Search

Abstract Immune deficits after CD19 chimeric antigen receptor (CAR) T‐cell therapy can be long‐lasting, predisposing patients to infections and non‐relapse mortality. In B‐cell non‐Hodgkin lymphoma (B‐NHL), the prognostic impact of immune reconstitution (IR) remains ill‐defined, and detailed cross‐product comparisons have not been performed to date. In this retrospective observational study, we longitudinally characterized lymphocyte subsets and immunoglobulin levels in 105 B‐NHL patients to assess patterns of immune recovery arising after CD19 CAR‐T. Three key IR criteria were defined as CD4+ T helper (TH) cells > 200/µL, any detectable B cells, and serum immunoglobulin G (IgG) levels >4 g/L. After a median follow‐up of 24.6 months, 38% of patients displayed TH cells, 11% showed any B cells, and 41% had IgG recovery. Notable product‐specific differences emerged, including deeper TH cell aplasia with CD28z‐ versus longer B‐cell aplasia with 41BBz‐based products. Patients with any IR recovery experienced extended progression‐free survival (PFS) (median 20.8 vs. 1.7 months, p

BackgroundThe pharmacoinvasive (PhI) strategy is the standard-of-care for ST-elevation myocardial infarction (STEMI) patients when primary percutaneous coronary intervention (pPCI) is unfeasible. Optimal timing for post-fibrinolytic PCI (lysis-PCI) remains elusive. Therefore, this study aimed to assess the clinical and economic impacts of early vs. delayed lysis-PCI in patients with STEMI.MethodsThis retrospective cohort study included 1,043 STEMI patients classified by lysis-PCI timing. The primary outcome was in-hospital major adverse cardiovascular events (4p-MACE), with secondary outcomes such as 3p-MACE, in-hospital mortality, and costs. Multivariable logistic regression models were used to assess the association between lysis-PCI timing and outcomes. Cost analyses were conducted from the perspective of Brazilian public healthcare system, with values converted to international dollars (Int$) for broader applicability.ResultsEvery 4-h delay in lysis-PCI was associated with a 44% reduction in in-hospital mortality [OR = 0.560 (95% CI: 0.381–0.775); p = 0.001] and a 13% decrease in 4p-MACE [OR = 0.877 (95% CI: 0.811–0.948); p = 0.001]. Additionally, 4-h delay in lysis-PCI was also associated with a significant reduction in in-hospital costs (Int$916.20 ± 99) and disease-induced years of productivity lost (β = −41.79 ± 151 years; p = 0.001). These significant trends remained consistent even after adjusting for confounders and applying propensity score matching. Older adults (aged ≥80) experienced an increase in 3p-MACE with earlier lysis-PCI.ConclusionDelaying lysis-PCI was found to be associated with reduced in-hospital cardiovascular adverse events and lower costs, particularly among older adults. Further research should develop evidence-based lysis-PCI protocols that optimize both clinical outcomes and cost-effectiveness.

Abstract Intramuscular fat (IMAT) infiltration, pathological adipose tissue that accumulates between muscle fibers, is a shared hallmark in a diverse set of diseases including muscular dystrophies and diabetes, spinal cord and rotator cuff injuries, as well as sarcopenia. While the mouse has been an invaluable preclinical model to study skeletal muscle diseases, they are also resistant to IMAT formation. To better understand this pathological feature, an adequate pre-clinical model that recapitulates human disease is necessary. To address this gap, we conducted a comprehensive in-depth comparison between three widely used mouse strains: C57BL/6J, 129S1/SvlmJ and CD1. We evaluated the impact of strain, sex and injury type on IMAT formation, myofiber regeneration and fibrosis. We confirm and extend previous findings that a Glycerol (GLY) injury causes significantly more IMAT and fibrosis compared to Cardiotoxin (CTX). Additionally, females form more IMAT than males after a GLY injury, independent of strain. Of all strains, C57BL/6J mice, both females and males, are the most resistant to IMAT formation. In regard to injury-induced fibrosis, we found that the 129S strain formed the least amount of scar tissue. Surprisingly, C57BL/6J of both sexes demonstrated complete myofiber regeneration, while both CD1 and 129S1/SvlmJ strains still displayed smaller myofibers 21 days post injury. In addition, our data indicate that myofiber regeneration is negatively correlated with IMAT and fibrosis. Combined, our results demonstrate that careful consideration and exploration are needed to determine which injury type, mouse model/strain and sex to utilize as preclinical model especially for modeling IMAT formation.

The Quiricó Formation is the richest fossiliferous unit of Sanfranciscana Basin. Its paleoasis context (a paleolake record inserted in a desert setting) includes vertebrates, diverse microfossils, and paleobotanical content. This study presents new findings regarding the fossil content of the formation, focusing on discoveries made at the Pripiri Geosite, located in Coração de Jesus municipality, northern part of the state of Minas Gerais, Brazil. Among the discoveries are poorly informative fragments of putative archosaurs and novel microfossiliferous taxa. The microfossil assemblage includes dinocysts and Euglenophyceae, both associated with microbial mat-forming cyanobacteria. These, in turn, comprise the filamentous Pseudoanabaena sp. and the coccoidal cells of Aphanocapsa sp. or Chroococus sp. These microbial mats are recurrent throughout the analyzed stratigraphic succession, primarily observed in sandstone bars featuring distinct evaporitic features. This assemblage provides ecological insights concerning the biota resistance to extreme climatic conditions, and their occurrence in oxygen-rich waters. The preservation of these cells in evaporite-rich rocks is also of great interest for astrobiological studies and the search for past life on Mars. Therefore, due to the importance of Pripiri locality, a quantitative analysis of its geodiversity and geoheritage value were performed, revealing that Pripiri comprises a geosite of national relevance in terms of its scientific, touristic and educational values. In this sense, some initiatives have been carried out to geoconservation and to promote the local geodiversity, aiming to contribute to the paleontological knowledge of the Early Cretaceous, as well as to the social and economic development of Coração de Jesus.

Objective: This study aimed to describe the histomorphologic characteristics of resected (unroofed) common wall tissue from repair of anomalous aortic origin of a coronary artery and to determine whether the histologic features correlate with clinical and imaging findings. Methods: The histology of resected tissue was analyzed and reviewed for the presence of fibrointimal hyperplasia, smooth muscle disarray, mucoid extracellular matrix accumulation, mural fibrosis, and elastic fiber disorganization and fragmentation using hematoxylin and eosin and special stains. Clinical, computed tomography imaging, and surgical data were correlated with the histopathologic findings. Results: Twenty specimens from 20 patients (age range, 7-18 years; 14 males) were analyzed. Anomalous aortic origin of a coronary artery involved the right coronary in 16 (80%), and a slit-like ostium was noted in 18 (90%). By computed tomography imaging, the median proximal coronary artery eccentricity index was 0.4 (range, 0.20-0.90). The median length of intramural course was 8.2 mm (range, 2.6-15.2 mm). The anomalous vessel was determined to be interarterial in 14 patients (93%, 15 had evaluable images). The median distance from a commissure was 2.5 mm above the sinotubular junction (STJ) (range: 2 mm below the STJ–14 mm above the STJ). Prominent histopathologic findings included elastic fiber alterations, mural fibrosis, and smooth muscle disarray. The shared wall of the aorta and intramural coronary artery is more similar to the aorta histologically. Mural fibrosis and elastic fiber abnormalities tended to be more severe in patients >10 years of age at the time of surgery, but this did not reach statistical significance. The extent of vascular changes did not appear to have a clear relationship with the imaging features. Conclusions: The findings confirm the aortic wall–like quality of the intramural segment of the coronary artery and the presence of pathologic alterations in the wall microstructure.

The intensification of production processes, resulting from the rise in pork production, contributes to environmental changes and increased interaction between humans, animals, and wildlife. This favorable scenario promotes the spread of potent viral species, such as PCV3, increasing the potential for the emergence of new pathogenic agents and variants. These changes in the epidemiology and manifestation of PCV3 highlight the need for enhanced understanding and control. The current literature presents challenges in the classification of PCV3, with different groups proposing diverse criteria. Establishing common terminology is crucial to facilitate comparisons between studies. While consensus among experts is valuable, new approaches must be transparent and comparable to existing literature, ensuring reproducible results and proper interpretation, and positively impacting public health. This study aims to review the literature on PCV3 infection, exploring its key aspects and highlighting unanswered questions.

Objective The objective of this study was to characterize physiochemical and nutrient profiles of feedstock and co-products from canola bio-oil processing that were impacted by source origin. The feedstocks and co-products (mash, pellet) were randomly collected from five different bio-oil processing plants with five different batches of samples in each bio-processing plant in Canada (CA) and China (CH). Methods The detailed chemical composition, energy profile, total digestible nutrient (TDN), protein and carbohydrate subfractions, and their degradation and digestion (CNCPS6.5) were determined. Results The results showed that TDN1x was similar in meals between CA and CH. CH meals and feedstock had higher, truly digestible crude protein (tdCP) and neutral detergent fiber (tdNDF) than CA while CA had higher truly digestible non-fiber carbohydrate (tdNFC). The metabolizable energy (ME3x), net energy (NELp3x, NEm3x, and NEg3x) were similar in meals between CA and CH. No differences were observed in energy profile of seeds between CA and CH. The protein and carbohydrate subfractions of seeds within CH were similar. The results also showed that pelleting of meals affected protein sub-fractionation of CA meals, except rapidly degradable fractions (PB1), rumen degradable (RDPB1) and undegrdable PB1 (RUPB1), and intestinal digestible PB1 (DIGPB1). Canola meals were different in the soluble (PA2) and slowly degradable fractions (PB2) between CA and CH. The carbohydrate fractions of intermediately degradable fraction (CB2), slowly degradable fraction (CB3), and undegradable fraction (CC) were different among CH meals. CH presented higher soluble carbohydrate (CA4) and lower CB2, and CC than CA meals. Conclusion The results indicated that although the seeds were similar within and between CA and CH, either oil-extraction process or meal pelleting seemed to have generated significantly different aspects in physiochemical and nutrient profiles in the meals. Nutritionists and producers need to regularly check nutritional value of meal mash and pellets for precision feeding.

Abstract Successful muscle regeneration relies on the interplay of multiple cell populations. However, the signals required for this coordinated intercellular crosstalk remain largely unknown. Here, we describe how the Hedgehog (Hh) signaling pathway controls the fate of fibro/adipogenic progenitors (FAPs), the cellular origin of intramuscular fat (IMAT) and fibrotic scar tissue. Using conditional mutagenesis and pharmacological Hh modulators in vivo and in vitro, we identify DHH as the key ligand that acts as a potent adipogenic brake by preventing the adipogenic differentiation of FAPs. Hh signaling also impacts muscle regeneration, albeit indirectly through induction of myogenic factors in FAPs. Our results also indicate that ectopic and sustained Hh activation forces FAPs to adopt a fibrogenic fate resulting in widespread fibrosis. In this work, we reveal crucial post-developmental functions of Hh signaling in balancing tissue regeneration and fatty fibrosis. Moreover, they provide the exciting possibility that mis-regulation of the Hh pathway with age and disease could be a major driver of pathological IMAT formation.

Abstract Facioscapulohumeral muscular dystrophy (FSHD) is caused by the epigenetic derepression of the 4q-linked D4Z4 macrosatellite repeat resulting in inappropriate expression of the D4Z4 repeat-encoded DUX4 gene in skeletal muscle. In 5% of FSHD cases, D4Z4 chromatin relaxation is due to germline mutations in one of the chromatin modifiers SMCHD1, DNMT3B or LRIF1. The mechanism of SMCHD1- and LRIF1-mediated D4Z4 repression is not clear. We show that somatic loss-of-function of either SMCHD1 or LRIF1 does not result in D4Z4 chromatin changes and that SMCHD1 and LRIF1 form an auxiliary layer of D4Z4 repressive mechanisms. We uncover that SMCHD1, together with the long isoform of LRIF1, binds to the LRIF1 promoter and silences LRIF1 expression. The interdependency of SMCHD1 and LRIF1 binding differs between D4Z4 and the LRIF1 promoter, and both loci show different transcriptional responses to either early developmentally or somatically perturbed chromatin function of SMCHD1 and LRIF1.

Objective This program aimed to reveal the association of feed intrinsic molecular structure with nutrient supply to animals from canola feedstocks and co-products from bio-oil processing. The special objective of this study was to quantify the relationship between molecular spectral feature and nutrient availability and develop nutrient prediction equation with vibrational molecular structure spectral profiles. Methods The samples of feedstock (canola oil seeds) and co-products (meals and pellets) from different bio-oil processing plants in Canada (CA) and China (CH) were submitted to this molecular spectroscopic technique and their protein and carbohydrate related molecular spectral features were associated with the nutritional results obtained through the conventional methods of analyses for chemical and nutrient profiles, rumen degradable and intestinal digestible parameters. Results The results showed that the spectral structural carbohydrates spectral peak area (ca. 1,487.8 to 1,190.8 cm−1) was the carbohydrate structure that was most significant when related to various carbohydrate parameters of canola meals (p0.50). And spectral total carbohydrate area (ca. 1,198.5 to 934.3 cm−1) was most significant when studying the various carbohydrate parameters of canola seeds (p0.50). The spectral amide structures (ca. 1,721.2 to 1,480.1 cm−1) were related to a few chemical and nutrient profiles, Cornell Net Carbohydrate and Protein System (CNCPS) fractions, truly absorbable nutrient supply based on the Dutch protein system (DVE/OEB), and NRC systems, and intestinal in vitro protein-related parameters in co-products (canola meals). Besides the spectral amide structures, α-helix height (ca. 1,650.8 to 1,643.1 cm−1) and β-sheet height (ca. 1,633.4 to 1,625.7 cm−1), and the ratio between them have shown to be related to many protein-related parameters in feedstock (canola oil seeds). Multi-regression analysis resulted in moderate to high R2 values for some protein related equations for feedstock (canola seeds). Protein related equations for canola meals and carbohydrate related equations for canola meals and seeds resulted in weak R2 and low p values (p

Objective Feed molecular structures can affect its availability to gastrointestinal enzymes which impact its digestibility and absorption. The molecular spectroscopy-attenuated total reflectance Fourier transform infrared vibrational spectroscopy (ATR-FTIR) is an advanced technique that measures the absorbance of chemical functional groups on the infrared region so that we can identify and quantify molecules and functional groups in a feed. The program aimed to reveal the association of intrinsic molecular structure with nutrient supply to animals from canola feedstocks and co-products from bio-oil processing. The objective of this study was to characterize special intrinsic carbohydrate and protein-related molecular structure spectral profiles of feedstock and co-products (meal and pellets) from bio-oil processing from two source origins: Canada (CA) and China (CH). Methods The samples of feedstock and co-products were obtained from five different companies in each country arranged by the Canola Council of Canada (CCC). The molecular structure spectral features were analyzed using advanced vibrational molecular spectroscopy-ATR-FTIR. The spectral features that accessed included: i) protein-related spectral features (Amide I, Amide II, α-helix, β-sheet, and their spectral intensity ratios), ii) carbohydrate-related spectral features (TC1, TC2, TC3, TC4, CEC, STC1, STC2, STC3, STC4, TC, and their spectral intensity ratios). Results The results showed that significant differences were observed on all vibrationally spectral features related to total carbohydrates, structural carbohydrates, and cellulosic compounds (p0.05) of co-products, where CH meals presented higher peaks of these structures than CA. Similarly, it was for the carbohydrate-related molecular structure of canola seeds where the difference between CA and CH occurred except for STC3 height, CEC and STC areas (p>0.05). The protein-related molecular structures were similar for the canola seeds from both countries. However, CH meals presented higher peaks of amide I, α-helix, and β-sheet heights, α-helix:β-sheet ratio, total amide and amide I areas (p

Sodium glucose cotransporter 2 inhibitors (SGLT2) have been increasingly pursued as a promising target for addressing residual cardiovascular risk. Prior trials demonstrated that SGLT2i not only promotes glucose-lowering, but also improves endothelial dysfunction, adiposity, fluid overload, and insulin sensitivity thus contributing to hemodynamic changes implicated in its cardiorenal benefits. The mechanisms in the effect of SGLT2i on blood pressure and their potential role in preventing cardiovascular events are hereby revised.

OBJECTIVES/GOALS: The objective of this research was to determine the associations of candidate genetic variants withdrug-induced long QT syndrome (diLQTS) risk, an adverse effect of over 150 FDA-approved drugsthat can lead to cardiac arrhythmias and sudden cardiac death. METHODS/STUDY POPULATION: This was a retrospective observational study of the genomic biobank at the University of Michigan Health System. Patients treated with a high-risk QT-prolonging drug and ECG measurements were included. The primary outcome was exaggerated prolongation of the QTc interval (i.e., >60 ms change from baseline and/or >500 ms absolute value) corrected using Bazett. We analyzed 3 genetic variants: KCNE1-D85N (rs1805128), SCN5A-G615E (rs12720452) and KCNE2-I57T (rs7415448) in the dominant genetic model. A Bonferroni-corrected p-value of 0.017 was considered statistically significant using logistic regression adjusted for clinical covariates. RESULTS/ANTICIPATED RESULTS: In total 6,083 self-reported white patients were included (12% event rate). The adjusted odd ratio for KCNE1-D85N was 2.24 (95%CI: 1.35-3.57; p=0.0011). The adjusted odds ratio forKCNE2-I57T was 1.40 (95%CI: 0.26-5.78, p=0.662). Only 4 total patients carried the SCN5A-G615E variant, and none of the carriers had prolonged QTc. DISCUSSION/SIGNIFICANCE: This is the largest study of candidate genetic variants in cardiac ion channels associated with the diLQTS risk. KCNE1-D85N was associated with diLQTS risk, while KCNE2-I57T was suggestive of a potential association. KCNE1-D85N should be considered in clinical guidelines as a risk factor of diLQTS.

This study aimed to assess (a) the biofilm producer ability and antimicrobial resistance profiles of Staphylococcus (Staph.) aureus and Streptococcus (Strep.) uberis isolated from cows with clinical mastitis (CM) and subclinical mastitis (SCM), and (b) the association between biofilm producer ability and antimicrobial resistance. We isolated a total of 197 Staph. aureus strains (SCM = 111, CM = 86) and 119 Strep. uberis strains (SCM = 15, CM = 104) from milk samples obtained from 316 cows distributed in 24 dairy herds. Biofilm-forming ability was assessed using the microplate method, while antimicrobial susceptibility was determined using the disk diffusion method against 13 antimicrobials. Among the isolates examined, 57.3% of Staph. aureus and 53.8% of Strep. uberis exhibited the ability to produce biofilm, which was categorized as strong, moderate, or weak. In terms of antimicrobial susceptibility, Staph. aureus isolates displayed resistance to penicillin (92.9%), ampicillin (50.8%), and tetracycline (52.7%). Conversely, Strep. uberis isolates exhibited resistance to penicillin (80.6%), oxacillin (80.6%), and tetracycline (37.8%). However, no significant correlation was found between antimicrobial resistance patterns and biofilm formation ability among the isolates.

Abstract Despite the successful deployment of efficacious vaccines and therapeutics, the development of novel vaccines for SARS-CoV-2 remains a major goal to increase vaccine doses availability and accessibility for lower income setting. We report here on the kinetics of Spike-specific humoral and T-cell response in young and old volunteers over 6 months follow-up after a single intramuscular administration of GRAd-COV2, a gorilla adenoviral vector-based vaccine candidate currently in phase-2 of clinical development. At all three tested vaccine dosages, Spike binding and neutralizing antibodies were induced and substantially maintained up to 3 months, to then contract at 6 months. Potent T-cell responses were readily induced and sustained throughout the study period, with only minor decline. No major differences in immune response to GRAd-COV2 vaccination were observed in the two age cohorts. In light of its favorable safety and immunogenicity, GRAd-COV2 is a valuable candidate for further clinical development and potential addition to the COVID-19 vaccine toolbox to help fighting SARS-CoV-2 pandemic.

Parkinson's disease (PD) is a chronic and progressive neurodegenerative disorder with a higher susceptibility to occur in men. Studies suggest that this susceptibility is related to the hormonal differences observed between men and women, being a risk factor for PD. In addition, testosterone supplementation has shown controversial results in animal models of PD and parkinsonian patients. This study evaluated the effect of chronic administration of testosterone propionate (TP) on motor behavior and neurochemical parameters in the reserpine-induced rat model of parkinsonism. Male Wistar rats received 15 injections of reserpine (RES – 0.1 mg/kg) every other day and were concomitantly treated with different doses (0.1, 1.0, or 5.0 mg/kg) of daily TP for 30 days. The rats were euthanized 48 h after the 15th injection of RES or vehicle. Brains were removed and subjected to Tyrosine hydroxylase (TH) immunohistochemistry. TP at 1.0 mg/kg reduced the damages caused by reserpine in the vacuous chewing and tong protrusion behaviors and prevented dopaminergic damage in the SNpc, VTA, and Striatum. TP at 5.0 mg/kg reduced the damages caused by reserpine in the catalepsy and tong protrusion behaviors, prevented the weight loss, and prevented dopaminergic damage in the VTA. Our results suggest that chronic administration of TP has a protective effect in a rat model of parkinsonism, improving motor alterations and dopamine depletion induced by RES.

Abstract Background Fragile X syndrome, the major cause of inherited intellectual disability among men, is due to deficiency of the synaptic functional regulator FMR1 protein (FMRP), encoded by the FMRP translational regulator 1 (FMR1) gene. FMR1 alternative splicing produces distinct transcripts that may consequently impact FMRP functional roles. In transcripts without exon 14 the translational reading frame is shifted. For deepening current knowledge of the differential expression of Fmr1 exon 14 along the rat nervous system development, we conducted a descriptive study employing quantitative RT-PCR and BLAST of RNA-Seq datasets. Results We observed in the rat forebrain progressive decline of total Fmr1 mRNA from E11 to P112 albeit an elevation on P3; and exon-14 skipping in E17–E20 with downregulation of the resulting mRNA. We tested if the reduced detection of messages without exon 14 could be explained by nonsense-mediated mRNA decay (NMD) vulnerability, but knocking down UPF1, a major component of this pathway, did not increase their quantities. Conversely, it significantly decreased FMR1 mRNA having exon 13 joined with either exon 14 or exon 15 site A. Conclusions The forebrain in the third embryonic week of the rat development is a period with significant skipping of Fmr1 exon 14. This alternative splicing event chronologically precedes a reduction of total Fmr1 mRNA, suggesting that it may be part of combinatorial mechanisms downregulating the gene’s expression in the late embryonic period. The decay of FMR1 mRNA without exon 14 should be mediated by a pathway different from NMD. Finally, we provide evidence of FMR1 mRNA stabilization by UPF1, likely depending on FMRP.

ABSTRACT: Diffusion MRI is the dominant non-invasive imaging method used to characterize white matter organization in health and disease. Increasingly, fiber-specific properties within a voxel are analyzed using fixels. While tools for conducting statistical analyses of fixel-wise data exist, currently available tools support only a limited number of statistical models. Here we introduce ModelArray, an R package for mass-univariate statistical analysis of fixel-wise data. At present, ModelArray supports linear models as well as generalized additive models (GAMs), which are particularly useful for studying nonlinear effects in lifespan data. In addition, ModelArray also aims for scalable analysis. With only several lines of code, even large fixel-wise datasets can be analyzed using a standard personal computer. Detailed memory profiling revealed that ModelArray required only limited memory even for large datasets. As an example, we applied ModelArray to fixel-wise data derived from diffusion images acquired as part of the Philadelphia Neurodevelopmental Cohort (n = 938). ModelArray revealed anticipated nonlinear developmental effects in white matter. Moving forward, ModelArray is supported by an open-source software development model that can incorporate additional statistical models and other imaging data types. Taken together, ModelArray provides a flexible and efficient platform for statistical analysis of fixel-wise data.

BackgroundAccurate measurement of ventricular volumes is an important clinical imaging goal. Three-dimensional echocardiography (3DEcho) is used increasingly as it is more available and less costly than cardiac magnetic resonance (CMR). For the right ventricle (RV), the current practice is to acquire 3DEcho volumes from the apical view. However, in some patients the RV may be better seen from the subcostal view. Therefore, this study compared RV volume measurements from the apical vs. the subcostal view, using CMR as a reference standard.MethodsPatients

Background Chimeric antigen receptor (CAR) T-cell therapies have demonstrated transformational outcomes in the treatment of B-cell malignancies, but their widespread use is hindered by technical and logistical challenges associated with ex vivo cell manufacturing. To overcome these challenges, we developed VivoVec, a lentiviral vector-based platform for in vivo engineering of T cells. UB-VV100, a VivoVec clinical candidate for the treatment of B-cell malignancies, displays an anti-CD3 single-chain variable fragment (scFv) on the surface and delivers a genetic payload that encodes a second-generation CD19-targeted CAR along with a rapamycin-activated cytokine receptor (RACR) system designed to overcome the need for lymphodepleting chemotherapy in supporting successful CAR T-cell expansion and persistence. In the presence of exogenous rapamycin, non-transduced immune cells are suppressed, while the RACR system in transduced cells converts rapamycin binding to an interleukin (IL)-2/IL-15 signal to promote proliferation.Methods UB-VV100 was administered to peripheral blood mononuclear cells (PBMCs) from healthy donors and from patients with B-cell malignancy without additional stimulation. Cultures were assessed for CAR T-cell transduction and function. Biodistribution was evaluated in CD34-humanized mice and in canines. In vivo efficacy was evaluated against normal B cells in CD34-humanized mice and against systemic tumor xenografts in PBMC-humanized mice.Results In vitro, administration of UB-VV100 resulted in dose-dependent and anti-CD3 scFv-dependent T-cell activation and CAR T-cell transduction. The resulting CAR T cells exhibited selective expansion in rapamycin and antigen-dependent activity against malignant B-cell targets. In humanized mouse and canine studies, UB-VV100 demonstrated a favorable biodistribution profile, with transduction events limited to the immune compartment after intranodal or intraperitoneal administration. Administration of UB-VV100 to humanized mice engrafted with B-cell tumors resulted in CAR T-cell transduction, expansion, and elimination of systemic malignancy.Conclusions These findings demonstrate that UB-VV100 generates functional CAR T cells in vivo, which could expand patient access to CAR T technology in both hematological and solid tumors without the need for ex vivo cell manufacturing.

Effective secondary response to antigen is a hallmark of immunological memory. However, the extent of memory CD8 T cell response to secondary boost varies at different times after a primary response. Considering the central role of memory CD8 T cells in long-lived protection against viral infections and tumors, a better understanding of the molecular mechanisms underlying the changing responsiveness of these cells to antigenic challenge would be beneficial. We examined here primed CD8 T cell response to boost in a BALB/c mouse model of intramuscular vaccination by priming with HIV-1 gag-encoding Chimpanzee adenovector, and boosting with HIV-1 gag-encoding Modified Vaccinia virus Ankara. We found that boost was more effective at day(d)100 than at d30 post-prime, as evaluated at d45 post-boost by multi-lymphoid organ assessment of gag-specific CD8 T cell frequency, CD62L-expression (as a guide to memory status) and in vivo killing. RNA-sequencing of splenic gag-primed CD8 T cells at d100 revealed a quiescent, but highly responsive signature, that trended toward a central memory (CD62L+) phenotype. Interestingly, gag-specific CD8 T cell frequency selectively diminished in the blood at d100, relative to the spleen, lymph nodes and bone marrow. These results open the possibility to modify prime/boost intervals to achieve an improved memory CD8 T cell secondary response.

Abstract Background We report a genomic surveillance of SARS-CoV-2 lineages circulating in Paraná, southern Brazil, from March 2020 to April 2021. Our analysis, based on 333 genomes, revealed that the first variants detected in the state of Paraná in March 2020 were the B.1.1.33 and B.1.1.28 variants. The variants B.1.1.28 and B.1.1.33 were predominant throughout 2020 until the introduction of the variant P.2 in August 2020 and a variant of concern (VOC), Gamma (P.1), in January 2021. The VOC Gamma, a ramification of the B.1.1.28 lineage first detected in Manaus (northern Brazil), has grown rapidly since December 2020 and was thought to be responsible for the deadly second wave of COVID-19 throughout Brazil. Methods The 333 genomic sequences of SARS-CoV-2 from March 2020 to April 2021 were generated as part of the genomic surveillance carried out by Fiocruz in Brazil Genomahcov Fiocruz. SARS-CoV-2 sequencing was performed using representative samples from all geographic areas of Paraná. Phylogenetic analyses were performed using the 333 genomes also included other SARS-CoV-2 genomes from the state of Paraná and other states in Brazil that were deposited in the GISAID. In addition, the time-scaled phylogenetic tree was constructed with up to 3 random sequences of the Gamma variant from each state in Brazil in each month of 2021. In this analysis we also added the sequences identified as the B.1.1.28 lineage of the Amazonas state and and the Gamma-like-II (P.1-like-II) lineage identified in different regions of Brazil. Results Phylogenetic analyses of the SARS-CoV-2 genomes that were previously classified as the VOC Gamma lineage by WHO/PANGO showed that some genomes from February to April 2021 branched in a monophyletic clade and that these samples grouped together with genomes recently described with the lineage Gamma-like-II. Additionally, a new mutation (E661D) in the spike (S) protein has been identified in nearly 10% of the genomes classified as the VOC Gamma from Paraná in March and April 2021.Finally, we analyzed the correlation between the lineage and the Gamma variant frequency, age group (patients younger or older than 60 years old) and the clinical data of 86 cases from the state of Paraná. Conclusions Our results provided a reliable picture of the evolution of the SARS-CoV-2 pandemic in the state of Paraná characterized by the dominance of the Gamma strain, as well as a high frequencies of the Gamma-like-II lineage and the S:E661D mutation. Epidemiological and genomic surveillance efforts should be continued to unveil the biological relevance of the novel mutations detected in the VOC Gamma in Paraná.

Background: Atrial fibrillation (AF) is the leading cause of ischemic stroke and treatment has focused on reducing this risk through anticoagulation. Direct Oral Anticoagulants (DOACs) are the first-line guideline-recommended therapy since they are as effective and overall safer than warfarin in preventing AF-related stroke. Although patients bleed less from DOACs compared to warfarin, bleeding remains the primary safety concern with this therapy.Hypothesis: Genetic variants known to modify the function of metabolic enzymes or transporters involved in the pharmacokinetics (PK) of DOACs could increase the risk of bleeding.Aim: To assess the association of eight, functional PK-related single nucleotide variants (SNVs) in five genes (ABCB1, ABCG2, CYP2J2, CYP3A4, CYP3A5) with the risk of bleeding from DOACs in non-valvular AF patients.Methods: A retrospective cohort study was carried out with 2,364 self-identified white non-valvular AF patients treated with either rivaroxaban or apixaban. Genotyping was performed with Illumina Infinium CoreExome v12.1 bead arrays by the Michigan Genomics Initiative biobank. The primary endpoint was a composite of major and clinically relevant non-major bleeding. Cox proportional hazards regression with time-varying analysis assessed the association of the eight PK-related SNVs with the risk of bleeding from DOACs in unadjusted and covariate-adjusted models. The pre-specified primary analysis was the covariate-adjusted, additive genetic models. Six tests were performed in the primary analysis as three SNVs are in the same haplotype, and thus p-values below the Bonferroni-corrected level of 8.33e-3 were considered statistically significant.Results: In the primary analysis, none of the SNVs met the Bonferroni-corrected level of statistical significance (all p > 0.1). In exploratory analyses with other genetic models, the ABCB1 (rs4148732) GG genotype tended to be associated with the risk of bleeding from rivaroxaban [HR: 1.391 (95%CI: 1.019–1.900); p = 0.038] but not from apixaban (p = 0.487).Conclusion: Eight functional PK-related genetic variants were not significantly associated with bleeding from either rivaroxaban or apixaban in more than 2,000 AF self-identified white outpatients.

Introdução: Criptococose é uma micose sistêmica causada por fungos do gênero Cryptococcus. É mais comum em indivíduos com HIV/AIDS, mas um aumento de casos é observado em pacientes onco-hematológicos. Objetivo: Descrever o caso de uma paciente com Linfoma Não Hodgkin de células do manto (LNH), diagnosticada com criptococose pulmonar durante quimioterapia (QT), submetida a transplante autólogo de células-tronco hematopoiéticas (auto-TCTH) após a segunda remissão completa e tratamento da doença fúngica. Resultados: Relato: Mulher, 52 anos, diagnosticada com LNH de células do manto em 2017, estágio clínico IV XSB. Iniciou QT com CHOP. Em fevereiro de 2018, no quarto ciclo de QT, foi realizado PET CT que visualizou nódulos sólidos irregulares por todo o parênquima pulmonar. Anatomopatológico de biópsia de nódulo no lobo superior direito evidenciou numerosas leveduras, com o isolamento de Cryptococcus neoformans em cultura. Antígeno sérico para Cryptococcus foi positivo. Foi iniciado tratamento com fluconazol em março de 2018. Após o diagnóstico infeccioso, o auto-TCTH foi contraindicado e a paciente teve remissão completa do LNH após 6 ciclos de QT. Houve redução dos nódulos pulmonares nos exames de imagem e negativação do antígeno sérico para Cryptococcus. Foi reduzida a dose de fluconazol para 300 mg/dia como terapia de manutenção. Em agosto de 2020, houve recidiva tardia do linfoma, foi iniciado nova QT com (R)-DHAOX e programado auto-TCTH sequencial. Em abril de 2021, houve a remissão completa do LNH de células do manto após término da QT, realizado auto-TCTH após condicionamento com BendaEAM. Nesse momento, apresentava antígeno sérico para Cryptococcus negativo e tomografia de crânio e tórax sem evidência de doença fúngica ativa. Recebeu profilaxia com dose única de ivermectina, cotrimoxazol até D-1, aciclovir, isoniazida (PPD 8 mm) e fluconazol 400 mg por dia. Foi mantido profilaxia secundária da criptococose com fluconazol por 6 meses. Conclusão: O caso foi tratado com sucesso antes do TCTH e não houve reativação da doença durante o período de neutropenia, nem após o transplante. Embora a Criptococose seja raramente observada em receptores de TCTH, a maioria dos casos se apresenta após o transplante. Não há recomendações na literatura quanto ao manejo no diagnóstico prévio ao transplante. Este relato traz a experiência de nosso serviço neste manejo e destaca a importância da avaliação do paciente no período pré-transplante e da profilaxia farmacológica adequada.

Abstract Background Healthcare workers have a 16 times greater risk of suffering workplace violence than workers in other sectors and around 50% experience workplace violence in the course of their career. The objective of this study is to explore the characteristics and circumstances of work-related killings of doctors. Methods Work-related homicides of doctors over the period 1988–2019 were identified retrospectively through the Italian national statistical agencies. Variables such as perpetrator, motive and location of the crime were obtained through forensic psychiatric work. After classification, the absolute and percent values of the main characteristics of the homicides were calculated. Results Over the period considered, 21 doctors were killed in Italy in connection with their professional activity. In 52% (n = 11) of cases, the killer was one of the doctor’s patients, in 29% (n = 6) of cases it was a patient’s relative, in 19% (n = 4) an occasional patient (first consultation). The location of the homicide was a community clinic in 48% (n = 10) of cases, the street in 19% (n = 4) of cases, the doctor’s home in 14% (n = 3), the hospital in 14% (n = 3) and the patient’s home in 5% (n = 1). In 57% (n = 12) of cases the perpetrator was not affected by any mental disorders. The motive for the homicide was revenge in 66.7% (n = 14) of cases; in 28.6% (n = 6) the revenge was preceded by stalking. Conclusions Doctors should be aware that the risk of being killed is not limited to hospital settings and that their patients’ family members might also pose a threat to them.

Abstract Background Considering many patients receive care from general hospitals, these healthcare institutions are uniquely situated to address mental and physical health needs. Little is documented, however, on the common current mental disorders diagnosed in patients receiving care in general hospital settings, especially in Puerto Rico. The objective of this study was to characterize the five most common current DSM-5 mental disorder diagnoses made in patients receiving non-psychiatric medical and surgical care from a general hospital system in southern Puerto Rico between January 2015 and December 2019. Methods Our clinical health psychology team provides integrated psychology consultation-liaison services to select clinical units in general hospitals across the southwestern region of Puerto Rico. The clinical team conducted routine standardized psychological evaluations at patients' bedside, arrived at a current DSM-5 diagnosis if warranted, and documented the diagnosis and other select variables. A retrospective study of cross-sectional data generated from the clinical team’s standardized evaluations of 5494 medical patients was implemented. Multinomial logistic regression analyses were used to assess the odds of being diagnosed with a current DSM-5 mental disorder during hospitalization. Results Overall, 53% of the entire sample was diagnosed with a mental disorder during hospitalization. Major depressive, neurocognitive, anxiety, substance-related and schizophrenia-spectrum disorders were the most frequently diagnosed. Interestingly, females were 23% less likely to have been diagnosed with major depressive disorder than males (aOR: 0.769, CI [0.650, 0.909], p = 0.002). This is to say males evidenced 1.30 higher odds of being diagnosed with depression compared to their female counterpart. Age, biological sex, civil status, employment status, monthly household income, previous mental disorder and history substance use/abuse history was differentially associated with receiving a current DSM-5 disorder. Conclusion The integration of clinical health psychology services within a general hospital facilitated our team’s work of identifying and treating co-occurring mental disorders among hospitalized patients receiving medical and surgical care. Future studies examining the opportunities and barriers of integrating clinical health psychology services within a general hospital’s administrative and clinical infrastructure for rapid identification and treatment of co-occurring mental disorders among medical patients is encouraged.

The treatment of saturated horse beds before they arrive at their final destination is necessary to avoid the risk of animal and environmental contamination. For this purpose, the composting process has great functionality due its to low cost, effectiveness, and operational ease. However, because of the nature of the materials used, this process can be long, and it is necessary to improve it to optimize composting cycles. This work aimed to isolate and identify fungi present in the compost piles of saturated equine bedding made with shavings and rice straw, identifying those with the greatest potential for cellulase production. Using specific cellulolytic media containing shavings or rice straw, seven strains were isolated. The total cellulase enzymatic activity of the isolates from the beds made with shavings was lower than that obtained from rice straw beds. Four strains showed high enzymatic potential for use in the shavings substrate (MA -6 2 f1, MA -6 2 f2, MA -7 9, and MA -7 10) and three for the rice straw substrate (PA -7 5, PA -7 7, and PA -7 10). The isolate PA -7 5 reached 0.376 IU mL−1, the best index among all the isolates. These isolates were identified as belonging to the Aspergillus fumigatus species.

Mycophenolate rapidly substituted azathioprine (AZA) in transplant immunosuppression regimens since the 1990s, when early clinical trials indicated better outcomes, although opposite results were also observed. However, none of these trials used the well-established optimization methods for AZA dosing, namely, thiopurine methyltransferase pharmacogenetics combined with monitoring of the thiopurine metabolites 6-thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine (6-MMP). Resistance to optimize AZA therapy remains today in transplant therapy, despite the fact that thiopurine metabolite testing is being used by other medical disciplines with evident improvement in clinical results. In a previous analysis, we found that active 6-TGN metabolites were not detectable in about 30% of kidney transplant patients under continuous use of apparently adequate azathioprine dosage, which demonstrates the need to monitor these metabolites for therapeutic optimization. Two of four case studies presented here exemplifies this fact. On the other hand, some patients have toxic 6-TGN levels with a theoretically appropriate dose, as seen in the other two case studies in this presentation, constituting one more important reason to monitor the AZA dose administered by its metabolites. This analysis is not intended to prove the superiority of one immunosuppressant over another, but to draw attention to a fact: there are thousands of patients around the world receiving an inadequate dose of azathioprine and, therefore, with inappropriate immunosuppression. This report is also intended to draw attention, to clinicians using thiopurines, that allopurinol co-therapy with AZA is a useful therapeutic pathway for those patients who do not adequately form active thioguanine metabolites.

Summary Aedes albopictus transmits several arboviral infections. In the absence of vaccines, control of mosquito populations is the only strategy to prevent vector‐borne diseases. As part of the search for novel, biological and environmentally friendly strategies for vector control, the isolation of new bacterial species with mosquitocidal activity represents a promising approach. However, new bacterial isolates may be difficult to grow and genetically manipulate. To overcome these limits, here we set up a system allowing the expression of mosquitocidal bacterial toxins in the well‐known genetic background of Bacillus subtilis. As a proof of this concept, the ability of B. subtilis to express individual or combinations of toxins of Bacillus thuringiensis israelensis (Bti) was studied. Different expression systems in which toxin gene expression was driven by IPTG‐inducible, auto‐inducible or toxin gene‐specific promoters were developed. The larvicidal activity of the resulting B. subtilis strains against second‐instar Ae. albopictus larvae allowed studying the activity of individual toxins or the synergistic interaction among Cry and Cyt toxins. The expression systems here presented lay the foundation for a better improved system to be used in the future to characterize the larvicidal activity of toxin genes from new environmental isolates.

Abstract Background Strain and synchrony can be calculated from a variety of software packages, but there is a paucity of data with inter-vendor comparisons in children. To test the hypothesis that different packages may affect results, independent of acquisition, we compared values obtained using two commercially available analysis tool (QLAB and TomTec), with several different settings. Methods The study population included 108 children; patients were divided into three groups: (1) normal cardiac structure and conduction; (2) ventricular paced rhythm; and (3) flattened ventricular septum (reflecting right ventricular pressure or volume load lesions). We analyzed the same image acquired from the apical 4-chamber (AP4) and short-axis at the mid-papillary level (SAXM) views in both QLAB (versions 10.5 and 10.8) and TomTec (version 1.2). In QLAB version 10.8, low, medium, and high quantification smoothness settings were employed. In TomTec, images were analyzed with both low and high frame rates. Tracking quality for each package was graded. AP4 and SAXM strain and synchrony values were recorded. A mixed-effects linear regression model was used, with main effect considered significant if the p-value was

Cryptococcosis is an invasive mycosis caused by Cryptococcus spp. that affects the lungs and the central nervous system (CNS). Due to the severity of the disease, it may occur concomitantly with other pathogens, as a coinfection. Pseudomonas aeruginosa (Pa), an opportunistic pathogen, can also cause pneumonia. In this work, we studied the interaction of C. gattii (Cg) and Pa, both in vitro and in vivo. Pa reduced growth of Cg by the secretion of inhibitory molecules in vitro. Macrophages previously stimulated with Pa presented increased fungicidal activity. In vivo, previous Pa infection reduced morbidity and delayed the lethality due to cryptococcosis. This phenotype was correlated with the decreased fungal burden in the lungs and brain, showing a delay of Cg translocation to the CNS. Also, there was increased production of IL-1β, CXCL-1, and IL-10, together with the influx of iNOS-positive macrophages and neutrophils to the lungs. Altogether, Pa turned the lung into a hostile environment to the growth of a secondary pathogen, making it difficult for the fungus to translocate to the CNS. Further, iNOS inhibition reverted the Pa protective phenotype, suggesting its important role in the coinfection. Altogether, the primary Pa infection leads to balanced pro-inflammatory and anti-inflammatory responses during Cg infection. This response provided better control of cryptococcosis and was decisive for the mild evolution of the disease and prolonged survival of coinfected mice in a mechanism dependent on iNOS.

OBJECTIVES/GOALS: Assess the association of PK-related single nucleotide variants (SNVs) with the risk of bleeding from DOACs in non-valvular AF patients. METHODS/STUDY POPULATION: A retrospective cohort study was carried out with 2,364 Caucasians with non-valvular AF and treated with rivaroxaban or apixaban. Patients were genotyped as part of the genomic biobank at the University of Michigan Health System. The primary endpoint was a composite of major and clinically relevant non-major (CRNM) bleeding. Cox proportional hazards regression with time-varying analysis assessed the association of 8 SNVs in 5 PK genes (ABCB1, ABCG2, CYP3A4, CYP3A5, CYP2J2) with the risk of bleeding from DOACs in unadjusted and covariate-adjusted models. Six tests were performed as 3 of the SNVs are in the same haplotype. P-values below the Bonferroni-corrected level of 8.33e-3 were considered statistically significant. RESULTS/ANTICIPATED RESULTS: A total of 412 (17.4%) major and CRNM bleeding events occurred over 2.27 ± 2.03 years of follow-up. None of the PK SNVs were significantly associated with bleeding risk on DOACs in both unadjusted and covariate-adjusted Cox regression models. DISCUSSION/SIGNIFICANCE: The effects of these eight genetic variants on exposure to DOACs may not be strong enough to translate into differences in clinical outcomes. Especially if the genetic inheritance underlying the risk of bleeding from DOACs is polygenic, reinforcing the need for further genomic studies on this subject.

Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disease associated with loss of upper and lower motor neurons in the primary motor cortex and spinal cord, respectively. Motor deficits are the main clinical features observed in patients with the disease. However, it has been suggested that the presence of neurotoxic factors in the cerebrospinal fluid (CSF) from ALS patients causes loss of motor neurons. The present study investigated the motor and histopathological changes induced by intracerebroventricular (ICV) infusion of CSF from ALS patients in middle aged male Wistar rats. Middle aged male rats were divided into three groups: (1) control group, animals injected with artificial CSF solution; (2) N-ALS group, animals injected with CSF from volunteers without neurological disease; and (3) ALS group, animals inoculated with CFS from a patient with definite ALS. After surgical and infusion procedures, animals were evaluated in different motor tests (grip strength; catalepsy and open field tests). Moreover, animals’ spinal cords were histologically investigated. We observed that ALS-CSF infusion reduced grip strength and led to motor changes and reduction in the number of motor neurons and glial cells in thoracic and lumbar regions of spinal cord. However, CSF N-ALS caused reduction of nerve and glial cells in the thoracic but not in the lumbar region. Our data suggest that ALS-CSF is associated with neurodegenerative mechanisms observed in ALS pathology.

Objetivo: Describir la ingesta alimentaria antes y después de las sesiones de teleorientación y teleseguimiento en pacientes con hipertensión arterial y diabetes mellitus. Materiales y métodos: Estudio de diseño cuasiexperimental en el cual se evaluó la ingesta alimentaria de 80 participantes, con diagnóstico de hipertensión arterial y diabetes mellitus, al inicio de la intervención y al finalizar las sesiones de teleorientación y teleseguimiento programadas. Resultados: Al empezar la intervención solo el 6,25 % obtuvo una valoración de ingesta alimentaria adecuada, la cual aumentó hasta 71,25 % al finalizar las sesiones de teleorientación y teleseguimiento programadas. Conclusiones: La implementación de un sistema de teleorientación y teleseguimiento nutricional en pacientes con hipertensión arterial y diabetes mellitus mejora la ingesta alimentaria, lo que permite confirmar la eficacia del uso de las TIC en las intervenciones sanitarias para el beneficio de la población.

Lipopolysaccharide (LPS) is a peculiar component of the outer membrane (OM) of many Gram-negative bacteria that renders these bacteria highly impermeable to many toxic molecules, including antibiotics. LPS is assembled at the OM by a dedicated intermembrane transport system, the Lpt (LPS transport) machinery, composed of seven essential proteins located in the inner membrane (IM) (LptB2CFG), periplasm (LptA), and OM (LptDE). Defects in LPS transport compromise LPS insertion and assembly at the OM and result in an overall modification of the cell envelope and its permeability barrier properties. LptA is a key component of the Lpt machine. It connects the IM and OM sub-complexes by interacting with the IM protein LptC and the OM protein LptD, thus enabling the LPS transport across the periplasm. Defects in Lpt system assembly result in LptA degradation whose stability can be considered a marker of an improperly assembled Lpt system. Indeed, LptA recruitment by its IM and OM docking sites requires correct maturation of the LptB2CFG and LptDE sub-complexes, respectively. These quality control checkpoints are crucial to avoid LPS mistargeting. To further dissect the requirements for the complete Lpt transenvelope bridge assembly, we explored the importance of LPS presence by blocking its synthesis using an inhibitor compound. Here, we found that the interruption of LPS synthesis results in the degradation of both LptA and LptD, suggesting that, in the absence of the LPS substrate, the stability of the Lpt complex is compromised. Under these conditions, DegP, a major chaperone–protease in Escherichia coli, is responsible for LptD but not LptA degradation. Importantly, LptD and LptA stability is not affected by stressors disturbing the integrity of LPS or peptidoglycan layers, further supporting the notion that the LPS substrate is fundamental to keeping the Lpt transenvelope complex assembled and that LptA and LptD play a major role in the stability of the Lpt system.

Inbreeding depression is the reduction in fitness and vigor resulting from mating of close relatives observed in many plant and animal species. The extent to which the genetic load of mutations contributing to inbreeding depression is due to large-effect mutations versus variants with very small individual effects is unknown and may be affected by population history. We compared the effects of outcrossing and self-fertilization on 18 traits in a landrace population of maize, which underwent a population bottleneck during domestication, and a neighboring population of its wild relative teosinte. Inbreeding depression was greater in maize than teosinte for 15 of 18 traits, congruent with the greater segregating genetic load in the maize population that we predicted from sequence data. Parental breeding values were highly consistent between outcross and selfed offspring, indicating that additive effects determine most of the genetic value even in the presence of strong inbreeding depression. We developed a novel linkage scan to identify quantitative trait loci (QTL) representing large-effect rare variants carried by only a single parent, which were more important in teosinte than maize. Teosinte also carried more putative juvenile-acting lethal variants identified by segregation distortion. These results suggest a mixture of mostly polygenic, small-effect partially recessive effects in linkage disequilibrium underlying inbreeding depression, with an additional contribution from rare larger-effect variants that was more important in teosinte but depleted in maize following the domestication bottleneck. Purging associated with the maize domestication bottleneck may have selected against some large effect variants, but polygenic load is harder to purge and overall segregating mutational burden increased in maize compared to teosinte.

This paper aims to analyse the criticalities and strengths of distance learning provided during this period of health emergency, by analysing the opinions of university students with disabilities and SLD. Specifically, we present the results of an ad hoc questionnaire administered to the students with disabilities and Specific Learning Disability (SLD) enrolled at the University of Calabria. Through this, we aimed to provide important insights, to be taken into account in the future, especially in view of the re-arrangement of the University’s teaching activities. Furthermore, we intended to highlight how universal design in the educational field should constitute a widespread and systematic working method for the creation of truly inclusive learning and relationship environments, also at a distance. Come immaginare una didattica universitaria post-Covid inclusiva. Il presente contributo si pone l’obiettivo di analizzare le criticità e i punti di forza della didattica a distanza (DaD) erogata durante questo periodo di emergenza sanitaria, analizzando le opinioni degli studenti e delle studentesse con disabilità e Disturbi Specifici dell’Apprendimento (DSA). In particolare, si presentano i risultati ottenuti tramite la somministrazione di un questionario ad hoc agli studenti e alle studentesse con disabilità e DSA iscritti presso l’Università della Calabria. In tal modo si auspica di fornire utili spunti di riflessione, dei quali tener conto in futuro, soprattutto in vista di una riorganizzazione delle attività didattiche dell’Ateneo. Si tenterà, inoltre, di mettere in luce come la progettazione universale in campo educativo debba costituire una modalità di lavoro diffusa e sistematica per la creazione di ambienti di apprendimento e relazione davvero inclusivi, anche a distanza.

With current drug treatments failing due to toxicity, low efficacy and resistance; leishmaniasis is a major global health challenge that desperately needs new validated drug targets. Inspired by activity of the natural chalcone 2',6'-dihydroxy-4'-methoxychalcone (DMC), the nitro-analogue, 3-nitro-2',4',6'- trimethoxychalcone (NAT22, 1c) was identified as potent broad spectrum antileishmanial drug lead. Structural modification provided an alkyne containing chemical probe that labelled a protein within the parasite that was confirmed as cytosolic tryparedoxin peroxidase (cTXNPx). Crucially, labelling is observed in both promastigote and intramacrophage amastigote life forms, with no evidence of host macrophage toxicity. Incubation of the chalcone in the parasite leads to ROS accumulation and parasite death. Deletion of cTXNPx, by CRISPR-Cas9, dramatically impacts upon the parasite phenotype and reduces the antileishmanial activity of the chalcone analogue. Molecular docking studies with a homology model of in-silico cTXNPx suggest that the chalcone is able to bind in the putative active site hindering access to the crucial cysteine residue. Collectively, this work identifies cTXNPx as an important target for antileishmanial chalcones.

In this exploratory mixed-methods analysis of students' perceptions of inclusion in introductory STEM courses for STEM majors, we asked students to rate inclusion in their class and to provide an open-text explanation of their rating. Analyzing 1930 qualitative responses resulted in a codebook containing academic, identity, and nonspecific categories. The majority of responses (>80%) cited academic factors such as interactions between students and instructors or course elements and policies. Most academic responses aligned with evidence-based teaching practices fostering inclusion, describing a range of strategies and policies instructors can implement to increase students' perceptions of inclusion. A small number of student responses indicated that their perception of the required knowledge background for the course impacted course inclusivity. Few differences in frequency distributions were found between subgroups examined (gender, race and ethnicity, self-reported inclusion score, and discipline). Additionally, tracking a subset of students (135) across three courses revealed that most (80%) cited different factors influencing their perception of inclusion in each course. This suggests students' perceptions of inclusive practices are complex, and most students recognize multiple factors that influence their inclusion. Overall, our findings suggest instructors can significantly influence students' perceptions of inclusion by using multiple inclusive teaching strategies and course policies.

ABSTRACT The inadequate agricultural management in the semiarid region of northeastern Brazil, associated with the fragility of most soils, has caused degradation of the quality of this resource. This study aimed to evaluate the quality of Quartzipsamments under different uses through the multivariate analysis of chemical and physical attributes. Five areas were selected: four of them with agricultural use (onion, banana, cassava, and corn) and the other with Caatinga. Soil samples were collected at depths of 0.00–0.10, 0.10–0.20, and 0.20–0.40 m for chemical and physical analyses. The data were submitted to descriptive analysis and then to the multivariate analysis using the principal component analysis (PCA) technique and Ward’s clustering method. PCA and clustering techniques allowed the identification of different production systems in subsurface layers, suggesting the effect of fertility on the differentiation of areas with onion and banana, as well as the clustering of cassava and corn cultivations. Physical attributes and sodium content of the surface layer of the area with onion showed a tendency of compaction and salinization process of this environment. In general, fertility was the main responsible for the improvement of soil quality with agricultural use when compared to the Caatinga.

Abstract Background Myocardial infarction (MI) elicits an intense acute inflammatory response that is essential for cardiac repair. However, an excessive inflammatory response also favors myocardial apoptosis, cardiac remodeling, and cardiovascular mortality. Omega-3 polyunsaturated fatty acids (ω-3) bear anti-inflammatory effects, which may mitigate the inflammatory response during MI. This study investigated whether ω-3 intake is associated with attenuation of the MI-related inflammatory response and cardiac remodeling. Methods ST-elevation MI (STEMI) patients (n = 421) underwent clinical, biochemical, nutritional, 3D echocardiogram, Cardiac Magnetic Resonance imaging (CMRi) at 30 days and 3D echocardiogram imaging at six months after the MI. Blood tests were performed at day one (D1) and day five (D5) of hospitalization. Changes in inflammatory markers (ΔD5-D1) were calculated. A validated food frequency questionnaire estimated the nutritional consumption and ω-3 intake in the last 3 months before admission. Results The intake of ω-3 below the median (

Objective To estimate the pooled prevalence of multimorbidity (≥2 non-communicable diseases in the same individual) among adults of the general population of Latin American and the Caribbean (LAC).Design Systematic review and meta-analysis.Data sources MEDLINE, Embase, Global Health, Scopus and LILACS up to 1 July 2020.Eligibility criteria for selecting studies The outcome was the prevalence of multimorbidity. Reports were selected whether they enrolled adult individuals (age ≥18 years) from the general population.Data extraction and synthesis Reviewers extracted relevant data and assessed risk of bias independently. A random-effects meta-analysis was conducted to report pooled prevalence estimates of multimorbidity; pooled estimates by pre-specified subgroups (eg, national studies) were also pursued.Results From 5830 results, we selected 28 reports, mostly from Brazil and 16 were based on a nationally representative sample. From the 28 selected reports, 26 were further included in the meta-analysis revealing a pooled multimorbidity prevalence of 43% (95% CI: 35% to 51%; I2: 99.9%). When only reports with a nationally representative sample were combined, the pooled prevalence was 37% (95% CI: 27% to 47%; I2: 99.9%). When the ascertainment of multimorbidity was based on self-reports alone, the pooled prevalence was 40% (95% CI: 31% to 48%; I2: 99.9%); this raised to 52% (95% CI: 33% to 70%; I2: 99.9%) for reports including self-reported and objective diagnosis.Conclusions Our results complement and advance those from global efforts by incorporating much more reports from LAC. We revealed a larger presence of multimorbidity in LAC than previously reported.PROSPERO registration number CRD42020196177.

Gram-negative bacteria have a complex cell envelope with two membranes and a periplasm containing the peptidoglycan layer. The outer membrane is firmly connected to the peptidoglycan by highly abundant proteins.

Neutrophils play an important role in the outcome of leishmaniasis, contributing either to exacerbating or controlling the progression of infection, a dual effect whose underlying mechanisms are not clear. We recently reported that CD4+ and CD8+ T cells, and dendritic cells of Leishmania amazonensis-infected mice present high expression of PD-1 and PD-L1, respectively. Given that the PD-1/PD-L1 interaction may promote cellular dysfunction, and that neutrophils could interact with T cells during infection, we investigated here the levels of PD-L1 in neutrophils exposed to Leishmania parasites. We found that both, promastigotes and amastigotes of L. amazonensis induced the expression of PD-L1 in the human and murine neutrophils that internalized these parasites in vitro. PD-L1-expressing neutrophils were also observed in the ear lesions and the draining lymph nodes of L. amazonensis-infected mice, assessed through cell cytometry and intravital microscopy. Moreover, expression of PD-L1 progressively increased in neutrophils from ear lesions as the disease evolved to the chronic phase. Co-culture of infected neutrophils with in vitro activated CD8+ T cells inhibits IFN-γ production by a mechanism dependent on PD-1 and PD-L1. Importantly, we demonstrated that in vitro infection of human neutrophils by L braziliensis induced PD-L1+ expression and also PD-L1+ neutrophils were detected in the lesions of patients with cutaneous leishmaniasis. Taken together, these findings suggest that the Leishmania parasite increases the expression of PD-L1 in neutrophils with suppressor capacity, which could favor the parasite survival through impairing the immune response.