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Ciguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus Gambierdiscus. Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrices, are numerous in the literature. However, little information on their effectiveness for nonpolar CTXs is available, yet these congeners, contributing to the risk of CP, are required for the establishment of effective food safety monitoring programs. An evaluation of six extraction/purification protocols, performed with CTX3C spiked on fish flesh and a neuroblastoma cell-based assay (CBA-N2a), revealed recoveries from 6 to 45%. This led to the development of an optimized 3-day protocol designed for a large number of samples, with CTX1B and CTX3C eluting in a single fraction and showing recoveries of 73% and 70%, respectively. In addition, a reduction in adverse matrix effects in the CBA-N2a analyses was demonstrated with naturally contaminated specimens, increasing the sensitivity of the method, which now meets the very low guidance level recommended by international agencies. However, efforts are still required to reduce the signal suppression observed in LC-MS/MS analysis. This optimized protocol contributes to the technological advancement of detection methods, promoting food safety and improving CP risk assessment in marine products. [ABSTRACT FROM AUTHOR]

In our recent study, we demonstrated using 7Li solid-state Nuclear Magnetic Resonance (ssNMR) and single-crystal X-ray diffraction that the cathode LiFeV2O7 possesses a defect associated with the positioning of vanadium atoms. We proposed that this defect could be the source of extra signals detected in the 7Li spectra. In this context, we now apply density functional theory (DFT) calculations to assign the experimental signals observed in 7Li NMR spectra of the pristine sample. The calculation results are in strong agreement with the experimental observations. DFT calculations are a useful tool to interpret the observed paramagnetic shifts and understand how the presence of disorder affects the spectra behavior through the spin-density transfer processes. Furthermore, we conducted a detailed study of the lithiated phase combining operando synchrotron powder X-ray diffraction (SPXRD) and DFT calculations. A noticeable volume expansion is observed through the first discharge cycle which likely contributes to the enhanced lithium dynamics in the bulk material, as supported by previously published ssNMR data. DFT calculations are used to model the lithiated phase and demonstrate that both iron and vanadium participate in the redox process. The unusual electronic structure of the V4+ exhibits a single electron on the 3dxy orbital perpendicular to the V–O–Li bond being a source of a negative Fermi contact shift observed in the 7Li NMR of the lithiated phase. [ABSTRACT FROM AUTHOR]

The development of ground-breaking Survival Motor Neuron (SMN) replacement strategies has revolutionized the field of Spinal Muscular Atrophy (SMA) research. However, the limitations of these therapies have now become evident, highlighting the need for the development of complementary targets beyond SMN replacement. To address these challenges, here we explored, in in vitro and in vivo disease models, Stathmin-2 (STMN2), a neuronal microtubule regulator implicated in neurodegenerative diseases like Amyotrophic Lateral Sclerosis (ALS), as a novel SMN-independent target for SMA therapy. Our findings revealed that STMN2 overexpression effectively restored axonal growth and outgrowth defects in induced pluripotent stem cell-(iPSC)-derived motor neurons (MNs) from SMA patients. Intracerebroventricular administration of adeno-associated virus serotype 9 (AAV9) carrying Stmn2 cDNA significantly ameliorated survival rates, motor functions, muscular and neuromuscular junction pathological features in SMA mice, mirrored by in vitro outcomes. Overall, this pioneering study not only provides insight into the therapeutic potential of STMN2 in SMA, but also suggests its broader applications for MN diseases, marking a substantial step forward in addressing the multifaceted challenges of neurological diseases treatment. [ABSTRACT FROM AUTHOR]

Soil microorganisms play a significant role in the dynamic regulation of organic matter in soils. To assess the influence of agricultural practices on soil functional profiling, we examined the effect of soil disturbance and plant sequence with different levels of mycotrophy on wheat microbiomes metabolism. Soil samples were analyzed with community-level physiological profiles (CLPP) using Biolog™ Ecoplates. The results of average well color development (AWCD) showed that the degree of mycotrophy of preceding crop and soil disturbance affected the soil microbiome, although no impact on Shannon Evenness Index was observed during the experiment. The Shannon–Wiener Diversity Index showed variations among the different preceding plants, but not in wheat analysis. The pattern of the C sources metabolism also changed differentially regarding plant type and soil disturbance during the experiment, being also different within the highly mycotrophic plants (legume and grass). In the legume, an increase in the metabolism of amine/amides and phenolic acids was observed, whilst in the grass, an increase in the metabolism of phosphate-carbons (P carbon) and carbohydrates was more evident. Principal component analysis showed that a grouping in the distinct phases of the experiment correlated with the widening of the metabolism of amino acids, carboxylic acids, and carbohydrates. The results indicate that soil functional community structure reflects soil agricultural practice conditions. Previous plant types and soil disturbance impacted the soil microbiome metabolic response (AWCD) in wheat, generating different patterns of carbon metabolism related to previous plant mycotrophy. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the influence of light‐curing time and increment thickness on the microhardness and degree of conversion (DC) of bulk fill composite resins with different application systems. Translucency parameter (TP) was also measured. Specimens of resin composites were prepared in a circular matrix using a single increment with different thicknesses (2, 4, and 6 mm) and light‐cured with distinct times (10, 20, and 40 s). The materials tested (n = 5 pergroup) were Filtek One (FO) bulk fill, Tetric N‐Ceram (TC) bulk fill, SonicFill 3 (SF), VisCalor (VC) bulk. After 24 h, Knoop microhardness (KN) was measured, and the bottom/top ratio (B/Tratio) was calculated. The DC was measured using Fourier transformed infrared (FTIR) spectroscopy. The TP was assessed in additional specimens with 1 mm thick (n = 5). The data were statistically analyzed with analysis of variance (ANOVA) and Tukey's tests (5%). Significant differences were observed for all factors, for both B/Tratio and DC (p < 0.05). The higher increment thickness and the lower curing times resulted in lower B/Tratio and DC means. The VC and TC resins exhibited the overall higher B/Tratio, and the highest TP. SF presented the lowest values of B/Tratio and DC, with compromised polymerization at 6 mm depth. TP (means ± SD) were FO (12.85 ± 1.0)1, SF (15.62 ± 0.73)2, TC (20.32 ± 0.49)3, and VC (20.53 ± 0.73)3. We concluded that the greater the thickness of the increment, the lower the DC. Higher light curing times resulted on increased DC of the tested composites. The higher translucent materials VC and TC showed the greater B/Tratio; and FO resin exhibited the higher DC values. [ABSTRACT FROM AUTHOR]

The overuse of antimicrobials in livestock has contributed to the emergence and selection of clinically relevant multidrug-resistant bacteria. In Brazil, there is no conclusive information on the occurrence of Escherichia coli producing extended-spectrum β-lactamase (ESβL) in cattle breeding, which is an important sector of agribusiness in this country. Herein, we investigated the presence of ESβL-positive E. coli strains in dairy cattle from a commercial farm with routine practice of therapeutic cephalosporins. Ninety-five rectal swab samples were collected from healthy dairy calves and cows under treatment with ceftiofur. Samples were screened for the presence of ESβL producers, and positive isolates were identified by MALDI-TOF, with subsequent screening for genes encoding ESβL variants by PCR and sequencing. The presence of ESβL (CTX-M-15)-producing E. coli was confirmed in calves, and lactating and dry cows. Most ESβL strains with genetic homologies ≥ 90% were grouped into two major PFGE clusters, confirming the suscessful expansion of clonally related lineages in animals from different lactating cycles, on the same property. Four representatives CTX-M-15-positive E. coli strains had their genomes sequenced, belonging to the clonal complex (CC) 23 and sequence type (ST) 90. A phylogeographical landscape of ST90 was performed revealing a global One Health linkage. Our results highlight the intestinal microbiota of dairy cattle as a hotspot for the spread of critical priority ESβL-producing E. coli and demonstrate that ST90 is an international clone genomically adapted to human and animal hosts, which deserve additional investigation to determine its zoonotic potential and impact in food chain. [ABSTRACT FROM AUTHOR]

Mesenchymal stromal cells (MSCs) and their released factors (secretome) are intriguing options for regenerative medicine approaches based on the management of inflammation and tissue restoration, as in joint disorders like osteoarthritis (OA). Production strategy may modulate cells and secretome fingerprints, and for the latter, the effect of serum removal by starvation used in clinical-grade protocols has been underestimated. In this work, the effect of starvation on the molecular profile of interleukin 1 beta (IL1β)-primed adipose-derived MSCs (ASCs) was tested by assessing the expression level of 84 genes related to secreted factors and 84 genes involved in defining stemness potential. After validation at the protein level, the effect of starvation modulation in the secretomes was tested in a model of OA chondrocytes. IL1β priming in vitro led to an increase in inflammatory mediators' release and reduced anti-inflammatory potential on chondrocytes, features reversed by subsequent starvation. Therefore, when applying serum removal-based clinical-grade protocols for ASCs' secretome production, the effects of starvation must be carefully considered and investigated. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a degenerative joint disorder characterized by the progressive deterioration of articular cartilage driven and sustained by catabolic and inflammatory processes that lead to pain and functional impairment. Adipose-derived stem cells (ASCs) have emerged as a promising therapeutic strategy for OA due to their regenerative potential, which mainly relies on the adaptive release of paracrine molecules that are soluble or encapsulated in extracellular vesicles (EVs). The biological effects of EVs specifically depend on their cargo; in particular, microRNAs (miRNAs) can specifically modulate target cell function through gene expression regulation. This study aimed to investigate the impact of collection site (abdominal vs. peri-trochanteric adipose tissue) and collection method (surgical excision vs. lipoaspiration) on the miRNAs profile in ASC-derived EVs and their potential implications for OA therapy. EV-miRNA cargo profiles from ASCs of different origins were compared. An extensive bioinformatics search through experimentally validated and OA-related targets, pathways, and tissues was conducted. Several miRNAs involved in the restoration of cartilage homeostasis and in immunomodulation were identified in all ASC types. However, EV-miRNA expression profiles were affected by both the tissue-harvesting site and procedure, leading to peculiar characteristics for each type. Our results suggest that adipose-tissue-harvesting techniques and the anatomical site of origin influence the therapeutic efficacy of ASC-EVs for tissue-specific regenerative therapies in OA, which warrants further investigation. [ABSTRACT FROM AUTHOR]

An 8‐year‐8‐month‐old whippet was investigated by magnetic resonance imaging for acute thoracolumbar discomfort and pelvic limb stiffness, which had progressed to severe ambulatory paraparesis and proprioceptive ataxia of the pelvic limbs in the 24 hours before presentation. An aggressive, extradural T10 vertebral mass‐like lesion was identified, with resultant marked compression of the spinal cord. The lesion was heterogeneously T2‐weighted hyperintense and moderately T1‐weighted hyperintense relative to the adjacent vertebrae and paravertebral musculature. Cytological analysis of the lesion was consistent with a melanotic melanoma, which was confirmed by histopathological analysis and immunohistochemistry. The vertebral column is an uncommon location for either primary or metastatic melanomas in both humans and veterinary species. This lesion was particularly unusual in that its imaging characteristics were not typical for melanotic melanoma. This case report illustrates the unusual anatomical and imaging features of this tumour and outlines the diagnostic considerations pertinent to this case. [ABSTRACT FROM AUTHOR]

Copyright of Revista Foco (Interdisciplinary Studies Journal) is the property of Revista Foco and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Beckwith–Wiedemann Syndrome (BWS) is an imprinting disorder characterized by overgrowth, stemming from various genetic and epigenetic changes. This study delves into the role of IGF2 upregulation in BWS, focusing on insulin-like growth factor pathways, which are poorly known in this syndrome. We examined the IGF2R, the primary receptor of IGF2, WNT, and autophagy/lysosomal pathways in BWS patient-derived lymphoblastoid cell lines, showing different genetic and epigenetic defects. The findings reveal a decreased expression and mislocalization of IGF2R protein, suggesting receptor dysfunction. Additionally, our results point to a dysregulation in the AKT/GSK-3/mTOR pathway, along with imbalances in autophagy and the WNT pathway. In conclusion, BWS cells, regardless of the genetic/epigenetic profiles, are characterized by alteration of the IGF2R pathway that is associated with the perturbation of the autophagy and lysosome processes. These alterations seem to be a key point of the molecular pathogenesis of BWS and potentially contribute to BWS's characteristic overgrowth and cancer susceptibility. Our study also uncovers alterations in the WNT pathway across all BWS cell lines, consistent with its role in growth regulation and cancer development. [ABSTRACT FROM AUTHOR]

The therapeutic effect of mesenchymal stromal cells (MSCs) has been described for a variety of disorders, including those affecting musculoskeletal tissues. In this context, the literature reports several data about the regenerative effectiveness of MSCs derived from bone marrow, adipose tissue, and an amniotic membrane (BMSCs, ASCs, and hAMSCs, respectively), either when expanded or when acting as clinical-grade biologic pillars of products used at the point of care. To date, there is no evidence about the superiority of one source over the others from a clinical perspective. Therefore, a reliable characterization of the tissue-specific MSC types is mandatory to identify the most effective treatment, especially when tailored to the target disease. Because molecular characterization is a crucial parameter for cell definition, the need for reliable normalizers as housekeeping genes (HKGs) is essential. In this report, the stability levels of five commonly used HKGs (ACTB, EF1A, GAPDH, RPLP0, and TBP) were sifted into BMSCs, ASCs, and hAMSCs. Adult and fetal/neonatal MSCs showed opposite HKG stability rankings. Moreover, by analyzing MSC types side-by-side, comparison-specific HKGs emerged. The effect of less performant HKG normalization was also demonstrated in genes coding for factors potentially involved in and predicting MSC therapeutic activity for osteoarthritis as a model musculoskeletal disorder, where the choice of the most appropriate normalizer had a higher impact on the donors rather than cell populations when compared side-by-side. In conclusion, this work confirms HKG source-specificity for MSCs and suggests the need for cell-type specific normalizers for cell source or condition-tailored gene expression studies. [ABSTRACT FROM AUTHOR]

Among the available therapeutics for the conservative treatment of osteoarthritis (OA), mesenchymal stromal cells (MSCs)-based products appear to be the most promising. Alongside minimally manipulated cell-based orthobiologics, where MSCs are the engine of the bioactive properties, cell expansion under good manufacturing practice (GMP) settings is actively studied to obtain clinical-grade pure populations able to concentrate the biological activity. One of the main characteristics of GMP protocols is the use of clinical-grade reagents, including the recently released serum-free/xeno-free (SFM/XFM) synthetic media, which differ significantly from the traditional reagents like those based on fetal bovine serum (FBS). As SFM/XFM are still poorly characterized, a main lack is the notion of reliable housekeeping genes (HKGs) for molecular studies, either standalone or in combination with standard conditions. Indeed, the aim of this work was to test the stability of five commonly used HKGs (ACTB, EF1A, GAPDH, RPLP0, and TBP) in adipose-derived MSCs (ASCs) cultivated in two commercially available SFM/XFM and to compare outcomes with those obtained in FBS. Four different applets widely recognized by the scientific community (NormFinder, geNorm, comparative ΔCt method, and BestKeeper) were used and data were merged to obtain a final stability order. The analysis showed that cells cultured in both synthetic media had a similar ranking for HKGs stability (GAPDH being best), albeit divergent from FBS expanded products (EF1A at top). Moreover, it was possible to identify specific HKGs for side by side studies, with EF1A/TBP being the most reliable normalizers for single SFM/XFM vs. FBS cultured cells and TBP the best one for a comprehensive analysis of all samples. In addition, stability of HKGs was donor-dependent. The normalization effect on selected genes coding for factors known to be involved in OA pathology, and whose amount should be carefully considered for the selection of the most appropriate MSC-based treatment, showed how HKGs choice might affect the perceived amount for the different media or donor. Overall, this work confirms the impact of SFM/XFM conditions on HKGs stability performance, which resulted similarly for both synthetic media analyzed in the study. [ABSTRACT FROM AUTHOR]

Hepatic injuries in COVID-19 are not yet fully understood and indirect pathways (without viral replication in the liver) have been associated with the activation of vascular mechanisms of liver injury in humans infected with SARS-CoV-2. Golden Syrian hamsters are an effective model for experimental reproduction of moderate and self-limiting lung disease during SARS-CoV-2 infection. As observed in humans, this experimental model reproduces lesions of bronchointerstitial pneumonia and pulmonary vascular lesions, including endotheliitis (attachment of lymphoid cells to the luminal surface of endothelium). Extrapulmonary vascular lesions are well documented in COVID-19, but such extrapulmonary vascular lesions have not yet been described in the Golden Syrian hamster model of SARS-CoV-2 infection. The study aimed to evaluate microscopic liver lesions in Golden Syrian hamsters experimentally infected with SARS-CoV-2. In total, 38 conventional Golden Syrian hamsters, divided into infected group (n=24) and mock-infected group (n=14), were euthanized at 2-, 3-, 4-, 5-, 7-, 14-, and 15-days post infection with SARS-CoV-2. Liver fragments were evaluated by histopathology and immunohistochemical detection of SARS-CoV-2 Spike S2 antigens. The frequencies of portal vein endotheliitis, lobular activity, hepatocellular degeneration, and lobular vascular changes were higher among SARS-CoV-2-infected animals. Spike S2 antigen was not detected in liver. The main results indicate that SARS-CoV-2 infection exacerbated vascular and inflammatory lesions in the liver of hamsters with pre-existing hepatitis of unknown origin. A potential application of this animal model in studies of the pathogenesis and evolution of liver lesions associated with SARS-CoV-2 infection still needs further evaluation. [ABSTRACT FROM AUTHOR]

Mitofusin-2 (MFN2) is an outer mitochondrial membrane protein essential for mitochondrial networking in most cells. Autosomal dominant mutations in the MFN2 gene cause Charcot–Marie–Tooth type 2A disease (CMT2A), a severe and disabling sensory-motor neuropathy that impacts the entire nervous system. Here, we propose a novel therapeutic strategy tailored to correcting the root genetic defect of CMT2A. Though mutant and wild-type MFN2 mRNA are inhibited by RNA interference (RNAi), the wild-type protein is restored by overexpressing cDNA encoding functional MFN2 modified to be resistant to RNAi. We tested this strategy in CMT2A patient-specific human induced pluripotent stem cell (iPSC)-differentiated motor neurons (MNs), demonstrating the correct silencing of endogenous MFN2 and replacement with an exogenous copy of the functional wild-type gene. This approach significantly rescues the CMT2A MN phenotype in vitro, stabilizing the altered axonal mitochondrial distribution and correcting abnormal mitophagic processes. The MFN2 molecular correction was also properly confirmed in vivo in the MitoCharc1 CMT2A transgenic mouse model after cerebrospinal fluid (CSF) delivery of the constructs into newborn mice using adeno-associated virus 9 (AAV9). Altogether, our data support the feasibility of a combined RNAi and gene therapy strategy for treating the broad spectrum of human diseases associated with MFN2 mutations. [ABSTRACT FROM AUTHOR]

Copyright of Revista Foco (Interdisciplinary Studies Journal) is the property of Revista Foco and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Several drug screening campaigns identified Calpeptin as a drug candidate against SARS-CoV-2. Initially reported to target the viral main protease (Mpro), its moderate activity in Mpro inhibition assays hints at a second target. Indeed, we show that Calpeptin is an extremely potent cysteine cathepsin inhibitor, a finding additionally supported by X-ray crystallography. Cell infection assays proved Calpeptin's efficacy against SARS-CoV-2. Treatment of SARS-CoV-2-infected Golden Syrian hamsters with sulfonated Calpeptin at a dose of 1 mg/kg body weight reduces the viral load in the trachea. Despite a higher risk of side effects, an intrinsic advantage in targeting host proteins is their mutational stability in contrast to highly mutable viral targets. Here we show that the inhibition of cathepsins, a protein family of the host organism, by calpeptin is a promising approach for the treatment of SARS-CoV-2 and potentially other viral infections. Reinke et. al. show that targeting host proteins like cathepsins is a promising strategy for treating SARS-CoV-2 and other viral infections due to their mutational stability in contrast to highly mutable viral targets. [ABSTRACT FROM AUTHOR]

An emergence of evidence suggests that severe COVID-19 is associated with an increased risk of developing breast and gastrointestinal cancers. The aim of this research was to assess the risk of heart tumors development in patients who have had COVID-19. Methods: A comparative analysis of 173 heart tumors was conducted between 2016 and 2023. Immunohistochemical examination with antibodies against spike SARS-CoV-2 was performed on 21 heart tumors: 10 myxomas operated before 2020 (the control group), four cardiac myxomas, one proliferating myxoma, three papillary fibroelastomas, two myxofibrosarcomas, one chondrosarcoma resected in 2022–2023. Immunohistochemical analysis with antibodies against CD34 and CD68 was also conducted on the same 11 Post-COVID period heart tumors. Immunofluorescent examination with a cocktail of antibodies against spike SARS-CoV-2/CD34 and spike SARS-CoV-2/CD68 was performed in 2 cases out of 11 (proliferating myxoma and classic myxoma). Results: A 1.5-fold increase in the number of heart tumors by 2023 was observed, with a statistically significant increase in the number of myxomas. There was no correlation with vaccination, and no significant differences were found between patients from 2016–2019 and 2021–2023 in terms of gender, age, and cardiac rhythm dis-orders. Morphological examination revealed the expression of spike SARS-CoV-2 in tumor cells, endothelial cells, and macrophages in 10 out of 11 heart tumors. Conclusion: The detection of SARS-CoV-2 persistence in endothelium and macrophages as well as in tumor cells of benign and malignant cardiac neoplasms, the increase in the number of these tumors, especially cardiac myxomas, after the pandemic by 2023 may indicate a trend toward an increased risk of cardiac neoplasms in COVID-19 patients, which re-quires further research on this issue and a search for new evidence. [ABSTRACT FROM AUTHOR]

Background: The so-called Low Cardiac Output Syndrome (LCOS) is one of the most common complications in pediatric patients with congenital heart disease undergoing corrective surgery. LCOS requires high concentrations of inotropes to support cardiac contractility and improve cardiac output, allowing for better systemic perfusion. To date, serum lactate concentrations and central venous oxygen saturation (ScVO2) are the most commonly used perfusion markers, but they are not completely reliable in identifying a state of global tissue hypoxia. The study aims to evaluate whether the venoarterial carbon dioxide difference/arterial-venous oxygen difference ratio [P(v-a)CO2/C(a-v)O2] can be a good index to predict the development of LCOS in the aforementioned patients, so as to treat it promptly. Methods: This study followed a population of 98 children undergoing corrective cardiac surgery from June 2018 to October 2020 at the Department of Cardiac Surgery of University Hospital Integrated Trust and their subsequent admission at the Postoperative Cardiothoracic Surgery Intensive Care Unit. During the study, central arterial and venous blood gas analyses were carried out before and after cardiopulmonary bypass (CPB) (pre-CPB and post-CPB), at admission to the intensive care unit, before and after extubation, and at any time of instability or modification of the patient's clinical and therapeutic conditions. Results: The data analysis shows that 46.9% of the children developed LCOS (in line with the current literature) but that there is no statistically significant association between the P(v-a)CO2/C(a-v)O2 ratio and LCOS onset. Despite the limits of statistical significance, however, a 31% increase in the ratio emerged from the pre-CPB phase to the post-CPB phase when LCOS is present. Conclusions: This study confirms a statistically significant association between the most used markers in adult patients (serum lactate concentration, ScVO2, and oxygen extraction ratio—ERO2) measured in the pre-CPB phase and the incidence of LCOS onset, especially in patients with hemodynamic instability before surgery. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effect of film-forming polymer solutions of different concentrations and pH values, either associated or not with sodium fluoride (F; 225 ppm F−), when applied during the initial stage of salivary pellicle formation, to prevent the dissolution of hydroxyapatite (HA), which was determined by the pH-stat method. Polyacrylic acid (PA), chitosan, sodium linear polyphosphate (LPP), polyvinyl methyl ether/maleic anhydride (PVM/MA), and propylene glycol alginate (PGA) were tested in three concentrations (lower, medium, and higher), two pH values (native or adjusted), and either associated or not with F. Distilled water, F, and stannous ion+fluoride (Sn/F; 225 ppm F− and 800 ppm Sn2+, as SnCl2) solutions were the controls, totalizing 63 groups. HA crystals were pretreated with human saliva for 1 min to allow pellicle formation, then immersed in the experimental solutions (1 min), and exposed to saliva for another 28 min. Subsequently, they were added to a 0.3% citric acid solution (pH = 3.8), connected to a pH-stat system that added aliquots of 28 μL 0.1 N HCl for a total reaction time of 5 min. Data were analyzed with one-way ANOVA and Tukey's tests (α = 0.05). For PA alone, the concentrations of 0.1% (native pH), 0.06%, and 0.08% (both pH adjusted) showed significantly lower HA dissolution than the negative control. PA concentrations of 0.1% and 0.08%, of both pH values, improved the effect of F against HA dissolution to a near-identical value as Sn/F. All solutions containing chitosan and LPP significantly reduced HA dissolution in comparison with the control. For chitosan, the concentration of 0.5% (in both pH values) improved the effect of F. LPP at 0.5% (native pH) and all associations of LPP with F outperformed the effect of F. Some PVM/MA solutions significantly reduced HA dissolution but PVM/MA could not improve the protection of F. PGA was incapable of reducing HA dissolution or improving F effect. It was concluded that chitosan, LPP, and some PA and PVM/MA solutions used alone were capable of reducing HA dissolution. Only PA, chitosan, and LPP were able to enhance fluoride protection, but for PA and chitosan, this was influenced by the polymer concentration. [ABSTRACT FROM AUTHOR]

Background: Multiple myeloma (MM) is an incurable plasma cell malignancy, accounting for approximately 1% of all cancers. Despite recent advances in the treatment of MM, due to the introduction of proteasome inhibitors (PIs) such as bortezomib (BTZ) and carfilzomib (CFZ), relapses and disease progression remain common. Therefore, a major challenge is the development of novel therapeutic approaches to overcome drug resistance, improve patient outcomes, and broaden PIs applicability to other pathologies. Methods: We performed genetic and drug screens to identify new synthetic lethal partners to PIs, and validated candidates in PI-sensitive and -resistant MM cells. We also tested best synthetic lethal interactions in other B-cell malignancies, such as mantle cell, Burkitt's and diffuse large B-cell lymphomas. We evaluated the toxicity of combination treatments in normal peripheral blood mononuclear cells (PBMCs) and bone marrow stromal cells (BMSCs). We confirmed the combo treatment' synergistic effects ex vivo in primary CD138+ cells from MM patients, and in different MM xenograft models. We exploited RNA-sequencing and Reverse-Phase Protein Arrays (RPPA) to investigate the molecular mechanisms of the synergy. Results: We identified lysine (K)-specific demethylase 1 (LSD1) as a top candidate whose inhibition can synergize with CFZ treatment. LSD1 silencing enhanced CFZ sensitivity in both PI-resistant and -sensitive MM cells, resulting in increased tumor cell death. Several LSD1 inhibitors (SP2509, SP2577, and CC-90011) triggered synergistic cytotoxicity in combination with different PIs in MM and other B-cell neoplasms. CFZ/SP2509 treatment exhibited a favorable cytotoxicity profile toward PBMCs and BMSCs. We confirmed the clinical potential of LSD1-proteasome inhibition in primary CD138+ cells of MM patients, and in MM xenograft models, leading to the inhibition of tumor progression. DNA damage response (DDR) and proliferation machinery were the most affected pathways by CFZ/SP2509 combo treatment, responsible for the anti-tumoral effects. Conclusions: The present study preclinically demonstrated that LSD1 inhibition could provide a valuable strategy to enhance PI sensitivity and overcome drug resistance in MM patients and that this combination might be exploited for the treatment of other B-cell malignancies, thus extending the therapeutic impact of the project. [ABSTRACT FROM AUTHOR]

Copyright of Revista Foco (Interdisciplinary Studies Journal) is the property of Revista Foco and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The aim of the study was to evaluate the association between salivary anti-Porphyromonas gingivalis IgA antibodies and the leprosy reaction. The levels of salivary anti - P. gingivalis IgA antibodies, together with salivary flow and pH were measured in individuals diagnosed with leprosy and associated with the development of the leprosy reaction. Saliva was collected from 202 individuals diagnosed with leprosy at a reference leprosy treatment center, 106 cases with the leprosy reaction and 96 controls without the leprosy reaction. Anti - P. gingivalis IgA was evaluated by indirect immunoenzyme assay. Non-conditional logistic regression analysis was employed to estimate the association between antibody levels and the leprosy reaction. There was a positive statistically significant association between the levels of anti - P. gingivalis IgA and the presence of the leprosy reaction, controlling for confounders: age, sex, level of education and alcoholic beverage consumption: ORajusted: 2.55; IC 95%: 1.34–4.87. Individuals with leprosy who had high levels of salivary anti - P. gingivalis IgA had approximately twice as many chances of developing the leprosy reaction. The findings suggest a possible relationship between salivary anti - P. gingivalis IgA antibodies and the leprosy reaction. Key Points: Oral manifestations may predispose to the leprosy reaction. Salivary anti - P. gingivalis IgA antibodies are associated with leprosy reaction. Marker for leprosy reaction in individuals with periodontitis. [ABSTRACT FROM AUTHOR]

The clinical manifestation study of post-acute sequelae of SARS-CoV-2 infection (PASC) has shown a lack of knowledge regarding its morphology and pathogenesis. The aim of this research was to investigate morphological manifestations of PASC in the myocardium. Materials and Methods: The study included 38 patients requiring endomyocardial biopsy (EMB) during the post-acute phase of coronavirus infection and a control group including patients requiring EMB prior to the SARS-CoV-2 pandemic. The patients' clinical and laboratory data were analyzed. Histological examination and immunohistochemistry (IHC) of the myocardial tissue was conducted with antibodies to CD3, CD68, HLA-DR, MHC1, C1q, VP1 enteroviruses, spike protein SARS-CoV-2, Ang1, von Willebrand factor (VWF), and VEGF. The morphometric analysis included counting the mean number of inflammatory infiltrate cells per mm2 and evaluating the expression of SARS-CoV-2 spike protein, HLA-DR, MHC1, C1q, Ang1, VWF, and VEGF using a scoring system. If the expression of SARS-CoV-2 spike protein was >3 points, an additional IHC test with antibodies to ACE2, CD16 as well as RT-PCR testing of the myocardial tissue were performed. For two patients, immunofluorescence tests of the myocardial tissue were performed using antibody cocktails to SARS-CoV-2 spike protein/CD16, SARS-CoV-2 spike protein/CD68, CD80/CD163. The statistical data analysis was carried out using the Python programming language and libraries such as NumPy, SciPy, Pandas, and Matplotlib. Results: The study demonstrated a significant increase in the number of CD68+ macrophages in the myocardium of PASC patients compared to patients who did not have a history of COVID-19 (p = 0.014 and p = 0.007 for patients with and without myocarditis, respectively), predominantly due to M2 macrophages. An increase in the number of CD68+ macrophages was more frequently observed in patients with shorter intervals between the most recent positive SARS-CoV-2 PCR test and the time of performing the EMB (r = −0.33 and r = −0.61 for patients with and without myocarditis, respectively). The expression scores of Ang1, VEGF, VWF, and C1q in PASC patients did not significantly differ from those in EMB samples taken before 2019. Conclusion: The myocardium of PASC patients demonstrated a significant increase in the number of CD68+ macrophages and a decrease in the expression of markers associated with angiopathy. No evidence of coronavirus-associated myocarditis was observed in any PASC patient. [ABSTRACT FROM AUTHOR]

In order to investigate the effectiveness of vibrational spectroscopy in the characterization of polymer piezoelectric materials, poly(vinylidene fluoride) (PVDF) samples are carefully studied. The characterizations are performed by using different Fourier transform infrared sampling techniques, i.e. transmission (FTIR), attenuated total reflectance (FTIR-ATR) and photoacoustic spectroscopy (FTIR-PAS). The attenuated total reflectance (ATR) and photoacoustic (PAS) sampling methods allow us to study the IR spectra for thick piezoelectric samples. To the best of our knowledge, it has been determined for the first time the relative fractions of the β phase, F(β), in PVDF by FTIR-PAS. [ABSTRACT FROM AUTHOR]

Artificial materials with piezoelectric properties have been increasingly investigated as potential candidates for replacing various living tissues. In this manuscript, results of studies on the synthesis and characterization PVDF/PLA blend filaments containing piezoelectric/ferroelectric PVDF β-phase are reported. SEM images reveal that all studied filaments are immiscible blends, while FTIR results reveal blends with a predominance of PVDF β-polar phase whose highest percentage (75%) was observed for the mass ratio of 80% of PVDF to 20% of PLA. This blend shows adequate characteristics for biomedical applications when processed by filament fabrication techniques. [ABSTRACT FROM AUTHOR]

Background: Few studies have compared COVID-19 patients from different waves. This study aims to conduct a clinical and morphological analysis of patients who died from COVID-19 during four waves. Methods: The study involved 276 patients who died from COVID-19 during four waves, including 77 patients in the first wave, 119 patients in the second wave, and 78 patients in the third wave. We performed a histological examination of myocardium samples from autopsies and additionally analyzed the samples by PCR. We conducted immunohistochemistry of the myocardium for 21 samples using antibodies against CD3, CD45, CD8, CD68, CD34, Ang1, VWF, VEGF, HLA-DR, MHC1, C1q, enteroviral VP1, and SARS-CoV-2 spike protein. We also did immunofluorescent staining of three myocardial specimens using VP1/SARS-CoV-2 antibody cocktails. Further, we ran RT-ddPCR analysis for 14 RNA samples extracted from paraffin-embedded myocardium. Electron microscopic studies of the myocardium were also performed for two samples from the fourth wave. Results: Among the 276 cases, active myocarditis was diagnosed in 5% (15/276). Of these cases, 86% of samples expressed VP1, and individual cells contained SARS-CoV-2 spike protein in 22%. Immunofluorescence confirmed the co-localization of VP1 and SARS-CoV-2 spike proteins. ddPCR did not confidently detect SARS-CoV-2 RNA in the myocardium in any myocarditis cases. However, the myocardium sample from wave IV detected a sub-threshold signal of SARS-CoV-2 by qPCR, but myocarditis in this patient was not confirmed. Electron microscopy showed several single particles similar to SARS-CoV-2 virions on the surface of the endothelium of myocardial vessels. A comparison of the cardiovascular complication incidence between three waves revealed that the incidence of hemorrhage (48 vs. 24 vs. 17%), myocardial necrosis (18 vs. 11 vs. 4%), blood clots in the intramural arteries (12 vs. 7 vs. 0%), and myocarditis (19 vs. 1 vs. 6%) decreased over time, and CD8-T-killers appeared. Immunohistochemistry confirmed the presence of endotheliitis in all 21 studied cases. Conclusions: This study compared myocardial damage in patients who died during three COVID-19 waves and showed a decrease in the incidence of endotheliitis complications (thrombosis, hemorrhage, necrosis) and myocarditis over time. However, the connection between myocarditis and SARS-CoV-2 infection remains unproven. [ABSTRACT FROM AUTHOR]

In newborn humans, and up to approximately 2 y of age, calvarial bone defects can naturally regenerate. This remarkable regeneration potential is also found in newborn mice and is absent in adult mice. Since previous studies showed that the mouse calvarial sutures are reservoirs of calvarial skeletal stem cells (cSSCs), which are the cells responsible for calvarial bone regeneration, here we hypothesized that the regenerative potential of the newborn mouse calvaria is due to a significant amount of cSSCs present in the newborn expanding sutures. Thus, we tested whether such regenerative potential can be reverse engineered in adult mice by artificially inducing an increase of the cSSCs resident within the adult calvarial sutures. First, we analyzed the cellular composition of the calvarial sutures in newborn and in older mice, up to 14-mo-old mice, showing that the sutures of the younger mice are enriched in cSSCs. Then, we demonstrated that a controlled mechanical expansion of the functionally closed sagittal sutures of adult mice induces a significant increase of the cSSCs. Finally, we showed that if a calvarial critical size bone defect is created simultaneously to the mechanical expansion of the sagittal suture, it fully regenerates without the need for additional therapeutic aids. Using a genetic blockade system, we further demonstrate that this endogenous regeneration is mediated by the canonical Wnt signaling. This study shows that controlled mechanical forces can harness the cSSCs and induce calvarial bone regeneration. Similar harnessing strategies may be used to develop novel and more effective bone regeneration autotherapies. [ABSTRACT FROM AUTHOR]

Introduction: Tyrosine kinase inhibitor (TKI) therapy has greatly improved the prognosis of patients with chronic myeloid leukemia (CML), improving the survival expectancy of patients with chronic phase (CP) CML to that of the general population. However, despite these advances, nearly 50% of patients with CP CML experience failure to respond to frontline therapy, and most fail to respond to the subsequent second-line TKI. Treatment guidelines for patients failing second-line therapy are lacking. This study aimed to determine the efficacy of TKIs as third-line therapy in a "real-world" clinical practice setting and identify factors favorably influencing the long-term outcomes of therapy. Methods: We have retrospectively analyzed the medical records of 100 patients with CP CML. Results: The median age of the patients was 51 (range, 21-88) years, and 36% of the patients were men. The median duration of the third-line TKI therapy was 22 (range, 1-147) months. Overall, the rate of achieving complete cytogenetic response (CCyR) was 35%. Among the four patient groups with different levels of responses at baseline, the best results were achieved in the groups with any CyR at the baseline of third-line therapy. Thus, ?CyR was reached in all 15 and 8/16 (50%) patients with partial cytogenetic response (PCyR) or minimal or minor CyR (mmCyR), respectively, whereas CCyR was detected only in 12/69 (17%) patients without any CyR at baseline (p < 0.001). Univariate regression analysis revealed that the factors negatively associated with CCyR achievement in thirdline TKI therapy were the absence of any CyR on first- or second-line TKI therapy (p < 0.001), absence of CHR prior to third-line TKI (p = 0.003), and absence of any CyR prior to third-line TKI (p < 0.001). During the median observation time from treatment initiation to the last visit [56 (4-180) months], 27% of cases progressed into accelerated phase or blast phase CML, and 32% of patients died. Discussion: Progression-free survival (PFS) and overall survival (OS) were significantly higher in patients with CCyR on third-line than in the group without CCyR on third-line therapy. At the last visit, third-line TKI therapy was ongoing in 18% of patients, with a median time of treatment exposure of 58 (range, 6-140) months; 83% of these patients had stable and durable CCyR, suggesting that patients without CHR at baseline and without CCyR at least by 12 months on third-line TKI should be candidates for allogeneic stem cell transplantation, third-generation TKIs, or experimental therapies. [ABSTRACT FROM AUTHOR]

The decision of premature culling cows directly impacts the profitability of dairy farms. A comprehensive characterization of the primary causes of culling reasons would greatly improve both management and selection objectives in dairy cattle breeding programs. Therefore, this study aimed to analyze the temporal frequencies of 34 culling reasons in Canadian Holstein cows. After data editing and quality control, records from 3,096,872 cows culled from 9,683 herds spread across Canada were used for the analyses covering the periods from 1996 to 2020. Reproductive issues were the main culling reason accounting for 23.02%, followed by milk production (20.82%), health (20.39%), conformation problems (13.69%), economic factors (13.10%), accidents (5.67%), age-related causes (1.67%), and workability (1.63%). Nearly fifty-eight percent of cows were culled after 47 months of age. The observed frequencies of culling due to economic factors were lower than expected from 1996 to 2014 and higher than expected between 2015 and 2020. Reproduction issues had the highest culling frequencies during fall (24.54%), winter (24.02%), and spring (22.51%), while health issues were the most frequent (22.51%) culling reason in the summer season. Health issues (25.50%) and milk production (27.71%) were the most frequent culling reasons in the provinces of Quebec and Ontario, respectively. Reproductive issues showed the highest frequency across climates based on the Köppen climate classification, except for Csb (Dry-summer subtropical or Mediterranean climate) and Bsk (Middle latitude steppe climate), which correspond to small regions in Canada, where production was the most frequent culling reason (29.42% and 21.56%, respectively). Reproductive and milk performance issues were the two main culling reasons in most ecozones, except in Boreal Shield and Atlantic Marine, where health issues had the highest frequencies (25.12 and 23.75%, respectively). These results will contribute to improving management practices and selective decisions to reduce involuntary culling of Holstein cows. [ABSTRACT FROM AUTHOR]

Hyaluronic acid (HA) is an exopolysaccharide extracted from several sources such as rooster combs, umbilical cords and microorganisms. A system that controls temperature, agitation and aeration of bacterial cultures could make the HA production autonomous. Therefore, HA of microbial origin is set to take over alternative methods of production. Furthermore, the use of different nutrient sources in the culture medium and the purification stage applied in the process can cause physicochemical alterations on the bioproduct. For instance, structural modifications that change the molecular weight of HA may alter its elastic and viscoelastic properties. As a result, HA synthesized by microbes has applications in pharmacology, biotechnology, and tissue engineering. Our aim here, is to show the vast range of applications by compiling articles and patents on the culture media or genetic modifications of microorganisms that synthesize HA. [ABSTRACT FROM AUTHOR]

In cropping systems, the choices adopted for the tillage system used and plants cultivated can strongly influence the soil microbial population and its functional profile. Arbuscular mycorrhizal fungi are an important component of soil microbiome and their mutualistic symbiosis with the majority of higher plants grant the latter a wide range of benefits. The extraradical mycelium developed by these fungi expands the volume of soil influenced and harbours a diversity of microbes establishing a distinct environment of complementary interactions. We assessed how growing plants with different levels of mycotrophy modifies the biological activity profile in the soil under Mn toxicity and whether this is modified by soil disturbance. Following mycotrophic plants, soil contained a more active microbiome than after the non‐mycotrophic plants, as expressed by higher values of soil basal respiration or dehydrogenase activity. Additionally, the count of phosphorus solubilizes and activity of phosphatase were greater after mycotrophic plants. Even among mycotrophic plants, different profiles of biological activity can be distinguished after growing a legume or grass. ERM disruption by soil disturbance decreased most of the parameters studied and for phosphatase activity and P solubilizers in a more significant way. These results indicate that even under Mn toxicity, the microbiome associated with AMF symbiosis following mycotrophic plants growth presented a higher biological activity and had a differential biological response towards the stress imposed by soil disturbance, when compared with the microbiome associated with non‐mycotrophic roots. [ABSTRACT FROM AUTHOR]

Multiple myeloma (MM) has a highly heterogeneous genetic background, which complicates its molecular tracking over time. Nevertheless, each MM patient's malignant plasma cells (PCs) share unique V(D)J rearranged sequences at immunoglobulin loci, which represent ideal disease biomarkers. Because the tumor-specific V(D)J sequence is highly expressed in bulk RNA in MM patients, we wondered whether it can be identified by single-cell RNA sequencing (scRNA-seq). To this end we analyzed CD138+ cells purified from bone marrow aspirates of 19 samples with PC dyscrasias by both a standard method based on bulk DNA and by an implementation of the standard 10x Genomics protocol to detect expressed V(D)J sequences. A dominant clonotype was easily identified in each sample, accounting on average for 83.65% of V(D)J-rearranged cells. Compared with standard methods, scRNA-seq analysis proved highly concordant and even more effective in identifying clonal productive rearrangements, by-passing limitations related to the misannealing of consensus primers in hypermutated regions. We next validated its accuracy to track 5 clonal cells with absolute sensitivity in a virtual sample containing 3180 polyclonal cells. This shows that single-cell V(D)J analysis may be used to find rare clonal cells, laying the foundations for functional single-cell dissection of minimal residual disease. [ABSTRACT FROM AUTHOR]

As a faster, reliable, and low cost technique, applicable to large samplings, near infrared (NIR) spectroscopy technology has been widely applied for highthroughput phenotyping in forest breeding programmes. The aim of this study was to develop multivariate models for estimating the chemical and physical properties of juvenile wood based on NIR signatures of milled wood. Moreover, two approaches, namely, external validation by clone and by age, were tested to validate the model for estimating extractive content. NIR spectra of wood specimens taken from three clones of Eucalyptus urophylla (one to six years old) grown in southern Brazil were used to calibrate and validate models for predicting the wood basic density, total extractives, ash content, holocellulose content, syringyl to guaiacyl ratio (S/G) and elementary components of the wood. PLS-R models were validated by an independent set of wood specimens and presented promising statistics for the estimating wood density (R2p = 0.768), extractives (R2p = 0.912), ash (R2p = 0.936) and carbon (R2p = 0.697) contents from NIR signatures measured in the milled wood of young trees. Furthermore, NIR models for estimating the extractive content of wood were validated using the clones or ages left out of the training sets. Most models presented satisfactory statistics (R2 > 90%) and could be applied to routine laboratory analyses or to select potential trees in Eucalyptus breeding programmes. [ABSTRACT FROM AUTHOR]

In the current agronomic context, the adoption of alternative forms of soil management is essential to increase crop yield. Agricultural sustainability requires practices that generate positive impacts and promote an increase in microbiome diversity as a tool to overcome adverse environmental conditions. An important ally is the indigenous arbuscular mycorrhizal fungi (AMF) that can improve plant growth and provide protection against abiotic stress such as metal toxicity. In a greenhouse experiment, this work studied the effect of wheat growth on several parameters of biological activity and functional microbiome in relation to wheat antecedent plant mycotrophy and soil disturbance under Mn stress. When the wheat was planted after highly mycotrophic plants and the soil was not previously disturbed, the results showed a 60% increase in wheat arbuscular colonization and a 2.5-fold increase in dry weight along with higher values of photosynthetic parameters and dehydrogenase activity. Conversely, soil disturbance before wheat planting increased the β-glucosidase activity and the count of manganese oxidizers, irrespectively of antecedent plant, and decreased drastically the wheat dry weight, the AMF colonization and the chlorophyll content compared to the undisturbed treatment. These findings suggest that not only the wheat growth but also the soil functional microbiome associated is affected by the antecedent type of plant and previous soil disturbance imposed. In addition, the improvement in wheat dry weight despite Mn toxicity may rely on shifts in biological activity associated to a well-established and intact ERM early developed in the soil. [ABSTRACT FROM AUTHOR]

Biomaterials that effectively act in biological systems, as in treatment and healing of damaged or lost tissues, must be able to mimic the properties of the body's natural tissues in its various aspects (chemical, physical, mechanical and surface). These characteristics influence cell adhesion and proliferation and are crucial for the success of the treatment for which a biomaterial will be required. In this context, the electrospinning process has gained prominence in obtaining fibers of micro- and nanometric sizes from polymeric solutions aiming to produce scaffolds for tissue engineering. In this manuscript, poly(vinylidene fluoride) (PVDF) was used as a polymeric matrix for the manufacture of piezoelectric scaffolds, exploring the formation of the β -PVDF piezoelectric phase. Micro- and nanometric hydroxyapatite (HA) particles were incorporated as a dispersed phase in this matrix, aiming to produce multifunctional composite membranes also with bioactive properties. The results show that it is possible to produce membranes containing micro- and nanofibers of the composite by the electrospinning process. The HA particles show good dispersion in the polymer matrix and predominance of β -PVDF phase. Also, the composite showed apatite growth on its surface after 21 days of immersion in simulated body fluid (SBF). Tests performed on human fibroblasts culture revealed that the electrospun membranes have low cytotoxicity attesting that the composite shows great potential to be used in biomedical applications as bone substitutions and wound healing. [ABSTRACT FROM AUTHOR]

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The post-Bionian paradigm in psychoanalysis invites us to listen to the session as a waking-dream-thought where unconscious-thinking-in progress is continuous. The hypothesis put forward here and illustrated using clinical material is that we can use the notion of day's residues as a metaphor to refer to the incoming narrative of the patient. Whatever the patient brings to the session can be conceived as "day's residues" in that they are potential instigators of waking-dream-thought in the session. This metaphor helps the analyst place brackets around the outside of the session, deconcretizing what apparently are hard facts, so that immediate contact is made to create a shared perspective, possibly producing in this session "food" for the mind. To create the waking-dream-thought of the session, the analyst may consider listening to the incoming narrative as a metaphor. This is not a new or different concept but a particular kind of elaboration on the metaphoric stance taken by psychoanalysts of all stripes; it is an elaboration that expands the ways we can describe the session and narrow the gap between talking about the session and the experience of the session itself. [ABSTRACT FROM AUTHOR]

We report on a 15‐year‐long outbreak of bovine tuberculosis (bTB) in wildlife from a Brazilian safari park. A timeline of diagnostic events and whole‐genome sequencing (WGS) of 21 Mycobacterium bovis isolates from deer and llamas were analyzed. Accordingly, from 2003 to 2018, at least 16 animals, from eight species, died due to TB, which is likely an underestimated number. In three occasions since 2013, the deer presented positive tuberculin tests, leading to the park closure and culling of all deer. WGS indicated that multiple M. bovis strains were circulating, with at least three founding introductions since the park inauguration in 1977. Using a previously sequenced dataset of 71 M. bovis genomes from cattle, we found no recent transmission events between nearby farms and the park based on WGS. Lastly, by discussing socio‐economic and environmental factors escaping current regulatory gaps that were determinant of this outbreak, we pledge for the development of a plan to report and control bTB in wildlife in Brazil. [ABSTRACT FROM AUTHOR]

Manganese (Mn) and aluminum (Al) toxicities are serious edaphic limitations to crop production in acidic soils. Excess Mn can be countered using a stress-adapted soil microbiota that establish symbiotic relationships with native plants. The arbuscular mycorrhizal fungi (AMF) associated with Lolium rigidum L. develop extraradical mycelia (ERM) that quickly colonize wheat and lead to greater shoot growth by promoting stress-evading mechanisms that are not yet completely explained. In the present study, wheat growth was assessed after 3 weeks on disturbed and undisturbed (intact ERM) acidic soil where the native non-mycotrophic Silene gallica L. or strongly mycotrophic L. rigidum were previously developed. The physiological and biochemical mechanisms responsible for increased growth were analyzed by assessing wheat leaf chlorophyll content, photosystem II quantum yield and performance index, enzymatic activity of ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol peroxidase (GPX), superoxide dismutase (SOD) and contents and subcellular localization of Mn, Mg, Si and K. The soil from native plants had a beneficial effect on shoot weight and chlorophyll levels. The highest benefits were obtained for wheat grown in soil with intact ERM associated with L. rigidum. In this condition, where earlier mycorrhization was favored, the Mn content decreased, alongside the content of Si, while the Mg/Mn ratio increased. Mn was redirected to the apoplast, while Si was redirected to the symplast. The activity of APX, GPX and SOD increased, probably due to increased metabolic growth (higher shoot weight and chlorophyll content). Understanding the mechanisms induced by native AMF responsible for increasing wheat performance can contribute to the establishment of sustainable approaches for crop production in acidic soils with Mn toxicity. The use of native plant AMF developers can improve the sustainable use of natural resources in the scope of greener agricultural practices. [ABSTRACT FROM AUTHOR]