Optimización de la protección en radiología y cardiología intervencionista pediatrica en América Latina y el Caribe (OPRIPALC) / Optimization of protection in pediatric interventional cardiology and radiology in Latin America and the Caribbean (OPRIPALC)

El objetivo del presente artículo ha sido describir el programa "Optimización de la Protección en Radiología Intervencionista Pediátrica en América Latina y el Caribe" (OPRIPALC) que nace el año 2018 como respuesta conjunta de la Organización Panamericana de la Salud y la Organización Mundial de la Salud, en cooperación con el Organismo Internacional de Energía Atómica, para colaborar con sus Estados miembros en asegurar que las exposiciones a la radiación de los pacientes pediátricos sean las mínimas necesarias durante los procedimientos intervencionistas. Actualmente, hay 18 centros de los siguientes 10 países que participan: Argentina, Brasil, Chile, Colombia, Costa Rica, Cuba, Ecuador, México, Perú y Uruguay. Para el desarrollo del programa se plantean una serie de objetivos, productos, actividades y resultados esperados. La puesta en marcha de la WEB de OPRIPALC ha significado un instrumento muy válido para seguir la información actualizada del programa. Un programa actualizado de formación en radioprotección para los profesionales implicados en el programa, se está realizando por medio de "webinars". Se deberá seguir actuando en la aplicación del programa de control de calidad básico para los equipos de rayos X participantes y validar los valores de los Niveles de Referencia para Diagnóstico (NRDs). Se propone formar un equipo de trabajo entre los Físicos Médicos y Tecnólogos Médicos participantes de OPRIPALC para implicarse en las pruebas de control básicas que todos los centros debieran realizar. Se han presentado algunos resultados iniciales de OPRIPALC en eventos científicos internacionales. Se está avanzando en proponer unos primeros valores sobre NRDs en procedimientos de intervencionismo cardiológico pediátrico por bandas de edad y peso. OPRIPALC es una de las pocas iniciativas de carácter regional para obtener valores de NRDs en procedimientos intervencionistas pediátricos. Se espera que tanto los valores de referencia como la metodología empleada en OPRIPALC, puedan ser utilizados en otras regiones del mundo.

The objective of this article has been to describe the program "Optimization of Protection in Pediatric Interventional Radiology in Latin America and the Caribbean" (OPRIPALC) that was born in 2018 as a joint response of the Pan American Health Organization and the World Organization of the Health, in cooperation with the International Atomic Energy Agency, to collaborate with its member states in ensuring that radiation exposures of pediatric patients are the minimum necessary during interventional procedures. Currently, there are 18 centers from the following 10 countries participating: Argentina, Brazil, Chile, Colombia, Costa Rica, Cuba, Ecuador, Mexico, Peru and Uruguay. For the development of the program, a series of objectives, products, activities and expected results are proposed. The launch of the OPRIPALC WEBSITE has been a very valid instrument for following up-to-date information on the program. An updated training program in radiation protection for the professionals involved in the program is being carried out through webinars. It should continue acting in the application of the basic quality control program for the participating X-ray equipment and validate the values of the Diagnostic Reference Levels (DRLs). It is proposed to form a work team among the OPRIPALC participating medical physicists to get involved in the basic control tests that all centers should carry out. Some initial results of OPRIPALC have been presented at international scientific events. Progress is being made in proposing first values on DRLs in pediatric cardiac intervention procedures by age and weight bands. OPRIPALC is one of the few regional initiatives to obtain DRLs values in pediatric interventional procedures. It is expected that both the reference values and the methodology used in OPRIPALC can be used in other regions of the world.

A Traceable DNA-Replicon Derived Vector to Speed Up Gene Editing in Potato: Interrupting Genes Related to Undesirable Postharvest Tuber Traits as an Example.

In potato (Solanum tuberosum L.), protoplast techniques are limited to a few genotypes; thus, the use of regular regeneration procedures of multicellular explants causes us to face complexities associated to CRISPR/Cas9 gene editing efficiency and final identification of individuals. Geminivirus-based replicons contained in T-DNAs could provide an improvement to these procedures considering their cargo capability. We built a Bean yellow dwarf virus-derived replicon vector, pGEF-U, that expresses all the editing reagents under a multi-guide RNA condition, and the Green Fluorescent Protein (GFP) marker gene. Agrobacterium-mediated gene transfer experiments were carried out on 'Yagana-INIA', a relevant local variety with no previous regeneration protocol. Assays showed that pGEF-U had GFP transient expression for up to 10 days post-infiltration when leaf explants were used. A dedicated potato genome analysis tool allowed for the design of guide RNA pairs to induce double cuts of genes associated to enzymatic browning (StPPO1 and 2) and to cold-induced sweetening (StvacINV1 and StBAM1). Monitoring GFP at 7 days post-infiltration, explants led to vector validation as well as to selection for regeneration (34.3% of starting explants). Plant sets were evaluated for the targeted deletion, showing individuals edited for StPPO1 and StBAM1 genes (1 and 4 lines, respectively), although with a transgenic condition. While no targeted deletion was seen in StvacINV1 and StPPO2 plant sets, stable GFP-expressing calli were chosen for analysis; we observed different repair alternatives, ranging from the expected loss of large gene fragments to those showing punctual insertions/deletions at both cut sites or incomplete repairs along the target region. Results validate pGEF-U for gene editing coupled to regular regeneration protocols, and both targeted deletion and single site editings encourage further characterization of the set of plants already generated.

CRISPR/Cas9 Targeted Editing of Genes Associated With Fungal Susceptibility in Vitis vinifera L. cv. Thompson Seedless Using Geminivirus-Derived Replicons.

The woody nature of grapevine (Vitis vinifera L.) has hindered the development of efficient gene editing strategies to improve this species. The lack of highly efficient gene transfer techniques, which, furthermore, are applied in multicellular explants such as somatic embryos, are additional technical handicaps to gene editing in the vine. The inclusion of geminivirus-based replicons in regular T-DNA vectors can enhance the expression of clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) elements, thus enabling the use of these multicellular explants as starting materials. In this study, we used Bean yellow dwarf virus (BeYDV)-derived replicon vectors to express the key components of CRISPR/Cas9 system in vivo and evaluate their editing capability in individuals derived from Agrobacterium-mediated gene transfer experiments of 'Thompson Seedless' somatic embryos. Preliminary assays using a BeYDV-derived vector for green fluorescent protein reporter gene expression demonstrated marker visualization in embryos for up to 33 days post-infiltration. A universal BeYDV-based vector (pGMV-U) was assembled to produce all CRISPR/Cas9 components with up to four independent guide RNA (gRNA) expression cassettes. With a focus on fungal tolerance, we used gRNA pairs to address considerably large deletions of putative grape susceptibility genes, including AUXIN INDUCED IN ROOT CULTURE 12 (VviAIR12), SUGARS WILL EVENTUALLY BE EXPORTED TRANSPORTER 4 (VviSWEET4), LESION INITIATION 2 (VviLIN2), and DIMERIZATION PARTNER-E2F-LIKE 1 (VviDEL1). The editing functionality of gRNA pairs in pGMV-U was evaluated by grapevine leaf agroinfiltration assays, thus enabling longer-term embryo transformations. These experiments allowed for the establishment of greenhouse individuals exhibiting a double-cut edited status for all targeted genes under different allele-editing conditions. After approximately 18 months, the edited grapevine plants were preliminary evaluated regarding its resistance to Erysiphe necator and Botrytis cinerea. Assays have shown that a transgene-free VviDEL1 double-cut edited line exhibits over 90% reduction in symptoms triggered by powdery mildew infection. These results point to the use of geminivirus-based replicons for gene editing in grapevine and other relevant fruit species.

A fast and efficient protocol for small RNA extraction in Japanese plum and other Prunus species

Background: Small ribonucleic acids represent an important repertoire of mobile molecules that exert key roles in several cell processes including antiviral defense. Small RNA based repertoire includes both small interfering RNA (siRNA) and microRNA (miRNA) molecules. In the Prunus genus, sharka disease, caused by the Plum pox virus (PPV), first occurred on European plum (Prunus domestica) and then spread over among all species in this genus and thus classified as quarantine pathogen. Next-generation sequencing (NGS) was used for the study of siRNA/miRNA molecules; however, NGS relies on adequate extraction protocols. Currently, knowledge of PPV-Prunus interactions in terms of siRNA populations and miRNA species is still scarce, and siRNA/miRNA extraction protocols are limited to species such as peach, almond, and sweet cherry. Results: We describe a reliable procedure for siRNA/miRNA purification from Prunus salicina trees, in which previously used protocols did not allow adequate purification. The procedure was based on a combination of commercially available RNA purification kits and specific steps that yielded high quality purifications. The resulting molecules were adequate for library construction and NGS, leading to the development of a pipeline for analysis of both siRNAs and miRNAs in the PPV­P. salicina interactions. Results showed that PPV infection led to altered siRNA profiles in Japanese plum as characterized by decreased 24-nt and increased 21- and 22-nt siRNAs. Infections showed miR164 and miR160 generation and increased miR166, miR171, miR168, miR319, miR157, and miR159. Conclusion: We propose this protocol as a reliable and reproducible small RNA isolation procedure for P. salicina and other Prunus species.

La calidad de los servicios de radiología en cinco países latinoamericanos / The quality of radiology services in five Latin American countries

OBJETIVO: Determinar la correlación entre ciertos indicadores de calidad para los servicios de imaginología y la certeza en la interpretación de los exámenes radiológicos para cuatro quejas frecuentes: las masas de la mama, el malestar del aparato digestivo, el dolor de espalda y los síntomas de la tuberculosis. MÉTODOS: Se evaluaron veintiséis servicios de radiología en Argentina, Bolivia, Colombia, Cuba y México. Se evaluaron los equipos de mamografía y de radiografía/fluoroscopia convencional usados en los servicios seleccionados utilizando protocolos comunes, hojas de especificaciones técnicas, instrumen-tos de prueba, maniquíes y sistemas de dosimetría calibrados. Los estudios se realizaron en establecimientos de complejidad media. Se obtuvo el consentimiento informado de todos los pacientes estudiados, y se garantizó la confidencialidad de los resultados. Se evaluaron y documentaron los siguientes parámetros: el tipo de establecimiento (público o privado); la población cubierta; el número de pacientes y exámenes; los equipos radiológicos, los de procesamiento de imágenes y los suministros; la educación y la capacitación del personal profesional y técnico; los programas de la garantía de la calidad y del mantenimiento preventivo, y la adherencia a las normas de seguridad radiológica. Se determinaron el funcionamiento de los equipos de rayos X, los receptores de la imagen y las procesadoras; las condiciones del cuarto oscuro y de la visualización de las imágenes; las dosis recibidas por los pacientes y la calidad de la imagen, usando parámetros uniformados en todos los casos. Los paneles independientes de radiólogos, reconocidos como expertos por la sociedad radiológica local, evaluaron la calidad de las imágenes clínicas obtenidas y realizaron una interpretación radiológica para cada paciente usando las mismas películas e historia clínica a disposición de los médicos especialistas en imaginología de la institución. El acuerdo entre...

OBJECTIVE: To determine the correlation between certain quality indicators for imaging services and the accurate interpretation of radiological exams for four frequent complaints: breast lumps, gastrointestinal discomfort, back pain, and symptoms of tuberculosis. METHODS: Twenty-six radiology services in Argentina, Bolivia, Colombia, Cuba, and Mexico were assessed. The mammography and conventional radiographic/fluoroscopic equipment used in selected services were evaluated utilizing common protocols, data sheets, testing instruments, phantoms, and calibrated dosimetry systems. The studies were performed in medium-complexity facilities. Informed consent was obtained from all patients studied, and the confidentiality of results was guaranteed. The following parameters were documented: type of facility (public vs. private); population covered; patient workload; radiological and image-processing equipment and supplies; education and training of professional and technical staff; quality assurance and preventive maintenance programs, and adherence to radiation safety standards. The performance of x-ray units, image receptors and processors; darkroom and image viewing conditions; patient doses and image quality, were determined using standardized parameters in all cases. Independent panels of radiologists, recognized as experts by the local radiological society, assessed the quality of the clinical images obtained and performed a radiological interpretation for each patient using the same films and clinical history available to the institution's imaging physicians. The agreement between the panel of expert's reports and those of local radiologists was taken as an indicator of the radiological diagnostic accuracy. RESULTS: Analyses were carried out of 366 mammograms, 343 radiological procedures for gastrointestinal complaints, 319 X-rays of the spinal column, and 157 chest radiographs. The agreement between the radiological interpretation of...

Bacteriological water quality along the Tijuana-Ensenada, Baja California, México shoreline.

This survey was part of a Binational Program (Mexico-United States) in microbiological water quality, with a goal to assess the shoreline bacteriological water quality from Tijuana to Ensenada, Mexico. Samples were collected at 29 sites (19 beaches and 10 outfalls), from the United States border to Punta Banda, Baja California, during summer (1998) and winter (1999). Total coliforms, fecal coliforms and enterococci were used as bacterial indicators. Standard methods were used for total and fecal coliforms, while the Enterolert quick method (IDEXX) was used for the enterococci. Compared with outfalls, the beaches exceeded water quality standards by a small percent, 25.3% in summer and 17% in winter. For outfalls, the percentage of shoreline that exceeded bacterial indicator thresholds had a minor value in summer (32.7%) than in winter (50%). Sites near wastewater discharges had the lowest quality and did not meet the microbiological water quality criteria for recreational use.

[The quality of radiology services in five Latin American countries]. / La calidad de los servicios de radiología en cinco países latinoamericanos.

OBJECTIVE: To determine the correlation between certain quality indicators for imaging services and the accurate interpretation of radiological exams for four frequent complaints: breast lumps, gastrointestinal discomfort, back pain, and symptoms of tuberculosis. METHODS: Twenty-six radiology services in Argentina, Bolivia, Colombia, Cuba, and Mexico were assessed. The mammography and conventional radiographic/fluoroscopic equipment used in selected services were evaluated utilizing common protocols, data sheets, testing instruments, phantoms, and calibrated dosimetry systems. The studies were performed in medium-complexity facilities. Informed consent was obtained from all patients studied, and the confidentiality of results was guaranteed. The following parameters were documented: type of facility (public vs. private); population covered; patient workload; radiological and image-processing equipment and supplies; education and training of professional and technical staff; quality assurance and preventive maintenance programs, and adherence to radiation safety standards. The performance of x-ray units, image receptors and processors; darkroom and image viewing conditions; patient doses and image quality, were determined using standardized parameters in all cases. Independent panels of radiologists, recognized as experts by the local radiological society, assessed the quality of the clinical images obtained and performed a radiological interpretation for each patient using the same films and clinical history available to the institution's imaging physicians. The agreement between the panel of expert's reports and those of local radiologists was taken as an indicator of the radiological diagnostic accuracy. RESULTS: Analyses were carried out of 366 mammograms, 343 radiological procedures for gastrointestinal complaints, 319 X-rays of the spinal column, and 157 chest radiographs. The agreement between the radiological interpretation of the panel of experts and of the local physician ranged from 70% to 100%, except in the case of spinal column films in Cuba (57.8%) and of mammograms in Mexico (33.3%), which the panel of experts found to be among those having the poorest quality. There was a significant positive correlation between the accuracy of the radiological interpretation and the quality of the radiological images. Image quality showed a positive correlation with the technicians' level of education and training. Studies performed in services that had automatic film processors and that complied with the indicators established for screen-film contact yielded better images and a higher proportion of studies with concordant results. More than 50% of the viewboxes did not satisfy the quality criteria for luminance and homogeneity. CONCLUSIONS: A good quality image is critical to achieving an accurate diagnosis. Emphasis should be placed on the continuing education of radiology technicians and on the acquisition and maintenance of adequate equipment and accessories, especially viewboxes, intensifying screens, and automatic film processors, given the impact they have on image quality.