Nuevas estrategias docentes en Histología. Más aprendizaje y menos enseñanza: Uso de microscopios virtuales e Historrelatos.

Current teaching at the University needs novel methodologies to increase students’ motivation. Here, we present two approaches to engage the student body to Human Histology subject at the University of Malaga. Virtual teaching was propelled by the COVID-19 crisis and confinement. The software for the study of histological/histopathological samples has become a valuable tool. Moreover, digital competences are in high demand within the biomedical field but students usually do not receive sufficient training. For these reasons, we have implemented the use of virtual microscopy (VM, Olympus), sharing 66 digitalized slides accessible under a username/password. VM provides real-time dynamic microscopy and offers an innovative experience at exceptionally high resolution. VM allows students to explore the samples online from anywhere, favoring autonomy and self-learning. Moreover, VM enables capturing specific tissue areas using these pictures to ask specific questions. On the other hand, transversal competences such as reading and writing skills, along with synthesis capability can be underdeveloped in our students. We initiated the activity of writing stories about histology contents (Histostories). Professional graphic designers from a webpage of scientific divulgation (masscience.com) illustrated the first story about erythrocytes. We conducted a survey among medical students to analyze the impact of this narration on their learning. Most of them welcome the initiative, considering it as an appropriate and enjoyable instrument for summarizing and revising the concepts. Immunity was among the topics more demanded between the students. Finally, we encouraged our students to write their own Histostories mentored by our teaching staff. These stories are shared through the virtual campus and on masscience website. So far, two medical students are collaborating with us in this experimental project that we expect it will bring more benefits to both readers and participants.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Epigenetic targets to enhance antitumor immune response through the induction of tertiary lymphoid structures

Tertiary lymphoid structures (TLS) are ectopic lymphoid aggregates found in sites of chronic inflammation such as tumors and autoimmune diseases. The discovery that TLS formation at tumor sites correlated with good patient prognosis has triggered extensive research into various techniques to induce their formation at the tumor microenvironment (TME). One strategy is the exogenous induction of specific cytokines and chemokine expression in murine models. However, applying such systemic chemokine expression can result in significant toxicity and damage to healthy tissues. Also, the TLS formed from exogenous chemokine induction is heterogeneous and different from the ones associated with favorable prognosis. Therefore, there is a need to optimize additional approaches like immune cell engineering with lentiviral transduction to improve the TLS formation in vivo. Similarly, the genetic and epigenetic regulation of the different phases of TLS neogenesis are still unknown. Understanding these molecular regulations could help identify novel targets to induce tissue-specific TLS in the TME. This review offers a unique insight into the molecular checkpoints of the different stages and mechanisms involved in TLS formation. This review also highlights potential epigenetic targets to induce TLS neogenesis. The review further explores epigenetic therapies (epi-therapy) and ongoing clinical trials using epi-therapy in cancers. In addition, it builds upon the current knowledge of tools to generate TLS and TLS phenotyping biomarkers with predictive and prognostic clinical potential.The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The work is funded by Instituto de Salud Carlos III through the projects PI18/01592 (Co-funded by the European Regional Development Fund/European Social Fund “A way to make Europe”/”Investing in your future”) and PI22/01816 (Co-funded by the European Union), Sociedad Española de Oncología Médica (SEOM21, SEOM23); Sistema Andaluz de Salud, through the projects SA 0263/2017, Nicolás Monardes, PI-0135-2018, PI-0121-2020 and RH-0090-2020; Consejería de Transformación económica, Industria, Conocimiento y Universidades through the projects CV20-62050 and ProyExcel_01002; Spanish Group of Melanoma (Award for Best Research Project 2020), Fundación Bancaria Unicaja through the project C19048, Asociación Española Contra el Cáncer, Talento Clínico (AECC 2020), and Andalusia-Roche Network Mixed Alliance in Precision Medical Oncology (AC20057), the Spanish Group of Melanoma (GEM23) and University of Malaga Research Plan (B1-2022_28)

New teaching strategies in Histology. More learning and less teaching

Las transformaciones de la educación Médica conciernen a la Histología, ocasionando reducciones en la carga docente y asignación de créditos, redefinición de competencias y objetivos de aprendizaje, así como una creciente orientación médica de sus contenidos y una disminución de las clases magistrales. Por ello se hace necesario el empleo de nuevos métodos que se aproximen más al aprendizaje que a la enseñanza, pero que no supongan ni un aumento de la carga docente del alumnado ni un incremento de los ya hipertrofiados planes de estudios. En la Facultad de Medicina de Málaga hemos implementado nuevas metodologías docentes: orientación médica , clase inversa, ABP, microscopia virtual, HistolCasts), HistolWord, Instagram, Historrelatos), y evaluación continua . Sobre estas actividades los estudiantes han mostrado un alto grado de participación y satisfacción, estimulando su interés y motivación por la Histología y mejorando el rendimiento académico. Adicionalmente, muchas de estas estrategias se pueden extrapolar a otras áreas de la educación médica, tanto para estudiantes como para residentes y formación continuada. Algunas de estas metodologías ya han sido ya presentadas y otras lo serán por algunos de mis compañeros. Me voy a referir a continuación al HistolWord. Se trata de una actividad de gamificación basada en el juego del pasapalabra, empleando términos histológicos. Se formaron 32 equipos de 5 estudiantes que compitieron en un sistema de eliminatorias desde dieciseisavos de final, confeccionándose 70 roscos de palabras. Todo se desarrollaba en un aula con casi 200 estudiantes, donde se leían las preguntas y se proyectaba el rosco, de manera que no solo participaban los dos equipos que se enfrentaban en ese momento, sino todos los presentes. Los estudiantes indicaron la utilidad de HistolWord como motivación para el estudio, complemento de las clases, revisión y aplicaciones médicas, existiendo una correlación positiva con las calificaciones.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Pathological, immunological and parasitological study of sheep vaccinated with the recombinant protein 14-3-3z and experimentally infected with Fasciola hepatica

In this study, the immunogenicity and protective capacity of a new recombinant vaccine candidate, the rFh14-3-3z protein was analysed in sheep experimentally challenged with Fasciola hepatica, in terms of fluke burden, faecal egg counts, hepatic damage and humoral immune response. Three groups of 8 animals each were used for study, group 1 was immunised with the rFh14-3-3z in Montanide adjuvant, whereas group 2 and 3 remained as adjuvant control and infection control groups, respectively. The parasitological analysis showed that no significant reduction in fluke burden, fluke size and faecal egg counts was detected. The extent of hepatic damage was very similar between groups. Nonetheless, animals immunised with the rFh14-3-3z protein induced the development of specific IgG1 and IgG2, being the IgG1 the predominant antibody; which confirms the immunogenicity of this protein in sheep. This is the first report of the 14-3-3z proteins as vaccine against the infection with F. hepatica.Finaciación de los siguientes Proyectos Nacionales: AGL2002-00644, AGL2015-67023-C2-1-R/2-R del Ministerio de Economía, Industria y Competitividad del Gobierno de España

Microbiome alterations and Alzheimer's Disease: modeling strategies with transgenic mice.

In the last decade, the role of the microbiota-gut-brain axis has been gaining momentum in the context of many neurodegenerative and metabolic disorders, including Alzheimer's disease (AD) and diabetes, respectively. Notably, a balanced gut microbiota contributes to the epithelial intestinal barrier maintenance, modulates the host immune system, and releases neurotransmitters and/or neuroprotective short-chain fatty acids. However, dysbiosis may provoke immune dysregulation, impacting neuroinflammation through peripheral-central immune communication. Moreover, lipopolysaccharide or detrimental microbial end-products can cross the blood-brain barrier and induce or at least potentiate the neuropathological progression of AD. Thus, after repeated failure to find a cure for this dementia, a necessary paradigmatic shift towards considering AD as a systemic disorder has occurred. Here, we present an overview of the use of germ-free and/or transgenic animal models as valid tools to unravel the connection between dysbiosis, metabolic diseases, and AD, and to investigate novel therapeutical targets. Given the high impact of dietary habits, not only on the microbiota but also on other well-established AD risk factors such as diabetes or obesity, consistent changes of lifestyle along with microbiome-based therapies should be considered as complementary approaches.Partial funding for open access charge: Universidad de Málaga/CBU

Th1/Th2 balance in the liver and hepatic lymph nodes of vaccinated and unvaccinated sheep during acute stages of infection with Fasciola hepatica

The expression of IFNγ and IL4 was quantified using q-PCR in the liver and hepatic lymph nodes (HLN) of sheep during early stages of infection with Fasciola hepatica (1, 3, 9 and 18days post-infection, dpi). A group of animals (Group 1) were vaccinated with Fasciola hepatica recombinant cathepsin L1 (FhCL1) in montanide 70 VG prior to infection, a second group (group 2) was used as infected control and a third (group 3) was used as uninfected control. To study vaccine efficacy three additional groups were sacrificed 19 weeks post-infection (group 4 immunized with CL1, group 5 with the adjuvant and group 6 was used as infected control). The vaccinated group did not show significant fluke reduction compared to the adjuvant group and infected control group. IL4 expression was observed to increase at 9 dpi and was further elevated at 18 dpi in the liver and HLN of vaccinated and infected control groups compared to the uninfected group. IFNγ expression exhibited different dynamics in the liver and HLN compared to IL4; thus, in the liver this cytokine increased at 9 dpi in the vaccinated and at 18 dpi in vaccinated and infected control groups, while in the HLN it decreased gradually and significantly from 1 dpi onwards. These results suggest that a marked Th2 polarization is present from 9 dpi in HLN and from 18 dpi in the liver. The increase of IFNγ in the liver may correspond with tissue damage response with granuloma formation. The FhCL1 vaccine did not alter the Th1/Th2 balance when compared to unvaccinated and infected sheep. The study of IFNγ and IL4 in the various tissue compartments in sheep could facilitate selection of new adjuvants inducing a strong Th1 response for a more rationale vaccine formulation.This work was supported by EU grants (FPVII-265862-PARAVAC, H2020-635408-PARAGONE) and National grant (AGL2015-67023-C2-1-R)

"Ab initio" synthesis of zeolites for preestablished catalytic reactions

[EN] Unlike homogeneous catalysts that are often designed for particular reactions, zeolites are heterogeneous catalysts that are explored and optimized in a heuristic fashion. We present a methodol. for synthesizing active and selective zeolites by using org. structure-¿directing agents that mimic the transition state (TS) of preestablished reactions to be catalyzed. In these zeolites, the pores and cavities could be generated approaching a mol.-¿recognition pattern. For disproportionation of toluene and isomerization of ethylbenzene into xylenes, the TSs are larger than the reaction products. Zeolite ITQ-¿27 showed high disproportionation activity, and ITQ-¿64 showed high selectivity for the desired para and ortho isomers. For the case of a product and TS of similar size, we synthesized a catalyst, MIT-¿1, for the isomerization of endo-¿dicyclopentane into adamantane.This work has been supported by the European Union through the European Research Council (grant ERC-AdG-2014-671093, SynCatMatch) and the Spanish government through the "Severo Ochoa Program" (grant SEV 2012-0267). The Electron Microscopy Service of the Universitat Politecnica de Valencia (UPV) is acknowledged for help with sample characterization. The Red Espanola de Supercomputacion (RES) and Centre de Calcul de la Universitat de Valencia are gratefully acknowledged for computational facilities and technical assistance. E.M.G. acknowledges "La Caixa-Severo Ochoa" International Ph.D. Fellowship (call 2015). We thank I. Millet for technical assistance and V. J. Margarit and A. Cantin for helpful discussions.Gallego-Sánchez, EM.; Portilla Ovejero, MT.; Paris-Carrizo, CG.; Leon Escamilla, EA.; Boronat Zaragoza, M.; Moliner Marin, M.; Corma Canós, A. (2017). "Ab initio" synthesis of zeolites for preestablished catalytic reactions. Science. 355(6329):1051-1054. https://doi.org/10.1126/science.aal0121S105110543556329Vermeiren, W., & Gilson, J.-P. (2009). Impact of Zeolites on the Petroleum and Petrochemical Industry. Topics in Catalysis, 52(9), 1131-1161. doi:10.1007/s11244-009-9271-8Climent, M. J., Corma, A., & Iborra, S. (2011). Heterogeneous Catalysts for the One-Pot Synthesis of Chemicals and Fine Chemicals. Chemical Reviews, 111(2), 1072-1133. doi:10.1021/cr1002084De Vos, D. E., & Jacobs, P. A. (2005). Zeolite effects in liquid phase organic transformations. Microporous and Mesoporous Materials, 82(3), 293-304. doi:10.1016/j.micromeso.2005.01.038Jacobs, P. A., Dusselier, M., & Sels, B. F. (2014). 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Study of peritoneal macrophage immunophenotype in sheep experimentally infected with Fasciola hepatica

During Fasciola hepatica infection, the parasite has the capability to modulate the host immune response towards a non-protector Th2 type instead of Th1. This type of immune response is closely related to the alternative activation of macrophages (M2 profile) as has been shown in vivo in murine models. In this study, an experiment was carried out in order to evaluate the expression of CD68, CD14, CD206 and iNOS in cells present in the peritoneal fluid of sheep during early stages of infection with F. hepatica (1, 3, 9 and 18 days post-infection, dpi) by immunocytochemistry. To the authors’ knowledge, this is the first report that studies the in vivo immunophenotype of macrophages from the peritoneal fluid of sheep infected with F. hepatica. Throughout the experiments the absolute number of leucocytes progressively increased, reaching its highest value at 18 dpi, mainly due to the increase of eosinophils. This immunocytochemical study had two purposes: 1) CD68 expression was assessed with Hansel counterstaining, to optimally identify peritoneal macrophages, eosinophils and lymphocytes; 2) expression of CD14, CD206 and iNOS was evaluated to identify alternative or classical pathways of macrophage activation. The results showed a significant increase in CD14 from day 3 dpi compared with the non-infected group. CD206 expression at all time-points showed a significant and dramatic increase in comparison with the uninfected group. On the other hand, iNOS expression showed little variation, and was significantly decreased at 18 dpi in comparison with the uninfected group. These results suggest that F. hepatica induces an alternative activation of peritoneal macrophages of sheep from the first day post-infection, which may facilitate parasite survival. This is the first report describing M2 activation of peritoneal macrophages in ruminants infected with F. hepatica.Financiación y miembro investigador en proyectos europeos H2020-SFS-2014-2-635408 PARAGONE - Vaccines for animal parasites” y un Proyecto Nacional INTERFAS (AGL2015-67023-C2-1-R9