Exosomal Osteoclast-Derived miRNA in Rheumatoid Arthritis: From Their Pathogenesis in Bone Erosion to New Therapeutic Approaches.

Rheumatoid arthritis (RA) is an autoimmune disease that causes inflammation, pain, and ultimately, bone erosion of the joints. The causes of this disease are multifactorial, including genetic factors, such as the presence of the human leukocyte antigen (HLA)-DRB1*04 variant, alterations in the microbiota, or immune factors including increased cytotoxic T lymphocytes (CTLs), neutrophils, or elevated M1 macrophages which, taken together, produce high levels of pro-inflammatory cytokines. In this review, we focused on the function exerted by osteoclasts on osteoblasts and other osteoclasts by means of the release of exosomal microRNAs (miRNAs). Based on a thorough revision, we classified these molecules into three categories according to their function: osteoclast inhibitors (miR-23a, miR-29b, and miR-214), osteoblast inhibitors (miR-22-3p, miR-26a, miR-27a, miR-29a, miR-125b, and miR-146a), and osteoblast enhancers (miR-20a, miR-34a, miR-96, miR-106a, miR-142, miR-199a, miR-324, and miR-486b). Finally, we analyzed potential therapeutic targets of these exosomal miRNAs, such as the use of antagomiRs, blockmiRs, agomiRs and competitive endogenous RNAs (ceRNAs), which are already being tested in murine and ex vivo models of RA. These strategies might have an important role in reestablishing the regulation of osteoclast and osteoblast differentiation making progress in the development of personalized medicine.

Analysis of Novel Immunological Biomarkers Related to Rheumatoid Arthritis Disease Severity.

Rheumatoid factor (RF) and anti-citrullinated protein antibodies (ACPAs) are the most frequently used rheumatoid arthritis (RA) diagnostic markers, but they are unable to anticipate the patient's evolution or response to treatment. The aim of this study was to identify possible severity biomarkers to predict an upcoming flare-up or remission period. To address this objective, sera and anticoagulated blood samples were collected from healthy controls (HCs; n = 39) and from early RA (n = 10), flare-up (n = 5), and remission (n = 16) patients. We analyzed leukocyte phenotype markers, regulatory T cells, cell proliferation, and cytokine profiles. Flare-up patients showed increased percentages of cluster of differentiation (CD)3+CD4- lymphocytes (p < 0.01) and granulocytes (p < 0.05) but a decreased natural killer (NK)/T lymphocyte ratio (p < 0.05). Analysis of leukocyte markers by principal component analysis (PCA) and receiver operating characteristic (ROC) curves showed that CD45RO+ (p < 0.0001) and CD45RA+ (p < 0.0001) B lymphocyte expression can discriminate between HCs and early RA patients, while CD3+CD4- lymphocyte percentage (p < 0.0424) and CD45RA+ (p < 0.0424), CD62L+ (p < 0.0284), and CD11a+ (p < 0.0185) B lymphocyte expression can differentiate between flare-up and RA remission subjects. Thus, the combined study of these leukocyte surface markers could have potential as disease severity biomarkers for RA, whose fluctuations could be related to the development of the characteristic pro-inflammatory environment.

Variable Intrinsic Expression of Immunoregulatory Biomarkers in Breast Cancer Cell Lines, Mammospheres, and Co-Cultures.

Advances in immunotherapy have increased interest in knowing the role of the immune system in breast cancer (BC) pathogenesis. Therefore, immune checkpoints (IC) and other pathways related to immune regulation, such as JAK2 and FoXO1, have emerged as potential targets for BC treatment. However, their intrinsic gene expression in vitro has not been extensively studied in this neoplasia. Thus, we evaluated the mRNA expression of tumor-cell-intrinsic CTLA-4, PDCD1 (PD1), CD274 (PD-L1), PDCD1LG2 (PD-L2), CD276 (B7-H3), JAK2, and FoXO1 in different BC cell lines, derived mammospheres, and co-cultures with peripheral blood mononuclear cells (PBMCs) by real-time quantitative polymerase chain reaction (qRT-PCR). Our results showed that intrinsic CTLA-4, CD274 (PD-L1), and PDCD1LG2 (PD-L2) were highly expressed in triple-negative cell lines, while CD276 was predominantly overexpressed in luminal cell lines. In contrast, JAK2 and FoXO1 were under-expressed. Moreover, high levels of CTLA-4, PDCD1 (PD1), CD274 (PD-L1), PDCD1LG2 (PD-L2), and JAK2 were found after mammosphere formation. Finally, the interaction between BC cell lines and peripheral blood mononuclear cells (PBMCs) stimulates the intrinsic expression of CTLA-4, PCDC1 (PD1), CD274 (PD-L1), and PDCD1LG2 (PD-L2). In conclusion, the intrinsic expression of immunoregulatory genes seems very dynamic, depending on BC phenotype, culture conditions, and tumor-immune cell interactions.

Gamificación educativa en el laboratorio de biología celular / Educational gamification in the cell biology laboratory

La incorporación de estrategias de gamificación en la docencia se ha descrito como una herramienta para aumentar la motivación y el compromiso de los alumnos con la materia. Bajo esta premisa, se ha desarrollado una experiencia de innovación educativa mediante la plataforma Kahoot! en la primera y última práctica de laboratorio de la asignatura de Biología Celular del Grado en Biología. Los participantes fueron 135 alumnos repartidos en 12 grupos de laboratorio, que se dividieron entre experimentales y controles. Todos los grupos resolvieron un cuestionario en papel acerca de los conceptos explicados en clase, al finalizar ambas prácticas (post-test), pero sólo aquellos grupos experimentales resolvían un cuestionario antes de la clase (pre-test). Antes de la primera práctica, los alumnos de los grupos experimentales respondieron al pre-test mediante el Kahoot! Sin embargo, para la última práctica algunos grupos lo resolvieron jugando al Kahoot! y otros, con papel y bolígrafo. Los resultados mostraron que aquellos alumnos que fueron seleccionados para jugar a Kahoot!, obtuvieron un mayor número de aciertos en el test realizado tras la sesión práctica (post-test) con respecto a aquellos que no resolvieron ningún pre-test o, que lo hicieron de un modo clásico. Por lo tanto, nuestros resultados sugieren que implementar la jugabilidad en la docencia incrementa considerablemente la motivación del alumnado debido, probablemente, a cambios fisiológicos experimentados por el cerebro durante el juego y a la creación de un clima positivo, que facilitan el proceso de aprendizaje.

SUMMARY: The incorporation of gamification strategies in teaching has been described as a tool to increase the motivation and engagement of students with the subject. Under this premise, an educational innovation experience has been developed using the Kahoot! platform in the first and last laboratory practice of the Cell Biology course of the Biology degree. The participants were 135 students divided into 12 laboratory groups, which were divided into experimental and control groups. All groups solved a questionnaire on paper about the concepts explained in class, at the end of both practices (post-test), but only the experimental groups solved a questionnaire before the class (pre-test). Before the first practice, students in the experimental groups answered the pre-test using Kahoot! However, for the last practice, some groups solved it by playing Kahoot! and others with pen and paper. The results showed that those students who were selected to play Kahoot! obtained a higher number of correct answers in the test performed after the practical session (post-test) than those who did not solve any pre- test or who did it in a classical way. Therefore, our results suggest that implementing gamification in teaching considerably increases student motivation, probably due to physiological changes experienced by the brain during the game and the creation of a positive climate, which facilitates the learning process.

Subcellular sorting of neuregulins controls the assembly of excitatory-inhibitory cortical circuits.

The assembly of specific neuronal circuits relies on the expression of complementary molecular programs in presynaptic and postsynaptic neurons. In the cerebral cortex, the tyrosine kinase receptor ErbB4 is critical for the wiring of specific populations of GABAergic interneurons, in which it paradoxically regulates both the formation of inhibitory synapses as well as the development of excitatory synapses received by these cells. Here, we found that Nrg1 and Nrg3, two members of the neuregulin family of trophic factors, regulate the inhibitory outputs and excitatory inputs of interneurons in the mouse cerebral cortex, respectively. The differential role of Nrg1 and Nrg3 in this process is not due to their receptor-binding EGF-like domain, but rather to their distinctive subcellular localization within pyramidal cells. Our study reveals a novel strategy for the assembly of cortical circuits that involves the differential subcellular sorting of family-related synaptic proteins.

Antioxidant Supplementation Modulates Neutrophil Inflammatory Response to Exercise-Induced Stress.

The aim of the present report was to evaluate the inflammatory response to a 2000-m running test considering neutrophil myeloperoxidase as an inflammatory marker, and to verify if supplements rich in antioxidants could modulate Post-test antioxidant and anti-inflammatory responses. To this end, a 21-day homogenization period was carried out with three groups: a control group, a supplemented group taking an almond beverage enriched with vitamins C and E and a third group consuming the same beverage but enriched with Lippia citriodora extract. At the end of this period, participants performed a 2000-m run, and blood samples were obtained the day before and immediately after the running test. Plasma and neutrophils were isolated. As a result, plasma creatine kinase and myoglobin increased, indicating Post-test muscle damage. Plasma oxidative markers were increased in all groups, except in the group supplemented with the almond beverage. Neutrophil antioxidant enzymes were significantly increased only in the control group, suggesting an antioxidant effect of the supplements provided in the other groups. Myeloperoxidase activity was significantly increased after the test in the control group, while increased enzyme levels were detected in plasma of the supplement groups. Therefore, antioxidant consumption seems to favour myeloperoxidase release. The connection of this observation with post-exercise recovery will require further investigation.

Changes in metabolic and inflammatory parameters in a type 1 diabetic patient performing extreme activities / Cambios en parámetros metabólicos e inflamatorios en un paciente diabético tipo 1 realizando actividades extremas

Background: physical activity in type 1 diabetic patients allows a better control of glycaemia and glycosylated hemoglobin, helps to maintain a residual endocrine pancreatic mass and optimizes subsequent insulin requirements. These improvements might be due in part to increases in anti-inflammatory cytokines that could help to minimize β-cell destruction. However, type, intensity and frequency of exercise for type 1 diabetic patients remain to be established. Case report: we present the case of a 48-year-old man diagnosed with type 1 diabetes at the age of 23. He is a professional alpinist and recently was recruited in a program of the Yuri Gagarin Cosmonaut Training Center (Russia) to be the first diabetic astronaut. Metabolic and inflammatory responses were assessed after performing two extreme activities. Discussion: well programmed extreme activities accompanied by a correct dietetic intervention can reduce the adverse metabolic and inflammatory processes that appear due to exercise and diabetes

Introducción: la actividad física en pacientes diabéticos tipo 1 permite un mejor control de la glucemia y hemoglobina glucosilada, ayuda a mantener una masa residual de páncreas endocrino y optimiza las necesidades de insulina. Estas mejoras podrían ser debidas en parte al incremento en citocinas antiinflamatorias que ayudarían a minimizar la destrucción de células β. Sin embargo, el tipo, la intensidad y la frecuencia de ejercicio para pacientes diabéticos tipo 1 no han sido establecidos. Caso clínico: presentamos el caso de un varón de 48 años de edad diagnosticado de diabetes tipo 1 a los 23. Es alpinista profesional y recientemente ha sido reclutado por el Centro de Entrenamiento para Cosmonautas Yuri Gagarin (Rusia) para ser el primer astronauta diabético. Hemos comparado respuestas metabólicas e inflamatorias tras realizar dos actividades extremas. Discusión: actividades extremas bien programadas y con una correcta intervención dietética pueden reducir la descompensación metabólica e inflamatoria causada por la combinación actividad-enfermedad

Changes in metabolic and inflammatory parameters in a type 1 diabetic patient performing extreme activities.

INTRODUCTION: Background: physical activity in type 1 diabetic patients allows a better control of glycaemia and glycosylated hemoglobin, helps to maintain a residual endocrine pancreatic mass and optimizes subsequent insulin requirements. These improvements might be due in part to increases in anti-inflammatory cytokines that could help to minimize ß-cell destruction. However, type, intensity and frequency of exercise for type 1 diabetic patients remain to be established. Case report: we present the case of a 48-year-old man diagnosed with type 1 diabetes at the age of 23. He is a professional alpinist and recently was recruited in a program of the Yuri Gagarin Cosmonaut Training Center (Russia) to be the first diabetic astronaut. Metabolic and inflammatory responses were assessed after performing two extreme activities. Discussion: well programmed extreme activities accompanied by a correct dietetic intervention can reduce the adverse metabolic and inflammatory processes that appear due to exercise and diabetes.

INTRODUCCIÓN: Introducción: la actividad física en pacientes diabéticos tipo 1 permite un mejor control de la glucemia y hemoglobina glucosilada, ayuda a mantener una masa residual de páncreas endocrino y optimiza las necesidades de insulina. Estas mejoras podrían ser debidas en parte al incremento en citocinas antiinflamatorias que ayudarían a minimizar la destrucción de células ß. Sin embargo, el tipo, la intensidad y la frecuencia de ejercicio para pacientes diabéticos tipo 1 no han sido establecidos. Caso clínico: presentamos el caso de un varón de 48 años de edad diagnosticado de diabetes tipo 1 a los 23. Es alpinista profesional y recientemente ha sido reclutado por el Centro de Entrenamiento para Cosmonautas Yuri Gagarin (Rusia) para ser el primer astronauta diabético. Hemos comparado respuestas metabólicas e inflamatorias tras realizar dos actividades extremas. Discusión: actividades extremas bien programadas y con una correcta intervención dietética pueden reducir la descompensación metabólica e inflamatoria causada por la combinación actividad-enfermedad.

Differences of Clonogenic Mesenchymal Stem Cells on Immunomodulation of Lymphocyte Subsets.

Mesenchymal stem cells (MSC) are a widely used population in cell therapy for their ability to differentiate into distinct tissues and more lately, for their immunomodulatory properties. However, the use of heterogeneous populations could be responsible for the nondesired outcomes reflected in the literature. Here, we analyse the different capacities of five one-cell-derived MSC clones to exert their immunomodulation ex vivo. We assessed proliferation assays in cocultures of MSC clones and purified cluster of differentiation (CD)3+, CD4+, or CD8+ lymphocytes; analysed the regulatory T (Treg) cells fold change rate; determined the effects on viability of peripheral blood mononuclear cells (PBMC); and also measured the coculture cytokine profiles (Th1/Th2). Conditioned media (CM) of different clones were also used to perform both proliferation assays and to analyse Treg fold change. The five clones analysed in this work were able to generate heterogeneous environments. Different clones inhibited proliferation of CD3+ and CD4+ lymphocytes, with different intensities. Surprisingly, all clones promoted proliferation of CD8+ lymphocytes. Different MSC clones and their CM were able to increase the number of Treg with different intensities. Finally, different clones also promoted different effects on the viability of PBMC treated with ultraviolet light. Considering all these data together, it seems that different clones, even from the same donor, can promote a wide spectrum of responses from anti-inflammatory to proinflammatory character. This fact may be important to standardise the design of personalized cell therapy protocols, thus diminishing the aforementioned undesired outcomes existing nowadays in this type of therapies.