RESUMEN
Objective: Rheumatoid arthritis causes inflammation, pain, and joint degradation, necessitating treatment with anti-inflammatory drugs and corticosteroids, posing various challenges. We aimed to evaluate the effects of photobiomodulation (PBM) at two different doses associated to platelet-rich plasma (PRP) in an in vivo model of induced acute arthritis in Wistar rats' knee. Methods: Eighty-four Wistar rats were assigned into seven groups, including animals treated with PBM and/or PRP. On day 0, arthritis was induced in sham and treated groups through the intra-articular injection of zymosan (200 µg). Twenty-four hours after induction, the PBM groups were treated with an AsGaAl laser, whereas the PRP-treated groups received intra-articular injections with a concentration of 8 × 105 platelets obtained from another four animals. After 3 days, the animals were euthanized, and the interleukin (IL)-6 and complement C3 gene and protein expression levels were analyzed. Statistical analysis was performed using the mean ± SD with analysis of variance and Tukey's posttest, with a significance level set at 5% (p < 0.05). Results: Synovial inflammation decreased in PBM-treated groups; however, PRP alone showed no significant difference. Gene expression analysis revealed a significant difference in IL-6 and C3 levels in the PBM and PBM+PRP-treated groups. Meanwhile, the PRP alone group exhibited significance for IL-6. Moreover, the PBM and PBM+PRP-treated groups showed a significant difference in C3 protein expression levels, whereas the PRP alone group showed no difference. Conclusion: The increase in cellular activity in the synovial membrane and the decrease protein expression levels are owing to the reduction in proinflammatory mediators following PBM therapy.
RESUMEN
Pyroptosis is a lytic cell death mode that helps limit the spread of infections and is also linked to pathology in sterile inflammatory diseases and autoimmune diseases1-4. During pyroptosis, inflammasome activation and the engagement of caspase-1 lead to cell death, along with the maturation and secretion of the inflammatory cytokine interleukin-1ß (IL-1ß). The dominant effect of IL-1ß in promoting tissue inflammation has clouded the potential influence of other factors released from pyroptotic cells. Here, using a system in which macrophages are induced to undergo pyroptosis without IL-1ß or IL-1α release (denoted Pyro-1), we identify unexpected beneficial effects of the Pyro-1 secretome. First, we noted that the Pyro-1 supernatants upregulated gene signatures linked to migration, cellular proliferation and wound healing. Consistent with this gene signature, Pyro-1 supernatants boosted migration of primary fibroblasts and macrophages, and promoted faster wound closure in vitro and improved tissue repair in vivo. In mechanistic studies, lipidomics and metabolomics of the Pyro-1 supernatants identified the presence of both oxylipins and metabolites, linking them to pro-wound-healing effects. Focusing specifically on the oxylipin prostaglandin E2 (PGE2), we find that its synthesis is induced de novo during pyroptosis, downstream of caspase-1 activation and cyclooxygenase-2 activity; further, PGE2 synthesis occurs late in pyroptosis, with its release dependent on gasdermin D pores opened during pyroptosis. As for the pyroptotic metabolites, they link to immune cell infiltration into the wounds, and polarization to CD301+ macrophages. Collectively, these data advance the concept that the pyroptotic secretome possesses oxylipins and metabolites with tissue repair properties that may be harnessed therapeutically.
Asunto(s)
Macrófagos , Oxilipinas , Piroptosis , Secretoma , Cicatrización de Heridas , Animales , Femenino , Humanos , Ratones , Caspasa 1/metabolismo , Movimiento Celular , Proliferación Celular , Ciclooxigenasa 2/metabolismo , Dinoprostona/biosíntesis , Dinoprostona/metabolismo , Fibroblastos/metabolismo , Fibroblastos/citología , Gasderminas/metabolismo , Inflamasomas/metabolismo , Interleucina-1beta , Lipidómica , Macrófagos/metabolismo , Macrófagos/citología , Ratones Endogámicos C57BL , Oxilipinas/metabolismo , Proteínas de Unión a Fosfato/metabolismo , Secretoma/metabolismo , Cicatrización de Heridas/fisiologíaRESUMEN
Acute systemic inflammation critically alters the function of the immune system, often promoting myelopoiesis at the expense of lymphopoiesis. In the thymus, systemic inflammation results in acute thymic atrophy and, consequently, impaired T-lymphopoiesis. The mechanism by which systemic inflammation impacts the thymus beyond suppressing T-cell development is still unclear. Here, we describe how the synergism between TL1A and IL-18 suppresses T-lymphopoiesis to promote thymic myelopoiesis. The protein levels of these two cytokines were elevated in the thymus during viral-induced thymus atrophy infection with murine cytomegalovirus (MCMV) or pneumonia virus of mice (PVM). In vivo administration of TL1A and IL-18 induced acute thymic atrophy, while thymic neutrophils expanded. Fate mapping with Ms4a3-Cre mice demonstrated that thymic neutrophils emerge from thymic granulocyte-monocyte progenitors (GMPs), while Rag1-Cre fate mapping revealed a common developmental path with lymphocytes. These effects could be modeled ex vivo using neonatal thymic organ cultures (NTOCs), where TL1A and IL-18 synergistically enhanced neutrophil production and egress. NOTCH blockade by the LY411575 inhibitor increased the number of neutrophils in the culture, indicating that NOTCH restricted steady-state thymic granulopoiesis. To promote myelopoiesis, TL1A, and IL-18 synergistically increased GM-CSF levels in the NTOC, which was mainly produced by thymic ILC1s. In support, TL1A- and IL-18-induced granulopoiesis was completely prevented in NTOCs derived from Csf2rb-/- mice and by GM-CSFR antibody blockade, revealing that GM-CSF is the essential factor driving thymic granulopoiesis. Taken together, our findings reveal that TL1A and IL-18 synergism induce acute thymus atrophy while promoting extramedullary thymic granulopoiesis in a NOTCH and GM-CSF-controlled manner.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos , Interleucina-18 , Timo , Miembro 15 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral , Animales , Interleucina-18/metabolismo , Timo/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Ratones , Miembro 15 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/metabolismo , Ratones Endogámicos C57BL , Granulocitos/metabolismo , Mielopoyesis , Neutrófilos/inmunología , Neutrófilos/metabolismo , Receptores Notch/metabolismo , Linfopoyesis , AtrofiaRESUMEN
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.
Asunto(s)
Interleucina-18 , Linfocitos , Humanos , Recién Nacido , Ratones , Animales , Linfocitos/metabolismo , Inmunidad Innata , Citocinas/metabolismo , Interleucina-12 , AtrofiaRESUMEN
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
Asunto(s)
Humanos , Femenino , Adulto , Diarrea/etiología , Enterocolitis/complicaciones , Enterocolitis/diagnóstico , Eosinofilia/complicaciones , Eosinofilia/diagnósticoRESUMEN
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.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Fosforilación , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Procesamiento Proteico-Postraduccional , Plantas Modificadas Genéticamente/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismoRESUMEN
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.
Asunto(s)
Enfermedades Cardiovasculares , Humanos , Masculino , Anciano , Femenino , Enfermedades Cardiovasculares/epidemiología , Enfermedades Cardiovasculares/prevención & control , Brasil , Estudios Transversales , Dieta , Dieta SaludableRESUMEN
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.
Asunto(s)
Enfermedades Cardiovasculares , Dieta , Brasil , AterosclerosisRESUMEN
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.
Asunto(s)
Doxorrubicina , Nanopartículas , Animales , Ratones , Línea Celular Tumoral , Doxorrubicina/química , Apoptosis , Nanopartículas/química , ADN/farmacología , Sistemas de Liberación de Medicamentos/métodos , Portadores de Fármacos/farmacología , Portadores de Fármacos/químicaRESUMEN
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.
Asunto(s)
Educación de Pregrado en Medicina , Obstetricia , Entrenamiento Simulado , Estudiantes de Medicina , Embarazo , Humanos , Masculino , Femenino , Educación de Pregrado en Medicina/métodos , Obstetricia/educación , Entrenamiento Simulado/métodos , Parto Obstétrico , Competencia ClínicaRESUMEN
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.
Asunto(s)
Plantas , Agregado de Proteínas , Plantas/metabolismo , Chaperonas Moleculares/metabolismo , Proteínas de Choque Térmico/metabolismo , Biosíntesis de ProteínasRESUMEN
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.
Asunto(s)
Hidroxibutiratos , Andamios del Tejido , Humanos , Andamios del Tejido/química , Ácido 3-Hidroxibutírico , Hidroxibutiratos/química , Ingeniería de Tejidos/métodos , Poliésteres/químicaRESUMEN
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.
RESUMEN
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.
Asunto(s)
Extractos Vegetales/farmacología , Annona/química , Antioxidantes/farmacología , Extractos Vegetales/toxicidad , Pruebas de Toxicidad/métodos , Hojas de la Planta/químicaRESUMEN
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)
Asunto(s)
Humanos , Masculino , Persona de Mediana Edad , Psoriasis/tratamiento farmacológico , Tuberculosis Latente/prevención & control , Isoniazida/uso terapéutico , Antituberculosos/uso terapéutico , Índice de Severidad de la Enfermedad , Inducción de Remisión , Estudios RetrospectivosRESUMEN
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.
Asunto(s)
Trampas Extracelulares , Inflamasomas , Neutrófilos , Proteínas de Unión a Fosfato , Proteínas Citotóxicas Formadoras de Poros , Piroptosis , Animales , Inflamasomas/metabolismo , Interleucina-1beta/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Ratones , Ratones Endogámicos C57BL , Mitógenos/metabolismo , NADP/metabolismo , NADPH Oxidasas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR , Neutrófilos/metabolismo , Proteínas de Unión a Fosfato/metabolismo , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Pirina/metabolismoRESUMEN
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.
Asunto(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , Genómica , Humanos , Técnicas de Diagnóstico Molecular/métodos , Mutación/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , SARS-CoV-2/genética , Sensibilidad y EspecificidadRESUMEN
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.
Asunto(s)
Cloroquina , Hidroxicloroquina , Cloroquina/química , Cloroquina/farmacología , ADN/química , Humanos , Hidroxicloroquina/química , Hidroxicloroquina/farmacología , Pinzas ÓpticasRESUMEN
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.
Asunto(s)
Evolución Biológica , Hepatocitos/metabolismo , Macrófagos/metabolismo , Proteogenómica , Animales , Núcleo Celular/metabolismo , Hígado Graso/genética , Hígado Graso/patología , Homeostasis , Humanos , Macrófagos del Hígado/metabolismo , Antígenos Comunes de Leucocito/metabolismo , Lípidos/química , Hígado/metabolismo , Linfocitos/metabolismo , Ratones Endogámicos C57BL , Modelos Biológicos , Células Mieloides/metabolismo , Obesidad/patología , Proteoma/metabolismo , Transducción de Señal , Transcriptoma/genéticaRESUMEN
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.