Retos y oportunidades en el estudio de vesículas extracelulares: contexto institucional a nivel mundial y situación actual en Colombia / Challenges and opportunities in the study of extracellular vesicles: Global institutional context and national state of the art

Resumen En la última década se ha incrementado el número de estudios y publicaciones sobre las vesículas extracelulares y los exosomas. En Colombia, ha habido interés y avances en su estudio, lo que se evidencia en el aumento de publicaciones y proyectos de investigación. Sin embargo, este es un campo de investigación aún en desarrollo, con desafíos analíticos y limitaciones técnicas, por lo cual, en el planteamiento de los proyectos de investigación y desarrollo, es necesario considerar cuál es el estado del campo científico a nivel mundial en cuanto a la nomenclatura y la clasificación de las vesículas extracelulares, las técnicas, recursos, requisitos y especificaciones de calidad y las instituciones que regulan el campo. La respuesta a esta pregunta permitirá desarrollar estudios que cumplan con los estándares internacionales, y las exigencias y recomendaciones institucionales. Sin embargo, la información científica disponible se encuentra dispersa y no todos los aspectos son tratados a cabalidad. En este actualización se condensa la información disponible y se presentan los términos oficiales para denominar las vesículas extracelulares y la nomenclatura aceptada actualmente, así como la evolución del campo, la homogenización de los parámetros experimentales, el establecimiento de autoridades científicas, instituciones y recursos, y las recomendaciones que se han generado a nivel mundial para el desarrollo de investigaciones en vesículas extracelulares, incluidos su aislamiento, caracterización y estudio funcional. Por último, se analiza el contexto nacional de una forma crítica, teniendo en cuenta las fortalezas institucionales, los errores usualmente cometidos, y las técnicas y tecnologías analíticas disponibles.

Abstract In the last decade, the number of studies and publications on extracellular vesicles (EV) and exosomes has boomed. Colombia has displayed interest and progress in their study as shown in the increase of research project publications and products. However, this research field is still developing and has its own analytical challenges and technical limitations. For planning research projects and developing EV studies it is necessary to consider what is the state of the scientific field worldwide concerning EV nomenclature and classification, available techniques, resources, requirements and quality specifications, and the institutions that regulate the field. Answering this question will elicit EV studies that comply with international standards and respond to institutional demands and recommendations. However, the scientific information available is scattered and not all the aspects are considered in full. In this update, the available information is condensed and the official terms and currently defined nomenclature is presented, as well as the evolution of the field, the homogenization of the experimental parameters, the establishment of scientific authorities, institutions, and resources, and the recommendations generated worldwide for their development and research including their isolation, characterization, and functional studies. Finally, I analyzed the national context in a critical way, considering institutional strengths, common mistakes, and available analytical techniques and technologies.

Analytical Performance of Wako and Sekisui Clinical Chemistry Assays on Hitachi LABOSPECT 008

BACKGROUND: We evaluated the analytical performance of Wako assays for albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), complement C3 and C4, calcium, creatine kinase (CK), C-reactive protein (CRP), direct bilirubin (DBIL), iron, gamma-glutamyl transferase (GGT), HDL cholesterol (HDLC), inorganic phosphorus (IP), LDL cholesterol (LDLC), total bilirubin (TBIL), total protein (TP), and uric acid (UA), as well as the performance of Sekisui assays for albumin, BUN, calcium, CRP, HDLC, IP, LDLC, TP, and UA by using Hitachi LABOSPECT 008 (Hitachi High-Tech Co., Japan). METHODS: Liquid Assayed Multiqual Control (Bio-Rad Laboratories, USA) and pooled patients' sera were analyzed for 20 days. Wako linearity material (Wako Pure Chemical Industries, Ltd., Japan) and Sysmex Interference Check A Plus kit (Sysmex Co., Japan) were used to test linearity and interference, respectively. Concentrations of the target analytes were measured using Hitachi LABOSPECT 008 in 100 residual patient specimens and compared to those in Pureauto S series reagent (Sekisui Medical, Japan), which were measured using Hitachi 7600 (Hitachi High-Tech Co., Japan). RESULTS: Total coefficients of variation (CVs) for the tested analytes were 0.91-9.26% in Wako and 1.04-7.46% in Sekisui assays. Linearity was demonstrated up to the highest concentration within the analytical range in all the assays except for Wako albumin and Sekisui TP. Wako and Sekisui albumin, BUN, CRP, HDLC, and LDLC assays, and in Wako C3, C4, calcium, and UA assays showed no interference with the test concentrations used. All the tested assays, except for Wako AST, LDLC, and TP, and Sekisui calcium and TP, demonstrated comparability with comparative method for at least one medical decision level. CONCLUSIONS: Our study results showed that the analytical performances of Wako and Sekisui chemistry assays evaluated using Hitachi LABOSPECT 008 had appropriate analytical performance for clinical use.

Caracterização analítica de sucos e néctares de laranja adoçados com sacarose e edulcorantes / Analytical characterization of orange juices and nectars sweetened with sucrose and sweet flavoring agents

Objetivo: Caracterizar físico-quimicamente, sucos não adoçados e néctares de laranja adoçados com sacarose ou edulcorantes, quanto ao seu pH, acidez titulável (AT) e teor de sólidos solúveis totais (SST) e avaliar a correlação desta última propriedade com as demais. Método: Analisaram-se alíquotas de três lotes de dois sucos de laranja e de dois néctares com adição de sacarose ou dois com edulcorantes. Água mineral foi empregada como controle. O teor de SST foi determinado em refratômetro de Abbé. O pH foi registrado em peagômetro digital, enquanto a AT foi quantificada titulando-se amostras das bebidas com NaOH 0,1 M até o alcance dos pHs 5,5 e 7,0. Os dados foram submetidos ao teste de correlação de Pearson, análise de regressão, análise de variância e teste de Tukey (alfa = 0,05). Resultados: Os teores de SST apresentaram forte correlação com a AT, sendo a relação entre elas do tipo quadrática. Embora os valores de pH não sejam dependentes da presença de sacarose ou edulcorantes, se as bebidas são isentas dos mesmos, significativamente maior quantidade de base foi necessária para que se atingissem os pHs 5,5 e 7,0. Conclusão: Sucos e néctares de laranja apresentaram valores semelhantes de pH, os quais não se correlacionaram com a presença de sacarose ou edulcorantes nas bebidas. A acidez titulável foi maior para o suco e menor para os néctares, independentemente do fato de possuírem sacarose ou edulcorantes em sua composição. A elevação do teor de sólidos solúveis totais não implicou em redução da acidez titulável das bebidas.

Objective: To characterize physically and chemically, non-sweetened orange juices and orange nectars sweetened with sucrose or sweet flavoring agents, with respect to their pH, titratable acidity (TA) and total soluble solids content (TSSC), as well as to evaluate the correlation of the latter property with the others. Method: Aliquots of three lots of two orange juices and two orange nectars containing sucrose and two containing sweet flavoring agents were evaluated. Mineral water was used as a control. The TSSC was determined using an Abbe refractometer. The pH was recorded using a digital pH meter, while TA was quantified by titrating samples of the beverages with 0.1 M NaOH until reaching pHs 5.5 and 7.0. Data were subjected to Pearson's correlation test, regression analysis, analysis of variance and Tukey's test (alpha=0.05). Results: TSSC values presented a strong correlation with TA, and these properties exhibited a quadratic relationship. Although the pH values were not dependent on the presence of sucrose or sweet flavoring agents, a significantly greater amount of base was necessary to reach pHs 5.5 and 7.0 in the beverages without sucrose or flavoring agents. Conclusion: Orange juices and nectars presented similar pH values, which was not associated with the presence of sucrose or sweet flavoring agents in the beverages. Higher TA values were obtained for the juice and lower for the nectars, regardless of containing sucrose or sweet flavoring agents. The increase of TSSC did not implicate in decrease of TA in the beverages.