Development of an accurate and rapid method for whole genome characterization of canine parvovirus.

Canine parvovirus is a highly contagious pathogen affecting domestic dogs and other carnivores globally. Monitoring CPV through continuous genomic surveillance is crucial for mapping variability and developing effective control measures. Here, we developed a method using multiplex-PCR-next-generation sequencing to obtain full-length CPV genomes directly from clinical samples. This approach utilizes tiling and tailed amplicons to amplify overlapping fragments of roughly 250 base pairs. This enables the creation of Illumina libraries by conducting two PCR reaction runs. We tested the assay in 10 fecal samples from dogs diagnosed with CPV and one CPV-2 vaccine strain. Furthermore, we applied it to a feline sample previously diagnosed with the feline panleukopenia virus. The assay provided 100 % genome coverage and high sequencing depth across all 12 samples. It successfully provided the sequence of the coding regions and the left and right non-translated regions, including tandem and terminal repeats. The assay effectively amplified viral variants from divergent evolutionary groups, including the antigenic variants (2a, 2b, and 2c) and the ancestral CPV-2 strain included in vaccine formulations. Moreover, it successfully amplified the entire genome of the feline panleukopenia virus found in cat feces. This method is cost-effective, time-efficient, and does not require lab expertise in Illumina library preparation. The multiplex-PCR-next-generation methodology facilitates large-scale genomic sequencing, expanding the limited number of complete genomes currently available in databases and enabling real-time genomic surveillance. Furthermore, the method helps identify and track emerging CPV viral variants, facilitating molecular epidemiology and control. Adopting this approach can enhance our understanding of the evolution and genetic diversity of Protoparvovirus carnivoran1.

Model of Liver Fibrosis Induction by Thioacetamide in Rats for Regenerative Therapy Studies.

Hepatic fibrosis is caused by chronic injury due to toxic, infectious, or metabolic causes, and it may progress to cirrhosis and hepatocellular carcinoma. There is currently no antifibrotic therapy authorized for human use; however, there are promising studies using cell therapies. There are also no animal models that exactly reproduce human liver fibrosis that can be used to better understand the mechanisms of its regression and identify new targets for treatment and therapeutic approaches. On the other hand, mesenchymal stem cells (MSC) have experimentally demonstrated fibrosis regression effects, but it is necessary to have an animal model of advanced liver fibrosis to evaluate the effect of these cells. The aim of this work was to establish a protocol for the induction of advanced liver fibrosis in rats using thioacetamide (TAA), which will allow us to perform trials using MSC as a possible therapy for fibrosis regression. For this purpose, we selected 24 female rats and grouped them into three experimental groups: the control group (G-I) without treatment and groups II (G-II) and III (G-III) that received TAA by intraperitoneal injection for 24 weeks. Then, 1 × 106/kg adipose mesenchymal stem cells (ASCs) were infused intravenously. Groups G-I and G-II were sacrificed 7 days after the last dose of ASC, and G-III was sacrificed 8 weeks after the last ASC infusion, all with xylazine/ketamine (40 mg/kg). The protocol used in this work established a model of advanced hepatic fibrosis as corroborated by METAVIR tests of the histological lesions; by the high levels of the markers α-SMA, CD68, and collagen type I; by functional alterations due to elevated markers of the hepatic lesions; and by alterations of the leukocytes, lymphocytes, and platelets. Finally, transplanted cells in the fibrous liver were detected. We conclude that TAA applied using the protocol introduced in this study induces a good model of advanced liver fibrosis in rats.

Association between PIK3CA Mutations in Blood and Tumor-Infiltrating Lymphocytes in Peruvian Breast Cancer Patients.

OBJECTIVE: To evaluate the relationship between circulating tumor DNA (ctDNA) presence and tumor features including tumor-infiltrating lymphocyte (TIL) levels in Peruvian breast cancer patients. MATERIALS AND METHODS: This was a prospective study conducted at the Instituto Nacional de Enfemedades Neoplasicas, Peru. We evaluated level of TIL and PIK3CA mutations in ctDNA. Clinical characteristics, including outcome data, were collected from the patient file. Survival was calculated from the date of blood sample drawn to the event time. Data collected were analyzed using SPSS software version 25. RESULTS: We analyzed plasma samples from 183 breast cancer patients. most cases were of Luminal-B (44.8%) phenotype and stage II (41.5%), and median stromal TIL was 30%. PIK3CA mutation in ctDNA was detected in 35% cases (most with E545K) and was associated with lower TIL level (p=0.04). PIK3CA in ctDNA tended to be associated with advanced stages (p=0.09) in the whole series and with higher recurrence rates (p=0.053) in the non-metastatic setting. Patients with presence of PIK3CA in ctDNA tended to have shorter survival (p=0.083). CONCLUSION: Presence of PIK3CA mutation in ctDNA was frequently found in our Peruvian breast cancer series, was associated with lower TIL levels and tended to predict poor outcomes.

First report of canine morbillivirus infection of adipose tissue-derived stem cells from dogs with distemper.

Background and Aim: Ribonucleic acid viruses remain latent in different cell types, including mesenchymal stem cells; however, the distemper virus remains undetected in these cells. This study aimed to determine whether adipose stem cells (ASCs) from dogs with distemper disease are infected with the canine morbillivirus (CM). Materials and Methods: Twelve dogs with the neurological phase of the disease and who were positive for CM by reverse transcription polymerase chain reaction (RT-PCR), were studied. ASCs from adipose tissue of the lesser omentum of these infected dogs were isolated and characterized. Direct fluorescence was used to detect the viral antigen in cell cultures. Flow cytometry and RT-PCR identified detectable quantities of the virus in two cultures, while electron microscopy confirmed the CM particles within ASCs. Results: This study revealed that ASCs of the omentum of dogs with distemper disease can be infected with CM, indicating their possible involvement in this virus latency and persistence. This suggests that its detection should be considered within the quality control process of stem cells intended for regenerative medicine. Conclusion: To the best of our knowledge, this is the first study that demonstrates that omentum ASCs from dogs with distemper disease can be infected with CM and may be involved in viral latency or persistence. Our study also suggests that the detection of CM should be considered within the quality control process of stem cells intended for regenerative medicine.

Biological characteristics of a sub-population of cancer stem cells from two triple-negative breast tumour cell lines.

Triple-negative breast tumours (TNBTs) make up 15-20% of all breast tumours. There is no treatment for them, and the role that cancer stem cells (CSCs) have in carcinogenesis is still unclear, so finding markers and therapeutic targets in CSC exosomes requires these cells to exist as a homogeneous cell population. The objective of this work was to determine differences in ultrastructural morphology, proliferative capacity, and mouse-xenotransplantation characteristics of the MDA-MB-231 and MDA-MB-436 TNBT cell lines with the CD44 high /CD24 low phenotype in order to study their exosomes. The results show that the CD44 high /CD24 low MBA-MB-231 cells had a population doubling time of 41.56 h, compared to 44.79 h in the MDA-MB-436 cell line. After magnetic immunoseparation, 18.75% and 14.56% of the stem cell population of the MDA-MB-231 and MDA-MB-436 cell lines, respectively, were of the CD44 high /CD24 low phenotype, which were expanded to reach purities of 80.4% and 87.6%. The same expanded lineage in both cell lines was shown to possess the pluripotency markers Nanog and Oct4. Under a scanning electron microscope, the CD44 high /CD24 low lineage of the MBA-MD-231 cell line formed groups of more interconnected cells than this lineage of the MBA-MD-436 line. A total of 16% of the mice inoculated with the CD44 high /CD24 low lineage of either cell line presented tumours of the breast, lung, and submandibular ganglia, in whose tissues variable numbers of inoculated cells were found 30 days post-inoculation. By magnetic immunoselection, it was possible to isolate in similar quantities and characterize, expand, and xenotransplant the CD44 high /CD24 low lineage of the MDA-MB-231 and MDA-MB-436 cell lines. The former cell line has greater proliferative capacity, the two lines differ under scanning electron microscopy in how they intercommunicate, and both cell lines induce new tumours in mice and persist at least 30 days post-inoculation in the transplanted animal so their exosomes would also be different.

Multidose intramuscular allogeneic adipose stem cells decrease the severity of canine atopic dermatitis: A pilot study.

AIM: The aim of this pilot study was to evaluate the therapeutic and safety performance of an intramuscular treatment protocol of multidose of allogeneic adipose stem cells (ASCs) isolated, characterized, and expanded ex vivo from a healthy canine donor. MATERIALS AND METHODS: Twelve dogs diagnosed with canine atopic dermatitis (CAD) were intramuscularly treated with 0.5×106 of cryopreserved ASCs from a healthy immunized young canine Ehrlichia canis free donor weekly for 6 weeks. Treatment efficacy was evaluated by the pruritus index and the CAD Lesion Index (CADLI) test. Safety and adverse effects were determined by injection site reaction, weight, blood chemistry, liver function, and whole blood count. RESULTS: Canine ASCs obtained from a donor met the minimum qualities required for this type of cells and showed viability of 90% after thawing. The efficacy of the CADLI score and the pruritus index in 12 dogs with atopic dermatitis was statistically significant efficacy. No adverse reactions were observed at the intramuscular application site, or in relation to animal weight, blood cell populations, or liver and renal function. CONCLUSION: These results suggest that intramuscular administration of cryopreserved ASCs to dogs with atopic dermatitis is a promising cellular therapeutic product for the relief of the symptoms of this disease; however, the duration of the effects obtained with this dose and with other doses should be evaluated, as well as possible immune reactions. As far as we know, this is the first report of the use of multiple intramuscular doses cryopreserved ASCs to treat atopic dermatitis.

Educación continuada en Radiología: Perspectivas hacia un modelo basado en competencias / Continuing Education Radiology: Perspectives towards a Model Based On Competences

La educación médica continuada hace referencia a la planeación, organización, desarrollo y ejecución de diversas actividades académicas, con el fin de actualizar los conocimientos y desarrollar competencias a lo largo de la vida profesional de los posgraduados. En la especialidad de radiología e imágenes diagnósticas, la renovación del conocimiento va de la mano de los adelantos tecnológicos y la mejor evidencia científica, lo que implica un alto dinamismo en la validez de los mismos. En el presente artículo se revisa cuál es el propósito y justificación de un programa de recertificación en radiología con miras al mejoramiento de la calidad y el desempeño profesional, se discuten diferentes modelos internacionales de formación continuada, así como las metodologías más eficientes para la adquisición de competencias, ajustado a las necesidades y perspectivas de aprendizaje del radiólogo en nuestro país

The continuing medical education (CME) refers to the planning, organization, development and implementation of various academic activities, in order to update knowledge and develop skills throughout the professional life of postgraduates. In the specialty of Radiology and Diagnostic Imaging, renewal of knowledge goes hand in hand with technological advances and the best scientific evidence, which implies a high dynamism in the validity thereof. In this article we review what the purpose and justification of a recertification program in radiology with a view to improving the quality and professional performance, different international models of continuing education are discussed as well as the most efficient methodologies for the acquisition of competencies, tailored to the learning needs and perspectives of the radiologist in our country

Diagnóstico prenatal de acretismo placentario: hallazgos y utilidad del ultrasonido y la resonancia magnética: Reporte de casos en el Hospital Universitario de la Fundación Santa Fe de Bogotá (Colombia) / Prenatal diagnosis of placental accretism: findings and role of ultrasound and magnetic resonance: Cases report from university hospital of Fundación Santa Fe de Bogotá (Colombia)

Objetivo: describir y revisar los hallazgos relevantes en imágenes diagnósticas del abordaje prenatal de pacientes con diagnóstico de acretismo placentario. Materiales y métodos: presentamos los casos de tres mujeres que fueron atendidas en un centro hospitalario de alta complejidad, con factores de riesgo para acretismo placentario y evaluación prenatal por ultrasonido (US) y resonancia magnética (RM), cuyo diagnóstico fue corroborado con estudio anatomopatológico. Se describen los aspectos clínicos, los hallazgos por imágenes y su correlación histológica. Se realizó una búsqueda de la literatura a través de PubMed, Lilacs y EBSCO, se incluyeron artículos originales, revisiones de tema y reportes de casos sobre la utilidad y los aspectos más importantes del US y la RM en el diagnóstico prenatal de acretismo placentario. Resultados: se incluyeron 11 artículos originales, 8 de revisión y 1 reporte caso. La placenta previa asociada a lagunas placentarias en ultrasonido y en RM a bandas hipointensas placentarias y el abombamiento del útero son considerados como los principales signos de acretismo. El US asociado a Doppler color tiene una sensibilidad de 97 % y una especificidad del 92 %. La RM puede definir el compromiso uterino y de los órganos vecinos, con una sensibilidad que varía entre el 77 y el 89,6 %, la especificidad es cercana al 92 %. Conclusiones: el abordaje prenatal precoz a través de las imágenes diagnósticas no invasivas brinda información de utilidad a los clínicos del compromiso y la extensión en la evaluación de pacientes con factores de riesgo para acretismo placentario.

Objective: To describe and review the relevant findings of the prenatal diagnostic imaging approach in patients placenta accreta diagnosis. Materials and methods: We present 3 cases of women with risk factors for placenta accreta assessed prenatally at a high-complexity hospital, using ultrasound (US) and magnetic resonance imaging (MRI), with histopathological confirmation of the diagnosis. Clinical and imaging findings are described, together with their histological correlation. A search of the literature was conducted through PubMed, LILACS, and Ebsco, including original papers, topic reviews and case reports on the use and more important aspects of US and MRI in the prenatal diagnosis of placenta accreta. Results: 11 Original articles, 8 Review articles and 1 case report from our country were included. Placenta accreta is an obstetric pathology that threatens the health of the mother and fetus. Placenta previa associated to placental gaps in US and intraplacental hypointense bands and uterine bulge in MRI are considered as the major signs of accretism. US with color Doppler has 97 % of sensitivity and 92 % of specificity, MRI defines uterine and neighboring organs compromise (s = 77-89.6 %, e = 92 %). Conclusion: Early prenatal approach through noninvasive diagnostic imaging plays a usefull role providing information about the compromise and extention in patients with risk factors for placental accretism.

Estrategias diagnósticas dirigidas al seguimiento de los pacientes con cáncer tratados con nuevas terapias moleculares / Diagnostic strategies aimed to monitor cancer patients treated with new molecular therapies

La evaluación objetiva de la respuesta a la terapia anticáncer es muy importante. La estandarización y la utilización de un lenguaje común es fundamental para poder comparar las terapias en diferentes escenarios. Se puede afirmar que existía una relación directa entre tamaño tumoral y la respuesta, axioma derivado de los criterios de la OMS desde 1979. Debido a las limitaciones de estos criterios, los criterios RECIST fueron creados en el 2000; ahora se reconoce que las imágenes diagnósticas son el pilar en el seguimiento de los pacientes con cáncer. Sin embargo, con la aparición de nuevas tecnologías ha llevado a nuevos criterios, como los RECIST 1.1 y los PERCIST, una vez se reconoció la deficiencia de aquellos creados en el 2000. Mientras tanto, se han diseñado nuevas terapias anticáncer con mecanismos de acción molecular, lo cual ha demostrado que la evaluación anatómica no es actualmente el único parámetro y se debe correlacionar con técnicas de perfusión y de función, como Doppler, resonancia magnética o TA C y la inclusión de nuevos criterios de seguimiento por PET (PERCIST). El principal objetivo de este artículo de revisión es reconocer de manera suficiente los estándares actuales en términos de criterios RECIST, su aplicación práctica, las limitaciones actuales y cómo los radiólogos oncólogos están progresando en la evaluación de los tumores según su comportamiento biológico y la terapia que el paciente esté recibiendo.

Imágenes de resonancia magnética como demostración de recurrencia de cáncer de seno posterior a mastectomía radical y reconstrucción con colgajo músculo-cutáneo transverso de recto abdominal: presentación de caso / Magnetic resonance imaging in the evaluation of recurrent breast cancer after radical mastectomy and transverse rectus abdominis musculocutaneous flap reconstruction: case report

La reconstrucción con colgajo TRAM (colgajo musculo-cutáneo transverso de recto abdominal) es una técnica quirúrgica ampliamente utilizada en pacientes que han sido sometidas a mastectomía radical. La recurrencia local de cáncer de seno se debe al tejido glandular remanente posterior a la mastectomía; su incidencia varía entre 3,8% y 10,6%, dependiendo del estadio de la enfermedad inicial, el subtipo histológico y su comportamiento biológico. Estas recurrencias pueden ser detectadas mediante el examen físico e imágenes diagnósticas, dentro de las cuales la resonancia magnética es la modalidad que ofrece la mejor diferenciación entre cambios posquirúrgicos esperados y recurrencias; sin embargo no se ha establecido un protocolo de seguimiento para estas pacientes. En este artículo se presenta un caso y se revisan las características imaginológicas propias de la reconstrucción como de una recidiva tumoral.

Tratamiento de las lesiones meniscales de acuerdo con la categorización morfológica: concordancia entre resonancia magnética y artroscopia / Treatment of meniscal lesions according to the morphological categorization: agreement between MRI and arthroscopy

Objetivo: determinar la concordancia entre resonancia magnética (RM) y artroscopia de rodilla como patrón de oro en la escogencia del tipo de tratamiento para una ruptura meniscal con base en su caracterización morfológica. Materiales y métodos: estudio de concordancia diagnóstica de conformidad que incluyó pacientes que consultaron a la Fundación Santa Fe con lesiones meniscales, a quienes se practicó RM con cortes axiales, y fueron llevados a artroscopia. Se correlacionaron posibles tratamientos de acuerdo con configuración de la ruptura según hallazgos imagenológicos y artroscópicos, y se determinó concordancia entre éstos. Resultados: el coeficiente Kappa calculado fue de 0,61. Conclusiones: la concordancia encontrada entre RM y artroscopia es buena permitiendo predecir el posible tratamiento de acuerdo con la configuración de la ruptura.

Adherencia al uso de sistema bi-rads en el reporte mamográfico y concordancia entre la clasificación específica en estudios de tamizaje en Colombia / Adherence to the Bi-Rads system in mammographic reports and concordance with specific classification in screening studies in Colombia

El artículo busca examinar la frecuencia del uso de BI-RADS en el reporte mamográfico después de seis años de la primera publicación en Colombia y posteriores esfuerzos de difusión en el país, así como identificar los tipos de errores más frecuentes en la concordancia entre la categoría BI-RADS asignada y las recomendaciones específicas de ésta, según lo estipulado por el Colegio Americano de Radiología. Se realizó un muestreo por conveniencia de reportes mamográficos de diferentes centros radiológicos del país, entre 1996 y 2004. Se encontró una adherencia significativa (87,21) a la utilización de la clasificación BI-RADS en el reporte mamográfico. La concordancia general encontrada fue discreta (60 por ciento)...