ASCL1 promotes Scrt2 expression in the neural tube.

ASCL1 is a transcription factor that directs neural progenitors towards lineage differentiation. Although many of the molecular mechanisms underlying its action have been described, several of its targets remain unidentified. We identified in the chick genome a putative enhancer (cE1) upstream of the transcription factor Scratch2 (Scrt2) locus with a predicted heterodimerization motif for ASCL1 and POU3F2. In this study, we investigated the role of ASCL1 and this enhancer in regulating the expression of the Scrt2 in the embryonic spinal cord. We confirmed that cE1 region interacted with the Scrt2 promoter. cE1 was sufficient to mediate ASCL1-driven expression in the neural tube through the heterodimerization sites. Moreover, Scrt2 expression was inhibited when we removed cE1 from the genome. These findings strongly indicate that ASCL1 regulates Scrt2 transcription in the neural tube through cE1.

Stability of Proximal Femoral Osteotomies in Pediatric Bone Models Fixed with Flexible Intramedullary Nails and Evaluated by the Finite Element Method.

Objective To evaluate the stability of osteotomies created in the subtrochanteric and trochanteric regions in a pediatric femur model fixed by flexible intramedullary rods. Method Tomographic sections were obtained from a pediatric femur model with two elastic titanium rods and converted to a three-dimensional model. This model created a mesh with tetrahedral elements according to the finite element method. Three virtual models were obtained, and osteotomies were performed in different regions: mediodiaphyseal, subtrochanteric, and trochanteric. A vertical load of 85N was applied to the top of the femoral head, obtaining the displacements, the maximum and minimum main stress, and the equivalent Von Mises stress on the implant. Results With the applied load, displacements were observed at the osteotomy site of 0.04 mm in the diaphyseal group, 0.5 mm in the subtrochanteric group, and 0.06 mm in the trochanteric group. The maximum stress in the diaphyseal, subtrochanteric, and trochanteric groups was 10.4 Pa, 7.52 Pa, and 26.4 Pa, respectively. That is around 40% higher in the trochanteric group in regards to the diaphyseal (control). The minimum stress of the bone was located in the inner cortical of the femur. The equivalent Von Mises stress on the implants occurred at osteotomy, with a maximum value of 27.6 Pa in the trochanteric group. Conclusion In both trochanteric and subtrochanteric osteotomies, fixation stability was often lower than in the diaphyseal model, suggesting that flexible intramedullary nails are not suitable implants for proximal femoral fixations.

Methyl N-{(1R)-2-[(meth-oxy-carbon-yl)-oxy]-1-phenyleth-yl}carbamate.

The title mol-ecule, C12H15NO5, is a methyl carbamate derivative obtained by reacting (R)-2-phenyl-glycinol and methyl chloro-formate, with calcium hydroxide as heterogeneous catalyst. Supra-molecular chains are formed in the [100] direction, based on N-H⋯O hydrogen bonds between the amide and carboxyl-ate groups. These chains weakly inter-act in the crystal, and the phenyl rings do not display significant π-π inter-actions.

Plastid genome of Chenopodium petiolare from Trujillo, Peru.

OBJECTIVES: The Peruvian Andean region is an important center for plant domestication. However, to date, there have been few genetic studies on native grain, which limits our understanding of their genetic diversity and the development of new genetic studies for their breeding. Herein, we revealed the plastid genome of Chenopodium petiolare to expand our knowledge of its molecular markers, evolutionary studies, and conservation genetics. DATA DESCRIPTION: Total genomic DNA was extracted from fresh leaves (voucher: USM < PER > :MHN333570). The DNA was sequenced using Illumina Novaseq 6000 (Macrogen Inc., Seoul, Republic of Korea) and reads 152,064 bp in length, with a large single-copy region of 83,520 bp and small single-copy region of 18,108 bp were obtained. These reads were separated by a pair of inverted repeat regions (IR) of 25,218 bp, and the overall guanine and cytosine (GC) was 37.24%. The plastid genome contains 130 genes (111 genes were unique and 19 genes were found duplicated in each IR region), including 86 protein-coding genes, 36 transfer RNA-coding genes, eight ribosomal RNA-coding genes, and 25 genes with introns (21 genes with one intron and four genes with two introns). The phylogenetic tree reconstructed based on single-copy orthologous genes and maximum likelihood analysis indicated that Chenopodium petiolare is most closely related to Chenopodium quinoa.

Cisplatin Toxicity Causes Neutrophil-Mediated Inflammation in Zebrafish Larvae.

Cisplatin is an antineoplastic agent used to treat various tumors. In mammals, it can cause nephrotoxicity, tissue damage, and inflammation. The release of inflammatory mediators leads to the recruitment and infiltration of immune cells, particularly neutrophils, at the site of inflammation. Cisplatin is often used as an inducer of acute kidney injury (AKI) in experimental models, including zebrafish (Danio rerio), due to its accumulation in kidney cells. Current protocols in larval zebrafish focus on studying its effect as an AKI inducer but ignore other systematic outcomes. In this study, cisplatin was added directly to the embryonic medium to assess its toxicity and impact on systemic inflammation using locomotor activity analysis, qPCR, microscopy, and flow cytometry. Our data showed that larvae exposed to cisplatin at 7 days post-fertilization (dpf) displayed dose-dependent mortality and morphological changes, leading to a decrease in locomotion speed at 9 dpf. The expression of pro-inflammatory cytokines such as interleukin (il)-12, il6, and il8 increased after 48 h of cisplatin exposure. Furthermore, while a decrease in the number of neutrophils was observed in the glomerular region of the pronephros, there was an increase in neutrophils throughout the entire animal after 48 h of cisplatin exposure. We demonstrate that cisplatin can have systemic effects in zebrafish larvae, including morphological and locomotory defects, increased inflammatory cytokines, and migration of neutrophils from the hematopoietic niche to other parts of the body. Therefore, this protocol can be used to induce systemic inflammation in zebrafish larvae for studying new therapies or mechanisms of action involving neutrophils.

The color purple: Purple urine bag syndrome: Identifying the phenomenon to ease unnecessary anxiety, fear, and distress.

OBJECTIVES: Purple urine bag syndrome (PUBS) is a condition that causes an intense purple discoloration of the urine, predominately in frail, dependent, and bedridden persons who are chronically catheterized and have urinary tract infections. Despite being considered a benign syndrome, PUBS can cause great anxiety, fear, and distress in health professionals, chronically ill persons, and caregivers or family members who provide care. METHODS: We report the case of a 98-year-old institutionalized woman with Alzheimer's dementia with a long-term urinary catheter who developed PUBS. RESULTS: Although alarming and distressing for the resident and the health-care team, PUBS was resolved by treating the underlying urinary tract infection and applying good genital hygiene and catheter replacement. SIGNIFICANCE OF RESULTS: Identifying PUBS and its clinical features and management proved to be significantly helpful in ameliorating the anxiety, fear, and distress around the phenomenon.

Aspirin 15cH has Different Effects on Morphology and Function of Lipopolysaccharide-Challenged RAW 264.7 Macrophages In Vitro Compared to a Pharmacological Dose of Aspirin.

INTRODUCTION: Aspirin is one of the most commonly used drugs worldwide. It is known to present antipyretic, anti-inflammatory and anti-thrombotic actions, making it extremely useful in a wide range of clinical contexts. Interestingly, homeopathically prepared Aspirin 15cH has been found to have a pro-thrombotic effect in rats, raising the hypothesis that Aspirin 15cH could also modulate the activity of inflammatory cells in different pathological processes. OBJECTIVE: Our objective was to assess what effect Aspirin 15cH has on RAW 264.7 macrophages in vitro. METHODS: The effects of Aspirin 15cH on biochemical and morphological activities of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages were evaluated. These effects were compared with unchallenged macrophages (negative control), untreated LPS-stimulated macrophages, macrophages treated with succussed water (vehicle control), or aspirin 200 µg/mL (pharmacological inhibitor of LPS activity). Cell morphology (adhered cell area and cytoskeleton arrangements), cell viability, toll-like receptor-4 (TLR-4) expression, and the production of nitric oxide, cytokines and intracellular reactive oxygen species were assessed. RESULTS: Aspirin 15cH reduced the number of cells expressing TLR-4 on the surface (p = 0.03) and induced a "columnar" morphology of macrophage pseudopods, indicating changes in cytoskeleton arrangement. When cells were treated with both Aspirin 15cH and LPS, cell morphology became heterogeneous, suggesting that sub-populations of cells had differing sensitivities to LPS or Aspirin 15cH. Exposure of the cells to LPS alone, succussed water or aspirin 200 µg/mL produced effects consistent with the literature. CONCLUSION: Aspirin 15cH, aspirin 200 µg/mL, LPS and succussed water appear to act as independent stimuli able to induce different patterns of macrophage response. Aspirin 15cH induced changes suggestive of M2 polarization of the macrophages (i.e., toward a wound healing or tissue repair, rather than inflammatory, phenotype). These preliminary findings need to be confirmed in further specific studies.

High glucose impairs human periodontal ligament cells migration through lowered microRNAs 221 and 222.

OBJECTIVE: To investigate the effects of miR-221 and miR-222 and high glucose on human periodontal ligament (PL) cells morphology, cytoskeleton, adhesion, and migration. BACKGROUND: Chronic hyperglycemia is common in uncontrolled diabetes mellitus (DM) and plays a central role in long-term DM complications, such as impaired periodontal healing. We have previously shown that high glucose increases apoptosis of human PL cells by inhibiting miR-221 and miR-222 and consequently augmenting their target caspase-3. However, other effects of miR-221/222 downregulation on PL cells are still unknown. METHODS: Cells from young humans' premolar teeth were cultured for 7 days under 5 or 30 mM glucose. Directional and spontaneous migration on fibronectin were studied using transwell and time-lapse assays, respectively. F-actin staining was employed to study cell morphology and the actin cytoskeleton. MiR-221 and miR-222 were inhibited using antagomiRs, and their expressions were evaluated by real-time RT-PCR. RESULTS: High glucose inhibited PL cells early adhesion, spreading, and migration on fibronectin. Cells exposed to high glucose showed reduced polarization, velocity, and directionality. They formed several simultaneous unstable and short-lived protrusions, suggesting impairment of adhesion maturation. MiR-221 and miR-222 inhibition also reduced migration, decreasing cell directionality but not significantly cell velocity. After miR-221 and miR-222 downregulation cells showed morphological resemblance with cells exposed to high glucose. CONCLUSION: High glucose impairs human PL cells migration potentially through a mechanism involving reduction of microRNA-221 and microRNA-222 expression. These effects may contribute to the impairment of periodontal healing, especially after surgery and during guided regeneration therapies.

SARS-CoV-2 Spike protein peptides displayed in the Pyrococcus furiosus RAD system preserve epitopes antigenicity, immunogenicity, and virus-neutralizing activity of antibodies.

Amongst the potential contribution of protein or peptide-display systems to study epitopes with relevant immunological features, the RAD display system stands out as a highly stable scaffold protein that allows the presentation of constrained target peptides. Here, we employed the RAD display system to present peptides derived from the SARS-CoV-2 Spike (S) protein as a tool to detect specific serum antibodies and to generate polyclonal antibodies capable of inhibiting SARS-CoV-2 infectivity in vitro. 44 linear S-derived peptides were genetically fused with the RAD scaffold (RAD-SCoV-epitopes) and screened for antigenicity with sera collected from COVID-19-infected patients. In a second step, selected RAD-SCoV-epitopes were used to immunize mice and generate antibodies. Phenotypic screening showed that some of these antibodies were able to recognize replicating viral particles in VERO CCL-81 and most notably seven of the RAD-SCoV-epitopes were able to induce antibodies that inhibited viral infection. Our findings highlight the RAD display system as an useful platform for the immunological characterization of peptides and a potentially valuable strategy for the design of antigens for peptide-based vaccines, for epitope-specific antibody mapping, and for the development of antibodies for diagnostic and therapeutic purposes.

FLEXIBLE INTRAMEDULLARY NAILS IN PEDIATRIC SUBTROCHANTERIC FEMUR FRACTURE: BIOMECHANICAL STUDY.

Objective: Evaluate the stability provided by two flexible intramedullary nails (FINs) in a simulation of fractures at the proximal levels in pediatric femur models. Methods: Two FINs were inserted in 18 synthetic models of pediatric femurs. Fractures were simulated at one of three levels, and the models were divided into the following groups (n=6): diaphysis (control), subtrochanteric and trochanteric. Flex-compression tests were performed with force up to 85 N. Relative stiffness and the average deformation was obtained. Torsion tests were performed by rotating the proximal fragment until 20°, to obtain the average torque. Results: At flex-compression, the set's average relative stiffness and average deformations were: 54.360x103 N/m and 1.645 mm in the control group, respectively. In the subtrochanteric group, the relative stiffness was 31.415x103 N/m (-42.2%) and the deformation was 2.424 mm (+47.3%) (p<0.05). For the trochanteric group, the relative stiffness was 30.912x103 N/m (+43.1%) and the deformation was 2.508 mm (+52.4%) (p<0.05). In torsion, the average torque was 1.410 Nm in the control group; 1.116 Nm in the subtrochanteric group (-20.8%), and 2.194 Nm in the trochanteric group (+55.6%) (p<0.05). Conclusion: FINs do not seem to be biomechanically competent for the treatment of proximal femoral fractures. Level of Evidence I; Therapeutic Studies - Investigating the results of treatment.

Objetivo: avaliar a estabilidade proporcionada por duas hastes intramedulares flexíveis na simulação de fraturas nos níveis proximais em modelos pediátricos de fêmur. Métodos: Duas hastes foram inseridas em 18 modelos sintéticos de fêmures pediátricos. As fraturas foram simuladas em um dos três níveis, e os modelos foram divididos nos seguintes grupos (n=6): diáfise(controle), subtrocantérico e trocantérico. Testes de flexão-compressão foram realizados com força de até 85N. A rigidez relativa e a deformação média foram obtidas. Os testes de torção foram realizados girando o fragmento proximal até 20°, para obter o torque médio. Resultados: Na flexo-compressão, a rigidez relativa média e as deformações médias do conjunto foram: 54,360x103 N/m e 1,645 mm no grupo controle, respectivamente. No grupo subtrocantérico a rigidez relativa foi de 31,415x103 N/m (-42,2%) e a deformação foi de 2,424 mm (+47,3%) (p<0,05). Para o grupo trocantérico a rigidez relativa foi de 30,912x103 N/m (+43,1%) e a deformação foi de 2,508 mm (+52,4%) (p<0,05). Na torção, o torque médio foi de 1.410 Nm no grupo controle; 1,116 Nm no grupo subtrocantérico (-20,8%) e 2,194 Nm no grupo trocantérico (+55,6%) (p<0,05). Conclusão: As hastes intramedulares flexíveis não parecem ser biomecanicamente competentes para o tratamento das fraturas proximais do fêmur. Nível de Evidência I; Estudos terapêuticos - Investigação dos resultados do tratamento.

Determination of Nutritional and Antioxidant Properties of Maya Nut Flour (Brosimum alicastrum) for Development of Functional Foods.

Maya nut (Brosimum alicastrum) is a novel food with high nutritional value. This research aimed to evaluate the nutritional and antioxidant properties of Maya nut flour (MNF) made from seeds dried by different methods (sun-dried and using hot air at 45 °C and 60 °C) to explore its incorporation into cookies and evaluate its nutritional and functional properties. The naturally sun-dried flour (NF) had the highest content of ash (3.64 ± 0.11 g/100 g), protein (6.35 ± 0.44 g/100 g), crude fiber (6.75 ± 0.29 g/100 g), and functional properties (water and oil absorption). The color of the flour was affected by the different drying methods. While the drying methods influenced the total polyphenolic content (TPC) and antioxidant activity (AA) of MNF, they did not affect the morphology of the native starch or generated important molecular-structural changes. The substitution of 60% of wheat flour with NF in the cookie's formula increased the protein and fiber content, whereas 20% substitution increased its AA. MNF is a source of protein, dietary fiber, micronutrients, and functional compounds that can enrich cookie formulations.

Interpreting Image-based Profiles using Similarity Clustering and Single-Cell Visualization.

Image-based profiling quantitatively assesses the effects of perturbations on cells by capturing a breadth of changes via microscopy. Here, we provide two complementary protocols to help explore and interpret data from image-based profiling experiments. In the first protocol, we examine the similarity among perturbed cell samples using data from compounds that cluster by their mechanisms of action. The protocol includes steps to examine feature-driving differences between samples and to visualize correlations between features and treatments to create interpretable heatmaps using the open-source web tool Morpheus. In the second protocol, we show how to interactively explore images together with the numerical data, and we provide scripts to create visualizations of representative single cells and image sites to understand how changes in features are reflected in the images. Together, these two tutorials help researchers interpret image-based data to speed up research. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Exploratory analysis of profile similarities and driving features Basic Protocol 2: Image and single-cell visualization following profile interpretation.

"Mãos de Conforto" (Hands of Comfort): A novel non-pharmacological intervention to ease agitation in elderly persons with dementia.

Behavioral symptoms associated with dementia, such as agitation, are frequent and associated with well-known negative consequences for patients, their carers, and their environment. Pharmacological treatments for agitation using sedatives and antipsychotics are known to have several undesirable side effects and modest efficacy. Non-pharmacological alternatives are recommended as first-line options for agitation in persons with dementia with few side effects, but there is limited evidence of efficacy. We developed a novel and simple non-pharmacological alternative for agitation in dementia residents based on a Brazilian intervention using warm water surgical gloves used in patients with COVID-19 in intensive care units during the pandemic. We coined it "Mãos de Conforto" - Hands of Comfort. We report a series of 7 cases in 3 residents with dementia who whore Hands of Comfort.

Plastid genome of Passiflora tripartita var. mollissima (poro-poro) from Huánuco, Peru.

Passiflora tripartita var. mollissima, known locally as poro-poro, is an important native fruit used in traditional Peruvian medicine with relevant agro-industrial and pharmaceutical potential for its antioxidant capacity for human health. However, to date, only a few genetic data are available, which limits exploring its genetic diversity and developing new genetic studies for its improvement. We report the poro-poro plastid genome to expand the knowledge of its molecular markers, evolutionary studies, molecular pathways, and conservation genetics. The complete chloroplast (cp) genome is 163,451 bp in length with a typical quadripartite structure, containing a large single-copy region of 85,525 bp and a small single-copy region of 13,518 bp, separated by a pair of inverted repeat regions (IR) of 32,204 bp, and the overall GC content was 36.87%. This cp genome contains 128 genes (110 genes were unique and 18 genes were found duplicated in each IR region), including 84 protein-coding genes, 36 transfer RNA-coding genes, eight ribosomal RNA-coding genes, and 13 genes with introns (11 genes with one intron and two genes with two introns). The inverted repeat region boundaries among species were similar in organization, gene order, and content, with a few revisions. The phylogenetic tree reconstructed based on single-copy orthologous genes and maximum likelihood analysis demonstrates poro-poro is most closely related to Passiflora menispermifolia and Passiflora oerstedii. In summary, our study constitutes a valuable resource for studying molecular evolution, phylogenetics, and domestication. It also provides a powerful foundation for conservation genetics research and plant breeding programs. To our knowledge, this is the first report on the plastid genome of Passiflora tripartita var. mollissima from Peru.

Flexible intramedullary nails in pediatric subtrochanteric femur fracture: biomechanical study / Hastes flexíveis na fixação de fratura subtrocantérica de fêmur pediátrico: análise biomecânica

ABSTRACT Objective Evaluate the stability provided by two flexible intramedullary nails (FINs) in a simulation of fractures at the proximal levels in pediatric femur models. Methods Two FINs were inserted in 18 synthetic models of pediatric femurs. Fractures were simulated at one of three levels, and the models were divided into the following groups (n=6): diaphysis (control), subtrochanteric and trochanteric. Flex-compression tests were performed with force up to 85 N. Relative stiffness and the average deformation was obtained. Torsion tests were performed by rotating the proximal fragment until 20°, to obtain the average torque. Results At flex-compression, the set's average relative stiffness and average deformations were: 54.360x103 N/m and 1.645 mm in the control group, respectively. In the subtrochanteric group, the relative stiffness was 31.415x103 N/m (-42.2%) and the deformation was 2.424 mm (+47.3%) (p<0.05). For the trochanteric group, the relative stiffness was 30.912x103 N/m (+43.1%) and the deformation was 2.508 mm (+52.4%) (p<0.05). In torsion, the average torque was 1.410 Nm in the control group; 1.116 Nm in the subtrochanteric group (-20.8%), and 2.194 Nm in the trochanteric group (+55.6%) (p<0.05). Conclusion FINs do not seem to be biomechanically competent for the treatment of proximal femoral fractures. Level of Evidence I; Therapeutic Studies - Investigating the results of treatment.

RESUMO Objetivo avaliar a estabilidade proporcionada por duas hastes intramedulares flexíveis na simulação de fraturas nos níveis proximais em modelos pediátricos de fêmur. Métodos Duas hastes foram inseridas em 18 modelos sintéticos de fêmures pediátricos. As fraturas foram simuladas em um dos três níveis, e os modelos foram divididos nos seguintes grupos (n=6): diáfise(controle), subtrocantérico e trocantérico. Testes de flexão-compressão foram realizados com força de até 85N. A rigidez relativa e a deformação média foram obtidas. Os testes de torção foram realizados girando o fragmento proximal até 20°, para obter o torque médio. Resultados Na flexo-compressão, a rigidez relativa média e as deformações médias do conjunto foram: 54,360x103 N/m e 1,645 mm no grupo controle, respectivamente. No grupo subtrocantérico a rigidez relativa foi de 31,415x103 N/m (-42,2%) e a deformação foi de 2,424 mm (+47,3%) (p<0,05). Para o grupo trocantérico a rigidez relativa foi de 30,912x103 N/m (+43,1%) e a deformação foi de 2,508 mm (+52,4%) (p<0,05). Na torção, o torque médio foi de 1.410 Nm no grupo controle; 1,116 Nm no grupo subtrocantérico (-20,8%) e 2,194 Nm no grupo trocantérico (+55,6%) (p<0,05). Conclusão As hastes intramedulares flexíveis não parecem ser biomecanicamente competentes para o tratamento das fraturas proximais do fêmur. Nível de Evidência I; Estudos terapêuticos - Investigação dos resultados do tratamento.

Effect of Gastrointestinal Digestion on the Bioaccessibility of Phenolic Compounds and Antioxidant Activity of Fermented Aloe vera Juices.

Plant-based beverages are enriched by the fermentation process. However, their biocompounds are transformed during gastrointestinal digestion, improving their bioaccessibility, which is of primary importance when considering the associated health benefits. This study aimed to evaluate the effect of in vitro gastrointestinal digestion on phenolic compound bioaccessibility and antioxidant activity of novel Aloe vera juices fermented by probiotic Enterococcus faecium and Lactococcus lactis. Aloe vera juices were digested using the standardized static INFOGEST protocol. During digestion, phenolic compounds and antioxidant activity (DPPH, ABTS, and FRAP) were accessed. The digestion process was seen to significantly increase the total phenolic content of the fermented Aloe vera juices. The fermentation of Aloe vera increased the bioaccessibility of juice biocompounds, particularly for kaempferol, ellagic acid, resveratrol, hesperidin, ferulic acid, and aloin. The phenolics released during digestion were able to reduce the oxidative radicals assessed by ABTS and FRAP tests, increasing the antioxidant action in the intestine, where they are absorbed. The fermentation of Aloe vera by probiotics is an excellent process to increase the bioavailability of beverages, resulting in natural added-value functional products.

Novel Bio-Functional Aloe vera Beverages Fermented by Probiotic Enterococcus faecium and Lactobacillus lactis.

Aloe vera has been medicinally used for centuries. Its bioactive compounds have been shown to be very effective in the treatment of numerous diseases. In this work, a novel functional beverage was developed and characterized to combine the health benefits of probiotic bacteria with the Aloe vera plant itself. Two Aloe vera juices were obtained by fermentation either by a novel isolated Enterococcus faecium or a commercial Lactococcus lactis. The extraction of Aloe vera biocompounds for further fermentation was optimized. Extraction with water plus cellulase enhanced the carbohydrates and phenolic compounds in the obtained extracts. The biotransformation of the bioactive compounds from the extracts during fermentation was assessed. Both probiotic bacteria were able to grow on the Aloe vera extract. Lactic acid and short-chain fatty acids (SCFA) together with fourteen individual phenolic compounds were quantified in the produced Aloe vera juice, mainly epicatechin, aloin, ellagic acid, and hesperidin. The amount of total phenolic compounds was maintained through fermentation. The antioxidant activity was significantly increased in the produced juice by the ABTS method. The novel produced Aloe vera juice showed great potential as a functional beverage containing probiotics, prebiotics, SCFA, and phenolic compounds in its final composition.

The Relationship between Metacomprehension and Reading Comprehension in Spanish as a Second Language / La relación entre la metacomprensión y la comprensión lectora en español como segunda lengua

The importance of reading proficiency in a second language (L2) is growing worldwide. Reading in a L2 involves many questions about the abilities of reading comprehension, including metacomprehension, a core reading ability in native speakers (L1) that allows them to be aware of their reading comprehension level and to regulate their learning. However, its link to L2 has not been sufficiently researched. This study examines the relationship between the reading comprehension scores of diverse texts and metacomprehension, assessed by the ECOMPLEC test and PROLEC-R reading task, in Portuguese students studying Spanish as a L2. Similar to the results found in L1 research, we found a significant relationship between reading comprehension and metacomprehension in expository and discontinuous texts: students with low reading comprehension scores overestimated their metacomprehension while metacomprehension abilities of students with high reading comprehension abilities were more accurate. These results provide a frame of reference for future studies on metacomprehension in L2.

El dominio de la lectura en una segunda lengua (L2) tiene cada vez más importancia a nivel global. La lectura en una L2 implica muchas cuestiones sobre las capacidades de comprensión lectora, como la metacomprensión, una habilidad de lectura fundamental en hablantes nativos (L1) que les permite ser conscientes de su nivel de comprensión lectora y les ayuda a regular su aprendizaje. Sin embargo, su papel en una L2 no ha sido lo suficientemente investigado. El presente estudio examina la relación entre las puntuaciones de comprensión lectora de varios tipos de textos y la metacomprensión, evaluada a través de la batería ECOMPLEC y la tarea de lectura de la batería PROLEC-R en estudiantes portugueses que estudian español como L2. De manera similar a los resultados encontrados en investigaciones en L1, hemos descubierto una relación significativa entre la comprensión lectora y la metacomprensión en textos expositivos y discontinuos: los estudiantes con bajas puntuaciones de comprensión lectora sobreestimaron su capacidad de metacomprensión, mientras que las habilidades de metacomprensión de los estudiantes con puntuaciones altas en comprensión lectora fueron más adecuadas. Los resultados proporcionan un marco de referencia para futuros estudios sobre metacomprensión en L2.

Effective Genome Editing in Leishmania (Viannia) braziliensis Stably Expressing Cas9 and T7 RNA Polymerase.

Until 2015, loss-of-function studies to elucidate protein function in Leishmania relied on gene disruption through homologous recombination. Then, the CRISPR/Cas9 revolution reached these protozoan parasites allowing efficient genome editing with one round of transfection. In addition, the development of LeishGEdit, a PCR-based toolkit for generating knockouts and tagged lines using CRISPR/Cas9, allowed a more straightforward and effective genome editing. In this system, the plasmid pTB007 is delivered to Leishmania for episomal expression or integration in the ß-tubulin locus and for the stable expression of T7 RNA polymerase and Cas9. In South America, and especially in Brazil, Leishmania (Viannia) braziliensis is the most frequent etiological agent of tegumentary leishmaniasis. The L. braziliensis ß-tubulin locus presents significant sequence divergence in comparison with Leishmania major, which precludes the efficient integration of pTB007 and the stable expression of Cas9. To overcome this limitation, the L. major ß-tubulin sequences, present in the pTB007, were replaced by a Leishmania (Viannia) ß-tubulin conserved sequence generating the pTB007_Viannia plasmid. This modification allowed the successful integration of the pTB007_Viannia cassette in the L. braziliensis M2903 genome, and in silico predictions suggest that this can also be achieved in other Viannia species. The activity of Cas9 was evaluated by knocking out the flagellar protein PF16, which caused a phenotype of immobility in these transfectants. Endogenous PF16 was also successfully tagged with mNeonGreen, and an in-locus complementation strategy was employed to return a C-terminally tagged copy of the PF16 gene to the original locus, which resulted in the recovery of swimming capacity. The modified plasmid pTB007_Viannia allowed the integration and stable expression of both T7 RNA polymerase and Cas9 in L. braziliensis and provided an important tool for the study of the biology of this parasite.

Loss of mTORC2 Activity in Neutrophils Impairs Fusion of Granules and Affects Cellular Metabolism Favoring Increased Bacterial Burden in Sepsis.

Sepsis is a complex infectious syndrome in which neutrophil participation is crucial for patient survival. Neutrophils quickly sense and eliminate the pathogen by using different effector mechanisms controlled by metabolic processes. The mammalian target of rapamycin (mTOR) pathway is an important route for metabolic regulation, and its role in neutrophil metabolism has not been fully understood yet, especially the importance of mTOR complex 2 (mTORC2) in the neutrophil effector functions. In this study, we observed that the loss of Rictor (mTORC2 scaffold protein) in primary mouse-derived neutrophils affects their chemotaxis by fMLF and their microbial killing capacity, but not the phagocytic capacity. We found that the microbicidal capacity was impaired in Rictor-deleted neutrophils because of an improper fusion of granules, reducing the hypochlorous acid production. The loss of Rictor also led to metabolic alterations in isolated neutrophils, increasing aerobic glycolysis. Finally, myeloid-Rictor-deleted mice (LysMRic Δ/Δ) also showed an impairment of the microbicidal capacity, increasing the bacterial burden in the Escherichia coli sepsis model. Overall, our results highlight the importance of proper mTORC2 activation for neutrophil effector functions and metabolism during sepsis.