDV infection. Although PRRSV RNA was rarely found via PCR from samples collected within the mill, PRRSV infection was observed in pigs given the chlorine dioxide with and without organic matter removal treatments and the organic matter removal plus heat treatment. Overall, all treatments reduced detectable RNA for all viruses between the inoculation step and the final decontamination step; however, PRRSV particles remained infectious following decontamination regardless of PCR results in a swine bioassay.

Are Clay Minerals Systematically the Products of Aqueous Alteration in Cosmic Bodies?

The formation of chondrite materials represents one of the earliest mineralogical processes in the solar system. Phyllosilicates are encountered at various stages of the chondrule formation, from the initial stages (IDP agglomerates) to the final steps (chondrule internal alteration). While typically linked to aqueous alteration, recent studies reveal that phyllosilicates could precipitate directly from residual fluids in post-magmatic or deuteric conditions and under a wide range of temperatures, pressures, water/rock ratios, and H2/H2O ratio conditions. This study re-examined the formation of hydrated phyllosilicates in chondrules and associated fine-grained rims (FGRs) using published petrographical, mineralogical, and chemical data on carbonaceous chondrites. Given that chondrules originate from the melting of interplanetary dust particles, the water liberated by the devolatilization of primary phyllosilicates, including clay minerals or ice melting, reduces the melting temperature and leads to water dissolution into the silicate melt. Anhydrous minerals (e.g., olivine and diopside) form first, while volatile and incompatible components are concentrated in the residual liquid, diffusing into the matrix and forming less porous FGRs. Serpentine and cronstedtite are the products of thermal metamorphic-like mineral reactions. The mesostasis in some lobated chondrules is composed of anhydrous and hydrous minerals, i.e., diopside and serpentine. The latter is probably not the alteration product of a glassy precursor but rather a symplectite component (concomitant crystallization of diopside and serpentine). If so, the symplectite has been formed at the end of the cooling process (eutectic-like petrographical features). Water trapped inside chondrule porosity can lead to the local replacement of olivine by serpentine without external water input (auto-alteration). In the absence of water, hydrated phyllosilicates do not crystallize, forming a different mineral assemblage.