Developing a practical guide for teaching histology: an evaluation of the didactic components

The Didactic Guide is a valuable tool complementing and making learning more dynamic. It is done using creative didactic strategies that simulate the presence of the tutor and generate a dialogue in order to offer students different possibilities to improve their understanding and self-discovery learning.This way the student is the protagonist of his own learning process. In this context, is highly important to consider the self discovery learning as a goal facilitating significant learning. The use of practical guides enables students to manage their own time, goals, techniques, contents and evaluation. In medical histology teaching several models of didactic guides could be use, and they normally include numerous activities, text, questionnaires, pictures, and drawings that may enhance the effectiveness of this tool in the learning process. In this work we have evaluated the usefulnes of different sections of a histology didactic guide in order to determine the key sections that enhance the learning process in human histology. For this purpose, a practical histology guide was designed with different sections: message text, theoretical text, objectives, drawings, pictures, clinical cases, games, blank spaces for self notes and drawing and final self evaluation questions. First, a simple questionnaire was applied in 90 students enrolled in histology practical seccions to analyze the student´s perceptions and preferences related to the histology guide. Finally, for all questionnaires average results and standard deviations were calculated for each option and all participants, as well for each gender, separately. Comparisons were done for drawings vs drawing blank spaces, teoric content vs notes blank space, drawings vs pictures and for each gender separately using Mann-Whitney non-parametrical test. Our findings revealed that visual strategies such as images and pictures were considered to be more useful for learning histology in the practical session. Similary, the students rated the self evaluation questions and blank spaces for self notes and drawing to be more attractive to the students. However, texts with theoretical information, messages, objectives, and clinical cases revealed to be less useful for the students in the learning process of medical histology. Moreover, statistically significant differences between theoretical content vs notes blank space was observed. All these results point out the importance of including pictures and drawings in the practical guide accompained of blank spaces that allow the development of creativity and autonomy that lead the students into a self discovery learning. Interestingly the students do not appreciate the presence of theoretical background in the practical guide as relevant information for their academic formation in the practical session

Information and Scientific Impact of Advanced Therapies in the Age of Mass Media: Altmetrics-Based Analysis of Tissue Engineering

This study was supported by CTS-115 (Tissue Engineering Research Group, University of Granada) from Junta de Andalucia, Spain, the Spanish State Research Agency through the project PID2019-105381GA-I00/AEI/10.13039/501100011033 (iScience) , a postdoctoral grant (RH-0145-2020) from the Andalusia Health System, and the European Union Fondo Europeo de Desarrollo Regional para la Inversion Territorial Integrada Grant for Cadiz Province (PI-0032-2017) . The authors thank Altmetric LLP (London, UK) for granting access to Altmetric Explorer for research purposes.Background: Tissue engineering (TE) constitutes a multidisciplinary field aiming to construct artificial tissues to regenerate end-stage organs. Its development has taken place since the last decade of the 20th century, entailing a clinical revolution. TE research groups have worked and shared relevant information in the mass media era. Thus, it would be interesting to study the online dimension of TE research and to compare it with traditional measures of scientific impact. Objective: The objective of this study was to evaluate the online dimension of TE documents from 2012 to 2018 using metadata obtained from the Web of Science (WoS) and Altmetric and to develop a prediction equation for the impact of TE documents from altmetric scores. Methods: We analyzed 10,112 TE documents through descriptive and statistical methods. First, the TE temporal evolution was exposed for WoS and 15 online platforms (news, blogs, policy, Twitter, patents, peer review, Weibo, Facebook, Wikipedia, Google, Reddit, F1000, Q&A, video, and Mendeley Readers). The 10 most cited TE original articles were ranked according to the normalized WoS citations and the normalized Altmetric Attention Score. Second, to better comprehend the TE online framework, correlation and factor analyses were performed based on the suitable results previously obtained for the Bartlett sphericity and Kaiser–Meyer–Olkin tests. Finally, the linear regression model was applied to elucidate the relation between academics and online media and to construct a prediction equation for TE from altmetrics data. Results: TE dynamic shows an upward trend in WoS citations, Twitter, Mendeley Readers, and Altmetric Scores. However, WoS and Altmetric rankings for the most cited documents clearly differ. When compared, the best correlation results were obtained for Mendeley Readers and WoS (ρ=0.71). In addition, the factor analysis identified 6 factors that could explain the previously observed differences between academic institutions and the online platforms evaluated. At this point, the mathematical model constructed is able to predict and explain more than 40% of TE WoS citations from Altmetric scores. Conclusions: Scientific information related to the construction of bioartificial tissues increasingly reaches society through different online media. Because the focus of TE research importantly differs when the academic institutions and online platforms are compared, basic and clinical research groups, academic institutions, and health politicians should make a coordinated effort toward the design and implementation of adequate strategies for information diffusion and population health education.CTS-115 (Tissue Engineering Research Group, University of Granada) from Junta de Andalucia, SpainSpanish Government PID2019-105381GA-I00/AEI/10.13039/501100011033Andalusia Health System RH-0145-2020European Union Fondo Europeo de Desarrollo Regional para la Inversion Territorial Integrada Grant for Cadiz Province PI-0032-201

Translational Applications of Artificial Intelligence and Machine Learning for Diagnostic Pathology in Lymphoid Neoplasms: A Comprehensive and Evolutive Analysis

Genomic analysis and digitalization of medical records have led to a big data scenario within hematopathology. Artificial intelligence and machine learning tools are increasingly used to integrate clinical, histopathological, and genomic data in lymphoid neoplasms. In this study, we identified global trends, cognitive, and social framework of this field from 1990 to 2020. Metadata were obtained from the Clarivate Analytics Web of Science database in January 2021. A total of 525 documents were assessed by document type, research areas, source titles, organizations, and countries. SciMAT and VOSviewer package were used to perform scientific mapping analysis. Geographical distribution showed the USA and People’s Republic of China as the most productive countries, reporting up to 190 (36.19%) of all documents. A third-degree polynomic equation predicts that future global production in this area will be three-fold the current number, near 2031. Thematically, current research is focused on the integration of digital image analysis and genomic sequencing in Non-Hodgkin lymphomas, prediction of chemotherapy response and validation of new prognostic models. These findings can serve pathology departments to depict future clinical and research avenues, but also, public institutions and administrations to promote synergies and optimize funding allocation.Andalusia Health System - RH-0145-2020EU FEDER ITI Grant for Cadiz Province PI-0032-201

Cell viability evaluation in tissue constructs. A preliminary study

Los autores agradecen a las Profesoras Allice Warley (King’s Co- llege, London) y María del Carmen Sánchez-Quevedo (Universidad de Granada) por el asesoramiento recibido en el desarrollo de las técnicas de microscopía electrónica analítica por energía dispersiva de rayos-X.Objetivo: Aplicar la microscopia electrónica analítica por energía dispersiva de rayos X a la evaluación de la viabilidad celular en constructos tisulares de fibrina agarosa y determinar la viabilidad celular en el seno del constructo Material y Métodos: Se evalúan fibroblastos aislados y fibroblastos en el seno del constructo a los 7 y 21 días con la técnica de Live/Dead y de microscopia electrónica analítica por energía dispersiva de rayos X. Resultados: El estudio demuestra que la aplicación de la técnica es factible y que la viabilidad es elevada en los constructos a los 21 días sin que existan signo de apoptosis en la población celular. Conclusión: La microscopia electrónica analítica por energía dispersiva de rayos X permite evaluar la viabilidad celular en los constructos tisulares y establecer un control de calidad más exigente para su aplicación terapéutica.Objective: Apply X-ray microanalytical electron microscopy to evaluate cell viability in fibrin agarose tissue constructs and determine cell viability within the construct. Material and Methods: Isolated fibroblasts and fibroblasts within the construct are evaluated at 7 and 21 days with the technique of Live/Dead and with X-ray microanalytical electron microscopy. Results: The study shows that the application of the technique is feasible that viability is high in the constructs at 21 days without sign of apoptosis in the cell population. Conclusion: X-ray microanalytical electron microscopy allows the evaluation of cell viability in tissue constructs and establish a more demanding quality control for therapeutic application

Evaluation of cell viability and apoptotic patterns in stem cells isolated from human dental pulp

Introducción: El desarrollo de sustitutos biológicos mediante Ingeniería Tisular requiere de la utilización de una fuente de células madre que, además de ser capaz de autorenovarse y diferenciarse, mantengan una funcionalidad y viabilidad óptima justo en el momento de su uso. El estudio de la viabilidad celular constituye un importante control de calidad, especialmente en aquellas poblaciones celulares con un alto potencial para su utilización en Ingeniería Tisular, como son las células madre de la pulpa dental (DPSC). El objetivo de este trabajo es la evaluación de la viabilidad durante los tres primeros subcultivos de células madre de la pulpa dental humana (hDPSC). Material y métodos: Se obtuvieron 3 subcultivos consecutivos de hDPSC de terceros molares humanos sanos (N = 3) mediante un proceso de digestión enzimática. La concentración intracelular de los iones sodio (Na), potasio (K), y cloro (Cl) fue determinada mediante microscopía analítica por energía dispersiva de rayos X (EPXMA). El porcentaje de células en apoptosis (fragmentación de DNA) fue determinado mediante el ensayo de fluorescencia TUNEL en el primer y tercer subcultivo. Resultados: En el segundo subcultivo se detectó un descenso significativo del potasio (p = 0,011) indicando un descenso en la viabilidad celular. En el tercer subcultivo, los niveles de cloro y sodio aumentaron de forma significativa (p = 0,010 y p = 0,002), generando un perfil iónico compatible con células en estado apoptótico. La fluorescencia a partir del ensayo TUNEL reveló 2,76 ± 1,80 % de células apoptóticas en el tercer subcultivo, mientras que en el primer subcultivo, dicha proporción fue de 0,55 ± 0,27%. Discusión: Las hDPSC se encuentran de forma nativa en la pulpa dental y estas condiciones nativas se pueden alterar cuando pasan a las condiciones de un medio de cultivo in vitro. Estas alteraciones pueden ser la respuesta a un proceso de adaptación. Dicha adaptación, junto con el estrés adicional generado por el tratamiento enzimático realizado para digerir la matriz extracelular, tiene como consecuencia una pérdida transitoria y leve de la viabilidad producida por un mecanismo de apoptosis. En resumen, los 3 primeros subcultivos de hDPSC deberán ser descartados para su uso en ingeniería tisular, por ser células en un estado de apoptosis activa.Introduction: The development of biological substitutes by Tissue Engineering needs the use of a stem cell source able not only to self-replicate and differentiate, but also to maintain optimal functionality and cell viability when used. The evaluation of cell viability is considered an important quality control, especially in cell lineages with high capabilities for being used in tissue engineering, as dental pulp stem cells (DPSC). The aim of this research is to evaluate the cell viability through three human dental pulp stem cells (hDPSC) subcultures. Materials and methodology:Three consecutive hDPSC subcultures were obtained from human sound third molars (N = 3) by enzymatic digestion. Intracellular ionic concentration of sodium (Na), potassium (K) and chlorine (Cl) was determined by electron probe X-ray microanalysis (EPXMA). Apoptotic cells percentage (DNA fragmentation) was determined by TUNEL fluorescence assay at first and third subculture Results: A significant decrease of potassium was detected at second subculture (p = 0,011), suggesting a loss of cell viability. At third subculture, chlorine and sodium statistically increased (p = 0,010 y p = 0,002), inducing an apoptotic-like ionic profile. Fluorescence from TUNEL assay revealed 2,76 ± 1,80 % of apoptotic cells at third subculture, while that ratio was 0,55 ± 0,27% at first subculture. Discussion: hDPSC are natively stored in the dental pulp and these native conditions may be altered by the in vitro cell culture conditions. In this regard, these alterations could be in response to an adaptative process. This adaptative process, besides cell stress as a consequence of an enzymatic treatment for the extracellular matrix digestion, produces a mild and temporary loss of cell viability by apoptosis. In summary, the first, second and third subculture of hDPSC should be discarded for the use in tissue engineering as they are apoptotic

Evaluation of the awareness of novel advanced therapies among family medicine residents in Spain

Advanced therapies are increasingly demanded by patients with the intent of treating some incurable conditions. Because family medicine professionals play an important role as health educators, their residency programs should incorporate new knowledge related to advanced therapies. To successfully implement these programs, how family medicine residents perceive these therapies should be investigated. The main components of perception, i.e. conceptual, procedural and attitudinal, refer to knowledge, skills and feelings, respectively. We designed a specific questionnaire to assess the components of perceptions of advanced therapies in 300 medical residents enrolled in the Spanish National Family Medicine Residency Program. Each component consisted of 4 or 5 topics and each topic contained 6 items. Respondents scored highest in the procedural component (average 4.12±1.00), followed by the attitudinal (3.94±1.07) and conceptual component (3.04±1.43). Differences among the three components were statistically significant (p<0.00017). Family medicine residents perceived that procedures to implement advanced therapies are well established, especially their application. However, they felt their cognitive background was insufficient to respond efficiently to the expectations generated by these new therapeutic tools, especially in the regulatory framework. High awareness of the risks and limitations of these treatments was reflected by residents’ preference for clinically tested therapies. Although they appropriately situated treatment with these therapies within hospital care, they associated the biofabrication of novel products with research centers, although these therapeutic tools can be produced in different facilities. These results are potentially useful for designing future training programs and health policies for family medicine residents, and suggest the need to implement specific training programs in advanced therapies at the conceptual, procedural and attitudinal level.This work was supported by CTS-115 (Tissue Engineering Group), Junta de Andalucia

Type I and type VII agarose characterization for use in tissue engineering

Objetivos: El objetivo del presente trabajo es el desarrollo de un nuevo protocolo para la inclusión de geles de agarosa en parafina que permita su estudio en histología, y la posterior caracterización de dos tipos distintos de agarosa, Tipo I y Tipo VII, para su uso en Ingeniería Tisular. Métodos: Se evaluaron distintos protocolos que permitiesen un correcto procesamiento de los geles de agarosa permitiendo obtener cortes de dichos geles incluidos en parafina. Una vez optimizado el protocolo, se realizó una caracterización de las agarosas tipo I y VII para su utilización en Ingeniería Tisular. Para ello se obtuvieron geles de agarosa de ambos tipos a concentraciones del 2% y 2.5%, y en cuyo interior había condrocitos. Se evaluó la permeabilidad de la membrana celular mediante la medida por espectrofotometría del ADN liberado al medio, el tamaño celular de los condrocitos cultivados y el estado celular mediante inmunohistoquímica frente a PCNA y Vimentina. También se realizó un estudio sobre la resistencia a la compresión que presentan los distintos geles con el paso del tiempo. Resultados: Tres de los protocolos propuestos hacían posible el procesamiento de los geles y permitieron la caracterización de los distintos tipos de agarosa. En este caso, el estudio de las agarosas tipo I y VII ha demostrado que los condrocitos crecen correctamente en ambos tipos de agarosas, pero que la tipo VII favorece el desarrollo de los mismos frente a la Tipo I. Además, la agarosa pierde elasticidad con el paso de los días soportando a la vez menos fuerza de compresión. Conclusiones: El Protocolo propuesto para el procesamiento y posterior inclusión de geles de agarosa en parafina en este trabajo permite realizar histología en los mismos, y ha permitido confirmar que la agarosa tipo VII favorece el desarrollo y supervivencia celular.Purpose: The objective of the present work is the development of a new protocol for the inclusion of paraffinembedded agarose gels to allow their study in histology and the subsequent characterization of two different types of agarose, Type I and Type VII, for their use in Tissue Engineering. Method: Different protocols were evaluated for a correct processing of the agarose gels allowing to obtain cuts of paraffin-embedded agarose gels. Once the protocol was optimized, a characterization of type I and VII agarose was performed for their use in tissue engineering. For this purpose, agarose gels of both types were obtained at concentrations of 2% and 2.5%, and in which chondrocytes were present. The permeability of the nuclear membrane was measured by spectrophotometry of the DNA released, the cell size of the cultured chondrocytes and the cellular state by immunohistochemistry against PCNA and Vimentin. A study on the compressive strength of different gels over time was also performed. Results: Three of the proposed protocols allowed the processing of the gels and allowed the characterization of the different types of agarose. In this case, the study of the type I and VII agarose has shown that chondrocytes grow correctly in both types of agarose, but type VII favors the development of the same against Type I. In addition, the agarose loses elasticity with the day pass while supporting less compressive force. Conclusions: The proposed protocol for the processing and subsequent inclusion of paraffin-agarose gels in this work allows the histology to be carried out in the same ones, and has allowed to confirm that type VII agarose favors the development and cellular survival

Histological assessment of nanostructured fibrin-agarose skin substitutes grafted in burnt patients. A time-course study

A previously developed fibrin-agarose skin model—UGRSKIN—showed promising clinical results in severely burnt patients. To determine the histological parameters associated to the biocompatibility and therapeutic effects of this model, we carried out a comprehensive structural and ultrastructural study of UGRSKIN grafted in severely burnt patients after 3 months of follow-up. The grafted epidermis was analogue to native human skin from day 30th onward, revealing well-structured strata with well-differentiated keratinocytes expressing CK5, CK8, CK10, claudin, plakoglobin, filaggrin, and involucrin in a similar way to controls, suggesting that the epidermis was able to mature and differentiate very early. Melanocytes and Langerhans cells were found from day 30th onward, together with a basement membrane, abundant hemidesmosomes and lack of rete ridges. At the dermal layer, we found an interface between the grafted skin and the host tissue at day 30th, which tended to disappear with time. The grafted superficial dermis showed a progressive increase in properly-oriented collagen fibers, elastic fibers and proteoglycans, including decorin, similarly to control dermis at day 60-90th of in vivo follow-up. Blood vessels determined by CD31 and SMA expression were more abundant in grafted skin than controls, whereas lymphatic vessels were more abundant at day 90th. These results contribute to shed light on the histological parameters associated to biocompatibility and therapeutic effect of the UGRSKIN model grafted in patients and demonstrate that the bioengineered skin grafted in patients is able to mature and differentiate very early at the epithelial level and after 60–90 days at the dermal level.Consejería de Salud y Familias, Junta de AndalucíaGrant/Award Numbers: PE- 0395-2019, PI-0458-2016Consejería de Transformación Económica, Industria,Conocimiento y Universidades, Grant/Award Number: B-CTS-450-UGR20Instituto de Salud Carlos III, Grant/Award Number: AC17/0001

Diagnostic evaluation of competences in a model of service-learning in the degree of Medicine

Objetivos: El objetivo de este trabajo es la evaluación diagnóstica de las competencias conceptuales, procedimentales y actitudinales en estudiantes de Medicina previo a la realización de un modelo de aprendizaje+servicio sobre hemodonación. Métodos: 161 estudiantes de Medicina participaron en este estudio realizando una encuesta de valoración de 15 competencias cocneptuales, procedimentales y actitudinales justo antes de la rotación por diferentes estaciones de aprendizaje + servicio en un Centro Regional de Transfusión Sanguínea. Las valoraciones fueron analizadas como evaluación diagnóstica previa. Resultados: 12 de las 15 competencias evaluadas obtuvieron una puntuación superior a 3, por lo que las compentecias previas fueron bastante buenas en el alumnado. Las comptencias de tipo actitudinal fueron las más valoradas (4.29 ± 0.54), siendo significativamente superiores a las procedimentales (3.50 ± 0.65); y estas, a su vez, superiores a las de tipo conceptual (2.78 ± 0.77). Las competencias conpcetuales fueron peor valiradas debido a la inclusión de algunas nociones técnicas que los estudiantes deberán aprender. Conclusiones: Los participantes en el estudio mostraron un nivel elevado de competencias a nivel procedimental y actitudinal previa a la relización de un aprendizaje+servicio sobre hemodonación. La adquisición de los nuevos conceptos permitirán reforzar aquellas actitudes y procedimientos que derivan de dichos conceptos, dotando al alumno de una educación sanitaria más completa que la provista por otros métodos pedagógicos tradicionales.Aim: The objective of this work is the diagnostic evaluation of conceptual, procedural and attitudinal skills in medical students previous to the realization of a model of service-learning on blood donation. Methods: 161 medical students participated in this study by conducting a survey to assess 15 conceptual, procedural and attitudinal competencies just before rotation through different servicelearning stations in a Regional Blood Transfusion Center. The assessments were analyzed as a previous diagnostic evaluation. Results: 12 of the 15 competencies evaluated obtained a score higher than 3, so that the previous competences were high in the students. Attitudinal skills were the most valued (4.29 ± 0.54), being significantly higher than procedural skills (3.50 ± 0.65); and these, in turn, were higher than conceptual skills (2.78 ± 0.77). The conceptual competences were less valued due to the inclusion of some technical notions that the students are still to learn. Conclusions: The participants in this study showed a high level of procedural and attitudinal competencies before the performance of a service-learning on blood donation. The acquisition of the new concepts will allow to strengthen those derived attitudes and procedures from those concepts, providing the student with a more complete health education than that provided by other traditional pedagogical methods