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PURPOSE: To evaluate the amount of bone matrix in autogenous block bone grafts that were fixed with or without perforation of the receptor bed. MATERIALS AND METHODS: Twelve rabbits received two 5-mm circular osteotomies each in the anterior parietal region. The bone was removed, perforated, and fixed by a titanium screw in the adjacent area, 3 mm from the border of the osteotomies. On the contralateral side, six perforations were made in the receptor site before the graft was fixated by the titanium screw. After 28 days, the animals were sacrificed and specimens were prepared for histologic study. Sections were analyzed with regard to the total area of the graft and the percentages of bone matrix in the graft, interface, and receptor bed. Analyses using the Wilcoxon test (P<.05) were performed. RESULTS: No statistically significant differences were found between the groups with or without perforations to the area of the graft (P=.85) or the percentage of hard tissue in the grafts (P=.53), the interface (P=.65), or the receptor bed (P=.18). CONCLUSION: Based upon this study that used a rabbit model with bone grafts placed in the parietal regions, cortical perforations of the receptor bone did not improve repair of bone grafts and did not increase the amount of bone matrix in the final grafted area. [ABSTRACT FROM AUTHOR]

Diethylpropion has been available in the market for treating obesity for over 50 years. Refined studies are lacking to fully elucidate its action spectrum. The aim of our study was to evaluate possible toxic effects of anorectic diethylpropion in Chinese hamster ovary (CHO) cells. Comet assay (detects breaks in the DNA strand), micronucleus test (detects clastogenic/aneugenic damage), and cell survival test (detects cytotoxic damage) were used to evaluate the toxic effects. In comet assay, we found that the damage scores with diethylpropion treatments at the concentrations of 20 and 40 μg/mL were more significant ( p < 0.05) than that of the negative control. When assessing the possible aneugenic and/or clastogenic damage caused by the drug in CHO cells, we found no difference ( p > 0.05) in the values of micronucleated cells when comparing different diethylpropion treatments and the negative control. Regarding the cell viability, for all the diethylpropion concentrations tested, higher values ( p < 0.05) of apoptosis were found compared with those of the negative control. In relation to the number of necrotic cells, no difference ( p > 0.05) was noted between the means of the three concentrations of diethylpropion evaluated and the negative control. In the experimental conditions, we conclude that diethylpropion has weak genotoxic and cytotoxic activities. [ABSTRACT FROM AUTHOR]

ETHNOPHARMACOLOGICAL RELEVANCE: Cordia ecalyculata Vell. and Echinodorus grandiflorus (Cham. & Schltdl.) Micheli are extensively used in Brazil as therapeutic preparations for indigenous groups and the general population. These plants have been used in the folk medicine as: tonic, diuretic, anti-inflammatory, appetite suppressants, for the treatment of snake bites, and weight loss. AIM OF THE STUDY: In this study, it was verified the possible cytotoxic and genotoxic effects of the crude extracts of. Cordia ecalyculata and Echinodorus grandiflorus, as well as their effectiveness in treating obesity. MATERIALS AND METHODS: The Micronucleus Test was used for the evaluation of possible clastogenic and aneugenic effects, and the Comet Assay was used for the evaluation of single-strand and double-strand DNA breaks. The cytotoxic effects of the crude extracts were verified by PCE/NCE ratio. Swiss mice (Mus musculus) were used as the experimental model. RESULTS: It was observed a significant (P<0.05) increase, dose-independent, in the average frequency of micronucleated erythrocytes in peripheral blood in mice treated with either the Cordia ecalyculata or Echinodorus grandiflorus extracts, in comparison with the negative control. There were no significant differences (P>0.05) in the frequency of micronucleated polychromatic erythrocytes for both extract treatment. We observed that treatment with the Cordia ecalyculata extract at concentrations of 1000 and 2000 mg/kg bw resulted in a PCE/NCE ratio that was larger (P<0.05) than the negative control. After 15 days of daily treatment, a dose of 2000 mg/kg bw of either phytotherapeutic did not reduce body mass gain or the amount of food consumed by Swiss mice when compared with the negative control (P>0.05). CONCLUSION: The results of this study allowed us to infer that the crude extracts of Cordia ecalyculata and Echinodorus grandiflorus do not display cytotoxic or genotoxic activities. However, they do possess weak clastogenic activity (without significance) on peripheral blood cells. Contrary to commonly held beliefs it was also found in this study that the extracts are not effective for obesity treatments. [ABSTRACT FROM AUTHOR]

Host-pathogen interactions are shaped by the metabolic status of both the host and pathogen. The host must regulate metabolism to fuel the immune response, while the pathogen must extract metabolic resources from the host to enable its own survival. In this study, we focus on the metabolic interactions of Mycobacterium abscessus with Drosophila melanogaster . We identify MAB&#95;1132c as an asparagine transporter required for pathogenicity in M. abscessus . We show that this requirement is specifically associated with damage to the host: flies infected with MAB&#95;1132c knockout bacteria, or with wild-type bacteria grown in asparagine-restricted conditions, are longer lived without showing a significant change in bacterial load. This is associated with a reduction in the host innate immune response, demonstrated by the decreased transcription of antimicrobial peptides as well as a significant reduction in the ability of the infection to disrupt systemic insulin signaling. Much of the increase in host survival during infection with asparagine-limited M. abscessus can be attributed to alterations in unpaired cytokine signaling. This demonstrates that asparagine transport in M. abscessus prior to infection is not required for replicative fitness in vivo but does significantly influence the interaction with the host immune responses., Competing Interests: Competing interests statement:The authors declare no competing interest.

The ability of plants to recover after stressful events is crucial for resuming growth and development and is a key trait when studying stress tolerance. However, there is a lack of information on the physiological responses and the time required to restore homeostasis after the stress experience. This study aimed to (i) enhance understanding of soybean photosynthesis performance during saline waterlogging and (ii) investigate the effects of this combined stress during the reoxygenation and recovery period. Soybean plants (cultivar PELBR10-6049 RR) were subjected to waterlogging, NaCl, or hypoxia + NaCl for 3 and 6 days. Afterward, plants were drained and allowed to recover for an additional two (short-term) and seven days (long-term). Compared to plants exposed to single stress, the combined hypoxia + NaCl treatment resulted in a lower net CO 2 assimilation rate, ФPSII, and levels of photosynthetic pigments during the waterlogging period. Furthermore, hypoxia + NaCl increased foliar electrolyte leakage during waterlogging. In response to short-term reoxygenation, these negative effects were amplified, while prolonged reoxygenation resulted in a slight increase in biomass accumulation. In conclusion, full recovery was not achieved under any condition during the reoxygenation periods tested. Notably, the brief reoxygenation phase imposed greater stress than the initial stress conditions for plants facing combined stress. Although extended recovery increased biomass accumulation, it remained lower in plants previously subjected to saline waterlogging., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Priming enables plants to respond more promptly, minimise damage, and survive subsequent stress events. Here, we aimed to assess the efficacy of priming and cross-priming in mitigating the stress caused by waterlogging and/or dehydration in soybeans (Glycine max ). Soybean plants were cultivated in a greenhouse in plastic pots in which soil moisture was maintained at pot capacity through irrigation. The first stress was applied in plants at the vegetative stage for 5days and involved either dehydration or waterlogging, depending on the treatment. Subsequently, the plants were irrigated or drained and maintained at pot capacity until the second stress. For the second stress, the conditions were repeated in plants at the reproductive stage. We then evaluated the levels of hydrogen peroxide (H2 O2 ), lipid peroxidation, total soluble sugars (TSS), amino acids, proline, and starch, and the activity of antioxidant, fermentative, and aminotransferase enzymes. Under waterlogging and dehydration, priming and cross-priming significantly increased the activity of antioxidant enzymes and the levels of TSS, amino acids, and proline while reducing H2 O2 concentration and lipid peroxidation. Under waterlogging, priming suppressed fermentative activity and increased carbohydrate content. This demonstrates that soybean plants activate their defence systems more promptly when subjected to priming.

Soil waterlogging and drought correspond to contrasting water extremes resulting in plant dehydration. Dehydration in response to waterlogging occurs due to impairments to root water transport, but no previous study has addressed whether limitations to water transport occur beyond this organ or whether dehydration alone can explain shoot impairments. Using common bean (Phaseolus vulgaris) as a model species, we report that waterlogging also impairs water transport in leaves and stems. During the very first hours of waterlogging, leaves transiently dehydrated to water potentials close to the turgor loss point, possibly driving rapid stomatal closure and partially explaining the decline in leaf hydraulic conductance. The initial decline in leaf hydraulic conductance (occurring within 24 h), however, surpassed the levels predicted to occur based solely on dehydration. Constraints to leaf water transport resulted in a hydraulic disconnection between leaves and stems, furthering leaf dehydration during waterlogging and after soil drainage. As leaves dehydrated later during waterlogging, leaf embolism initiated and extensive embolism levels amplified leaf damage. The hydraulic disconnection between leaves and stems prevented stem water potentials from declining below the threshold for critical embolism levels in response to waterlogging. This allowed plants to survive waterlogging and soil drainage. In summary, leaf and stem dehydration are central in defining plant impairments in response to waterlogging, thus creating similarities between waterlogging and drought. Yet, our findings point to the existence of additional players (likely chemicals) partially controlling the early declines in leaf hydraulic conductance and contributing to leaf damage during waterlogging., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Tendons are a frequent site of injury, which greatly impairs the movement and locomotion of patients. Regrettably, injuries at the tendon frequently require surgical intervention, which leads to a long path to recovery. Moreover, the healing of tendons often involves the formation of scar tissue at the site of injury with poor mechanical properties and prone to re-injury. Tissue engineering carries the promise of better and more effective solutions to the improper healing of tendons. Lately, the field of regenerative medicine has seen a significant increase in the focus on the potential use of non-coding RNAs (e.g., siRNAs, miRNAs, and lncRNAs) as molecular tools for tendon tissue engineering. This class of molecules is being investigated due to their ability to act as epigenetic regulators of gene expression and protein production. Thus, providing a molecular instrument to fine-tune, reprogram, and modulate the processes of tendon differentiation, healing, and regeneration. This review focuses particularly on the latest advances involving the use of siRNAs, miRNAs, and lncRNAs in tendon tissue engineering applications., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission., (Copyright © 2024 Peniche Silva, Balmayor and van Griensven.)

Aims and Background: The current report details transition of outsourced conventional dialysis therapy in the ICU services to an in-house prolonged intermittent renal replacement therapy (PIRRT) service model as a quality improvement project using the Tablo Hemodialysis System, Outset Medical, Inc. The goals were aimed at maintaining or improving clinical outcomes, while also reducing dialysis-related nursing staff burden and dialysis-related treatment costs., Methods: A descriptive comparative analysis was conducted of renal replacement therapy (RRT) of ≥6 hours in duration performed in the 1 year prior and 1 year after the ICU's in-house program launch using a PIRRT model including sequential 24-h treatments when medically necessary., Results: Overall, there were 145 intensive care unit (ICU) stays among 145 patients with 13,641 h of conventional ICU dialysis in the year prior to program transition. In the year post, there were 116 ICU stays among 116 patients with 5,098 h of PIRRT. By employing a PIRRT and sequential 24-h treatment strategy vs. the prior outsourced model, the mean dialysis treatment hours per patient were reduced (Pre, 94.1 h with 214 treatment starts; Post, 43.9 h with 370 treatment starts), increasing ICU nurse productivity by 50.2 h per patient. Overall, ICU length of stay and ICU mortality declined post-service transition by 4.8 days and 9.8 percentage points (pp), respectively, overall, and in the non-COVID subset by 1.6 days and 3.1 pp, respectively., Conclusions: Insourcing RRT with an innovative technology that can provide both PIRRT and 24-h sequential treatments can maintain or improve clinical outcomes in critically ill patients requiring RRT in the ICU, while reducing dialysis-related costs.

Main Conclusion: Saline and wet environments stress most plants, reducing growth and yield. Halophytes adapt with ion regulation, energy maintenance, and antioxidants. Understanding these mechanisms aids in breeding resilient crops for climate change. Waterlogging and salinity are two abiotic stresses that have a major negative impact on crop growth and yield. These conditions cause osmotic, ionic, and oxidative stress, as well as energy deprivation, thus impairing plant growth and development. Although few crop species can tolerate the combination of salinity and waterlogging, halophytes are plant species that exhibit high tolerance to these conditions due to their morphological, anatomical, and metabolic adaptations. In this review, we discuss the main mechanisms employed by plants exposed to saline waterlogging, intending to understand the mechanistic basis of their ion homeostasis. We summarize the knowledge of transporters and channels involved in ion accumulation and exclusion, and how they are modulated to prevent cytosolic toxicity. In addition, we discuss how reactive oxygen species production and cell signaling enhance ion transport and aerenchyma formation, and how plants exposed to saline waterlogging can control oxidative stress. We also address the morphological and anatomical modifications that plants undergo in response to combined stress, including aerenchyma formation, root porosity, and other traits that help to mitigate stress. Furthermore, we discuss the peculiarities of halophyte plants and their features that can be leveraged to improve crop yields in areas prone to saline waterlogging. This review provides valuable insights into the mechanisms of plant adaptation to saline waterlogging thus paving the path for future research on crop breeding and management strategies., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Open pit mining can cause loss in different ecosystems, including damage to habitats of rare and endemic species. Understanding the biology of these species is fundamental for their conservation, and to assist in decision-making. Sporobolus multiramosus is an annual grass endemic to the Amazon canga ecosystems, which comprise rocky outcrop vegetation covering one of the world's largest iron ore reserves. Here, we evaluated whether nitric oxide aids S. multiramosus in coping with water shortages and examined the physiological processes behind these adaptations. nitric oxide application improved the water status, photosynthetic efficiency, biomass production, and seed production and germination of S. multiramosus under water deficit conditions. These enhancements were accompanied by adjustments in leaf and root anatomy, including changes in stomata density and size and root endodermis thickness and vascular cylinder diameter. Proteomic analysis revealed that nitric oxide promoted the activation of several proteins involved in the response to environmental stress and flower and fruit development. Overall, the results suggest that exogenous nitric oxide has the potential to enhance the growth and productivity of S. multiramosus . Enhancements in seed productivity have significant implications for conservation initiatives and can be applied to seed production areas, particularly for the restoration of native ecosystems.

The resurgent navigation project known as the Hidrovia Paraguay-Paraná threatens the integrity of the Pantanal, the world's largest wetland, which is considered a biome of its own. Intensive barge navigation is intended to transport crops (soybean, sugar, corn) and cement, iron and manganese from areas of production in Brazil, Paraguay and Bolivia to the oceanic ports of the Plata River. This short communication assembles the information available on the potential impacts of the planned deepening of the natural channel of the Paraguay River in its ~700 km-long upper section. These river channel alterations would disconnect the river from its floodplain, shorten the inundation period, and shrink the wetland area, resulting in severe degradation of the globally outstanding biological and cultural diversity of the Pantanal. The river sediments are mostly sandy and would require perpetual dredging. The reaches needing the most intensive dredging are those of the highest ecological value, protected as a National Park, UNESCO World Heritage and Biosphere Reserves, and various Indigenous reserves and Ramsar sites. Climate change is projected to increase the occurrence of low water periods. Between 2019 and 2021, navigation was impossible even in the already-deepened reaches of the Paraguay River between Corumbá and Asuncion during long periods of the year. Thus, despite considerable financial and technical efforts, the success of the navigation project is doubtful, whereas enormous environmental, cultural, and social impacts can be anticipated. For these reasons, the Brazilian government had already turned down the project in 2000. We suggest alternative, less impactful modes of transport of commodities, e.g., via railway., (Copyright © 2023. Published by Elsevier B.V.)

The Bureau International des Poids et Mesures (BIPM) is developing a new transfer instrument to extend its centralized services for assessing the international equivalence of radioactive standards to new radionuclides. A liquid scintillation counter using the triple/double coincidence ratio method is being studied and tested in the CCRI(II)-P1.Co-60 pilot study. The pilot study, involving 13 participating laboratories with primary calibration capabilities, validated the approach against the original international reference system based on ionization chambers, which has been in operation since 1976. The results are in agreement and an accuracy suitable for purpose, below 5×10 -4 , is achieved. The pilot study also reveals an issue when impurities emitting low-energy electrons are present in the standard solution, which have a different impact on liquid scintillation counting compared to other primary measurement methods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The highly conserved angiosperm immune receptor HOPZ-ACTIVATED RESISTANCE1 (ZAR1) recognises the activity of diverse pathogen effector proteins by monitoring the ZED1-related kinase (ZRK) family. Understanding how ZAR1 achieves interaction specificity for ZRKs may allow for the expansion of the ZAR1-kinase recognition repertoire to achieve novel pathogen recognition outside of model species. We took advantage of the natural diversity of Arabidopsis thaliana kinases to probe the ZAR1-kinase interaction interface and found that A. thaliana ZAR1 (AtZAR1) can interact with most ZRKs, except ZRK7. We found evidence of alternative splicing of ZRK7, resulting in a protein that can interact with AtZAR1. Despite high sequence conservation of ZAR1, interspecific ZAR1-ZRK pairings resulted in the autoactivation of cell death. We showed that ZAR1 interacts with a greater diversity of kinases than previously thought, while still possessing the capacity for specificity in kinase interactions. Finally, using AtZAR1-ZRK interaction data, we rationally increased ZRK10 interaction strength with AtZAR1, demonstrating the feasibility of the rational design of a ZAR1-interacting kinase. Overall, our findings advance our understanding of the rules governing ZAR1 interaction specificity, with promising future directions for expanding ZAR1 immunodiversity., (© 2023 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Hospitals and nursing home wards are areas prone to the propagation of infections and are of particular concern regarding the spreading of dangerous viruses and multidrug-resistant bacteria (MDRB). MDRB infections comprise approximately 20% of cases in hospitals and nursing homes. Healthcare textiles, such as blankets, are ubiquitous in hospitals and nursing home wards and may be easily shared between patients/users without an adequate pre-cleaning process. Therefore, functionalizing these textiles with antimicrobial properties may considerably reduce the microbial load and prevent the propagation of infections, including MDRB. Blankets are mainly comprised of knitted cotton (CO), polyester (PES), and cotton-polyester (CO-PES). These fabrics were functionalized with novel gold-hydroxyapatite nanoparticles (AuNPs-HAp) that possess antimicrobial properties, due to the presence of the AuNPs' amine and carboxyl groups, and low propensity to display toxicity. For optimal functionalization of the knitted fabrics, two pre-treatments, four different surfactants, and two incorporation processes were evaluated. Furthermore, exhaustion parameters (time and temperature) were subjected to a design of experiments (DoE) optimization. The concentration of AuNPs-HAp in the fabrics and their washing fastness were critical factors assessed through color difference (ΔE). The best performing knitted fabric was half bleached CO, functionalized using a surfactant combination of Imerol ® Jet-B (surfactant A) and Luprintol ® Emulsifier PE New (surfactant D) through exhaustion at 70 °C for 10 min. This knitted CO displayed antibacterial properties even after 20 washing cycles, showing its potential to be used in comfort textiles within healthcare environments.

MicroRNAs (miRNAs) are short non-coding RNA sequences with the ability to inhibit the expression of a target mRNA at the post-transcriptional level, acting as modulators of both the degenerative and regenerative processes. Therefore, these molecules constitute a potential source of novel therapeutic tools. In this study, we investigated the miRNA expression profile that presented in enthesis tissue upon injury. For this, a rodent enthesis injury model was developed by creating a defect at a rat's patellar enthesis. Following injury, explants were collected on days 1 ( n = 10) and 10 ( n = 10). Contra lateral samples ( n = 10) were harvested to be used for normalization. The expression of miRNAs was investigated using a "Fibrosis" pathway-focused miScript qPCR array. Later, target prediction for the aberrantly expressed miRNAs was performed by means of the Ingenuity Pathway Analysis, and the expression of mRNA targets relevant for enthesis healing was confirmed using qPCRs. Additionally, the protein expression levels of collagens I, II, III, and X were investigated using Western blotting. The mRNA expression pattern of EGR1 , COL2A1 , RUNX2 , SMAD1 , and SMAD3 in the injured samples indicated their possible regulation by their respective targeting miRNA, which included miR-16, -17, -100, -124, -133a, -155 and -182. Furthermore, the protein levels of collagens I and II were reduced directly after the injury (i.e., day 1) and increased 10 days post-injury, while collagens III and X showed the opposite pattern of expression.

Nitrate (NO3 - ) nutrition is known to mitigate the damages caused by individual stresses of waterlogging and salinity. Here, we investigated the role of NO3 - in soybean plants exposed to these stresses in combination. Nodulated soybean cultivated under greenhouse conditions and daily fertilised with a nutrient solution without nitrogen were subjected to the following treatments: Water, NO3 - , NaCl, and NaCl+NO3 - . Then, plants were exposed to waterlogging (6days) and drainage (2days). Compared to plants exposed to isolated stress, the saline waterlogging resulted in higher concentrations of H2 O2 , O2 ˙- , and lipid peroxidation at the whole-plant level, mainly during drainage. Furthermore, saline waterlogging increased fermentation and the concentrations of Na+ and K+ in roots and leaves both during waterlogging and drainage. NO3 - supplementation led to augments in NO3 - and NO levels, and stimulated nitrate reductase activity in both organs. In addition, NO3 - nutrition alleviated oxidative stress and fermentation besides increasing the K+ /Na+ ratio in plants exposed to saline waterlogging. In conclusion, NO3 - supplementation is a useful strategy to help soybean plants overcome saline waterlogging stress. These findings are of high relevance for agriculture as soybean is an important commodity and has been cultivated in areas prone to saline waterlogging.

Prevention of bacterial adhesion is a way to reduce and/or avoid biofilm formation, thus restraining its associated infections. The development of repellent anti-adhesive surfaces, such as superhydrophobic surfaces, can be a strategy to avoid bacterial adhesion. In this study, a polyethylene terephthalate (PET) film was modified by in situ growth of silica nanoparticles (NPs) to create a rough surface. The surface was further modified with fluorinated carbon chains to increase its hydrophobicity. The modified PET surfaces presented a pronounced superhydrophobic character, showing a water contact angle of 156° and a roughness of 104 nm (a considerable increase comparing with the 69° and 4.8 nm obtained for the untreated PET). Scanning Electron Microscopy was used to evaluate the modified surfaces morphology, further confirming its successful modification with nanoparticles. Additionally, a bacterial adhesion assay using an Escherichia coli expressing YadA, an adhesive protein from Yersinia so-called Yersinia adhesin A, was used to assess the anti-adhesive potential of the modified PET. Contrarily to what was expected, adhesion of E. coli YadA was found to increase on the modified PET surfaces, exhibiting a clear preference for the crevices. This study highlights the role of material micro topography as an important attribute when considering bacterial adhesion.

The normal cellular prion protein (PrP C ) and its infectious conformer, PrP Sc , possess a disproportionately greater amount of methionines than would be expected for a typical mammalian protein. The thioether of methionine can be readily oxidized to the corresponding sulfoxide, which means that oxidation of methionine can be used to map the surface of the conformation of PrP C or PrP Sc , as covalent changes are retained after denaturation. We identified a set of peptides (TN M K, M LGSA M SR, LLGSA M SR, P M IHFGNDWEDR, EN M NR, EN M YR, I M ER, MM ER, M IER, VVEQ M CVTQYQK, and VVEQ M CITQYQR) that contains every methionine in sheep, cervid, mouse, and bank vole PrP. Each is the product of a tryptic digestion and is suitable for a multiple reaction monitoring (MRM) based analysis. The peptides chromatograph well. The oxidized and unoxidized peptides containing one methionine readily separate. The unoxidized, two singly oxidized, and doubly oxidized forms of the M LGSA M SR and MM ER peptides are also readily distinguishable. This approach can be used to determine the surface exposure of each methionine by measuring its oxidation after reaction with added hydrogen peroxide.

In sheep, the transmissibility and progression of scrapie, a sheep prion (PrP Sc ) disease, is strongly dependent upon specific amino acid polymorphisms in the natively expressed prion protein (PrP C ). Sheep expressing PrP C with lysine (K) polymorphism at position 171 (K171) are partially resistant to oronasal dosing of classical sheep scrapie. In addition, scrapie infected sheep expressing the K171 polymorphism show a longer incubation period compared to sheep homozygous (glutamine (Q)) at position 171. Quantitating the amount of the K171 polymorphism in a sheep scrapie sample can provide important information on the composition of PrP Sc . A tryptic peptide, 159 R.YPNQVYYRPVDK.Y 172 , derived from the digestion of 171K recombinant PrP, was identified as an analyte peptide suitable for a multiple reaction monitoring-based analysis. This method, using 15 N-labeled analogs and another internal peptide from the proteinase K-resistant core, permits the simultaneous quantitation of the total amount of PrP and the proportion of K171 polymorphism in the sample. Background molecules with similar retention times and transitions were present in samples from scrapie-infected sheep. Proteinase K digestion followed by ultracentrifugation-based isolation or phosphotungstic acid-based isolation were employed to minimize the contribution of those background molecules, making this approach suitable for quantitating the amount of the K171 polymorphism in heterozygous scrapie infected sheep.

The most devastating fungal disease of peaches and nectarines is brown rot, caused by Monilinia spp. Among the many plant responses against biotic stress, plant terpenoids play essential protective functions, including antioxidant activities and inhibition of pathogen growth. Herein, we aimed to characterize the expression of terpenoid biosynthetic genes in fruit tissues that presented different susceptibility to brown rot. For that, we performed artificial inoculations with Monilinia laxa at two developmental stages (immature and mature fruit) of two nectarine cultivars ('Venus' -mid-early season cultivar - and 'Albared' -late season cultivar-) and in vitro tests of the key compounds observed in the transcriptional results. All fruit were susceptible to M. laxa except for immature 'Venus' nectarines. In response to the pathogen, the mevalonic acid (MVA) pathway of the 'Venus' cultivar was highly induced in both stages rather than the methylerythritol phosphate (MEP) pathway, being the expression of some MEP-related biosynthetic genes [e.g., PROTEIN FARNESYLTRANSFERASE (PpPFT), and 3S-LINALOOL SYNTHASE (PpLIS)] different between stages. In 'Albared', both stages presented similar responses to M. laxa for both pathways. Comparisons between cultivars showed that HYDROXYMETHYLGLUTARYL-CoA REDUCTASE (PpHMGR1) expression levels were common in susceptible tissues. Within all the terpenoid biosynthetic pathway, linalool- and farnesal-related pathways stood out for being upregulated only in resistant tissues, which suggest their role in mediating the resistance to M. laxa. The in vitro antifungal activity of linalool and farnesol (precursor of farnesal) revealed fungicidal and fungistatic activities against M. laxa, respectively, depending on the concentration tested. Understanding the different responses between resistant and susceptible tissues could be further considered for breeding or developing new strategies to control brown rot in stone fruit., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Postharvest fungal pathogens benefit from the increased host susceptibility that occurs during fruit ripening. In unripe fruit, pathogens often remain quiescent and unable to cause disease until ripening begins, emerging at this point into destructive necrotrophic lifestyles that quickly result in fruit decay. Here, we demonstrate that one such pathogen, Botrytis cinerea, actively induces ripening processes to facilitate infections and promote disease in tomato (Solanum lycopersicum). Assessments of ripening progression revealed that B. cinerea accelerated external coloration, ethylene production, and softening in unripe fruit, while mRNA sequencing of inoculated unripe fruit confirmed the corresponding upregulation of host genes involved in ripening processes, such as ethylene biosynthesis and cell wall degradation. Furthermore, an enzyme-linked immunosorbent assay (ELISA)-based glycomics technique used to assess fruit cell wall polysaccharides revealed remarkable similarities in the cell wall polysaccharide changes caused by both infections of unripe fruit and ripening of healthy fruit, particularly in the increased accessibility of pectic polysaccharides. Virulence and additional ripening assessment experiments with B. cinerea knockout mutants showed that induction of ripening depends on the ability to infect the host and break down pectin. The B. cinerea double knockout Δbc polygalacturonase1 Δbc polygalacturonase2 lacking two critical pectin degrading enzymes was incapable of emerging from quiescence even long after the fruit had ripened at its own pace, suggesting that the failure to accelerate ripening severely inhibits fungal survival on unripe fruit. These findings demonstrate that active induction of ripening in unripe tomato fruit is an important infection strategy for B. cinerea., (© The Author(s) 2022. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

In this study, we tested whether waterlogging priming at the vegetative stage would mitigate a subsequent waterlogging event at the reproductive stage in soybean [Glycine max (L.) Merr.]. Plants (V3 stage) were subjected to priming for 7days and then exposed to waterlogging stress for 5days (R2 stage) with non-primed plants. Roots and leaves were sampled on the fifth day of waterlogging and the second and fifth days of reoxygenation. Overall, priming decreased the H2 O2 concentration and lipid peroxidation in roots and leaves during waterlogging and reoxygenation. Priming also decreased the activity of antioxidative enzymes in roots and leaves and increased the foliar concentration of phenols and photosynthetic pigments. Additionally, priming decreased fermentation and alanine aminotransferase activity during waterlogging and reoxygenation. Finally, priming increased the concentration of amino acids, sucrose, and total soluble sugars in roots and leaves during waterlogging and reoxygenation. Thus, primed plants were higher and more productive than non-primed plants. Our study shows that priming alleviates oxidative stress, fermentation, and carbohydrate consumption in parallel to increase the yield of soybean plants exposed to waterlogging and reoxygenation.

We propose a mathematical model for the transmission dynamics of SARS-CoV-2 in a homogeneously mixing non constant population, and generalize it to a model where the parameters are given by piecewise constant functions. This allows us to model the human behavior and the impact of public health policies on the dynamics of the curve of active infected individuals during a COVID-19 epidemic outbreak. After proving the existence and global asymptotic stability of the disease-free and endemic equilibrium points of the model with constant parameters, we consider a family of Cauchy problems, with piecewise constant parameters, and prove the existence of pseudo-oscillations between a neighborhood of the disease-free equilibrium and a neighborhood of the endemic equilibrium, in a biologically feasible region. In the context of the COVID-19 pandemic, this pseudo-periodic solutions are related to the emergence of epidemic waves. Then, to capture the impact of mobility in the dynamics of COVID-19 epidemics, we propose a complex network with six distinct regions based on COVID-19 real data from Portugal. We perform numerical simulations for the complex network model, where the objective is to determine a topology that minimizes the level of active infected individuals and the existence of topologies that are likely to worsen the level of infection. We claim that this methodology is a tool with enormous potential in the current pandemic context, and can be applied in the management of outbreaks (in regional terms) but also to manage the opening/closing of borders., (© 2021 Elsevier Inc. All rights reserved.)

Homeless substance users are particularly hard to treat. In this pilot study, we evaluated the acceptability and feasibility of incorporating Contingency Management (CM) into a public Abstinent-Contingent Housing (ACH) treatment program developed to treat currently homeless crack cocaine users. A total of 21 homeless crack cocaine users were randomized to receive 12 weeks of ACH alone (n = 9) or ACH plus CM (ACH + CM) (n = 12). Twelve treatment providers in the ACH treatment program were trained to deliver the CM intervention. CM was rated as relatively (41.7%) or very (58.3%) easy to understand and relatively (50%) or very (50%) easy to conduct by the ACH treatment providers. On a 10-point Likert scale, providers rated the importance of incorporating CM into public treatment programs for crack cocaine at M = 8.3 (SD = 2). Participants exposed to CM rated as relatively (33.3%) or very (66.7%) easy to understand. One hundred percent liked receiving the intervention "a lot," and 78.9% believed it helped them achieve and maintain crack cocaine abstinence. Finally, compared to the ACH condition, the ACH + CM condition was consistently associated with better treatment retention and cocaine use outcome measures, yelling small to large effect sizes. However, possibly due to the small sample size, most of these differences did not achieve statistical significance. CM was well integrated into the ACH treatment program and was well accepted by both the providers and participants, suggesting the feasibility of incorporating CM into a public treatment program for homeless crack cocaine users from low- and middle-income countries. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Host behavioural changes are among the most apparent effects of infection. 'Sickness behaviour' can involve a variety of symptoms, including anorexia, depression, and changed activity levels. Here, using a real-time tracking and behavioural profiling platform, we show that in Drosophila melanogaster, several systemic bacterial infections cause significant increases in physical activity, and that the extent of this activity increase is a predictor of survival time in some lethal infections. Using multiple bacteria and D. melanogaster immune and activity mutants, we show that increased activity is driven by at least two different mechanisms. Increased activity after infection with Micrococcus luteus, a Gram-positive bacterium rapidly cleared by the immune response, strictly requires the Toll ligand spätzle. In contrast, increased activity after infection with Francisella novicida, a Gram-negative bacterium that cannot be cleared by the immune response, is entirely independent of both Toll and the parallel IMD pathway. The existence of multiple signalling mechanisms by which bacterial infections drive increases in physical activity implies that this effect may be an important aspect of the host response., Competing Interests: The authors have declared that no competing interests exist.

Background: Immediately after a fracture occurs, a fracture hematoma (fxH) is formed. This fxH plays an important role in fracture healing and, under normal circumstances, aids in generating an environment in which a wide variety of cells orchestrate processes involved in fracture healing. MicroRNAs (miRNAs) may influence these processes. The aim of this study was therefore to determine the miRNA expression signature of human fxH in normal fracture healing and examine the potential influence of clinical parameters on these expression levels., Methods: fxH was harvested from 61 patients (mean age 52 ± 19; 32 ♀ ) during fracture surgery. miRNAs were isolated, transcribed and pooled for qPCR array analysis and validation. Qiagen fibrosis- and inflammation qPCR arrays were used based on an extensive literature study related to fracture healing and osteogenesis. Additionally, miRNA targets were determined., Results: From the array data, a selection of the twenty most regulated miRNAs, 10 up- and 10 down regulated, was validated in the study population. The expression levels of seven out of these twenty miRNAs were correlated to several clinical parameters. The time interval between trauma and surgery showed to influence the expression of three miRNAs, three other miRNAs were expressed in a patient age dependent manner and one miRNA was associated with the severity of trauma., Conclusion: This study portrayed the role and importance of miRNAs in human fxH, linked to key processes in fracture healing. Seven miRNAs showed to be involved in multiple processes that are important in the fracture healing cascade, such as angiogenesis, mineralisation and cellular differentiation. In silico target analysis revealed 260 mRNA targets for 14 out of the 20 validated miRNAs., The Translational Potential of This Article: These data broaden our view on the potential diagnostic and therapeutic implications of miRNAs in fracture healing., Competing Interests: There are no conflicts of interest to declare., (© 2022 The Authors.)

The interphase between tendon and bone consists of a highly specialised tissue called enthesis. Typically, the enthesis is described as a succession of four different zones: tendon, non-mineralised fibrocartilage, mineralised fibrocartilage and bone. However, the microstructure of the entheses, cellular composition and mechanical properties vary depending on their anatomical location. The present study aimed to characterise three of the most relevant sites of enthesis injury in a rat model: the patellar tendon, the Achilles tendon and the supraspinatus enthesis, in terms of biomechanics, histology and genetic expression. The patellar enthesis presented the highest ultimate load and lowest stiffness of the three, while the supraspinatus was the weakest and stiffest. The histological characterisation revealed key differences at the insertion site for each enthesis. The patellar enthesis showed a large cartilaginous area at the tendon-to-bone interphase whilst this interphase was smaller in the supraspinatus entheses samples. Furthermore, the Achilles tendon enthesis displayed a more abrupt transition from tendon to bone. Additionally, each enthesis exhibited a particular and distinct pattern of expression of tenogenic, chondrogenic and osteogenic markers. This study provided valuable insights for a better understanding of the three entheses at relevant anatomical sites. Moreover, the larger cross-sectional area of the patellar enthesis, the strong mechanical properties and the easier surgical access to this location led to the conclusion that the patellar tendon enthesis site could be most suitable for the development of a preclinical model for general enthesis regeneration studies in rats.

New point-of-care (POC) diagnosis of bacterial infections are imperative to overcome the deficiencies of conventional methods, such as culture and molecular methods. In this study, we identified new aptamers that bind to the virulence factor Yersinia adhesin A (YadA) of Yersinia enterocolitica using cell-systematic evolution of ligands by exponential enrichment (cell-SELEX). Escherichia coli expressing YadA on the cell surface was used as a target cell. After eight cycles of selection, the final aptamer pool was sequenced by high throughput sequencing using the Illumina Novaseq platform. The sequencing data, analyzed using the Geneious software, was aligned, filtered and demultiplexed to obtain the key nucleotides possibly involved in the target binding. The most promising aptamer candidate, Apt1, bound specifically to YadA with a dissociation constant (Kd) of 11 nM. Apt1 was used to develop a simple electrochemical biosensor with a two-step, label-free design towards the detection of YadA. The sensor surface modifications and its ability to bind successfully and stably to YadA were confirmed by cyclic voltammetry, impedance spectroscopy and square wave voltammetry. The biosensor enabled the detection of YadA in a linear range between 7.0 × 104 and 7.0 × 107 CFU mL−1 and showed a square correlation coefficient >0.99. The standard deviation and the limit of detection was ~2.5% and 7.0 × 104 CFU mL−1, respectively. Overall, the results suggest that this novel biosensor incorporating Apt1 can potentially be used as a sensitive POC detection system to aid the diagnosis of Y. enterocolitica infections. Furthermore, this simple yet innovative approach could be replicated to select aptamers for other (bacterial) targets and to develop the corresponding biosensors for their detection.

Antibiotic resistance is a severe global health threat and hence demands rapid action to develop novel therapies, including microscale drug delivery systems. Herein, a hierarchical microparticle system is developed to achieve bacteria-activated single- and dual-antibiotic drug delivery for preventing methicillin-resistant Staphylococcus aureus (MRSA) bacterial infections. The designed system is based on a capsosome structure, which consists of a mesoporous silica microparticle coated in alternating layers of oppositely charged polymers and antibiotic-loaded liposomes. The capsosomes are engineered and shown to release their drug payloads in the presence of MRSA toxins controlled by the Agr quorum sensing system. MRSA-activated single drug delivery of vancomycin and synergistic dual delivery of vancomycin together with an antibacterial peptide successfully kills MRSA in vitro. The capability of capsosomes to selectively deliver their cargo in the presence of bacteria, producing a bactericidal effect to protect the host organism, is confirmed in vivo using a Drosophila melanogaster MRSA infection model. Thus, the capsosomes serve as a versatile multidrug, subcompartmentalized microparticle system for preventing antibiotic-resistant bacterial infections, with potential applications to protect wounds or medical device implants from infections., (© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Anthocyanins have been associated with beneficial effects on human health. Cancer has been one of the main public health issues due to its aggressiveness and high mortality rate. This systematic review aimed to address recent research (from January 2000 to September 2021) on the anticancer activity of anthocyanins assessed by in vitro assays. The selected studies revealed that anthocyanins have anticancer potential by inhibiting cancer cell viability and proliferation, controlling cell cycle, and promoting apoptosis., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The use of multiphasic scaffolds to treat injured tendon-to-bone entheses has shown promising results in vitro. Here, we used two versions of a biphasic silk fibroin scaffold to treat an enthesis defect created in a rat patellar model in vivo. One version presented a mixed transition between the bony and the tendon end of the construct (S-MT) while this transition was abrupt in the second version (S-AT). At 12 weeks after surgery, the S-MT scaffold promoted better healing of the injured enthesis, with minimal undesired ossification of the insertion area. The expression of tenogenic and chondrogenic markers was sustained for longer in the S-MT-treated group and the tangent modulus of the S-MT-treated samples was similar to the native tissue at 12 weeks while that of the S-AT-treated enthesis was lower. Our study highlights the important role of the transition zone of multiphasic scaffolds in the treatment of complex interphase tissues such as the tendon-to-bone enthesis.

Objective: To evaluate the relationship between crack/cocaine use and domestic violence perpetration from the perspective of substance users' families. A secondary retrospective cross-sectional study, with 3162 family members of crack/cocaine users seeking treatment in the Recomeço Família Programme in São Paulo/Brazil was undertaken. Family members of crack/cocaine users reported that their relatives were more involved in domestic violence such as stealing (money and objects) at home [Odds Ratio Adjusted ORA = 2.17 (CI 95% 1.87; 2.53)], the family gave money to the user to buy drugs [ORA = 1.27 (1.08; 1.48)], and having problems with the judiciary [ORA = 1.48 (CI 95% 1.28; 1.71)]. Relatives of snorted cocaine users reported that there was physical and interpersonal violence, such as fathers being assaulted [ORA = 2.50 (CI 95% 1.08; 5.82)], assaulted someone else [ORA = 1.86 (CI 95% 1.32; 2.60)], threats of violence fights, arguments when the family talk about problematic drug use [ORA = 1.50 (CI 95% 1.13; 1.96)] and threatened some family members [ORA = 1.52 (CI 95% 1.14; 2.04)]. In this sample, there was a connection between crack/cocaine use and the perpetuation of domestic violence, corroborating with important implications for public policies, substance use treatment and prevention of domestic violence interventions.

Chronic wasting disease (CWD) is a prion disease of cervids (deer, elk, moose, etc.). It spreads readily from CWD-contaminated environments and among wild cervids. As of 2022, North American CWD has been found in 29 states, four Canadian provinces and South Korea. The Scandinavian form of CWD originated independently. Prions propagate their pathology by inducing a natively expressed prion protein (PrP C ) to adopt the prion conformation (PrP Sc ). PrP C and PrP Sc differ solely in their conformation. Like other prion diseases, transmissible CWD prions can arise spontaneously. The CWD prions can respond to selection pressures resulting in the emergence of new strain phenotypes. Annually, 11.5 million Americans hunt and harvest nearly 6 million deer, indicating that CWD is a potential threat to an important American food source. No tested CWD strain has been shown to be zoonotic. However, this may not be true for emerging strains. Should a zoonotic CWD strain emerge, it could adversely impact the hunting economy and game meat consumers., Competing Interests: The author declares no competing financial interest., (Not subject to U.S. Copyright. Published 2022 by American Chemical Society.)

The study aimed to analyze the agronomic performance of 11 safflower genotypes using adaptability and stability methods, while identifying safflower genotypes with stable behavior and a high grain yield in different environments of the Brazilian Cerrado. Ten lines and a cultivar of safflower were evaluated in four environments in the Brazilian conditions. Our results revealed the genotypes P30, P35, P9, P11, and P31 to be superior for grain yield and P43, P7, P11, and P31 to be superior for oil content. The lowest Wricke index, an indication of genotype stability, was observed for P9 (0.41%), which is considered the most stable genotype, followed by P35 (1.29%) and P31 (1.98%). For the predictability of the behavior of genotypes in the environments, P7 (80.85%), P35 (86.10%), P31 (85.90%), and P9 (97.42%) were considered predictable genotypes. The genotypes P11 (1045.6 kg ha -1 and 19.7%) and P21 (952.7 kg ha -1 and 20.6%) are recommended for cultivation in this region, considering both their grain yield and oil content. Safflower is viable to use out of season in the Brazilian Cerrado. The crop can generate profits for farmers and be used for oil production in periods of uncertain corn production.

We report a case of a 28-year-old man lost to follow-up with chronic hypoxaemia and a history of an uncorrected tricuspid hypoplasia, perimembranous ventricular septal defect (VSD) and pulmonary stenosis. Given this initial diagnosis, chronic hypoxaemia was deemed to result from right-to-left shunt through the VSD driven by elevated right ventricle pressures. However, the further investigation identified an inferior sinus venosus atrial septal defect, unveiling the true mechanism behind the clinical scenario. The patient was submitted to surgical correction with clinical improvement. This case illustrates the defiant evaluation of this type of atrial septal defect through echocardiography and underlines the importance of a multimodal evaluation to reach an accurate diagnosis and optimal management., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

Iron toxicity is a major challenge faced by plants in hypoxic soils; however, the consequences of such combined stress for soybean (Glycine max) remain to be determined. Here we assessed the physiological responses of soybean plants exposed to hypoxia and a high concentration of iron. Soil-grown plants cultivated in a greenhouse until the vegetative stage were transferred to a hydroponic system containing nutrient solution and subjected to two oxygen conditions (normoxia (6.2 mg L -1 ) and hypoxia (0.33 mg L -1 )) and two iron concentrations (Fe-EDTA) (0.09 and 1.8 mM) for 72 h. During hypoxia, high concentrations of iron in the nutrient solution resulted in increased iron accumulation in roots and leaves. Under this condition, the concentrations of zinc, nitrogen, potassium, and calcium decreased in the roots, while the concentration of nitrogen and magnesium decreased in the leaves. Additionally, during hypoxia, the higher concentration of iron led to an increase in the activity of the antioxidant enzymes in roots and leaves, while decreased the levels of the photosynthetic pigments, leaf gas exchange, and plant growth. In conclusion, high iron concentration in the root medium results in a considerably more severe damage condition to soybean plants under hypoxia compared to plants grown under low iron availability., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Large segmental osseous defects heal poorly. Recombinant, human bone morphogenetic protein-2 (rhBMP-2) is used clinically to promote bone healing, but it is applied at very high doses that cause adverse side effects and raise costs while providing only incremental benefit. We describe a previously unexplored, alternative approach to bone regeneration using chemically modified messenger RNA (cmRNA). An optimized cmRNA encoding BMP-2 was delivered to critical-sized femoral osteotomies in rats. The cmRNA remained orthotopically localized and generated BMP locally for several days. Defects healed at doses ≥25 μg of BMP-2 cmRNA. By 4 weeks, all animals treated with 50 μg of BMP-2 cmRNA had bridged bone defects without forming the massive callus seen with rhBMP-2. Moreover, such defects recovered normal mechanical strength quicker and initiated bone remodeling faster. cmRNA regenerated bone via endochondral ossification, whereas rhBMP-2 drove intramembranous osteogenesis; cmRNA provides an innovative, safe, and highly translatable technology for bone healing.

A prion's pathogenic character is enciphered in its conformation, which also defines the chemical environments of its amino acids. Differences in chemical environments influence the reactivity of amino acid side chains, in a conformation-dependent manner. Chemical oxidation of susceptible methionines would identify those methionines on the surface of a prion, which would reveal conformation-dependent information. We identified a set of methionine-containing peptides derived from the tryptic, chymotryptic, or tryptic/chymotryptic digestion of recombinant prion protein and the Sc237 strain of hamster-adapted scrapie. We developed a multiple reaction monitoring-based method of quantifying the extent of the methionine oxidation in those peptides. This approach can be used to define a prion's conformation and to distinguish among prion strains, which is an important component of food safety., Competing Interests: The authors declare no competing financial interest., (Not subject to U.S. Copyright. Published 2022 by American Chemical Society.)

Background: Smoked cocaine (i.e., crack use) is a severe health problem in Brazil, with the country being reported as having the largest crack market in the world. The objective of this study was to evaluate the effectiveness of incorporating Contingency Management targeting cocaine abstinence into a public treatment program in Brazil., Methods: Single-blind randomized controlled trial conducted at Unidade Recomeço Helvétia (URH), a public ambulatory treatment program for persons who use crack and live in the "Crackland" region in downtown São Paulo, Brazil. In total, 98 treatment-seeking individuals who use crack were randomized to one of two treatment conditions. Participants allocated to the control condition (n = 48) received 12 weeks of the standard treatment provided at URH. Participants allocated to the experimental condition (n = 50) received the same treatment in combination with Contingency Management (URH+CM). In URH+CM, participants were provided with vouchers with monetary value for submission of negative cocaine urinalysis twice weekly., Results: Compared to the URH group, the URH+CM group was significantly more likely to submit a negative cocaine urinalysis during treatment, with odds ratios ranging from 4.17 to 6.78, depending on how missing data was accounted for (p<0.01). Participants receiving Contingency Management also had higher odds of achieving three or more weeks of continuous abstinence (OR= 8.07; 95% CI [2.48, 26.24]), achieved longer durations of abstinence (B = 2.14; 95% CI [0.67, 3.61]), submitted a higher percentage of negative urinalysis (B = 19.85; 95% CI [6.89, 32.82]), and were retained in treatment for a longer period (B = 3.00; 95% CI [1.04, 4.97]), compared to those receiving URH alone (p<0.01 for all)., Conclusions: The incorporation of Contingency Management was effective in promoting cocaine abstinence and treatment retention. The large-scale dissemination of Contingency Management may be an effective strategy to treat Brazilians with cocaine use disorders., Competing Interests: Declarations of Interest McPherson and Roll have received research funding from the Bristol-Myers Squibb Foundation, Managed Health Connections, LLC, Ringful Health, LLC, and Pillsy, Inc. McPherson has also received research funding from the Orthopedic Specialty Institute and consulted for Consistent Care, LLC, and the United States Department of Justice. These funds are in no way related to the investigation reported here. Miguel, Simões, Yamauchi, Madruga, Smith, da Silva, Laranjeira, McDonell, and Mari have no financial relationships with commercial interests., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: The cellular prion protein (PrP C ) is a membrane-bound, multifunctional protein mainly expressed in neuronal tissues. Recent studies indicate that the native trafficking of PrP C can be misused to internalize misfolded amyloid beta and α-synuclein (aSyn) oligomers., Objectives: We define PrP C 's role in internalizing misfolded aSyn in α-synucleinopathies and identify further involved proteins., Methods: We performed comprehensive behavioral studies on four transgenic mouse models (ThySyn and ThySynPrP00, TgM83 and TgMPrP00) at different ages. We developed PrP C -(over)-expressing cell models (cell line and primary cortical neurons), used confocal laser microscopy to perform colocalization studies, applied mass spectrometry to identify interactomes, and determined disassociation constants using surface plasmon resonance (SPR) spectroscopy., Results: Behavioral deficits (memory, anxiety, locomotion, etc.), reduced lifespans, and higher oligomeric aSyn levels were observed in PrP C -expressing mice (ThySyn and TgM83), but not in homologous Prnp ablated mice (ThySynPrP00 and TgMPrP00). PrP C colocalized with and facilitated aSyn (oligomeric and monomeric) internalization in our cell-based models. Glimepiride treatment of PrP C -overexpressing cells reduced aSyn internalization in a dose-dependent manner. SPR analysis showed that the binding affinity of PrP C to monomeric aSyn was lower than to oligomeric aSyn. Mass spectrometry-based proteomic studies identified clathrin in the immunoprecipitates of PrP C and aSyn. SPR was used to show that clathrin binds to recombinant PrP, but not aSyn. Experimental disruption of clathrin-coated vesicles significantly decreased aSyn internalization., Conclusion: PrP C 's native trafficking can be misused to internalize misfolded aSyn through a clathrin-based mechanism, which may facilitate the spreading of pathological aSyn. Disruption of aSyn-PrP C binding is, therefore, an appealing therapeutic target in α-synucleinopathies. © 2021 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society., (© 2021 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.)

Controlling an epidemiological model is often performed using optimal control theory techniques for which the solution depends on the equations of the controlled system, objective functional and possible state and/or control constraints. In this paper, we propose a model-free control approach based on an algorithm that operates in 'real-time' and drives the state solution according to a direct feedback on the state solution that is aimed to be minimized, and without knowing explicitly the equations of the controlled system. We consider a concrete epidemic problem of minimizing the number of HIV infected individuals, through the preventive measure pre-exposure prophylaxis (PrEP) given to susceptible individuals. The solutions must satisfy control and mixed state-control constraints that represent the limitations on PrEP implementation. Our model-free based control algorithm allows to close the loop between the number of infected individuals with HIV and the supply of PrEP medication 'in real time', in such a manner that the number of infected individuals is asymptotically reduced and the number of individuals under PrEP medication remains below a fixed constant value. We prove the efficiency of our approach and compare the model-free control solutions with the ones obtained using a classical optimal control approach via Pontryagin maximum principle. The performed numerical simulations allow us to conclude that the model-free based control strategy highlights new and interesting performances compared with the classical optimal control approach.