Wound healing across the animal kingdom: Crosstalk between the immune system and the extracellular matrix

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Arenas Gómez, Claudia M., Sabin, K. Z., & Echeverri, K. Wound healing across the animal kingdom: Crosstalk between the immune system and the extracellular matrix. Developmental Dynamics, (2020): 1-13, doi:10.1002/dvdy.178.Tissue regeneration is widespread in the animal kingdom. To date, key roles for different molecular and cellular programs in regeneration have been described, but the ultimate blueprint for this talent remains elusive. In animals capable of tissue regeneration, one of the most crucial stages is wound healing, whose main goal is to close the wound and prevent infection. In this stage, it is necessary to avoid scar formation to facilitate the activation of the immune system and remodeling of the extracellular matrix, key factors in promoting tissue regeneration. In this review, we will discuss the current state of knowledge regarding the role of the immune system and the interplay with the extracellular matrix to trigger a regenerative response.The research in the Echeverri lab is supported NIH NCID R01 to Karen Echeverri and start‐up funds from the MBL. Keith Z. Sabin has been supported by an NIH T32 GM113846 grant

A de novo reference transcriptome for Bolitoglossa vallecula, an Andean mountain salamander in Colombia

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Arenas Gomez, C. M., Woodcock, M. R., Smith, J. J., Voss, S. R., & Delgado, J. P. A de novo reference transcriptome for Bolitoglossa vallecula, an Andean mountain salamander in Colombia. Data in Brief, 29, (2020): 105256, doi:10.1016/j.dib.2020.105256.The amphibian order Caudata, contains several important model species for biological research. However, there is need to generate transcriptome data from representative species of the primary salamander families. Here we describe a de novo reference transcriptome for a terrestrial salamander, Bolitoglossa vallecula (Caudata: Plethodontidae). We employed paired-end (PE) illumina RNA sequencing to assemble a de novo reference transcriptome for B. vallecula. Assembled transcripts were compared against sequences from other vertebrate taxa to identify orthologous genes, and compared to the transcriptome of a close plethodontid relative (Bolitoglossa ramosi) to identify commonly expressed genes in the skin. This dataset should be useful to future comparative studies aimed at understanding important biological process, such as immunity, wound healing, and the production of antimicrobial compounds.This work was funded by a research grant from COLCIENCIAS 569 (GRANT 027-2103) and CODI (Programa Sostenibilidad) 2013–2014 of the University of Antioquia. A PhD fellowship to the first author, Claudia Arenas was funded by the COLCIENCIAS 567 Grant. We thank the lab of Juan Fernando Alzate from the University of Antioquia for their help in developing our bioinformatic methodological approach. We thank Andrea Gómez and Melisa Hincapie for their help in animal collection and husbandry

A Tree Frog (\u3cem\u3eBoana pugnax\u3c/em\u3e) Dataset of Skin Transcriptome for the Identification of Biomolecules with Potential Antimicrobial Activities

Increases in the prevalence of multiply resistant microbes have necessitated the search for new molecules with antimicrobial properties. One noteworthy avenue in this search is inspired by the presence of native antimicrobial peptides in the skin of amphibians. Having the second highest diversity of frogs worldwide, Colombian anurans represent an extensive natural reservoir that could be tapped in this search. Among this diversity, species such as Boana pugnax (the Chirique-Flusse Treefrog) are particularly notable, in that they thrive in a diversity of marginal habitats, utilize both aquatic and arboreal habitats, and are members of one of few genera that are known to mount a robust immunological response against the fungus Batrachochytrium dendrobatidis, which has decimated the population of frogs worldwide. To search for molecules with potential antimicrobial activity, we have assembled and annotated a reference transcriptome from the skin of four wild captured B. pugnax from Antioquia, Colombia. Analysis of potential antimicrobial and immunological components was performed using ontology analyses, we identified several antimicrobial chemokines with particularly strong potential for exhibiting broadscale antimicrobial activities, as well as several genes related to rapid alteration of transcriptional (KRAB zinc finger protein) and phosphorylation (MAPK) responses to exogenous stressors. We also found eight families of transmembrane transport proteins, including sodium, potassium and voltage-dependent calcium channels, which will be invaluable in future studies aimed at more precisely defining the diversity and function of cationic antimicrobial peptides with alpha-helical structures. These data highlight the utility of frogs such as Boana pugnax in the search of new antimicrobial molecules. Moreover, the molecular datasets presented here allow us to expand our knowledge of this species and illustrate the importance of preserving the vast potential of Colombian biodiversity for the identification of useful biomolecules

Skin substitutes developed by tissue engineering

ABSTRACT: The skin is a complex tissue vulnerable to different processes that may alter its structure, integrity and functionality, among them: burns, chronic wounds and various congenital diseases. Technological advances in biomaterials manufacture and cell culture have allowed the production of skin substitutes thus providing an alternative therapy for some of these complications. This review aims to update the general aspects, composition, future prospects and implementation of the most common skin substitutes currently available in the international market. Some experiences of the Tissue Engineering and Cellular Therapy Group (GITTC), at the University of Antioquia, Faculty of Medicine, in Medellín, Colombia, are also presented.RESUMEN: La piel es un tejido complejo vulnerable a procesos que alteran su estructura, integridad y funcionalidad como, por ejemplo, quemaduras, heridas crónicas y diversas enfermedades congénitas. Los avances tecnológicos en la fabricación de biomateriales y en el cultivo de células han permitido la producción de sustitutos cutáneos que han sido una alternativa terapéutica para algunas de estas complicaciones. Esta revisión pretende actualizar los aspectos generales, composición, perspectivas futuras y de aplicación de los principales sustitutos cutáneos que se ofrecen actualmente en el mercado internacional. Igualmente, presentará algunas experiencias del Grupo de Ingeniería de Tejidos y Terapias Celulares (GITTC) de la Facultad de Medicina de la Universidad de Antioquia

Treatment of cutaneous or mucocutaneous leishmaniasis with anphotericin b liipid formulation, experience in a cohort of patients of the Military Hospital

El Tratamiento de la leishmaniasis cutánea en la población militar es un reto terapéutico para el personal médico de las fuerzas armadas debido a la aparición de cepas resistentes y fracaso del tratamiento a varios regímenes. Realizamos un estudio de series de casos retrospectivo en los pacientes que recibieron anfotericina B formulación lípidica en el HMC para la Leishmaniasis cutánea y mucocutánea con fracaso terapéutico a tratamiento convencional durante enero a diciembre de 2011. Se incluyeron un total de 70 pacientes de diferentes departamentos, que recibieron un curso completo de anfotericina b formulación lipídica (Amphocil ®). Los resultados mostraron que sesenta (85.7%) de setenta pacientes curaron con el primer ciclo de tratamiento. Diez pacientes (14.3%) no cumplieron criterios de curación y requirieron un segundo curso de tratamiento, de los cuales dos presentaron persistencia de las lesiones después de terminar el tratamiento. Fiebre y flebitis ocurrieron en 40% y 33% respectivamente y 4,2% presento toxicidad renal. Finalmente la formulación lipidica de la anfotericina B es una alternativa terapéutica en pacientes con leishmaniasis cutánea y mucocutánea con fracaso terapéutico a los esquemas convencionales con glucantime, pentamidina o anfotericina B desoxicolato .hospital militar centralTreatment of cutaneous leishmaniasis in the military population is a major therapeutic challenge for the medical staff of the Armed Forces because of the emergence of resistant strains and treatment failure to various regimens that some patients present. We performed a retrospective case series study in patients who received amphotericin B lipid formulation on the HMC for cutaneous and mucocutaneous Leishmaniasis with conventional therapeutic failure management schemes during January to December 2011. A total of 70 patients from different departments, which received a full course of amphotericin B lipid formulation (Amphocil ®) were included. The results showed that Sixty (85.7%) out of seventy patients cured with the first cycle of treatment. Ten patients (14.3%) did not meet healing criteria and required a second course of treatment, of which two had persistence of lesions after ending treatment. Fever and phlebitis occurred in 40% and 33% respectively and 4.2% had renal toxicity. Finally Amphotericin B lipid formulation is a therapeutic alternative in patients with cutaneous and mucocutaneous leishmaniasis with therapeutic failure to conventional schemes with glucantime, pentamidine and/or amphotericin B deoxycholat

Sustitutos cutáneos desarrollados por ingeniería de tejidos = Skin substitutes developed by tissue engineering

La piel es un tejido complejo vulnerable a procesos que alteran su estructura, integridad y funcionalidad como, por ejemplo, quemaduras, heridas crónicas y diversas enfermedades congénitas. Los avances tecnológicos en la fabricación de biomateriales y en el cultivo de células han permitido la producción de sustitutos cutáneos que han sido una alternativa terapéutica para algunas de estas complicaciones. Esta revisión pretende actualizar los aspectos generales, composición, perspectivas futuras y de aplicación de los principales sustitutos cutáneos que se ofrecen actualmente en el mercado internacional. Igualmente, presentará algunas experiencias del Grupo de Ingeniería de Tejidos y Terapias Celulares (GITTC) de la Facultad de Medicina de la Universidad de Antioquia

Human gut homeostasis and regeneration: the role of the gut microbiota and its metabolites

peer-reviewedThe healthy human gut is a balanced ecosystem where host cells and representatives of the gut microbiota interact and communicate in a bidirectional manner at the gut epithelium. As a result of these interactions, many local and systemic processes necessary for host functionality, and ultimately health, take place. Impairment of the integrity of the gut epithelium diminishes its ability to act as an effective gut barrier, can contribute to conditions associated to inflammation processes and can have other negative consequences. Pathogens and pathobionts have been linked with damage of the integrity of the gut epithelium, but other components of the gut microbiota and some of their metabolites can contribute to its repair and regeneration. Here, we review what is known about the effect of bacterial metabolites on the gut epithelium and, more specifically, on the regulation of repair by intestinal stem cells and the regulation of the immune system in the gut. Additionally, we explore the potential therapeutic use of targeted modulation of the gut microbiota to maintain and improve gut homeostasis as a mean to improve health outcomes.Enterprise Ireland and the European Union’s Horizon 2020 research and innovation programm

Analysis of High Molecular Mass Compounds from the Spider Pamphobeteus verdolaga Venom Gland. A Transcriptomic and MS ID Approach

Nowadays, spider venom research focuses on the neurotoxic activity of small peptides. In this study, we investigated high-molecular-mass compounds that have either enzymatic activity or housekeeping functions present in either the venom gland or venom of Pamphobeteus verdolaga. We used proteomic and transcriptomic-assisted approaches to recognize the proteins sequences related to high-molecular-mass compounds present in either venom gland or venom. We report the amino acid sequences (partial or complete) of 45 high-molecular-mass compounds detected by transcriptomics showing similarity to other proteins with either enzymatic activity (i.e., phospholipases A2, kunitz-type, hyaluronidases, and sphingomyelinase D) or housekeeping functions involved in the signaling process, glucanotransferase function, and beta-N-acetylglucosaminidase activity. MS/MS analysis showed fragments exhibiting a resemblance similarity with different sequences detected by transcriptomics corresponding to sphingomyelinase D, hyaluronidase, lycotoxins, cysteine-rich secretory proteins, and kunitz-type serine protease inhibitors, among others. Additionally, we report a probably new protein sequence corresponding to the lycotoxin family detected by transcriptomics. The phylogeny analysis suggested that P. verdolaga includes a basal protein that underwent a duplication event that gave origin to the lycotoxin proteins reported for Lycosa sp. This approach allows proposing an evolutionary relationship of high-molecular-mass proteins among P. verdolaga and other spider species

Análise imunofenotípica de amostras de medula óssea normal: aplicações no controle de qualidade em laboratórios de citometria.

Objective. To describe a standardized flow cytometry protocol for the relative and absolute quantification of hematopoietic cell subpopulations from normal bone marrow, and to evaluate the expression of different lineage-specific cell markers with a reactivity associated to cell differentiation to be used as part of the routine quality control in cytometry laboratories. Materials and methods. The immunophenotypical analysis of different cell subpopulations was done with samples from normal bone marrow using a panel of monoclonal and polyclonal antibodies useful in the characterization of acute leukemias with four different fluorescences, by means of a protocol that combines cell labeling of membrane and cytoplasm antigens. Expression analysis was done in terms of mean fluorescence intensity (MFI). Fluorescent beads at a known concentration were added for calculating the absolute count of cells.  Results. The antibody panel used allowed the identification and quantification of different normal leukocyte subpopulations of lymphatic and myeloid origin, including CD34+ stem cells and more differentiated cell populations in the granulocytic, monocytic, and erythroid cell lines. We established reference values for cell populations and cell marker expression ranges as part of routine quality control of cytometry laboratories. Conclusion. Immunophenotypic patterns identified as well as absolute and relative reference values for the different normal leukocyte populations from bone marrow can be used by cytometry laboratories as a basis for establishing reference parameters in phenotypic analyses of hematologic neoplasia. Key words: multiparametric flow cytometry, immunophenotype, hematologic neoplasia, normal bone marrow, reference values, quality control.Objetivo.Describir un protocolo estandarizado mediante citometría de flujo para cuantificar en términos absolutos y relativos distintas subpoblaciones celulares de médula ósea normal y analizar la expresión de diferentes marcadores celulares específicos de linaje cuya reactividad está asociada a la diferenciación celular para ser usado como parte del control de calidad de rutina en los laboratorios de citometría. Materiales y métodos. El análisis inmunofenotípico de distintas subpoblaciones celulares se realizó en muestras de MO normal empleando un panel de anticuerpos monoclonales y policlonales útiles para la caracterización fenotípica de leucemias agudas en 4 fluorescencias distintas, con un protocolo que combina marcaje celular de antígenos de membrana y de citoplasma.El análisis de expresión se realizó en términos de intensidad media de fluorescencia. Para el cálculo de recuentos absolutos se adicionaron esferas fluorescentes de concentración conocida. Resultados. El panel de anticuerpos utilizado permitió la identificación y cuantificación de las distintas subpoblaciones leucocitarias normales de origen linfoide y mieloide incluyendo células precursoras CD34+, y poblaciones celulares más diferenciadas incluidas en las líneas granulocítica, monocítica y eritroide. Se establecieron los valores de referencia de las poblaciones celulares y los rangos de expresión de los diferentes marcadores celulares importantes como parte del control de calidad de rutina en los laboratorios de citometría.Conclusión.Los patrones inmunofenotípicos identificados y la determinación de los valores absolutos y relativos de referencia de las distintas poblaciones leucocitarias normales en MO podrán ser utilizados por los laboratorios de citometría como modelo para establecer parámetros de referencia en el análisis fenotípico de neoplasias hematológicas. Palabras clave: citometría de flujo multiparamétrica, inmunofenotipo, neoplasias hematológicas, médula ósea normal, valores de referencia, control de calidad.Objetivo. Descrever um protocolo padronizado por citometria de fluxo para quantificar em termos absolutos e relativos diferentessubpopulações de células de medula óssea normal e analisar a expressão de diferentes marcadores celulares de linhagem específica, cujareatividade é associada com a diferenciação celular para ser usado como parte do controle de qualidade de rotina nos laboratórios decitometria de fluxo. Materiais e métodos. A análise imunofenotípica das subpopulações de células foi realizada em amostras de MOnormais utilizando um painel de anticorpos monoclonais e policlonais úteis para a caracterização fenotípica de leucemia aguda em quatrofluorescências, com um protocolo que combina rotulagem celular de antígeno de membrana celular e de citoplasma. A análise de expressãofoi realizada em termos de intensidade média de fluorescência. Para calcular a recontagem absoluta foram adicionadas esferas fluorescentesde concentração conhecida. Resultados. O painel de anticorpos utilizado permitiu a identificação e quantificação das subpopulações deleucócitos normais de origem linfóide e mielóide incluindo as células precursoras CD34+, e populações de células mais diferenciadasincluídas nas linhas granulocítica, monocítica e eritróide. Foram estabelecidos os valores de referência das populações celulares e osintervalos de expressão dos diferentes marcadores celulares importantes como parte da rotina de controle de qualidade em laboratórios decitometria. Conclusão. Os padrões imunofenotípicos identificados e a determinação dos valores absolutos e relativos de referência dasdiferentes populações de leucócitos normais em MOM podem ser utilizados pelos laboratórios de citometria como um modelo paraestabelecer parâmetros de referencia na análise fenotípica de neoplasias hematológicas.Palavras-chave: citometria de fluxo multiparâmetro, imunofenótipo, neoplasias hematológicas, medula óssea normal, valores de referência,controle de qualidade

Análise imunofenotípica de amostras de medula óssea normal: aplicações no controle de qualidade em laboratórios de citometria.

Objective. To describe a standardized flow cytometry protocol for the relative and absolute quantification of hematopoietic cell subpopulations from normal bone marrow, and to evaluate the expression of different lineage-specific cell markers with a reactivity associated to cell differentiation to be used as part of the routine quality control in cytometry laboratories. Materials and methods. The immunophenotypical analysis of different cell subpopulations was done with samples from normal bone marrow using a panel of monoclonal and polyclonal antibodies useful in the characterization of acute leukemias with four different fluorescences, by means of a protocol that combines cell labeling of membrane and cytoplasm antigens. Expression analysis was done in terms of mean fluorescence intensity (MFI). Fluorescent beads at a known concentration were added for calculating the absolute count of cells.  Results. The antibody panel used allowed the identification and quantification of different normal leukocyte subpopulations of lymphatic and myeloid origin, including CD34+ stem cells and more differentiated cell populations in the granulocytic, monocytic, and erythroid cell lines. We established reference values for cell populations and cell marker expression ranges as part of routine quality control of cytometry laboratories. Conclusion. Immunophenotypic patterns identified as well as absolute and relative reference values for the different normal leukocyte populations from bone marrow can be used by cytometry laboratories as a basis for establishing reference parameters in phenotypic analyses of hematologic neoplasia. Key words: multiparametric flow cytometry, immunophenotype, hematologic neoplasia, normal bone marrow, reference values, quality control.Objetivo.Describir un protocolo estandarizado mediante citometría de flujo para cuantificar en términos absolutos y relativos distintas subpoblaciones celulares de médula ósea normal y analizar la expresión de diferentes marcadores celulares específicos de linaje cuya reactividad está asociada a la diferenciación celular para ser usado como parte del control de calidad de rutina en los laboratorios de citometría. Materiales y métodos. El análisis inmunofenotípico de distintas subpoblaciones celulares se realizó en muestras de MO normal empleando un panel de anticuerpos monoclonales y policlonales útiles para la caracterización fenotípica de leucemias agudas en 4 fluorescencias distintas, con un protocolo que combina marcaje celular de antígenos de membrana y de citoplasma.El análisis de expresión se realizó en términos de intensidad media de fluorescencia. Para el cálculo de recuentos absolutos se adicionaron esferas fluorescentes de concentración conocida. Resultados. El panel de anticuerpos utilizado permitió la identificación y cuantificación de las distintas subpoblaciones leucocitarias normales de origen linfoide y mieloide incluyendo células precursoras CD34+, y poblaciones celulares más diferenciadas incluidas en las líneas granulocítica, monocítica y eritroide. Se establecieron los valores de referencia de las poblaciones celulares y los rangos de expresión de los diferentes marcadores celulares importantes como parte del control de calidad de rutina en los laboratorios de citometría.Conclusión.Los patrones inmunofenotípicos identificados y la determinación de los valores absolutos y relativos de referencia de las distintas poblaciones leucocitarias normales en MO podrán ser utilizados por los laboratorios de citometría como modelo para establecer parámetros de referencia en el análisis fenotípico de neoplasias hematológicas. Palabras clave: citometría de flujo multiparamétrica, inmunofenotipo, neoplasias hematológicas, médula ósea normal, valores de referencia, control de calidad.Objetivo. Descrever um protocolo padronizado por citometria de fluxo para quantificar em termos absolutos e relativos diferentessubpopulações de células de medula óssea normal e analisar a expressão de diferentes marcadores celulares de linhagem específica, cujareatividade é associada com a diferenciação celular para ser usado como parte do controle de qualidade de rotina nos laboratórios decitometria de fluxo. Materiais e métodos. A análise imunofenotípica das subpopulações de células foi realizada em amostras de MOnormais utilizando um painel de anticorpos monoclonais e policlonais úteis para a caracterização fenotípica de leucemia aguda em quatrofluorescências, com um protocolo que combina rotulagem celular de antígeno de membrana celular e de citoplasma. A análise de expressãofoi realizada em termos de intensidade média de fluorescência. Para calcular a recontagem absoluta foram adicionadas esferas fluorescentesde concentração conhecida. Resultados. O painel de anticorpos utilizado permitiu a identificação e quantificação das subpopulações deleucócitos normais de origem linfóide e mielóide incluindo as células precursoras CD34+, e populações de células mais diferenciadasincluídas nas linhas granulocítica, monocítica e eritróide. Foram estabelecidos os valores de referência das populações celulares e osintervalos de expressão dos diferentes marcadores celulares importantes como parte da rotina de controle de qualidade em laboratórios decitometria. Conclusão. Os padrões imunofenotípicos identificados e a determinação dos valores absolutos e relativos de referência dasdiferentes populações de leucócitos normais em MOM podem ser utilizados pelos laboratórios de citometria como um modelo paraestabelecer parâmetros de referencia na análise fenotípica de neoplasias hematológicas.Palavras-chave: citometria de fluxo multiparâmetro, imunofenótipo, neoplasias hematológicas, medula óssea normal, valores de referência,controle de qualidade