Advances in ex vivo expansion of hematopoietic stem and progenitor cells for clinical applications.

As caretakers of the hematopoietic system, hematopoietic stem cells assure a lifelong supply of differentiated populations that are responsible for critical bodily functions, including oxygen transport, immunological protection and coagulation. Due to the far-reaching influence of the hematopoietic system, hematological disorders typically have a significant impact on the lives of individuals, even becoming fatal. Hematopoietic cell transplantation was the first effective therapeutic avenue to treat such hematological diseases. Since then, key use and manipulation of hematopoietic stem cells for treatments has been aspired to fully take advantage of such an important cell population. Limited knowledge on hematopoietic stem cell behavior has motivated in-depth research into their biology. Efforts were able to uncover their native environment and characteristics during development and adult stages. Several signaling pathways at a cellular level have been mapped, providing insight into their machinery. Important dynamics of hematopoietic stem cell maintenance were begun to be understood with improved comprehension of their metabolism and progressive aging. These advances have provided a solid platform for the development of innovative strategies for the manipulation of hematopoietic stem cells. Specifically, expansion of the hematopoietic stem cell pool has triggered immense interest, gaining momentum. A wide range of approaches have sprouted, leading to a variety of expansion systems, from simpler small molecule-based strategies to complex biomimetic scaffolds. The recent approval of Omisirge, the first expanded hematopoietic stem and progenitor cell product, whose expansion platform is one of the earliest, is predictive of further successes that might arise soon. In order to guarantee the quality of these ex vivo manipulated cells, robust assays that measure cell function or potency need to be developed. Whether targeting hematopoietic engraftment, immunological differentiation potential or malignancy clearance, hematopoietic stem cells and their derivatives need efficient scaling of their therapeutic potency. In this review, we comprehensively view hematopoietic stem cells as therapeutic assets, going from fundamental to translational.

Fibulin-2 is an extracellular matrix inhibitor of oligodendrocytes relevant to multiple sclerosis.

Impairment of oligodendrocytes and myelin contributes to neurological disorders including multiple sclerosis (MS), stroke, and Alzheimer's disease. Regeneration of myelin (remyelination) decreases the vulnerability of demyelinated axons, but this repair process commonly fails with disease progression. A contributor to inefficient remyelination is the altered extracellular matrix (ECM) in lesions, which remains to be better defined. We have identified fibulin-2 (FBLN2) as a highly upregulated ECM component in lesions of MS and stroke and in proteome databases of Alzheimer's disease and traumatic brain injury. Focusing on MS, the inhibitory role of FBLN2 was suggested in the experimental autoimmune encephalomyelitis (EAE) model, in which genetic FBLN2 deficiency improved behavioral recovery by promoting the maturation of oligodendrocytes and enhancing remyelination. Mechanistically, when oligodendrocyte progenitors were cultured in differentiation medium, FBLN2 impeded their maturation into oligodendrocytes by engaging the Notch pathway, leading to cell death. Adeno-associated virus deletion of FBLN2 in astrocytes improved oligodendrocyte numbers and functional recovery in EAE and generated new myelin profiles after lysolecithin-induced demyelination. Collectively, our findings implicate FBLN2 as a hitherto unrecognized injury-elevated ECM, and a therapeutic target, that impairs oligodendrocyte maturation and myelin repair.

Additive Cytotoxic and Colony-Formation Inhibitory Effects of Aspirin and Metformin on PI3KCA-Mutant Colorectal Cancer Cells.

Human malignancies are one of the major health-related issues throughout the world and are anticipated to rise in the future. Despite huge investments made in anticancer drug development, limited success has been obtained and the average number of FDA approvals per year is declining. So, an increasing interest in drug repurposing exists. Metformin (MET) and aspirin (ASP) possess anticancer properties. This work aims to test the effect of these two drugs in combination on colorectal cancer (CRC) cells in vitro. The effects of MET and/or ASP on cell proliferation, viability, migratory ability, anchorage-independent growth ability (colony formation), and nutrient uptake were determined in two (HT-29 and Caco-2) human CRC cell lines. Individually, MET and ASP possessed antiproliferative, cytotoxic, and antimigratory effects and reduced colony formation in HT-29 cells (BRAF- and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PI3KCA)-mutant), although MET did not affect either 3H-deoxy-D-glucose or 14C-butyrate uptake and lactate production, and ASP caused only a small decrease in 14C-butyrate uptake. Moreover, in these cells, the combination of MET and ASP resulted in a tendency to an increase in the cytotoxic effect and in a potentiation of the inhibitory effect on colony formation, although no additive antiproliferative and antimigratory effects, and no effect on nutrient uptake and lactate production were observed. In contrast, MET and ASP, both individually and in combination, were almost devoid of effects on Caco-2 cells (BRAF- and PI3KCA-wild type). We suggest that inhibition of PI3K is the common mechanism involved in the anti-CRC effect of both MET, ASP and their combination and, therefore, that the combination of MET + ASP may especially benefit PI3KCA-mutant CRC cases, which currently have a poor prognostic.

construção de uma proposta de educação continuada sobre vacinação infantil para agentes comunitários de saúde / Construction of a continuing education proposal on childhood vaccination for community health agents / Construcción de una propuesta de educación continua sobre vacunación infantil para agentes comunitarios de salud

O objetivo deste estudo foi relatar a experiência do desenvolvimento de uma proposta de educação continuada executada pela equipe de enfermagem com agentes comunitários de saúde (ACS), considerando como foco a vacinação infantil. Trata-se de um relato de experiência elaborado e vivenciado por enfermeiro e acadêmico de enfermagem em novembro de 2019, com a participação de duas equipes pertencentes a uma Unidade Básica de Saúde do município de Arapiraca (AL). Através da metodologia de ensino da problematização por meio do Arco de Maguerez, foram realizadas oficinas e ações educativas para os ACS, a partir das necessidades relatadas por esses profissionais em relação à vacinação infantil. Tais atividades proporcionaram maior aproximação com a realidade vivenciada por quem ocupa a posição de educador e incentivador do cuidado da saúde durante a visita domiciliar. Além disso, enfatizaram a importância do enfermeiro enquanto responsável pela educação continuada da equipe. Verificou-se ainda que o diálogo aberto e horizontal, de modo empático, proporcionou a construção de uma ferramenta eficaz, de fácil manuseio e de baixo custo, que gerou uma melhor memorização e segurança não somente para os ACS nas visitas domiciliares, mas também nas atividades de outros profissionais atuantes no campo da promoção da saúde infantil.

This study reports the experience of developing an educational product based on continued education carried out by a nursing team with community health agents (CHA) considering childhood vaccination as the focus. This is an experience report that was prepared and experienced by a nurse and nursing student in November 2019, with the participation of two teams belonging to a basic health unit in the municipality of Arapiraca, Alagoas, Brazil. Using the problematization teaching methodology by Arco de Marguerez, workshops and educational actions were carried out for the CHA based on the needs they reported regarding childhood vaccination, which provided a greater approximation with the reality experienced by those in the position of educators and encouragers of health care during home visits. Furthermore, it emphasized the importance of nurses as responsible for the team's continuing education. It was also verified that open and horizontal dialogue empathetically provided the construction of an effective, easy to use, low cost tool, bringing better memorization and safety not only for community agents during home visits but also in activities from other professionals working in the field of child health promotion.

El objetivo de este estudio fue reportar la experiencia de desarrollo de una propuesta de educación continua realizada por el equipo de enfermería con agentes comunitarios de salud (ACS), considerando como enfoque la vacunación infantil. Este es un reporte de experiencia elaborado y vivido por un enfermero y un estudiante de enfermería en noviembre de 2019, con la participación de dos equipos de una unidad básica de salud de la ciudad de Arapiraca, Brasil. Utilizando la metodología de enseñanza de problematización del Arco de Maguerez, se realizaron talleres y acciones educativas para ACS a partir de las necesidades reportadas por estos profesionales con relación a la vacunación infantil. Estas actividades proporcionaron una mayor aproximación a la realidad vivida por quienes ocupan la posición de educador y motivador del cuidado de la salud durante la visita domiciliaria. Además, destacaron la importancia del enfermero como responsable de la formación continua del equipo. También se constató que el diálogo abierto y horizontal, de manera empática, propició la construcción de una herramienta efectiva, fácil de usar y de bajo costo, que generó una mejor memorización y seguridad no solo para los ACS durante las visitas domiciliarias, sino también en las actividades de otros profesionales que trabajan en el campo de la promoción de la salud infantil.

Ruptured Intracranial Aneurysm in a 60-Day-Old Infant: An Extreme Case.

The prevalence of aneurysms in children is low when compared to adults, being even rarer in the first year of life. They can be secondary to infections, traumatic brain injury, autoimmune diseases, or connective tissue diseases. Dissecting etiology is rare. A 60-day-old female infant, previously healthy, presented to the emergency department (ED) with irritability and loss of appetite since the preceding day, a fever of one-hour duration, and vomiting. Laboratory analysis revealed a hemoglobin level of 6.5 g/dL, without elevation of inflammatory markers. In the ED, she experienced two episodes, with a one-hour interval, of clonic movements of the upper eyelid and right upper limb, along with conjugate gaze deviation to the same side, which resolved after intravenous diazepam. Levetiracetam was initiated after the second episode. The anterior fontanelle became progressively tense. Brain computed tomography (CT) showed a voluminous intraparenchymal and subarachnoid hemorrhage with an aneurysm at the bifurcation of the left middle cerebral artery (MCA). Initially, an endovascular approach was tried but was not successful due to technical problems. Consequently, a Vaso-CT scan was performed that confirmed a dissecting aneurysm/pseudoaneurysm (8 mm × 10 mm × 10 mm) of the left MCA, originating from the upper wall of the M1 segment. Next, she underwent microsurgical exclusion of the aneurysm using microclips. Post-surgery brain CT showed acute ischemia in the entire MCA region. Follow-up angiography showed complete exclusion of the aneurysm. She evolved to grade 3 monoparesis of the upper limb at the six-month interval follow-up, which has been gradually improving with physical rehabilitation. The next-generation sequencing (NGS) panel for aneurysms and arterial dissections did not detect any pathogenic variants. Clinical presentation of cerebral aneurysms in infants can be subtle, and a high index of suspicion is required in cases of irritability, altered consciousness, seizures, bulging fontanelle, and motor deficits. Early detection is of utmost importance as it is associated with moderate mortality. Surgical treatment with the use of clips proved to be effective in this case.

Secretome of bone marrow mesenchymal stromal cells cultured in a dynamic system induces neuroprotection and modulates microglial responsiveness in an α-synuclein overexpression rat model.

BACKGROUND AIMS: Parkinson's disease (PD) is the second most common neurodegenerative disorder. The etiology of the disease remains largely unknown, but evidence have suggested that the overexpression and aggregation of alpha-synuclein (α-syn) play key roles in the pathogenesis and progression of PD. Mesenchymal stromal cells (MSCs) have been earning attention in this field, mainly due to their paracrine capacity. The bioactive molecules secreted by MSCs, i.e. their secretome, have been associated with enhanced neuronal survival as well as a strong modulatory capacity of the microenvironments where the disease develops. The selection of the appropriate animal model is crucial in studies of efficacy assessment. Given the involvement of α-syn in the pathogenesis of PD, the evidence generated from the use of animal models that develop a pathologic phenotype due to the action of this protein is extremely valuable. Therefore, in this work, we established an animal model based on the viral vector-mediated overexpression of A53T α-syn and studied the impact of the secretome of bone marrow mesenchymal stromal cells MSC(M) as a therapeutic strategy. METHODS: Adult male rats were subjected to α-syn over expression in the nigrostriatal pathway to model dopaminergic neurodegeneration. The impact of locally administered secretome treatment from MSC(M) was studied. Motor impairments were assessed throughout the study coupled with whole-region (striatum and substantia nigra) confocal microscopy evaluation of histopathological changes associated with dopaminergic neurodegeneration and glial cell reactivity. RESULTS: Ten weeks after lesion induction, the animals received secretome injections in the substantia nigra pars compacta (SNpc) and striatum (STR). The secretome used was produced from bone marrow mesenchymal stromal cells MSC(M) expanded in a spinner flask (SP) system. Nine weeks later, animals that received the viral vector containing the gene for A53T α-syn and treated with vehicle (Neurobasal-A medium) presented dopaminergic cell loss in the SNpc and denervation in the STR. The treatment with secretome significantly reduced the levels of α-syn in the SNpc and protected the dopaminergic neurons (DAn) within the SNpc and STR. CONCLUSIONS: Our results are aligned with previous studies in both α-syn Caenorhabditis elegans models, as well as 6-OHDA rodent model, revealing that secretome exerted a neuroprotective effect. Moreover, these effects were associated with a modulation of microglial reactivity supporting an immunomodulatory role for the factors contained within the secretome. This further supports the development of new studies exploring the effects and the mechanism of action of secretome from MSC(M) against α-syn-induced neurotoxicity.

Dual production of human mesenchymal stromal cells and derived extracellular vesicles in a dissolvable microcarrier-based stirred culture system.

BACKGROUND & AIMS: Cell therapies based on mesenchymal stromal cells (MSCs) have gained an increasing therapeutic interest in the context of multiple disorders. Nonetheless, this field still faces important challenges, particularly concerning suitable manufacturing platforms. Here, we aimed at establishing a scalable culture system to expand umbilical cord-derived Wharton's jelly MSC (MSC(WJ)) and their derived extracellular vesicles (EVs) by using dissolvable microcarriers combined with xeno(geneic)-free culture medium. METHODS: MSC(WJ) isolated from three donors were cultured at a starting density of 1 × 106 cells per spinner flask, i.e., 2.8 × 103 cells per cm2 of dissolvable microcarrier surface area. After a 6-day expansion period of MSC(WJ), extracellular vesicles (EVs) were produced for 24 h. RESULTS: Taking advantage of an intermittent agitation regimen, we observed high adhesion rates to the microcarriers (over 90% at 24 h) and achieved 15.8 ± 0.7-fold expansion after 6 days of culture. Notably, dissolution of the microcarriers was achieved through a pectinase-based solution to recover the cell product, reducing the hurdles of downstream processing. MSC identity was validated by detecting the characteristic MSC immunophenotype and by multilineage differentiation assays. Considering the growing interest in MSC-derived EVs, which are known to be mediators of the therapeutic features of MSC, this platform also was evaluated for EV production. Upon a 24-h period of conditioning, secreted EVs were isolated by ultrafiltration followed by anion-exchange chromatography and exhibited the typical cup-shaped morphology, small size distribution (162.6 ± 30.2 nm) and expressed EV markers (CD63, CD9 and syntenin-1). CONCLUSIONS: Taken together, we established a time-effective and robust scalable platform that complies with clinical-grade standards for the dual production of MSC(WJ) and their derived EV.

Altered purinergic P2X7 and A2B receptors signaling limits macrophage-mediated host defense in schistosomiasis.

BACKGROUND: The occurrence of co-infections during schistosomiasis, a neglected tropical disease, with other parasites have been reported suggesting an impaired host immune defense. Macrophage purinergic P2X7 receptor (P2X7R) play an important role against intracellular pathogens. Therefore, we investigated the P2X7R-mediated phagocytosis and killing capacity of Leishmania amazonensis by macrophages during schistosomiasis in vitro and in vivo. METHODS: Swiss and C57BL/6 (Wild type) and P2X7R-/- were randomized in two groups: control (uninfected) and Schistosoma mansoni-infected. Alternatively, control Swiss and S. mansoni-infected mice were also infected with L. amazonensis. RESULTS: The pre-treatment of macrophages with the P2X7R antagonist (A74003) or TGF-ß reduced the phagocytosis index, mimicking the phenotype of cells from S. mansoni-infected mice and P2X7R-/- mice. Apyrase also reduced the phagocytosis index corroborating the role of ATP to macrophage activation. Moreover, l-arginine-nitric oxide pathway was compromised, which could explain the reduced killing capacity in response to ATP in vitro and in vivo. We found an increased extracellular nucleotide (ATP, ADP and AMP) hydrolysis along with an increased frequency of F4/80+ CD39+ macrophages from the S. mansoni-infected group. Moreover, the content of adenosine in the cell supernatant was higher in the S. mansoni-infected group in relation to controls. Schistosomiasis also increased the expression of macrophage adenosine A2BR. In good accordance, both ADA and the selective A2BR antagonist restored the phagocytosis index of macrophages from S. mansoni-infected group. CONCLUSIONS: Altogether, the altered P2X7R and A2BR signaling limits the role of macrophages to host defense against L. amazonensis during schistosomiasis, potentially contributing to the pathophysiology and clinically relevant co-infections.

Evaluation of the theranostic potential of [64Cu]CuCl2 in glioblastoma spheroids.

BACKGROUND: Glioblastoma is an extremely aggressive malignant tumor with a very poor prognosis. Due to the increased proliferation rate of glioblastoma, there is the development of hypoxic regions, characterized by an increased concentration of copper (Cu). Considering this, 64Cu has attracted attention as a possible theranostic radionuclide for glioblastoma. In particular, [64Cu]CuCl2 accumulates in glioblastoma, being considered a suitable agent for positron emission tomography. Here, we explore further the theranostic potential of [64Cu]CuCl2, by studying its therapeutic effects in advanced three-dimensional glioblastoma cellular models. First, we established spheroids from three glioblastoma (T98G, U373, and U87) and a non-tumoral astrocytic cell line. Then, we evaluated the therapeutic responses of spheroids to [64Cu]CuCl2 exposure by analyzing spheroids' growth, viability, and cells' proliferative capacity. Afterward, we studied possible mechanisms responsible for the therapeutic outcomes, including the uptake of 64Cu, the expression levels of a copper transporter (CTR1), the presence of a cancer stem cell population, and the production of reactive oxygen species (ROS). RESULTS: Results revealed that [64Cu]CuCl2 is able to significantly reduce spheroids' growth and viability, while also affecting cells' proliferation capacity. The uptake of 64Cu, the presence of cancer stem-like cells and the production of ROS were in accordance with the therapeutic response. However, expression levels of CTR1 were not in agreement with uptake levels, revealing that other mechanisms could be involved in the uptake of 64Cu. CONCLUSIONS: Overall, our results further support [64Cu]CuCl2 potential as a theranostic agent for glioblastoma, unveiling potential mechanisms that could be involved in the therapeutic response.

Recommendations from the Brazilian Society of Anesthesiology (SBA) for difficult airway management in pediatric care

Abstract Difficult airway management in pediatrics during anesthesia represents a major challenge, requiring a careful approach, advanced technical expertise, and accurate protocols. The task force of the Brazilian Society of Anesthesiology (SBA) presents a report containing updated recommendations for the management of difficult airways in children and neonates. These recommendations have been developed based on the consensus of a panel of experts, with the objective of offering strategies to overcome challenges during airway management in pediatric patients. Grounded in evidence published in international guidelines and expert opinions, the report highlights crucial steps for the appropriate management of difficult airways in pediatrics, encompassing assessment, preparation, positioning, pre-oxygenation, minimizing trauma, and, paramountly, the maintenance of arterial oxygenation. The report also delves into additional strategies involving the use of advanced tools, such as video laryngoscopy, flexible intubating bronchoscopy, and supraglottic devices. Emphasis is placed on the simplicity of implementing the outlined recommendations, with a focus on the significance of continuous education, training through realistic simulations, and familiarity with the latest available technologies. These practices are deemed essential to ensure procedural safety and contribute to the enhancement of anesthesia outcomes in pediatrics.

Cross-cultural adaptation of the Scale of Perception of Respect for and Maintenance of the Dignity of the Inpatient (CuPDPH) to Brazilian Portuguese and its psychometric properties—A multicenter cross-sectional study

Abstract Objective To adapt the Scale of Perception of Respect for and Maintenance of the Dignity of the Inpatient (CuPDPH) to the Brazilian language and culture and to assess its psychometric properties. Results The scale was evaluated by 15 experts, and 239 patients from three tertiary hospitals in Rio de Janeiro. All participants signed a consent form. Data have shown adequacy of the model (KMO=0.839, Bartlett's test of sphericity: χ2(171) = 2241.3, p = 0.000010), good adjusted content validity (CVCa ≥ 0.90), internal consistency and reliability, such as α = 0.927. Discussion CuPDPH is a rating scale on observable professional attitudes. Illnesses change lives and impose adaptation to a new situation, perceived as depersonalization, leading patients to try to regain control of their lives. Patients expressed "ill will" to fill out the scale. Psychiatric patients' scale filling time was higher than others. A sample from three Rio de Janeiro third-level hospitals may not reflect the country's population; also, this adaptation may not comprise all linguistic variations of Brazilian Portuguese and Portuguese-speaking countries. Conclusion The Portuguese version of the Scale of Perception of Respect for and Maintenance of the Dignity of the Inpatient (CuPDPH), a 19-item, six-component version is a reliable instrument to measure the perception of internal medicine, surgical, and psychiatric patients on the maintenance of their dignity in Rio de Janeiro, Brazil. This knowledge could be used in advancing research on patients' perception of dignity, as well as professional ethical competencies, staff-patient relationship skills, and leadership development in medical and other healthcare professional education.

Recommendations from the Brazilian Society of Anesthesiology (SBA) for difficult airway management in pediatric care.

Difficult airway management in pediatrics during anesthesia represents a major challenge, requiring a careful approach, advanced technical expertise, and accurate protocols. The task force of the Brazilian Society of Anesthesiology (SBA) presents a report containing updated recommendations for the management of difficult airways in children and neonates. These recommendations have been developed based on the consensus of a panel of experts, with the objective of offering strategies to overcome challenges during airway management in pediatric patients. Grounded in evidence published in international guidelines and expert opinions, the report highlights crucial steps for the appropriate management of difficult airways in pediatrics, encompassing assessment, preparation, positioning, pre-oxygenation, minimizing trauma, and, paramountly, the maintenance of arterial oxygenation. The report also delves into additional strategies involving the use of advanced tools, such as video laryngoscopy, flexible intubating bronchoscopy, and supraglottic devices. Emphasis is placed on the simplicity of implementing the outlined recommendations, with a focus on the significance of continuous education, training through realistic simulations, and familiarity with the latest available technologies. These practices are deemed essential to ensure procedural safety and contribute to the enhancement of anesthesia outcomes in pediatrics.

Inquérito epidemiológico para residentes do entorno da empresa polo petroquímico de Capuava no município de São Paulo / Epidemiological survey for residents surrounding the Capuava petrochemical hub company in the city of São Paulo

A poluição atmosférica é medida pela quantidade de substâncias poluentes presentes no ar, que o torna impróprio ou nocivo a saúde. Dentre as fontes com elevado potencial poluidor, encontram-se as indústrias petroquímicas, e estudos indicam maior prevalência de hipotireoidismo nas regiões próximas a um polo petroquímico. A partir de denúncias dos moradores da região próxima do Polo Petroquímico de Capuava (PPC), foi instaurada uma Comissão Parlamentar de Inquérito (CPI) pela Câmara Municipal de São Paulo, para investigação, e solicitado a realização de um inquérito epidemiológico. Objetivo: verificar se há maior prevalência de doença tireoidiana nas áreas próximas ao PPC comparando com áreas sem a exposição à emissão de poluentes por esse tipo de fonte fixa. Método: O inquérito epidemiológico foi realizado por sorteio aleatório em duas etapas, sendo cinco áreas trabalhadas com entrevista e levantamento de dados de saúde, exame dos entrevistados que apresentavam três ou mais sintomas e/ou referiram doença tireoidiana, e avaliação médica. Resultados: foram respondidos 3.674 questionários com 879 residentes encaminhados para coleta de exames, onde 654 concluíram todas as etapas passando em consulta médica. Desses, 101 residentes da área de exposição e 65 residentes da área controle receberam diagnóstico de doença tireoidiana. Quando analisada a população total entrevistada, a prevalência da doença tireoidiana foi de 4,6 na área de exposição e 4,4 na área controle. Já considerando apenas a população testada, a prevalência da doença na população foi de 23,7% e 28,5% respectivamente. Em relação a percepção de poluentes, 77% dos residentes da área de exposição referiram pelo menos uma queixa relacionada e emissão de fumaça, odor, poeira, fuligem ou ruído decorrentes das atividades executadas pelo PPC. Conclusão: o estudo demonstrou não haver uma diferença significativa entre as áreas, os resultados mostraram que a probabilidade de desenvolver doença tireoidiana não teve diferença significativa entre as áreas, entretanto, deve ser considerado o grande incômodo referido pela maioria dos entrevistados residentes na área de exposição. Novos estudos devem ser conduzidos nas áreas próximas a polos industriais para levantamento de dados de saúde e qualidade do ar.

Desempenho de pré-escolares em vocabulário e habilidades preditivas no ensino híbrido / Preschool performance in vocabulary and predictive skills in blended learning / Desempeño de preescolares en vocabulario y habilidades predictivas en aprendizaje combinado

Introdução: O ensino remoto limitou a vivência de experiências concretas em espaços coletivos de formação de conhecimentos e intensificou a vulnerabilidade infantil quanto à educação formal, em especial as crianças em fase inicial de alfabetização. A partir deste panorama, tornou-se fundamental investigar o desempenho de escolares em busca de informações sobre a repercussão ocasionada pela pandemia de COVID-19 ao desenvolvimento infantil. Objetivo: Verificar o desempenho de pré-escolares em vocabulário e habilidades preditivas no ensino híbrido, frente aos impactos da pandemia de COVID-19. Método: Participaram deste estudo 12 escolares, com idade entre 5 a 6 anos, pertencentes ao terceiro período da educação infantil. Para a avaliação foram selecionados o Protocolo de Identificação Precoce dos Problemas de Leitura, e os Testes de Vocabulário Auditivo e de Vocabulário Expressivo. Resultados: Os dados obtidos demonstraram significância estatística para as provas de Vocabulário Receptivo em relação ao Expressivo, Nomeação Automática Rápida e Conhecimento do Alfabeto, Segmentação Silábica e Produção de Rima, acertos para Vocabulário Expressivo e Conhecimento do Alfabeto, assim como para a quantidade de erros em Vocabulário Expressivo e para a Nomeação Automática Rápida. Conclusão: O sucesso almejado para o ingresso no 1º ano do Ensino Fundamental dependerá das variações individuais de cada aluno quanto ao desempenho dos aspectos avaliados neste trabalho e à qualidade dos estímulos recebidos. (AU)

Introduction: Remote teaching limited the experience of concrete experiences in collective spaces for the formation of knowledge and intensified children's vulnerability in terms of formal education, especially children in the initial phase of literacy. From this panorama, it became essential to investigate the performance of students in search of information about the impact caused by the pandemic of COVID-19 on child development. Objective: To verify the performance of preschoolers in vocabulary and predictive skills in hybrid teaching, given the impacts of the COVID-19 pandemic. Method: Participated in this study 12 schoolchildren, aged between 5 and 6 years, belonging to the third period of early childhood education. For the evaluation were selected the Protocol for Early Identification of Reading Problems, and the Auditory Vocabulary and Expressive Vocabulary Tests. Results: The data obtained showed statistical significance for the tests of Receptive Vocabulary in relation to Expressive, Rapid Automatic Naming and Alphabet Knowledge, Syllabic Segmentation and Rhyme Production, correct answers for Expressive Vocabulary and Alphabet Knowledge, as well as for the number of errors in Expressive Vocabulary and for the Automatic Rapid Naming. Conclusion: The desired success for entering the 1st year of Elementary School will depend on the individual variations of each student regarding the performance of the aspects evaluated in this work and the quality of the stimuli received. (AU)

Introducción: La enseñanza a distancia limitó la vivencia de experiencias concretas en espacios colectivos de formación de saberes e intensificó la vulnerabilidad de los niños frente a la educación formal, especialmente de los niños en fase inicial de alfabetización. A partir de este panorama, se hizo imprescindible investigar el desempeño de los escolares en busca de información sobre el impacto que la pandemia de la COVID-19 provocó en el desarrollo infantil. Objetivo: Verificar el desempeño de preescolares en vocabulario y habilidades predictivas en aprendizaje semipresencial, ante los impactos de la pandemia del COVID-19. Método: Participaron en este estudio 12 escolares, con edades comprendidas entre los 5 y 6 años, pertenecientes al tercer ciclo de educación infantil. Para la evaluación se seleccionaron el Protocolo para la Identificación Temprana de Problemas de Lectura, y las Pruebas de Vocabulario Auditivo y Vocabulario Expresivo. Resultados: Los datos obtenidos mostraron significancia estadística para las pruebas de Vocabulario Receptivo en relación con Expresivo, Denominación Automática Rápida y Conocimiento del Alfabeto, Segmentación Silábica y Producción de Rimas, aciertos para Vocabulario Expresivo y Conocimiento del Alfabeto, así como para el número de errores en Vocabulario Expresivo y para Nomenclatura Automática Rápida. Conclusión: El éxito deseado para el ingreso al 1° año de la Enseñanza Fundamental dependerá de las variaciones individuales de cada estudiante en cuanto al desempeño de los aspectos evaluados en este trabajo y la calidad de los estímulos recibidos. (AU)

Metal-amino acid complexes (Zn, Se, Cu, Fe, and Mn) enhance immune response, antioxidant capacity, liver function enzymes, and expression of cytokine genes in Nile Tilapia reared under field conditions.

OBJECTIVES: In the present study, we investigated the effects of partial or total replacement of dietary inorganic trace materials (ITM) with metal-amino acid complexes (MAACs) on immune response, antioxidant capacity, cytokine genes expression, and overall health status of Nile Tilapia Oreochromis niloticus reared in in-pond hapas (mesh cages) under field conditions. METHODS: All-male Nile Tilapia with an average initial weight of about 90 g fish-1 were stocked into the hapas, in quadruplicates, at a density of 60 fish per hapa (30 fish m-3 ). The fish in each hapa were acclimatized to culture conditions and test diets for 1 week. Water quality parameters, including temperature (°C), pH, dissolved oxygen (mg L-1 ), and total ammonia (mg L-1 ), were monitored regularly. The MAACs were formulated to contain the same mineral concentrations found in the inorganic premix. Metal-amino acid complexes replaced the ITM premix at 0 (control), 25, 50, 75, and 100% levels (Table 1). The diets were fed to caged Nile Tilapia twice a day, for 80 days, at a daily rate of 3% of their body weights. The fish were weighed every 10 days, and the daily rations were readjusted. At harvest, fish in each hapa were collected, counted, and weighed collectively. Five fish from each hapa were rapidly anesthetized, and used for chemical and physiological analyses, including antioxidant and immune response analyses, liver function analysis, and gene expression. RESULT: The activities of lysozyme, respiratory burst (%), alternative complement (ACH50), phagocytic cells, phenoloxidase, superoxide dismutase, glutathione peroxidase, and liver function enzymes were all improved with increasing supplemental MAACs up to the 50% substitution level. The expression of cytokine genes, including interferon, tumor necrosis factor, interleukin-1, serum alkaline phosphatase, the integrated microbial genomes, chloramphenicol acetyltransferase, and transforming growth factor genes, was significantly upregulated in fish fed on MAAC-supplemented diets compared with the control group. The 50% MAAC level produced the highest upregulation of these genes, whereas gene expression decreased with increasing dietary MAAC levels to 75% and 100%. The quadratic regression analysis indicated that about 60-65% MAAC is required for maximum promotion of immunological and oxidative stress responses, gene expression, and overall health status of Nile Tilapia reared in in-pond hapas under field conditions. CONCLUSION: Replacement of the ITM premix with a MAAC premix enhanced the immune and antioxidant responses of Nile Tilapia.

Demandas da Psicologia Escolar no Período de COVID-19: uma Revisão Sistemática / Demands of School Psychology in the Period of COVID-19: a Systematic Review / Demandas de la Psicología Escolar en el Periodo de la COVID-19: una Revisión Sistemática

Esta pesquisa investigou as demandas educacionais presentes no período da pandemia da COVID-19 no âmbito da Psicologia Escolar. Realizou-se um estudo de revisão sistemática, registrada sob o número CRD42021218696 no International Prospective Register of Ongoing Systematic Reviews (PROSPERO). Foi realizada uma busca abrangente nas bases de dados Educacional Resources Information Center (ERIC), Web of Science, PsycINFO, Embase e Pubmed. A revisão contemplou o período de dezembro de 2019 a abril de 2021. As buscas resultaram na seleção de 16 produções científicas, considerando o país de origem da publicação, delineamento das pesquisas, principais resultados e demandas encontradas. Os resultados indicaram impactos do ensino online: dificuldades relativas à transição para a modalidade remota e ao acesso aos recursos tecnológicos e à internet, insuficiente treinamento pedagógico, vulnerabilidade social e alteração comportamental dos estudantes. Defende-se o trabalho do psicólogo escolar como agente mediador de relações e processos de ensino, aprendizagem e desenvolvimento humano, tensionados na modalidade remota.

This research investigated the educational demands present in the period of the COVID-19 pandemic in the field of School Psychology. A systematic review study was conducted, registered under CRD42021218696 in the International Prospective Register of Ongoing Systematic Reviews (PROSPERO). A comprehensive search on the Educational Resources Information Center (ERIC), Web of Science, PsycINFO, Embase, and Pubmed databases was performed. The review covered the period from December 2019 to April 2021. The queries have led to the selection of 16 scientific publications, considering its country of origin, research design, main findings, and demands found. Results indicate impacts of online teaching: difficulties related to the transition to remote mode and access to technological resources and to the internet, scarce pedagogical training, social vulnerability, and behavioral change in students. The work of the school psychologist is defended as a mediator agent for relationships and processes of teaching, learning, and human development, which are tensioned in remote mode.

Este estudio investigó las demandas educativas presentes en el período de la pandemia de COVID-19 en el campo de la Psicología Escolar. Se realizó un estudio de revisión sistemática, registrado con el número CRD42021218696 en el Registro Internacional Prospectivo de Revisiones Sistemáticas en Curso (PROSPERO). Se realizó una búsqueda exhaustiva en las bases de datos Educacional Resources Information Center (ERIC), Web of Science, PsycINFO, Embase y Pubmed. La revisión abarcó el periodo comprendido entre diciembre de 2019 y abril de 2021. Las búsquedas resultaron en la selección de 16 producciones científicas, teniendo en cuenta el país de origen de la publicación, el diseño de la investigación, los principales resultados y las demandas encontradas. Los resultados indican los impactos de la enseñanza en línea: dificultades relativas a la transición a la modalidad remota y al acceso a los recursos tecnológicos y a Internet, entrenamiento pedagógico insuficiente, vulnerabilidad social y alteración del comportamiento de los estudiantes. Se defiende la labor del psicólogo escolar como agente mediador de las relaciones y procesos de enseñanza, aprendizaje y desarrollo humano, tensionados en la modalidad a distancia.

Optimized operation of a controlled stirred tank reactor system for the production of mesenchymal stromal cells and their extracellular vesicles.

The therapeutic effects of human mesenchymal stromal cells (MSC) have been attributed mostly to their paracrine activity, exerted through small-secreted extracellular vesicles (EVs) rather than their engraftment into injured tissues. Currently, the production of MSC-derived EVs (MSC-EVs) is performed in laborious static culture systems with limited manufacturing capacity using serum-containing media. In this work, a serum-/xenogeneic-free microcarrier-based culture system was successfully established for bone marrow-derived MSC cultivation and MSC-EV production using a 2  l-scale controlled stirred tank reactor (STR) operated under fed-batch (FB) or fed-batch combined with continuous perfusion (FB/CP). Overall, maximal cell numbers of (3.0 ± 0.12) × 108 and (5.3 ± 0.32) × 108 were attained at Days 8 and 12 for FB and FB/CP cultures, respectively, and MSC(M) expanded under both conditions retained their immunophenotype. MSC-EVs were identified in the conditioned medium collected from all STR cultures by transmission electron microscopy, and EV protein markers were successfully identified by Western blot analysis. Overall, no significant differences were observed between EVs isolated from MSC expanded in STR operated under the two feeding approaches. EV mean sizes of 163 ± 5.27 nm and 162 ± 4.44 nm (p > 0.05) and concentrations of (2.4 ± 0.35) × 1011 EVs/mL and (3.0 ± 0.48) × 1011 EVs/mL (p > 0.05) were estimated by nanoparticle tracking analysis for FB and FB/CP cultures, respectively. The STR-based platform optimized herein represents a major contribution toward the development of human MSC- and MSC-EV-based products as promising therapeutic agents for Regenerative Medicine settings.

Neurodifferentiation and Neuroprotection Potential of Mesenchymal Stromal Cell-Derived Secretome Produced in Different Dynamic Systems.

Parkinson's disease (PD) is the second most common neurodegenerative disorder and is characterized by the degeneration of the dopamine (DA) neurons in the substantia nigra pars compacta, leading to a loss of DA in the basal ganglia. The presence of aggregates of alpha-synuclein (α-synuclein) is seen as the main contributor to the pathogenesis and progression of PD. Evidence suggests that the secretome of mesenchymal stromal cells (MSC) could be a potential cell-free therapy for PD. However, to accelerate the integration of this therapy in the clinical setting, there is still the need to develop a protocol for the large-scale production of secretome under good manufacturing practices (GMP) guidelines. Bioreactors have the capacity to produce large quantities of secretomes in a scalable manner, surpassing the limitations of planar static culture systems. However, few studies focused on the influence of the culture system used to expand MSC, on the secretome composition. In this work, we studied the capacity of the secretome produced by bone marrow-derived mesenchymal stromal cells (BMSC) expanded in a spinner flask (SP) and in a Vertical-Wheel™ bioreactor (VWBR) system, to induce neurodifferentiation of human neural progenitor cells (hNPCs) and to prevent dopaminergic neuron degeneration caused by the overexpression of α-synuclein in one Caenorhabditis elegans model of PD. Results showed that secretomes from both systems were able to induce neurodifferentiation, though the secretome produced in the SP system had a greater effect. Additionally, in the conditions of our study, only the secretome produced in SP had a neuroprotective potential. Lastly, the secretomes had different profiles regarding the presence and/or specific intensity of different molecules, namely, interleukin (IL)-6, IL-4, matrix metalloproteinase-2 (MMP2), and 3 (MMP3), tumor necrosis factor-beta (TNF-ß), osteopontin, nerve growth factor beta (NGFß), granulocyte colony-stimulating factor (GCSF), heparin-binding (HB) epithelial growth factor (EGF)-like growth factor (HB-EGF), and IL-13. Overall, our results suggest that the culture conditions might have influenced the secretory profiles of cultured cells and, consequently, the observed effects. Additional studies should further explore the effects that different culture systems have on the secretome potential of PD.