Study of the migration of Fasciola hepatica juveniles across the intestinal barrier of the host by quantitative proteomics in an ex vivo model.

Fasciola hepatica is a trematode parasite that infects animals and humans causing fasciolosis, a worldwide-distributed disease responsible for important economic losses and health problems. This disease is of growing public health concern since parasite isolates resistant to the current treatment (triclabendazole) have increasingly been described. F. hepatica infects its vertebrate host after ingestion of the encysted parasite (metacercariae), which are found in the water or attached to plants. Upon ingestion, newly excysted juveniles of F. hepatica (FhNEJ) emerge in the intestinal lumen and cross the intestinal barrier, reach the peritoneum and migrate to the biliary ducts, where adult worms fully develop. Despite the efforts made to develop new therapeutic and preventive tools, to date, protection against F. hepatica obtained in different animal models is far from optimal. Early events of host-FhNEJ interactions are of paramount importance for the infection progress in fasciolosis, especially those occurring at the host-parasite interface. Nevertheless, studies of FhNEJ responses to the changing host environment encountered during migration across host tissues are still scarce. Here, we set-up an ex vivo model coupled with quantitative SWATH-MS proteomics to study early host-parasite interaction events in fasciolosis. After comparing tegument and somatic fractions from control parasites and FhNEJ that managed to cross a mouse intestinal section ex vivo, a set of parasite proteins whose expression was statistically different were found. These included upregulation of cathepsins L3 and L4, proteolytic inhibitor Fh serpin 2, and a number of molecules linked with nutrient uptake and metabolism, including histone H4, H2A and H2B, low density lipoprotein receptor, tetraspanin, fatty acid binding protein a and glutathione-S-transferase. Downregulated proteins in FhNEJ after gut passage were more numerous than the upregulated ones, and included the heath shock proteins HSP90 and alpha crystallin, amongst others. This study brings new insights into early host-parasite interactions in fasciolosis and sheds light on the proteomic changes in FhNEJ triggered upon excystment and intestinal wall crossing, which could serve to define new targets for the prevention and treatment of this widespread parasitic disease.

Recognition Pattern of the Fasciola hepatica Excretome/Secretome during the Course of an Experimental Infection in Sheep by 2D Immunoproteomics.

Excretory/secretory products released by helminth parasites have been widely studied for their diagnostic utility, immunomodulatory properties, as well as for their use as vaccines. Due to their location at the host/parasite interface, the characterization of parasite secretions is important to unravel the molecular interactions governing the relationships between helminth parasites and their hosts. In this study, the excretory/secretory products from adult worms of the trematode Fasciola hepatica (FhES) were employed in a combination of two-dimensional electrophoresis, immunoblot and mass spectrometry, to analyze the immune response elicited in sheep during the course of an experimental infection. Ten different immunogenic proteins from FhES recognized by serum samples from infected sheep at 4, 8, and/or 12 weeks post-infection were identified. Among these, different isoforms of cathepsin L and B, peroxiredoxin, calmodulin, or glutathione S-transferase were recognized from the beginning to the end of the experimental infection, suggesting their potential role as immunomodulatory antigens. Furthermore, four FhES proteins (C2H2-type domain-containing protein, ferritin, superoxide dismutase, and globin-3) were identified for the first time as non-immunogenic proteins. These results may help to further understand host/parasite relationships in fasciolosis, and to identify potential diagnostic molecules and drug target candidates of F. hepatica.

Potential Influence of Helminth Molecules on COVID-19 Pathology.

In recent months, the parasitology research community has been tasked with investigation of the influence of parasite coinfection on coronavirus disease 2019 (COVID-19) outcomes. Herein, we share our approach to analyze the effect of the trematode Fasciola hepatica as a modulator of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and of COVID-19 pathology.

Cartilage regeneration using a novel autologous growth factors-based matrix for full-thickness defects in sheep.

PURPOSE: To investigate the chondrogenic-regenerative properties of a novel autologous-made matrix composed of hyaline cartilage chips combined with a growth factors-based clot for full-thickness defects in sheep. METHODS: A full-thickness, 8-mm diameter cartilage defect was created in the weight-bearing area of the medial femoral condyle in 6 sheep. Treatment consisted of surgical implantation of an autologous-based matrix of hyaline cartilage chips combined with a clot of plasma poor in platelets and intraarticular injection of plasma rich in growth factors. Outcome measures at 1, 3 and 6 months included macroscopic International Cartilage Repair Society (ICRS) score, histological and immunohistochemical analysis for collagen expression, and transmission electron microscopy study. RESULTS: The 6-month macroscopic evaluation showed nearly normal (11.1 ± 0.7) cartilage repair assessment. The ICRS score was significantly higher at 6 months compared to 3 months (5.5 ± 1.3; p < 0.0001) and 1 (1.1 ± 0.4; p < 0.0001) month. At 6 months, hyaline cartilage tissue filling the defect was observed with adequate integration of the regenerated cartilage at the surrounding healthy cartilage margin. At 6 months, mature chondrons and cartilage matrix contained collagen fibers with masked fibrillary structure, and the expression of collagen in the newly formed cartilage was similar in intensity and distribution pattern compared to the healthy adjacent cartilage. CONCLUSIONS: This novel treatment enhanced chondrogenesis and regenerated hyaline cartilage at 6 months with nearly normal macroscopic ICRS assessment. Histological analysis showed equivalent structure to mature cartilage tissue in the defect and a collagen expression pattern in the newly formed cartilage similar to that found in adjacent healthy articular cartilage. The present technique may have clinical application for chondral injuries in humans because this procedure is cheap (no need for allograft, or expensive instrumentation/biomaterials/techniques), easy and fast-performing through a small arthrotomy, and safe (no rejection possibility because the patients' own tissue, cells, and plasma are used).

La región del cuello del caballo: connotaciones anátomo-aplicativas: una revisión / The region of the neck of the horse: anatomic applicative connotations: a review

La anatomía topográfica veterinaria es el estudio de las relaciones entre las diferentes partes del organismo y es uno de los métodos de estudio anatómico de los animales, es subsiguiente, al otro tipo de estudio, que es de carácter descriptivo, referido a la forma y estructura orgánica mediante una clasificación de órganos por aparatos y sistemas. La región anatómica del cuello es una importante área anatómica y fisiológica por sus múltiples connotaciones anatomo-clínicas. Se plantea como objetivo una revisión anatómica de la región del cuello del caballo y sus connotaciones anatomo-aplicadas. Se realizó una extensa revisión de la literatura de la anatomía del cuello del caballo así como sus connotaciones anatomo-aplicativas, las cuales se describen detalladamente. En la actualidad la región dorsal del cuello ha sido ampliamente estudiada en los caballos debido a su correlación con el síndrome metabólico equino y obesidad en el caballo. En conclusión la región topográfica del cuello representa anatómicamente una gran significación clínica en el caballo por las estructuras y accidentes anatómicos.

Veterinary topographic anatomy is the study of relationships between different parts of the body and is one of the methods of anatomical study of animals; it is subsequent to the other study, which is descriptive based on the shape and structure rated by organic organs organ systems. The anatomical neck region is an important anatomical and physiological area because of its many anatomical and clinical implications. The aim of this study was to a review of an anatomical region of the horse's neck and applied anatomical connotations. An extensive literature review of the anatomy of the horse's neck and its applicative anatomical connotations, which are described in detail, was conducted. Currently the dorsal neck region has been extensively studied in horses due to their correlation with equine metabolic syndrome and obesity in the horse. In conclusion topographic anatomical neck region represents a great clinical significance in the horse by accident and anatomical structures.

The Sigma class glutathione transferase from the liver fluke Fasciola hepatica.

BACKGROUND: Liver fluke infection of livestock causes economic losses of over US$ 3 billion worldwide per annum. The disease is increasing in livestock worldwide and is a re-emerging human disease. There are currently no commercial vaccines, and only one drug with significant efficacy against adult worms and juveniles. A liver fluke vaccine is deemed essential as short-lived chemotherapy, which is prone to resistance, is an unsustainable option in both developed and developing countries. Protein superfamilies have provided a number of leading liver fluke vaccine candidates. A new form of glutathione transferase (GST) family, Sigma class GST, closely related to a leading Schistosome vaccine candidate (Sm28), has previously been revealed by proteomics in the liver fluke but not functionally characterised. METHODOLOGY/PRINCIPAL FINDINGS: In this manuscript we show that a purified recombinant form of the F. hepatica Sigma class GST possesses prostaglandin synthase activity and influences activity of host immune cells. Immunocytochemistry and western blotting have shown the protein is present near the surface of the fluke and expressed in eggs and newly excysted juveniles, and present in the excretory/secretory fraction of adults. We have assessed the potential to use F. hepatica Sigma class GST as a vaccine in a goat-based vaccine trial. No significant reduction of worm burden was found but we show significant reduction in the pathology normally associated with liver fluke infection. CONCLUSIONS/SIGNIFICANCE: We have shown that F. hepatica Sigma class GST has likely multi-functional roles in the host-parasite interaction from general detoxification and bile acid sequestration to PGD synthase activity.

Características de las solicitudes de acreditación de actividades de formación continuada de las profesiones sanitarias en una Comunidad Autónoma / Characteristics of accreditation applications for continuing education activities of the health care proffessionals in a Regional Community

Objetivos. Conocer las características generales de las solicitudes de acreditación de actividades de formación continuada dirigidas a profesionales sanitarios en la Comunidad Autónoma de Madrid, dentro del sistema de acreditación de formación continuada del Sistema Nacional de Salud establecido en España. Proponer actuaciones futuras que contribuyan a mejorar la calidad de las actividades de formación continuada. Métodos. Estudio descriptivo retrospectivo transversal. Se incluyeron todas las solicitudes de acreditación de actividades docentes de formación continuada dirigidas a profesionales sanitarios, para las que se solicitó acreditación en la Comunidad de Madrid, desde 2002 a 2004, en una base de datos y se estudiaron 9 variables, que se analizaron con el programa informático SPSS 11.0. Resultados. Se han recibido 6.869 solicitudes, con un número total de horas programadas de 188.027 y un total de alumnos de 342.361. Existe un gran incremento de solicitudes en el período considerado. Las actividades más frecuentes son presenciales (93,9%), en forma de cursos (71,4%), de entre 21 y 40 alumnos (51,3%), contenido centrado en la práctica clínica (77,5%), de menos de 21 horas lectivas de duración (68,1%), dirigidas en exclusiva a una sola profesión (71,7%) que suele ser Medicina (60%) y solicitadas por entidades privadas (60,4%). Las organizaciones empresariales son las solicitantes más frecuentes (24,4%). Se acreditó el 91,6% de las actividades. Conclusiones. La acreditación de actividades de formación continuada está cobrando gran importancia. El análisis de las características de estas actividades puede permitir conocer y mejorar los diversos aspectos que ayuden a garantizar un incremento en la calidad de la formación continuada ofertada. En este sentido creemos necesario: a) desarrollar y consolidar un cuerpo de conocimientos comunes en todo el sistema de acreditación; b) aumentar la coordinación entre todas las estructuras implicadas en dicho sistema, y c) avanzar en la realización de auditorías y en la acreditación de centros de formación continuada

Objectives. To know the general characteristics of the applications for accreditation of continuing education activities for healthcare professionals in the "Madrid Community" within the Spanish continuing education accreditation system of the Spanish National Healthcare system. To propose future lines of work that contribute to the improvement in the quality of these activities. Methods. Cross-sectional retrospective descriptive study. All requests for accreditation of continuing education activities for healthcare professionals received in the "Madrid Community" (Madrid, Spain) between 2002 and 2004 have been included in a database where 9 different variables were analyzed with SPSS 11.0. Results. We received 6,869 applications with 188,027 programmed hours and 342,361 students. There is an increase in the number of applications during the period studied. The most frequent activities are those in which the student has to be present in the classroom (93.9%), courses (71.4%), those with 21 to 40 students (51.3%), focused on clinical practice (77.5%), with less than 21 hours (68.1%), aimed at a single profession (usually medicine -60%-) (71.7%) and requested by private organizations (60.4%). Companies are the most frequent organizations that request accreditation (24.4%). A total of 91.6% of activities were accredited. Conclusions. Accreditation of continuing education activities has become important for health professionals. The analysis of the characteristics of these activities makes it possible to know and improve the different aspects that guarantee an increase in the quality of continuing education offered. According to the above, we consider that the following is necessary: a) to develop and consolidate a common knowledge pool for the whole accreditation system; b) to increase coordination between all the different structures within the accreditation system, and c) to make progress in the auditing and accreditation of continuing education institutions