Type 1 immunity enables neonatal thymic ILC1 production.

Acute thymic atrophy occurs following type 1 inflammatory conditions such as viral infection and sepsis, resulting in cell death and disruption of T cell development. However, the impact type 1 immunity has on thymic-resident innate lymphoid cells (ILCs) remains unclear. Single-cell RNA sequencing revealed neonatal thymic-resident type 1 ILCs (ILC1s) as a unique and immature subset compared to ILC1s in other primary lymphoid organs. Culturing murine neonatal thymic lobes with the type 1 cytokines interleukin-12 (IL-12) and IL-18 resulted in a rapid expansion and thymic egress of KLRG1+CXCR6+ cytotoxic ILC1s. Live imaging showed the subcapsular thymic localization and exit of ILC1s following IL-12 + IL-18 stimulation. Similarly, murine cytomegalovirus infection in neonates resulted in thymic atrophy and subcapsular localization of thymic-resident ILC1s. Neonatal thymic grafting revealed that type 1 inflammation enhances the homing of cytokine-producing thymus-derived ILC1s to the liver and peritoneal cavity. Together, we show that type 1 immunity promotes the expansion and peripheral homing of thymic-derived ILC1s.

Gastroenterocolite eosinofílica - uma causa rara de diarreia / Eosinophilic gastroenterocolitis - a rare cause of diarrhea

Introdução: Colite eosinofílica é uma doença crônica rara. Pode ser idiopática ou estar associada à alergias alimentares e à doenças atópicas. Assim, a incidência é, muitas vezes, indefinida devido ao seu subdiagnóstico. Objetivo: Relatar o caso de uma paciente com colite eosinofílica e dermatite atópica. Método: Trata-se de um estudo tipo Relato de Caso utilizando dados do prontuário clínico, de uma paciente jovem que evoluiu com quadro de dor abdominal e diarreia sanguinolenta e que, após investigação por meio de exames laboratoriais, diversos exames de imagem, o resultado anatomopatológico evidenciou Ileíte eosinofílica. Resultados: O tratamento consistiu em mudança alimentar e uso de budesonida por via oral, havendo regressão do quadro e melhora da dermatite atópica. Conclusão: É necessário excluir a Colite Eosinofílica secundária, pois pode ser uma manifestação à diversas patologias, já que tem apresentação clínica inespecífica

Introducion: Eosinophilic colitis is a rare chronic disease. It may be idiopathic or associated with food allergies and atopic diseases. Thus, the incidence is often undefined due to underdiagnosis. Objective: To report the case of a patient with eosinophilic colitis and atopic dermatitis. Method: This is a case report study using data from the clinical record, of a young patient who developed abdominal pain and bloody diarrhea and who, after investigation through laboratory tests, several imaging tests, the result pathology showed eosinophilic ileitis. Results: Treatment consisted of dietary changes and oral budesonide, resulting in regression of the condition and improvement in atopic dermatitis. Conclusion: It is necessary to exclude secondary Eosinophilic Colitis, as it can be a manifestation of several pathologies, as it has a nonspecific clinical presentation

Introducción: La colitis eosinofílica es una enfermedad crónica rara. Puede ser idiopática o estar asociada a alergias alimentarias y enfermedades atópicas. Por tanto, la incidencia suele estar indefinida debido a un infradiagnóstico. Objetivo: Reportar el caso de un paciente con colitis eosinofílica y dermatitis atópica. Método: Se trata de un estudio de reporte de caso, utilizando datos de la historia clínica, de un paciente joven que desarrolló dolor abdominal y diarrea sanguinolenta y quien, luego de la investigación mediante pruebas de laboratorio, varias pruebas de imagen, el resultado de la patología arrojó ileítis eosinofílica. Resultados: El tratamiento consistió en cambios dietéticos y budesonida oral, lo que resultó en regresión del cuadro y mejoría de la dermatitis atópica. Conclusión: Es necesario excluir la Colitis Eosinofílica secundaria, ya que puede ser manifestación de varias patologías, al tener una presentación clínica inespecífica

Factors associated with diet quality among Brazilian individuals with cardiovascular diseases.

BACKGROUND: An individual's dietary pattern contributes in different ways to the prevention and control of recurrent cardiovascular events. However, the quality of the diet is influenced by several factors. The present study aimed to evaluate the quality of the diet of individuals with cardiovascular diseases and determine whether there is an association between sociodemographic and lifestyle factors. METHODS: This is a cross-sectional study carried out with individuals with atherosclerosis (coronary artery disease, cerebrovascular disease or peripheral arterial disease) recruited from 35 reference centres for the treatment of cardiovascular disease in Brazil. Diet quality was assessed according to the Modified Alternative Healthy Eating Index (mAHEI) and stratified into tertiles. For comparing two groups, the Mann-Whitney or Pearson's chi-squared tests were used. However, for comparing three or more groups, analysis of variance or Kruskal-Wallis was used. For the confounding analysis, a multinomial regression model was used. p < 0.05 was considered statistically significant. RESULTS: In total, 2360 individuals were evaluated: 58.5% male and 64.2% elderly. The median (interquartile range [IQR]) of the mAHEI was 24.0 (20.0-30.0), ranging from 0.4 to 56.0 points. When comparing the odds ratios (ORs) for the low (first tertile) and medium (second tertile) diet quality groups with the high-quality group (third tertile), it was observed that there was an association between diet quality with a family income of 1.885 (95% confidence intervals [CI] = 1.302-2.729) and 1.566 (95% CI = 1.097-2.235), as well as physical activity of 1.391 (95% CI = 1.107-1.749) and 1.346 (95% CI = 1.086-1.667), respectively. In addition, associations were observed between diet quality and region of residence. CONCLUSIONS: A low-quality diet was associated with family income, sedentarism and geographical area. These data are extremely relevant to assist in coping with cardiovascular disease because they enable an assessment of the distribution of these factors in different regions of the country.

Phase separation-based visualization of protein-protein interactions and kinase activities in plants.

Protein activities depend heavily on protein complex formation and dynamic posttranslational modifications, such as phosphorylation. The dynamic nature of protein complex formation and posttranslational modifications is notoriously difficult to monitor in planta at cellular resolution, often requiring extensive optimization. Here, we generated and exploited the SYnthetic Multivalency in PLants (SYMPL)-vector set to assay protein-protein interactions (PPIs) (separation of phases-based protein interaction reporter) and kinase activities (separation of phases-based activity reporter of kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary PPIs among cytoplasmic and nuclear proteins in plant cells via a robust image-based readout. Moreover, we applied the SYMPL toolbox to develop an in vivo reporter for SNF1-related kinase 1 activity, allowing us to visualize tissue-specific, dynamic SnRK1 activity in stable transgenic Arabidopsis (Arabidopsis thaliana) plants. The SYMPL cloning toolbox provides a means to explore PPIs, phosphorylation, and other posttranslational modifications with unprecedented ease and sensitivity.

Factors associated with diet quality among Brazilian individuals with cardiovascular diseases

BACKGROUND: An individual's dietary pattern contributes in different ways to the prevention and control of recurrent cardiovascular events. However, the quality of the diet is influenced by several factors. The present study aimed to evaluate the quality of the diet of individuals with cardiovascular diseases and determine whether there is an association between sociodemographic and lifestyle factors. METHODS: This is a cross-sectional study carried out with individuals with atherosclerosis (coronary artery disease, cerebrovascular disease or peripheral arterial disease) recruited from 35 reference centres for the treatment of cardiovascular disease in Brazil. Diet quality was assessed according to the Modified Alternative Healthy Eating Index (mAHEI) and stratified into tertiles. For comparing two groups, the Mann-Whitney or Pearson's chi-squared tests were used. However, for comparing three or more groups, analysis of variance or Kruskal-Wallis was used. For the confounding analysis, a multinomial regression model was used. p < 0.05 was considered statistically significant. RESULTS: In total, 2360 individuals were evaluated: 58.5% male and 64.2% elderly. The median (interquartile range [IQR]) of the mAHEI was 24.0 (20.0-30.0), ranging from 0.4 to 56.0 points. When comparing the odds ratios (ORs) for the low (first tertile) and medium (second tertile) diet quality groups with the high-quality group (third tertile), it was observed that there was an association between diet quality with a family income of 1.885 (95% confidence intervals [CI] = 1.302-2.729) and 1.566 (95% CI = 1.097-2.235), as well as physical activity of 1.391 (95% CI = 1.107-1.749) and 1.346 (95% CI = 1.086-1.667), respectively. In addition, associations were observed between diet quality and region of residence. CONCLUSIONS: A low-quality diet was associated with family income, sedentarism and geographical area. These data are extremely relevant to assist in coping with cardiovascular disease because they enable an assessment of the distribution of these factors in different regions of the country.

Obstetric simulation for undergraduate medical education: how to improve students' self-confidence and expectation according to gender.

OBJECTIVE: The aim of this study was to evaluate the effects of obstetric simulation training on undergraduate medical students to improve their self-confidence. METHODS: Fifth-year undergraduate medical students were invited to a 2-week course of simulation in obstetrics during their clerkship. The sessions included were as follows: (1) care for the second and third periods of childbirth, (2) partograph analysis and pelvimetry, (3) premature rupture of membranes at term, and (4) diagnosis and management of third-trimester bleeding. Before the first session and at the end of the training period, a questionnaire about self-confidence in obstetric procedures and skills was applied. RESULTS: A total of 115 medical students were included, of whom 60 (52.2%) were male and 55 (47.8%) were female. Comparing initial and final scores, the median results of the subscales "comprehension and preparation" (18 vs. 22, p<0.001), "knowledge of procedures" (14 vs. 20, p<0.001), and "expectation" (22 vs. 23, p<0.01) were significantly higher at the end of the training period in all items of the questionnaire than in the beginning. Differences were found based on the students' gender, i.e., female students had a significantly higher sum of scores than the male students in the initial subscale for "expectation" (median, 24 vs. 22, p<0.001) and "interest" (median, 23 vs. 21, p=0.032), and a higher sum of scores in the subscale for "expectation" (median, 23 vs. 21, p=0.010) in the final questionnaire. CONCLUSION: Obstetric simulation enhances the improvement of students' self-confidence in understanding both the physiology of childbirth and the obstetric care procedures. Further studies are needed to understand the influence of gender on obstetric care.

Virus-Like Particle-Encapsulated Doxorubicin Enters and Kills Murine Tumor Cells Differently from Free Doxorubicin.

The use of nanoparticles as chemotherapeutic carriers has been suggested as a way to overcome a range of side effects associated with classical cancer treatment such as poor selectivity and tumor resurgence. Obtaining precise control of the size and shape of therapeutic nanoparticles is crucial to optimize the targeting of tumor sites. In this work, it is shown that a previously developed system of polypeptide encapsulating individual DNA molecules, that forms rod-shaped nanoparticles of precisely controlled aspect ratio, can be loaded with the DNA-intercalating chemotherapeutic drug doxorubicin (DOX). It is characterized the size and shape of the DOX loaded-Virus-Like DNA Particles (DOX-VLDP) and shown that in this system the DOX payload does not leak out. Through in vitro cell studies, it is shown that DOX-VLDP is internalized by melanoma tumor cells (B16F10 cells) in a delayed and endocytosis-dependent way culminating in increased cytotoxicity and selectivity to tumor cells in comparison with free DOX. In addition, it is found that DOX-VLDP trigger apoptosis and autophagy pathways in treated cells. Taken together, the data on the DOX-VLDP nanoparticles shows that they kill cancer cells differently from free DOX.

Cell Behavior Changes and Enzymatic Biodegradation of Hybrid Electrospun Poly(3-hydroxybutyrate)-Based Scaffolds with an Enhanced Piezoresponse after the Addition of Reduced Graphene Oxide.

This is the first comprehensive study of the impact of biodegradation on the structure, surface potential, mechanical and piezoelectric properties of poly(3-hydroxybutyrate) (PHB) scaffolds supplemented with reduced graphene oxide (rGO) as well as cell behavior under static and dynamic mechanical conditions. There is no effect of the rGO addition up to 1.0 wt% on the rate of enzymatic biodegradation of PHB scaffolds for 30 d. The biodegradation of scaffolds leads to the depolymerization of the amorphous phase, resulting in an increase in the degree of crystallinity. Because of more regular dipole order in the crystalline phase, surface potential of all fibers increases after the biodegradation, with a maximum (361 ± 5 mV) after the addition of 1 wt% rGO into PHB as compared to pristine PHB fibers. By contrast, PHB-0.7rGO fibers manifest the strongest effective vertical (0.59 ± 0.03 pm V-1 ) and lateral (1.06 ± 0.02 pm V-1 ) piezoresponse owing to a greater presence of electroactive ß-phase. In vitro assays involving primary human fibroblasts reveal equal biocompatibility and faster cell proliferation on PHB-0.7rGO scaffolds compared to pure PHB and nonpiezoelectric polycaprolactone scaffolds. Thus, the developed biodegradable PHB-rGO scaffolds with enhanced piezoresponse are promising for tissue-engineering applications.

Detection and Quantification of Protein Aggregates in Plants.

Protein quality control is an important aspect of stress recovery. It maintains protein homeostasis through a machinery of regulatory proteins such as chaperones and proteases. When the system recognizes accumulation of misfolded or aggregated proteins, the cell recruits a set of regulatory proteins to initiate protein quality control. To understand the dynamics of stress-mediated aggregate protein formation and recovery in plants, robust methods aimed at detecting and measuring such protein aggregates are needed. This will help us to deepen our understanding of protein quality control mechanisms in plants.

Obstetric simulation for undergraduate medical education: how to improve students' self-confidence and expectation according to gender

SUMMARY OBJECTIVE: The aim of this study was to evaluate the effects of obstetric simulation training on undergraduate medical students to improve their self-confidence. METHODS: Fifth-year undergraduate medical students were invited to a 2-week course of simulation in obstetrics during their clerkship. The sessions included were as follows: (1) care for the second and third periods of childbirth, (2) partograph analysis and pelvimetry, (3) premature rupture of membranes at term, and (4) diagnosis and management of third-trimester bleeding. Before the first session and at the end of the training period, a questionnaire about self-confidence in obstetric procedures and skills was applied. RESULTS: A total of 115 medical students were included, of whom 60 (52.2%) were male and 55 (47.8%) were female. Comparing initial and final scores, the median results of the subscales "comprehension and preparation" (18 vs. 22, p<0.001), "knowledge of procedures" (14 vs. 20, p<0.001), and "expectation" (22 vs. 23, p<0.01) were significantly higher at the end of the training period in all items of the questionnaire than in the beginning. Differences were found based on the students' gender, i.e., female students had a significantly higher sum of scores than the male students in the initial subscale for "expectation" (median, 24 vs. 22, p<0.001) and "interest" (median, 23 vs. 21, p=0.032), and a higher sum of scores in the subscale for "expectation" (median, 23 vs. 21, p=0.010) in the final questionnaire. CONCLUSION: Obstetric simulation enhances the improvement of students' self-confidence in understanding both the physiology of childbirth and the obstetric care procedures. Further studies are needed to understand the influence of gender on obstetric care.

Estudo comparativo dos perfis fitoquímicos e citotóxicos do extrato alcoólico de folhas de graviola (annona muricata) e fruta do conde (annona squamosa) / Comparative study of the phytochemical and cytotoxic profiles of the alcoholic extract of soursop (annona muricata) and custard apple (annona squamosa) leaves / Estudio comparativo de los perfiles fitoquímicos y citotóxicos del extracto alcohólico de hojas de guanábana (annona muricata) y del conde (annona squamosa)

Introdução: As plantas apresentam potencial medicinal devido aos metabólitos secundários e possuem compostos com alto potencial anti-inflamatório, com possibilidades terapêuticas. Das espécies vegetais de interesse destacamos e comparamos a graviola (Annona muricata) e a fruta do conde (Annona squamosa). Objetivo: Comparar o potencial do extrato alcoólico bruto de folhas da graviola e fruta do conde em ensaios fitoquímicos e citotóxicos. Método: Os extratos de folhas de graviola e fruta do conde foram obtidos por percolação, com o uso de 20g de folhas secas e trituradas com 100 ml de álcool de cereais. Na padronização dos extratos foram utilizadas análises de idenficação de componentes químicos, os taninos, flavonoides, saponinas e alcaloides, por meio de difrentes reações químicas e os antioxidantes foram analisados pela atividade do DPPH. A citotoxicidade das diferentes concentrações dos extratos também foram analisadas por hemólise e pelo teste da membrana corioalantide (CAM). Resultados: As análises fitoquímicas indicaram a presença de alcaloides, taninos, flavonoides e saponinas para ambos os extratos com alta capacidade antioxidante. Os testes de hemólise e CAM mostraram ausência de citotoxicidade na concentração de 2% e 4% para ambos os extratos e elevada citotoxicidade em 10% para o extrato de graviola. Conclusão: O extrato alcoólico das folhas da graviola e fruta do conde mostraram importante perfil antioxidante e com compostos anti-inflamatórios com pouca diferença entre os extratos o que estimula estudos futuros e continuidade para exploração anti-inflamatória.

Introduction: Plants have medicinal potential due to secondary metabolites and have compounds with high antiinflammatory potential, with therapeutic possibilities. Of the plant species of interest we highlight and compare the graviola (Annona Muricata) and the fruit of the count (Annona squamosa). Objective: To compare the potential of crude alcoholic extract of soursop and conde fruit leaves in phytochemical and cytotoxic assays. Method: The leaves extracts of soursop and fruit of the count were obtained by percolation, with the use of 20g of dry leaves and crushed with 100 ml of grain alcohol. In the standardization of the extracts were used idenfication analyzes of chemical components, tannins, flavonoids, saponins and alkaloids, through different chemical reactions and antioxidants were analyzed by the activity of DPPH. The cytotoxicity of the different concentrations of the extracts were also analyzed by hemolysis and by the corioalantide membrane test (CAM). Results: Phytochemical analysis indicated the presence of alkaloids, tannins, flavonoids and saponins for both extracts with high antioxidant capacity. The hemolysis and CAM tests showed absence of cytotoxicity at the concentration of 2% and 4% for both extracts and high cytotoxicity in 10% for graviola extract. Conclusion: The alcoholic extract of the leaves of soursop and fruit of the count showed important antioxidant profile and with anti-inflammatory compounds with little difference between the extracts which stimulates future studies and continuity for anti-inflammatory exploration.

Introducción: Las plantas tienen potencial medicinal debido a los metabolitos secundarios y poseen compuestos con alto potencial antiinflamatorio, además de ser utilizadas en tratamientos terapéuticos. De las especies vegetales de interés, destacamos y comparamos la guanábana (Annona muricata) y la fruta del conde (Annona squamosa). Objetivo: Comparar el potencial del extracto alcohólico crudo de hojas de guanábana y chirimoya en ensayos fitoquímicos y citotóxicos. Método: El extracto de hojas de guanábana y fruta del conde se obtuvo por percolación, utilizando 20g de hojas secas y trituradas con 100 ml de alcohol de grano. En la estandarización de los extractos se utilizaron análisis para identificar componentes químicos, taninos, flavonoides, saponinas y alcaloides, a través de diferentes reacciones químicas y los antioxidantes fueron analizados por la actividad de DPPH. También se analizó la citotoxicidad de diferentes concentraciones de extractos por hemólisis y por la prueba de membrana de corioalantida (CAM). Resultados: Los análisis fitoquímicos indicaron la presencia de alcaloides, taninos y flavonoides y saponinas para ambos extractos con alta capacidad antioxidante. Las pruebas de hemólisis y CAM mostraron ausencia de citotoxicidad a una concentración del 4% y alta citotoxicidad a partir del 10% también para ambos extractos. Conclusión: El extracto alcohólico de hojas de guanábana y chirimoya mostró importantes perfiles antioxidantes y con compuestos antiinflamatorios, lo que estimula futuros estudios y continuidad para la exploración antiinflamatoria.

Psoríase vulgar em remissão após profilaxia com isoniazida para tuberculose latente: relato de caso / Psoriasis vulgar in remission after prophylaxis with isoniazid for latent tuberculosis: case report / Psoriasis vulgar en remisión tras profilaxis con isoniazida para tuberculosis latente: reporte de caso

Introdução: A psoríase é uma patologia sistêmica, multifatorial e com lesões cutâneas bem características. Objetivo: Investigar a possibilidade de existir uma correlação entre a infecção latente pelo Mycobacterium tuberculosis e a patogênese da Psoríase, a fim de contribuir com a terapêutica de casos de Psoríase grave ou refratários. Material e Método: Relato de caso de psoríase vulgar não responsiva aos tratamentos de primeira linha, em que a Isoniazida para ILTB induziu à remissão sustentada do quadro clínico. Baseada no caso, foi realizada uma revisão sobre o uso da Isoniazida no tratamento da ILTB nos pacientes com psoríase e que obtiveram regressão das lesões psoriáticas. O estudo utilizou dados da plataforma Pubmed. Resultados: Dos 50 artigos encontrados, 08 foram selecionados para leitura completa. Foram descritos apenas quatro casos como este na literatura internacional. Os achados desse estudo mostram que a possibilidade de existir uma correlação entre a infecção latente pelo Mycobacterium tuberculosis e a patogênese da Psoríase, a fim de contribuir com a terapêutica de casos de Psoríase grave ou refratários, ainda é incerta. Conclusão: Pesquisas mais detalhadas são necessárias para elucidar o real mecanismo dos anti tuberculínicos (especialmente a Isoniazida) no tratamento da Psoríase, quanto a possibilidade de existir uma correlação entre a infecção pelo Mycobacterium tuberculosis e a patogênese da Psoríase. Se isto for comprovado, teremos um grande incremento no arsenal terapêutico destinado aos pacientes com Psoríase grave ou refratários (AU)

Introduction: Psoriasis is a systemic pathology, multifactorial and with well characteristic skin lesions. Objective: To investigate the possibility of a correlation between latent infection by Mycobacterium tuberculosis and the pathogenesis of Psoriasis, in order to contribute to the therapy of severe or refractory Psoriasis cases. Material and Method: Case report of vulgar psoriasis not responsive to first-line treatments, in which Isoniazid for LTBI induced sustained remission of the clinical picture. Based on the case, a review was performed on the use of Isoniazid in the treatment of LTBI in patients with psoriasis and who obtained regression of psoriatic lesions. The study used data from the Pubmed platform. Results: Of the 50 articles found, 08 were selected for full reading. Only four cases like this have been described in the international literature. The findings of this study show that the possibility of a correlation between latent infection by Mycobacterium tuberculosis and the pathogenesis of Psoriasis, in order to contribute to the therapy of severe or refractory Psoriasis cases, is still uncertain. Conclusion: More detailed research is needed to elucidate the real mechanism of anti tuberculins (especially Isoniazid) in the treatment of Psoriasis, the possibility of a correlation between infection by Mycobacterium tuberculosis and the pathogenesis of Psoriasis. If this is proven, we will have a large increase in the therapeutic arsenal for patients with severe or refractory psoriasis (AU)

Introducción: La psoriasis es una patología sistémica, multifactorial, con lesiones cutáneas muy características. Objetivo: Investigar la posibilidad de una correlación entre la infección latente por Mycobacterium tuberculosis y la patogenia de la Psoriasis, con el fin de contribuir al tratamiento de casos severos o refractarios de Psoriasis. Material y Método: Reporte de caso de psoriasis vulgar sin respuesta a tratamientos de primera línea, en el cual Isoniazida para ITBL indujo remisión sostenida del cuadro clínico. En base al caso se realizó una revisión sobre el uso de Isoniazida en el tratamiento de la ITBL en pacientes con psoriasis y que lograron la regresión de las lesiones psoriásicas. El estudio utilizó datos de la plataforma Pubmed. Resultados: De los 50 artículos encontrados, 08 fueron seleccionados para lectura completa. Solo cuatro casos como este han sido descritos en la literatura internacional. Los hallazgos de este estudio muestran que la posibilidad de una correlación entre la infección latente por Mycobacterium tuberculosis y la patogenia de la Psoriasis, para contribuir al tratamiento de casos severos o refractarios de Psoriasis, es aún incierta. Conclusión: Se necesita una investigación más detallada para dilucidar el mecanismo real de los fármacos antituberculínicos (especialmente la Isoniazida) en el tratamiento de la Psoriasis, en cuanto a la posibilidad de una correlación entre la infección por Mycobacterium tuberculosis y la patogenia de la Psoriasis. De demostrarse esto, tendremos un gran incremento en el arsenal terapéutico destinado a pacientes con psoriasis severa o refractaria (AU)

GSDMD drives canonical inflammasome-induced neutrophil pyroptosis and is dispensable for NETosis.

Neutrophils are the most prevalent immune cells in circulation, but the repertoire of canonical inflammasomes in neutrophils and their respective involvement in neutrophil IL-1ß secretion and neutrophil cell death remain unclear. Here, we show that neutrophil-targeted expression of the disease-associated gain-of-function Nlrp3A350V mutant suffices for systemic autoinflammatory disease and tissue pathology in vivo. We confirm the activity of the canonical NLRP3 and NLRC4 inflammasomes in neutrophils, and further show that the NLRP1b, Pyrin and AIM2 inflammasomes also promote maturation and secretion of interleukin (IL)-1ß in cultured bone marrow neutrophils. Notably, all tested canonical inflammasomes promote GSDMD cleavage in neutrophils, and canonical inflammasome-induced pyroptosis and secretion of mature IL-1ß are blunted in GSDMD-knockout neutrophils. In contrast, GSDMD is dispensable for PMA-induced NETosis. We also show that Salmonella Typhimurium-induced pyroptosis is markedly increased in Nox2/Gp91Phox -deficient neutrophils that lack NADPH oxidase activity and are defective in PMA-induced NETosis. In conclusion, we establish the canonical inflammasome repertoire in neutrophils and identify differential roles for GSDMD and the NADPH complex in canonical inflammasome-induced neutrophil pyroptosis and mitogen-induced NETosis, respectively.

Molecular detection of omicron SARS-CoV-2 variant is achieved by RT-LAMP despite genomic mutations.

BACKGROUND: Severe acute respiratory syndrome coronavirus (SARS-CoV-2) omicron variant was first detected in South Africa in November 2021. Since then, the number of cases due to this variant increases enormously every day in different parts of the world. Mutations within omicron genome may impair the molecular detection resulting in false negative results during Coronavirus disease 19 (COVID-19) diagnosis. OBJECTIVES: To verify if colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting N and E genes would work efficiently to detect omicron SARS-CoV-2 variant and its sub-lineages. METHODS: SARS-CoV-2 reverse transcription quantitative polymerase chain reaction (RT-qPCR) positive samples were sequenced by next generation DNA sequencing. The consensus sequences generated were submitted to Pangolin tool for SARS-CoV-2 lineage identification. RT-LAMP reactions were performed at 65ºC/30 min targeting N and E. FINDINGS: SARS-CoV-2 omicron can be detected by RT-LAMP targeting N and E genes despite the genomic mutation of this more transmissible lineage. Omicron SARS-CoV-2 sub-lineages were tested and efficiently detected by RT-LAMP. We demonstrated that this test is very sensitive in detecting omicron variant, with LoD as low as 0.4 copies/µL. MAIN CONCLUSIONS: Molecular detection of omicron SARS-CoV-2 variant and its sub-lineages can be achieved by RT-LAMP despite the genomic mutations as a very sensitive surveillance tool for COVID-19 molecular diagnosis.

New Insights into the Mechanism of Action of the Drug Chloroquine: Direct Interaction with DNA and Cytotoxicity.

Chloroquine (CLQ) and hydroxychloroquine (HCLQ) are compounds largely employed in the treatment of various human diseases for decades. Nevertheless, a number of intrinsic details concerning their mechanisms of action, especially at the molecular level, are still unknown or have presented controversial results in the literature. Using optical tweezers, here, we investigate at the single-molecule level the molecular mechanism of action of the drug CLQ in its intrinsic interaction with the double-stranded (ds)DNA molecule, one of its targets inside cells, determining the binding modes and the physicochemical (binding) parameters of the interaction. In particular, we show that the ionic strength of the surrounding medium strongly influences such interaction, changing even the main binding mode. In addition, the cytotoxicity of CLQ against three different cell lines was also investigated here, allowing one to evaluate and compare the effect of the drug on the cell viability. In particular, we show that CLQ is highly cytotoxic at a very low (a few micromolar) concentration range for all cell lines tested. These results were rigorously compared to the equivalent ones obtained for the closely related compound hydroxychloroquine (HCLQ), allowing a critical comparison between the action of these drugs at the molecular and cellular levels.

Antibodies directed towards neuraminidase restrict influenza virus replication in primary human bronchial epithelial cells.

Influenza neuraminidase (NA) is implicated in various aspects of the virus replication cycle and therefore is an attractive target for vaccination and antiviral strategies. Here we investigated the potential for NA-specific antibodies to interfere with A(H1N1)pdm09 replication in primary human airway epithelial (HAE) cells. Mouse polyclonal anti-NA sera and a monoclonal antibody could block initial viral entry into HAE cells as well as egress from the cell surface. NA-specific polyclonal serum also reduced virus replication across multiple rounds of infection. Restriction of virus entry correlated with the ability of the serum or monoclonal antibody to mediate neuraminidase inhibition (NI). Finally, human sera with NI activity against the N1 of A(H1N1)pdm09 could decrease H6N1 virus infection of HAE cells, highlighting the potential contribution of anti-NA antibodies in the control of influenza virus infection in humans.

Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.

The liver is the largest solid organ in the body, yet it remains incompletely characterized. Here we present a spatial proteogenomic atlas of the healthy and obese human and murine liver combining single-cell CITE-seq, single-nuclei sequencing, spatial transcriptomics, and spatial proteomics. By integrating these multi-omic datasets, we provide validated strategies to reliably discriminate and localize all hepatic cells, including a population of lipid-associated macrophages (LAMs) at the bile ducts. We then align this atlas across seven species, revealing the conserved program of bona fide Kupffer cells and LAMs. We also uncover the respective spatially resolved cellular niches of these macrophages and the microenvironmental circuits driving their unique transcriptomic identities. We demonstrate that LAMs are induced by local lipid exposure, leading to their induction in steatotic regions of the murine and human liver, while Kupffer cell development crucially depends on their cross-talk with hepatic stellate cells via the evolutionarily conserved ALK1-BMP9/10 axis.

Perceptions, feelings, and the routine of older adults during the isolation period caused by the COVID-19 pandemic: a qualitative study in four countries.

OBJECTIVE: To evaluate the knowledge, routine, and perception of older adults from four countries about dealing with COVID-19 in the social isolation period. METHODS: Qualitative study with semistructured interviews. Older persons (≥60 years old), who lived in the urban centers of four different countries (Brazil, United States, Italy, and Portugal), were asked about the changes experienced during the pandemic, new habits or restrictions during isolation, sources of social and emotional support, and their knowledge about COVID-19 pandemic. Data was transcribed, codified, and submitted to content analysis. RESULTS: Twenty-five older persons (10 from Brazil, 5 from Italy, 5 from Portugal, and 5 from the United States) were interviewed. Participants reported feeling restricted in their daily life activities and emotional instability. Described adaptations in habits, coping strategies, and greater understanding of the diseased based on information available in the media. There was homogeneity in the statements of the elderly, showing that the pandemic affected them in a similar way, even though they lived in different cultures and contexts. CONCLUSION: The social isolation caused by the COVID-19 pandemic changed the structure for the performance of many occupations, having an impact in the perception social participation and wellbeing of elders. This data can aid health professionals to outline strategies to deal with the impact of the social isolation in older persons.

Vivência de cuidadores informais na transição de papéis após o processo de desospitalização / Vivencia de cuidadores informales en la transición de roles después del proceso de desospitalización / Informal caregivers' experience in role transition after dehospitalization

RESUMO Objetivo: apreender as vivências de cuidadores informais de pessoas dependentes no processo de transição de papéis após a desospitalização. Método: estudo descritivo, exploratório, qualitativo, realizado com cuidadores informais de pessoas dependentes que participaram de um protocolo de instrumentalização para alta. A coleta de dados ocorreu com 10 participantes da região sul do país, entre julho e agosto de 2021, por meio de entrevistas conduzidas no domicílio. Para a organização dos dados, utilizou-se o software IRAMUTEQ®. A análise se ancorou na Teoria das Transições. Resultados: os resultados abordaram o processo de transição situacional de cuidadores informais, emergindo três classes finais: Classe 1 - Dificuldades no cotidiano de cuidados com a pessoa dependente; Classe 2 - Significados atribuídos à transição de papéis; e Classe 3 - (Des)Continuidade do cuidado após a alta hospitalar. Considerações finais: pôde-se apreender que as vivências foram permeadas por fatores inibidores, de modo que os significados atribuídos a essa experiência se associaram às dificuldades com a transição para o papel de cuidador. O enfermeiro desempenhou papel de facilitador no processo de transição situacional, ao acompanhá-los, orientá-los e capacitá-los para o desenvolvimento de novas habilidades.

RESUMEN Objetivo: comprender las vivencias de cuidadores informales de personas dependientes en el proceso de transición de roles después de la desospitalización. Método: estudio descriptivo, exploratorio, cualitativo, realizado con cuidadores informales de personas dependientes que participaron de un protocolo de instrumentalización para el alta. La recolección de datos ocurrió con 10 participantes de la región sur del país, entre julio y agosto de 2021, por medio de entrevistas realizadas en el domicilio. Para la organización de los datos se utilizó el software IRAMUTEQ®. El análisis se basó en la Teoría de las Transiciones. Resultados: los resultados trataron el proceso de transición situacional de cuidadores informales, surgiendo tres clases finales: Clase 1 - Dificultades en el cotidiano de cuidados a la persona dependiente; Clase 2 - Significados atribuidos a la transición de roles; y Clase 3 - (Des)Continuidad del cuidado después del alta hospitalaria. Consideraciones finales: se pudo comprender que las vivencias están marcadas por factores inhibidores, de modo que los significados atribuidos a esa experiencia se asociaron a las dificultades con la transición para el rol de cuidador. El enfermero desempeñó un papel de facilitador en el proceso de transición situacional, al acompañarlos, orientarlos y capacitarlos para el desarrollo de nuevas habilidades.

ABSTRACT Objective: to understand the experiences of informal caregivers of dependent people in the role transition process after dehospitalization. Method: a descriptive, exploratory, qualitative study carried out with informal caregivers of dependent people who participated in a protocol of instrumentation for discharge. Data collection took place with 10 participants from southern Brazil, between July and August 2021, through interviews conducted at home. For data organization, we used IRAMUTEQ®. Analysis was anchored in the Transition Theory. Results: the results addressed the process of situational transition of informal caregivers, emerging three final classes: Class 1 - Difficulties in the daily care of dependent people; Class 2 - Meanings given to role transitions; and Class 3 - (Dis)continuity of care after hospital discharge. Final considerations: it was possible to understand that the experiences were permeated by inhibiting factors so that the meanings given to this experience were associated with the difficulties with the transition to the role of caregiver. Nurses played the role of facilitator in the situational transition process, by accompanying them, guiding them and enabling them to develop new skills.

Molecular detection of omicron SARS-CoV-2 variant is achieved by RT-LAMP despite genomic mutations

BACKGROUND Severe acute respiratory syndrome coronavirus (SARS-CoV-2) omicron variant was first detected in South Africa in November 2021. Since then, the number of cases due to this variant increases enormously every day in different parts of the world. Mutations within omicron genome may impair the molecular detection resulting in false negative results during Coronavirus disease 19 (COVID-19) diagnosis. OBJECTIVES To verify if colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting N and E genes would work efficiently to detect omicron SARS-CoV-2 variant and its sub-lineages. METHODS SARS-CoV-2 reverse transcription quantitative polymerase chain reaction (RT-qPCR) positive samples were sequenced by next generation DNA sequencing. The consensus sequences generated were submitted to Pangolin tool for SARS-CoV-2 lineage identification. RT-LAMP reactions were performed at 65ºC/30 min targeting N and E. FINDINGS SARS-CoV-2 omicron can be detected by RT-LAMP targeting N and E genes despite the genomic mutation of this more transmissible lineage. Omicron SARS-CoV-2 sub-lineages were tested and efficiently detected by RT-LAMP. We demonstrated that this test is very sensitive in detecting omicron variant, with LoD as low as 0.4 copies/µL. MAIN CONCLUSIONS Molecular detection of omicron SARS-CoV-2 variant and its sub-lineages can be achieved by RT-LAMP despite the genomic mutations as a very sensitive surveillance tool for COVID-19 molecular diagnosis.