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Catarina Costa Moura,1,2 Marcela A Segundo,1 José das Neves,3,4 Salette Reis,1 Bruno Sarmento3,4 1REQUIMTE, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Porto, Portugal; 2Faculty of Engineering, University of Porto, Porto, Portugal; 3CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Instituto de Ciências da Saúde-Norte, Gandra PRD, Portugal; 4INEB – Instituto de Engenharia Biomédica, University of Porto, Porto, Portugal Background: Rheumatoid arthritis (RA) is an autoimmune disease with severe consequences for the quality of life of sufferers. Regrettably, the inflammatory process involved remains unclear, and finding successful therapies as well as new means for its early diagnosis have proved to be daunting tasks. As macrophages are strongly associated with RA inflammation, effective diagnosis and therapy may encompass the ability to target these cells. In this work, a new approach for targeted therapy and imaging of RA was developed based on the use of multifunctional polymeric nanoparticles. Methods: Poly(lactic-co-glycolic acid) nanoparticles were prepared using a single emulsion-evaporation method and comprised the co-association of superparamagnetic iron oxide nanoparticles (SPIONs) and methotrexate. The nanoparticles were further functionalized with an antibody against the macrophage-specific receptor, CD64, which is overexpressed at sites of RA. The devised nanoparticles were characterized for mean particle size, polydispersity index, zeta potential, and morphology, as well as the association of SPIONs, methotrexate, and the anti-CD64 antibody. Lastly, the cytotoxicity of the developed nanoparticles was assessed in RAW 264.7 cells using standard MTT and LDH assays.Results: The nanoparticles had a mean diameter in the range of 130–200 nm and zeta potential values ranging from -32 mV to -16 mV. Association with either methotrexate or SPIONs did not significantly affect the properties of the nanoparticles. Conjugation with the anti-CD64 antibody, in turn, caused a slight increase in size and surface charge. Transmission electron microscopy confirmed the association of SPIONs within the poly(lactic-co-glycolic acid) matrix. Both anti-CD64 and methotrexate association were confirmed by Fourier transform infrared spectroscopy, and quantified yielding values as high as 36% and 79%, respectively. In vitro toxicity studies confirmed the methotrexate-loaded nanosystem to be more effective than the free drug.Conclusion: Multifunctional anti-CD64-conjugated poly(lactic-co-glycolic acid) nanoparticles for the combined delivery of methotrexate and SPIONs were successfully prepared and characterized. This nanosystem has the potential to provide a new theranostic approach for the management of RA. Keywords: FcγRI, methotrexate, poly(lactic-co-glycolic acid), superparamagnetic iron oxide nanoparticles, targeted drug delivery

Maria João Gomes,1 Susana Martins,2,3 Domingos Ferreira,2 Marcela A Segundo,1 Salette Reis1 1REQUIMTE, Laboratory of Applied Chemistry, Department of Chemical Sciences, Faculty of Pharmacy, 2Laboratory of Pharmaceutical Technology, Department of Drug Sciences, Faculty of Pharmacy, University of Porto, Porto, Portugal; 3Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense, Denmark Abstract: Alopecia is a dermatological disorder, commonly known as hair loss, which affects up to half of the Caucasian male population by middle age, and almost all (95%) Caucasian men by old age. Considering that alopecia affects so many people and that there is currently no scientifically proven treatment with few side effects, new drug-delivery systems able to improve alopecia therapy are urgently required. With this purpose in mind, the present study aimed to develop lipid nanoparticles (nanostructured lipid carriers) with the ability to incorporate and deliver anti-alopecia active compounds (minoxidil and finasteride) into the dermis and hair follicles. Lipid nanoparticles, prepared by ultrasonication method, showed mean particle sizes around 200 nm, which is sufficient for reaching the dermis and hair follicles, and zeta potential values around -30 mV, which indicates good physical stability. Over 28 days of storage, no significant variations in these parameters were observed, which indicates that all nanoformulations are stable in storage over that period. Cryo-scanning electron microscope measurements showed that all the lipid nanoparticles exhibited a spherical shape and a smooth surface regardless of their composition. Differential scanning calorimetry studies allowed the determination of phase transition temperatures and confirmed the recrystallization of the lipid nanoparticles (recrystallization index between 11% and 86%). A high loading efficiency was achieved for finasteride (between 70% and 90%), while less than 30% was achieved for minoxidil nanoparticles, over 28 days. Controlled release assays in physiological conditions demonstrated that nanoparticles loaded with minoxidil yielded a prolonged release, as desired. Penetration assays through pig ear skin demonstrated that nanoparticles loaded with minoxidil and finasteride had low levels of penetration. These results suggest that the proposed novel formulation presents several good characteristics indicating their suitability for dermal delivery of anti-alopecia active compounds. Keywords: nanostructured lipid carriers (NLC), alopecia, anti-alopecia therapy, minoxidil, finasteride

The Peruvian Amazonian native cacao faces ongoing challenges that significantly undermine its productivity. Among them, frosty pod rot disease and cadmium accumulation result in losses that need for effective and environmentally safe strategies, such as those based on bacteria. To explore the biological resources in the cacao soil, a descriptive study was conducted to assess the diversity of culturable bacteria across three production districts in the Amazonas region: La Peca, Imaza, and Cajaruro. The study also focused on the functional properties of these bacteria, particularly those related to the major issues limiting cacao cultivation. For this purpose, 90 native bacterial isolates were obtained from the cacao rhizosphere. According to diversity analysis, the community was composed of 19 bacterial genera, with a dominance of the Bacillaceae family and variable distribution among the districts. This variability was statistically supported by the PCoA plots and is related to the pH of the soil environment. The functional assessment revealed that 56.8% of the isolates showed an antagonism index greater than 75% after 7 days of confrontation. After 15 days of confrontation with Moniliophthora roreri, 68.2% of the bacterial population demonstrated this attribute. This capability was primarily exhibited by Bacillus strains. On the other hand, only 4.5% were capable of removing cadmium, highlighting the biocontrol potential of the bacterial community. In addition, some isolates produced siderophores (13.63%), solubilized phosphate (20.45%), and solubilized zinc (4.5%). Interestingly, these traits showed an uneven distribution, which correlated with the divergence found by the beta diversity.Our results revealed a diverse bacterial community inhabiting the Amazonian cacao rhizosphere, showcasing crucial functional properties related to the biocontrol of M. roreri. The information generated serves as a significant resource for the development of further biotechnological tools that can be applied to native Amazonian cacao.

The diversity and floristic composition of a primeval forest was studied, located in the district of Yambrasbamba–Bongará–Amazonas, delimiting a 1 ha area, and at an altitude of 1890 m.a.s.l. All individuals with diameter at breast height (DBH) ≥ 10 cm were inventoried. The plant diversity in the area was measured and a description of its composition and floristic structure was made. The following were recorded: a total of 640 trees distributed in 39 families, 60 genera and 152 species. The value of the Simpson’s index (D) was 0.974 and that of the Shannon–Wiener index was 4.264, indicating that the species had a high abundance of individuals. In turn, Fisher’s alpha value (α) was 23.744, indicating a regular diversity in montane forests in relation to different altitudinal gradients. The families with the highest number of individuals were Melastomataceae, Rubiaceae, Euphorbiaceae, Phyllanthaceae, and Lauraceae. The most abundant species were Alchornea acutifolia Müll.Arg. with 47 individuals (7.34%), Chimarrhis glabriflora Ducke with 39 individuals (6.09%), Hieronyma alchorneoides Allemão with 39 individuals (6.09%), and Cyathea lasiosora (Kuhn) Domin with 33 individuals (5.16%). A comparative analysis was carried out of plots of montane and premontane forests, and the studied plot presented had the third-highest register of families and genera, behind the plots studied in the provinces of Oxapampa and Chanchamayo.

This work aimedto design and manufacture a prosthetic prototype using additive manufacturing technology, or 3D printing, for a dog that has suffered an amputation. This project began out of the need that dogs that suffered amputations and develop muscular dystrophies and other health problems have, but prosthetics are not commonly found on the market. Alternatively, a prosthesis that meets the animal's needswas designed in computer-aided design (CAD) software. Later, the element was subjected to a static analysis through simulation. Then itwas printed,and the prototype was assembled, getting a functional and comfortable element that underwent performance tests with the benefiteddog, which presented optimal performance in terms of comfort and functionality. It is concluded that the realization of prosthetics through additive manufacturing technology is not only possible but also an alternative that reduces weight, costs, and time, besides providing enormous ease of design and manufacturing.

Yield in cherimoya (Annona cherimola Mill.) is affected by various factors reducing fertilization and fruit production such as: the protogynous dichogamy of the flowers, deficient natural pollination, and low pollen quality. The objective of this study was to determine in vitro the differences in pollen quality of nine ecotypes and one cultivar of A. cherimola from pollen samples collected at the female and male phenological stages of flowering. The percentage of pollen germination and the length of pollen tubes was evaluated in vitro at 2 and 24 hours after incubation at 21°C, in two phenological stages of the flowers during bloom in 2018 and 2019 seasons. The germination medium contained: 10% sucrose, 1% agar, 0.005% boric acid, and 0.025% calcium nitrate. Fabulosa and Austro ecotypes and the Fino de Jete cultivar had the highest pollen germination in the male phenological stage, after 2 and 24 hours in both seasons. Fabulosa had the highest percentage of germination in the female stage in both seasons. The ideal phenological stage for pollen collection was at anther dehiscence. Austro and Fabulosa ecotypes had the longest pollen tubes after 2 and 24 hours, in both phenological stages for both seasons. Fabulosa and Austro ecotypes, and Fino de Jete cultivar, showed promising pollen characteristics for hand pollination of cherimoya.

La prevención de la violencia escolar constituye un tema relevante, en el orden educativo y social, dado por la necesidad de contar con ambientes de formación cada vez más seguros y desarrolladores, capaces de fomentar la civilidad y la armonía, como recursos para enfrentar con prosocialidad las problemáticas que surgen en las relaciones con los demás, significando el lugar que corresponde a la orientación educativa en la prevención de la violencia. El presente trabajo tiene como objetivo el análisis de acciones orientadoras, para prevenir la violencia escolar en los estudiantes de bachillerato, en el contexto ecuatoriano, partiendo de un diagnóstico realizado con una muestra de 230 estudiantes y 20 docentes de Bachillerato, empleando como técnicas la Entrevista a márgenes: profundidad, el Grupo de discusión y el Cuestionario sobre intimidación y maltrato entre iguales de entre iguales, de Mora-Merchán y Mora (1995). Los resultados del diagnóstico realizado permitieron constar la existencia de violencia escolar en la institución educativa posibilitando la delimitación de los principales ejes de trabajo en la práctica pedagógica de cada jornada escolar y extraescolar para propiciar las transformaciones necesarias. PALABRAS CLAVE: violencia; orientación educativa; prevención de la violencia. System of guiding actions to prevent school violence in high school students ABSTRACT The prevention of school violence is a relevant issue, in the educational and social order, given by the need to have increasingly safe training environments and developers, capable of promoting civility and harmony, as resources to face with prosociality the problems that arise in relationships with others, signifying the place that corresponds to educational guidance in the prevention of violence. The present work aims to analyze guiding actions, to prevent school violence in high school students, in the Ecuadorian context, based on a diagnosis made with a sample of 230 students and 20 high school teachers, using as techniques the In-depth interview. The Discussion Group and the Questionnaire on intimidation and mistreatment between equals of equals, by Mora-Merchán and Mora (1995). The results of the diagnosis made allowed the existence of school violence in the educational institution to be confirmed, enabling the delimitation of the main lines of work in the pedagogical practice of each school and extracurricular day to promote the necessary transformations. KEYWORDS: violence; educational orientation; violence prevention.

Los sistemas silvopastoriles (SSP) son una alternativa para la producción ganadera sostenible. Por este motivo, el presente estudio se desarrolló con el objetivo de evaluar parámetros productivos y nutricionales del componente forrajero (CF) en distintos arreglos silvopastoriles con Alnus acuminata y su comparación con sistemas a campo abierto. Se estableció un diseño de bloques completos al azar, para lo cual, fueron seleccionadas 16 parcelas con características de homogeneidad en edad y tipo de CF. Se evaluó la composición florística, clasificación funcional de las especies herbáceas, biomasa, materia seca y composición nutricional. Los resultados obtenidos registraron la presencia de 22 especies, predominando la familia Poaceae (8 especies), asimismo se encontró que los arreglos silvopastoriles presentan el mayor porcentaje de especies deseables, situación contraria a lo ocurrido en los sistemas a campo abierto. Por otro lado, los parámetros productivos y nutricionales, mostraron diferencias significativas (P

El presente trabajo tuvo por objetivo evaluar la influencia de sustratos en el crecimiento y desarrollo de tomate (Lycopersicum esculentum Mill) cultivado bajo un sistema hidropónico en invernadero. Se instaló bajo un diseño completo al azar (DCA), con cuatro tratamientos: T1 (75% cascarilla de arroz + 25% arena, T2 (75% cascarilla de arroz carbonizada + 25% arena, T3 (75 % Aserrín + 25% arena, T4 (40% cascarilla de arroz carbonizada + 35% aserrín + 25% arena); 8 repeticiones y 32 plantas. Se encontró que el T2, favoreció el incremento de altura (139.13 cm), diámetro de tallo (10.95 mm), número de tomates de primera calidad (15.25) y rendimiento (3.03 kg/planta); no obstante, el T3 y T1 mostraron baja influencia en dichas variables. Se concluye que, el tomate obtiene mayores características morfológicas y fisiológicas en sustrato (cascarilla carbonizada + arena); el aserrín como sustrato no es ideal para su crecimiento y desarrollo.

Pancreatic cancer has a dismal prognosis and only a few treatment options are available. In the European Union, pancreatic cancer classifies as a rare disease, allowing drug developers to apply for orphan medicinal product (OMP) designation. The aim of this study was to provide more detail on OMPs for pancreatic cancer. All applications for OMP designation submitted to the EMA between 2000 and 2019 were identified. For each medicinal product that received an OMP designation, the mode of drug action, use of protocol assistance, and current life cycle status was determined. Fifty-two medicinal products received an OMP designation. At the time of submission, eighteen OMPs were at the non-clinical and 34 OMPs were at the clinical stage of development. At least fourteen kinds of mode of action were explored in the condition. For eighteen out of 52 OMPs protocol assistance was sought. At the time of data analysis, one OMP received marketing authorisation and 24 OMPs were ongoing in development. Many medicinal products for pancreatic cancer received an OMP designation and the majority of these products was already in the clinical stage of development. Nonetheless, the success rate of OMPs for pancreatic cancer that reach the market is low, and increasing this rate is something to aspire. Fortunately, development is still ongoing for a part of the OMPs, and a few developers are planning to submit a marketing authorisation application in the near future. This however does not guarantee success, as pancreatic cancer remains a difficult disease to treat. Developers are advised to make optimal use of incentives such as protocol assistance, establishing (early) dialogue between regulators and drug developers and to agree on important topics such as clinical trial design.

Cacao plant cadmium accumulation has become a major concern, especially for small Amazonian producers. A sustainable alternative to address its toxicity is the use of cadmium removal bacteria. In this regard, 138 rhizosphere isolates from cacao were examined. Supported by their phenotypic characterization and in vitro cadmium tolerance, three hypertolerant bacteria were selected and identified as members of the Bacillus (S1C2, R1C2) and Pseudomonas (V3C3) genera. They were able to grow normally and reduce the cadmium content under in vitro conditions. However, only S1C2 and R1C2 evidenced to employ intracellular Cd2+ accumulation, suggesting the variability of bacterial detoxification mechanisms. Their bioremediation capacity for Theobroma cacao CCN51 was also analyzed. Surprisingly, we found high detectable levels of Cd2+ in the non-cadmium supplemented control, suggesting an extra source of cadmium in the pot. Moreover, despite their cadmium reduction performance under in vitro conditions, they exerted highly variable outcomes on stem cadmium accumulation. While S1C2 and R1C2 showed a considerable reduction of Cd content in cacao stems, the strain V3C3 did not show any effect on Cd content. This highlights the complexity of the plant–bacteria interactions and the importance of the in vivo test for the selection of promising PGPR bacteria. Overall, our results suggest the cadmium alleviation potential and promising prospects of native Bacillus strains associated with Amazonian cacao.

Based on a review of literature related to the biological diversity of the inter-Andean Seasonally Dry Tropical Forest of the Marañón valley (ISDTFM), located in the regions of Cajamarca, Amazonas, Piura, La Libertad, Ancash and Huánuco (Perú), this study highlights the richness of endemic species of woody flora and tetrapod fauna, as well as addressing their threats and state of conservation. In fact, we report 143 woody flora endemics, most notably the species, Coursetia cajamarcana, Parkinsonia peruviana and Pradosia argentea, due to its Critically Endangered (CR) by the IUCN. Regarding tetrapod fauna endemic, there are four species of amphibians, 28 of reptiles, 22 of birds and six of mammals; outstanding among them Synallaxis maranonica due to its Critically Endangered (CR). A well-known problem with this forest is that it has been severely threatened by the expansion of the agricultural frontier and access routes, a situation that could become even more exacerbated if energy and extraction activities are developed. On the other hand, only 1.01% of all these forests, are protected under Protected Natural Areas (PNA). For this reason, it is essential to have this information in order to prioritise its conservation and protection.

Since the implementation of the EU Orphan Regulation in 2000, the Committee for Orphan Medicinal Products at the European Medicines Agency has been evaluating the benefits of proposed orphan medicines vs. satisfactory treatment methods. This type of evaluation is foreseen in the Orphan Regulation as the orphan designation criterion called the “significant benefit.” In this article, based on 20 years of experience, we provide a commentary explaining what is considered a satisfactory method of treatment in the context of the EU Orphan Regulation and for the purpose of the assessment of significant benefit. We discuss the challenges posed by continuously changing clinical practise, which is associated with the increasing number of treatment options, evolving nature of medicinal therapeutic indications and our understanding of them.

Twenty years of orphan regulation in Europe have now elapsed, with almost 2,400 orphan designated medicinal products and more than 190 orphan products authorised in the EU. Alongside the evolution in understanding of rare diseases, considerable regulatory knowledge has also been accumulated regarding the level of evidence that would support inclusion of products into the framework. This article reviews publications and regulatory documents pertaining to orphan medicinal product designation in the EU and discusses the general expectations in submitted applications as reflected in the current regulatory practise. Important elements to recommend granting a European orphan designation are the key considerations of orphan condition, medical plausibility, seriousness, and prevalence, while significant benefit is also assessed when there are authorised medicinal products for the sought indication. This review attempts to clarify the specific concepts currently used in that regard and discusses how the available data can be used to justify the criteria for designation. Moving away from theoretical expectations or assumptions, it stresses that the applications have to be complemented with nosological and epidemiological justifications pertaining to the proposed condition, as well as relevant data in specific non-clinical in vivo models or in affected patients to support inclusion into the orphan scheme.

Se realizaron soldaduras TIG en tuberías de acero inoxidable AISI 304 purgadas con argón, con 999, 500, 200, 100, 50, 25 y 10 ppm de oxígeno atmosférico en la cámara de purga para determinar si esto influye en la calidad superficial de la zona afectada por el calor (ZAC) en la parte interna de la tubería. En este estudio se realizaron ensayos mecánicos, análisis metalográfico y morfológico. Se determinó que los niveles de decoloración en la ZAC están directamente influenciados por la cantidad de oxigeno atmosférico presente en la purga, por lo que con cantidades entre 50 a 10 ppm se evidencia decoloraciones de acuerdo a las normas AWS D18.1 y AWS D18.2. Se evidenció buena resistencia en la junta soldada debido a que la falla se da en el metal base y no en la ZAC que no ha tenido influencia de la decoloración superficial que corresponde a 10 ppm.

This paper reports the synthesis and characterization of Bi1-xLuxFeO3 (x = 0.00, 0.02 and 0.04) produced by solid-state reaction, in order to evaluate the influence of lutetium on the structural and magnetic properties of bismuth ferrite (BiFeO3). The samples were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX) and magnetic analysis by vibrating sample magnetometer (VSM) in temperature range from 50 to 320 K. The obtained results allowed to confirm the formation of crystalline materials of rhombohedral structure, space-group R3c (161), with defined morphology and particle sizes between 2.25 and 4.5 μm. The Lu3+ insertion in structure generated an increasing in magnetization, purity of BiFeO3 and a decrease in the synthesis temperature compared with the reported in the literature.

En el beneficio del café se genera grandes volúmenes de aguas mieles con alto contenido de materia orgánica, ocasionando impactos al medio ambiente. Es por ello que el objetivo del presente trabajo fue evaluar tres sistemas de tratamiento de aguas mieles de café. Se comparó un filtro un físico (arena, carbón, piedra de río), un filtro biológico con jacinto acuático (Eichhornia crassipes) y lenteja de agua (Lemna minor L.) y, un sistema con coagulantes naturales con moringa (Moringa oleífera L.) y tuna (Opuntia ficus-indica). Se realizaron mediciones cada 15 días (entre mayo y junio de 2019), los datos se procesaron en el software R. Los resultados explican una función discriminante lineal de 0,9355, indicando que el filtro físico y el filtro biológico lograron reducir los parámetros de sólidos disueltos totales (SDT) de 3,266 mg.L-1 a 2,55 mg.L-1 y fosfatos (PO43¯) de 2,763 mg.L-1 a 0,975 mg.L-1 existiendo diferencia significativa entre los tratamientos.

Introducción: Los agroecosistemas de C. arabica bajo sombra son de los cultivos que se consideran no dañinos para la biodiversidad, sin embargo, dada la importancia ecológica que tienen en el resguardo de muchas especies son escasos los estudios en este cultivo en la zona. Objetivo: Proporcionar información sobre la diversidad y la fluctuación de insectos asociados al agroecosistema de café bajo sombra, mediante un estudio faunístico. Métodos: Los insectos se recolectaron sobre diez plantas de C. arabica divididas en tres estratos (bajo, medio y alto) cada 15 días durante los meses de junio a octubre del 2018, utilizando la red de golpeo, el aspirador y trampas Pitfall. Resultados: Recolectamos 1, 923 especímenes pertenecientes a 40 géneros, 35 familias y ocho órdenes. Hemiptera fue el orden con más riqueza de familias registradas (9) y más géneros (13). Coleoptera registró la mayor abundancia con 538 (27,98%). Apis sp y Canthon sp fueron los géneros con la mayor abundancia ambos con 108 individuos. La mayor riqueza, abundancia y diversidad se registró en el estrato medio con 35 especies, 756 individuos y con 1D= 28,53, 2D=25,56. La máxima abundancia de los insectos coincide con la precipitación y humedad alta, y con los valores de temperatura baja. La curva de acumulación se estabilizó a partir del muestreo cinco. Conclusión: Presentamos el primer estudio sobre la entomofauna en un agroecosistemas de C. arabica con manejo orgánico, mostrando que alberga una alta diversidad y que es importante para el refugio y la conservación de insectos benéficos.

La Universidad Regional Autónoma de los Andes –UNIANDES- es una institución de educación superior con extensiones en diferentes provincias del Ecuador. Dentro de las funciones que debe cumplir una universidad ecuatoriana está la vinculación con la sociedad; todas las actividades de carácter administrativo relacionadas a dicha función se encuentran centralizadas en el Departamento de Vinculación de UNIANDES en la matriz Ambato. Dicho departamento planteó a la Carrera de Sistemas de UNIANDES, la necesidad de contar con un sistema informático que gestione la información de los proyectos de vinculación que desarrolla dicha institución universitaria. Este sistema debía proporcionar a sus usuarios la posibilidad de acceder al mismo independientemente de la plataforma que usen (PC, teléfono inteligente, tableta); por lo cual se consideró el uso de la tecnología “diseño web adaptable” (responsive web design).Para el desarrollo del software se pensó en las metodologías de desarrollo ágil y de ellas la utilizada fue SCRUM. El software desarrollado permitió al Departamento de Vinculación contar con una herramienta informática adecuada para gestionar la información de sus proyectos. PALABRAS CLAVE: Aplicación web; diseño web adaptable; SCRUM; gestión de proyectos. WEB APPLICATION FOR THE MANAGEMENT OF LINKAGE PROJECTS WITH THE SOCIETY AT THE UNIANDES ABSTRACT UNIANDES university is a higher education institution with extensions in different provinces of Ecuador. One funtion the ecuadorian university is connection with the society. UNIANDES've got "vinculación con la sociedad" department in the Ambato city, All administrative activities about conecctions with the society are centralized in this department. Department needed a computer system to manage the information of projects developed by the university. The system must working on the all platforms (PC, smartphone, tablet). Therefore, the use of "adaptive web design" technology was considered. For the development of the software it was thought about the agile development methodologies and of them the one used was SCRUM. The software developed allowed the Linking Department to have an adequate computer tool to manage the information of their projects. KEYWORDS: Web application; adaptive web design; SCRUM; Project management.

Background: The yield of almonds [Prunus dulcis (Mill.) D.A. Webb] could be low due to climatic problems and any factor improving kernel size and weight, such as the use of plant bioregulators (PBRs), should be beneficial. Results: Three plant bioregulators: 24-epibrassinolide (BL), gibberellic acid (GA3) and kinetin (KN) were applied at three spray concentrations to Non Pareil and Carmel cultivars, at two phenological stages during bloom, in the 2014 and 2015 seasons. The results showed significant differences (P

El presente trabajo tuvo como objetivo identificar los principales géneros de nematodos fitoparásitos asociados al cultivo de piña ( Ananas comosus ) y sus respectivos porcentajes de ocurrencia (frecuencias) y densidades poblacionales promedio. Se analizaron muestras de suelo y raíces colectadas de 28 campos de piña en la provincia de Rodríguez de Mendoza, ubicadas entre 1796 y 1936 m.s . n.m. Las muestras fueron procesadas por duplicado mediante el método de Baerman modificado en bandeja, utilizan do 50 cc. de suelo y 5 g de raíces. En las muestras de suelo, se identificaron once géneros de nematodos fitoparásitos asociados al cultivo de piña. El género Helicotylenchus fue encontrado en todos los campos evaluados y presentó mayores promedios poblaci onales en muestras de suelo y raíces (24 , 5 y 3 , 9 respectivamente). El presente trabajo constituye el primer estudio de identificación de nematodos fitoparásitos en el cultivo de piña en el departamento de Amazonas y nos brinda información que permitirá est ablecer futuras estrategias para el control de éstos nematodos fitoparásitos .

Los bosques de la Amazonia peruana han experimentado importantes transformaciones desde mediados del siglo pasado. Este trabajo evaluó los Cambios de Cobertura y Uso del Suelo (CCUS) en la provincia de Rodríguez de Mendoza, mediante los métodos de clasificación supervisada de máxima probabilidad e interpretación visual interdependiente de imágenes del satélite Landsat, entre los períodos 1987–2001 y 2001–2016. Se construyeron matrices de tabulación cruzada y se calcularon tasas e índices anuales de cambio. Los resultados muestran una pérdida acumulada de 918,59 km2 de cobertura boscosa. La intensidad de CCUS y la tasa de deforestación fueron mayores en el segundo período de análisis. Se evidenció que las altas concentraciones de pérdida de cobertura boscosa están próximas a la red vial e hídrica. Las principales causas de la pérdida de bosques fueron la actividad ganadera y la expansión agrícola migratoria a pequeña explotación, favorecidas por la accesibilidad de infraestructuras de transporte.

The forests of the Peruvian Amazon have undergone important transformations since the middle of the last century. This work evaluated the changes of Land Cover/Land Use (LC/LU) in the province of Rodríguez de Mendoza (Peruvian Amazon), through the methods of supervised classification of maximum probability and interdependent visual interpretation of images of the Landsat satellite, between the 1987–2001 and 2001–2016. Cross-tabulation matrices were constructed and annual change rates were calculated. The results show an accumulated loss of 918,59 km2 of forest cover. The intensity of LC/LU and the deforestation rate were higher in the second period of analysis. It was evidenced that the high concentrations of loss of forest cover are close to the road and water network. The main causes of the loss of forests were the livestock activity and the migratory agricultural expansion to the small farm, favored by the accessibility of the transport infrastructures.

Peruvian aquaculture, specifically trout production, has had significant growth over the past ten years. However, the establishment and expansion of small-scale aquaculture have been carried out without considering the suitability of the land. In Peru, such land suitability studies have yet to be reported. Therefore, a methodological framework is presented for inland aquaculture, which may be replicated, with the necessary complements, for the entire Fisheries and Aquaculture sector in Peru. This work modeled the suitability of land for sustainable rainbow trout aquaculture in the Molinopampa district (Peru). Fifteen key criteria (socioeconomic, environmental and physicochemical) were identified for the proper development of fish activity. These were mapped using Remote Sensing (RS) and Geographic Information Systems (GIS). The Analytical Hierarchy Process (AHP) was applied to build peer-to-peer comparison matrices and weight the importance of the criteria. The weighted overlay of maps (criteria) made it possible to determine that 4.26%, 23.03% and 69.73% of the territory is “very suitable”, “moderately suitable” and “marginally suitable”, respectively, for the development of aquaculture. The implementation of this methodology will contribute to more effective investment planning and efforts, both by the government and by private initiative.

Background: In commercial almond [Prunus dulcis (Mill.) D.A. Webb] orchards in Chile, the percentage of fruit set is low and commonly reaches 5–30%. As almond trees bloom during a cool period and also suffer from self-incompatibility, any factor that can improve pollination, pollen germination and pollen tube growth or extend the effective pollination period, such as the application of plant bioregulators (PBRs), should be beneficial for fruit production. Results: Three plant bioregulators (PBRs): brassinolide (BL), gibberellic acid (GA3) and kinetin (KN) were evaluated for pollen germination and pollen tube growth in vitro, as well as for fruit set in almond cultivars Non Pareil and Carmel, in central Chile, during the 2013 and 2014 seasons. For pollen germination in vitro, the BL concentration of 10 mg L-1 had the highest value in Non Pareil (97.7%), after 4 h germination in 2014 growing season (the control was 90.9%). KN at a concentration of 50 μL L-1 induced the longest pollen tube growth of 1243.4 μm in Carmel after 8 h germination in 2013 (the control was 917.7 μm). In Non Pareil, the highest percentage of fruit set (31.0%) was achieved in 2014 by spraying during bloom at pink bud stage with KN at 50 μL L-1 (the control was 16.7%). Conclusions: A significant favorable effect of the tested PBRs was observed in pollen germination and pollen tube growth in vitro, as well as on fruit set in Non Pareil and Carmel almonds.

Taking advantage of the excellent qualities of the studied coals from the coalfields in Cundinamarca and terms of the types of coking ovens, for excellent performance and quality that allows them to be very competitive.The international market is very demanding in terms of the quality of coke and CRI and CSR indexes, that is why coals laboratories were implemented and improved the system of sampling and analysis of laboratories for more agile and modern. Mixtures were made with the test methodology and empirical methods combining swelling index and rank cals were the theree main types: high, medium and low volatile. But with studies and techological change and enviromental control, process is managed, allowing the coke industry to serve as a development center for the promotion of industries in the central area of this country. In the study the whole process is presented beginning with the explotaitation of coking coals, the care that must be taken into account to mainttain good quality, transport and enviromental measures, the process of crushing and grinding to prepare the optimal mix and laboratory tests necessary to maintain quality control, the types of ovens in Boyacá Colombia, yields and advances in the se technologies, and finally, the system shutdown process. All of the above was addressed in order to add value to the product with the fulfillment of the international market standards.

In 2008, the European Union introduced the Advanced Medicines Regulation aiming to improve regulation of advanced therapy medicinal products (ATMPs). We applied the ATMPs classification definitions in this Regulation to understand the link of this emerging group of medicinal products and the use of the Orphan Regulation. A total of 185 products that can be classified as ATMPs based on this Regulation have been submitted for orphan designation. Prior to its introduction in 2008, 4.5% of the products submitted for orphan designation met these criteria. This percentage went up to 15% after 2008. We analyzed several parameters associated with active ATMP ODDs focusing on sponsor type and EU-Member State origin, therapeutic area targeted, and ATMP classification [i.e., somatic cell therapy medicinal product, tissue-engineered product (TEP), or gene therapy medicinal product (GTMP)] and the use of regulatory services linked to incentives such as the use of protocol assistance (PA) and other Committees [Committee for Advanced Therapies (CAT) and the Pediatric Committee]. The aim here was to gain insight on the use of different services. The UK submits the largest number of ATMPs for ODD representing ~30% of the total to date. Few submissions have been received from central and Eastern European Member States as well as some of the larger Member States such as Germany (3.6%). ATMPs ODDs were primarily GTMPs (48.7%) and SCTMPs (43.3%). TEPs only represented 8% of all submissions for this medicinal class. This is different from non-ODDs ATMPs where GTMPs make only 20% of ATMPs. A total of 11.7% of ATMP ODDs had received formal CAT classification. A total of 29.8% of all orphan drug (OD) ATMPs requested PA. A total of 71.8% did not have an agreed pediatric investigation plan (PIP). Four products (Glybera one PA; Zalmoxis two; Holoclar one; Strimvelis three) have received a marketing authorization (MAA) and a 10-year market exclusivity. Strimvelis also completed their PIP, which was compliant and received the additional 2-year extension to their 10-year market exclusivity. One OD ATMP (Cerepro) received a negative opinion for MAA. The use of services linked to incentives offered by different legislations for ATMP ODDs is low, indicating a need for increasing awareness.

El estudio pretendió conocer el impacto de las Tecnologías de la Comunicación (TICS) en las relaciones interpersonales de estudiantes de dos universidades en la ciudad de Montería, Colombia.La muestra fueconformada por 130 universitarios. En su concreción, se utilizó el enfoque cuantitativo de investigación con diseño descriptivo,y la encuesta como técnica de recogida de datos. El estudio mostró que el 98.5% de los universitariosposeen teléfono móvil y acceso a la internet, como tecnologías de comunicación predominante; asimismo, los jóvenes afirmaronque sus relaciones sociales cambiaron a causa de las redes sociales y una minoría prefiere la comunicación cara a cara. También se observa la relación entre tecnología y la presencia desíntomas de malestar psicológico como ansiedad, irritabilidad, desesperación y estrés. AbstractThe study sought to determine the impact of communication technologies in interpersonal relations of students from two universities in the city of Monteria;Colombia. In the sample there were 130 university students. The quantitative approach with a descriptive design research and a survey and data collection technique was used. The study showed that 98.5% of college students have mobile phone and internet as predominant communication technologies; They said their social relations changed through social networks and only a minority prefer face to face communication, the relationship between technology and the presence of symptoms of psychological distress such as anxiety, irritability, despair and stress is also observed.

Se señalan algunos usos y abusos extendidos del diagnóstico y de la clasificación y se propone prudencia y estudio a la hora de postular la existencia de nuevas estructuras clínicas, además de la neurosis y la psicosis.

A sequential injection system for determination of sulfate in wines was developed. It was based on the formation of BaSO4, measured turbidimetrically at 420 nm. As sample dilution and acidification were required, both sample and acid were sent to a mixing chamber placed in a lateral port of the selection valve; after thorough mixing, up to six aliquots could be drawn and analyzed. A linear calibration curve was established using 10% v/v ethanol standards with concentrations between 300 and 1500 mg L-1 of K2SO4. The results obtained by the present methodology were statistically comparable with those furnished by the reference procedure, with relative deviations < 3.5%. The sample frequency was about 5 samples per h with relative standard deviations (RSD) < 10%.

A series of phosphatidylethanolamine fluorescent probes head-labelled with 3-carboxycoumarin was prepared by an improved bioconjugation approach through continuous flow synthesis. The established procedure, supported by a design of experiment (DoE) set-up, resulted in a significant reduction in the reaction time compared to the conventional batch method, in addition to a minor yield increase. The characterization of these probes was enhanced by an in-depth molecular dynamics (MD) study of the behaviour of a representative probe of this family, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine labelled with 3-carboxycoumarin (POPE-COUM), in bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine (SLPC) 2:1, mimicking the composition of the egg yolk lecithin membranes recently used experimentally by our group to study POPE-COUM as a biomarker of the oxidation state and integrity of large unilamellar vesicles (LUVs). The MD simulations revealed that the coumarin group is oriented towards the bilayer interior, leading to a relatively internal location, in agreement with what is observed in the nitrobenzoxadiazole fluorophore of commercial head-labelled NBD-PE probes. This behaviour is consistent with the previously stated hypothesis that POPE-COUM is entirely located within the LUVs structure. Hence, the delay on the oxidation of the probe in the oxygen radical absorbance capacity (ORAC) assays performed is related with the inaccessibility of the probe until alteration of the LUV structure occurs. Furthermore, our simulations show that POPE-COUM exerts very little global and local perturbation on the host bilayer, as evaluated by key properties of the unlabelled lipids. Together, our findings establish PE-COUM as suitable fluorescent lipid analogue probes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The presence of endocrine-disrupting compounds (EDCs) in water poses a significant threat to human and animal health, as recognized by regulatory agencies throughout the world. The Yeast Estrogen Screen (YES) assay is an excellent method to evaluate the presence of these compounds in water due to its simplicity and capacity to assess the bioaccessible forms/fractions of these compounds. In the presence of a compound with estrogenic activity, Saccharomyces cerevisiae cells, containing a lacZ reporter gene encoding the enzyme β-galactosidase, are induced, the enzyme is synthesised, and released to the extracellular medium. In this work, a YES-based approach encompassing the use of a lacZ reporter gene modified strain of S. cerevisiae, microcarriers as solid support, and a fluorescent substrate, fluorescein di-β-d-galactopyranoside, is proposed, allowing for the assessment of EDCs' presence after only 2 h of incubation. The proposed method provided an EC50 of 0.17 ± 0.03 nM and an LLOQ of 0.03 nM, expressed as 17β-estradiol. The assessment of different EDCs provided EC50 values between 0.16 and 1.2 × 10 3  nM. After application to wastewaters, similar results were obtained for EDCs screening, much faster, compared to the conventional 45 h spectrophotometric procedure using a commercial kit, showing potential for onsite high-throughput screening of environmental contamination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Around two million tons of olive oil are produced in Europe annually, with Portugal being among the top five European olive oil-producing countries. Olive oil production results in a substantial amount of waste in the form of olive leaves. These discarded olive leaves contain valuable phenolic compounds with antioxidant, anti-inflammatory, hypoglycaemic, neuroprotective, and antiproliferative properties. Due to their richness in polyphenols with health-promoting properties, olive leaves can be considered a potential functional food ingredient. Thus, sustainable practices for reusing olive leaf waste are in demand. In this study, the polyphenolic content in olive leaves from different Portuguese locations was determined using HPLC-UV-Vis after defining the best fit-for-purpose liquid extraction strategy. The differences in the in vitro antioxidant activity in these samples were determined by several methodologies based on radical scavenging (against 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS), 2,2-diphenyl-2-picrylhydrazyl (DPPH), and peroxyl radical (ORAC)) and on reducing properties (cupric-reducing antioxidant capacity (CUPRAC), and Folin-Ciocalteu assay (FC)), to unveil the relationship between the profile and quantity of polyphenols with antioxidant mechanisms and their capacity. At last, the stability of extracted compounds upon lyophilization and exposition to surrogate biological fluids was assessed, envisioning the future incorporation of olive leaves extracted compounds in food products.

Soil properties influence greatly the status of vine plants which consequently influences the quality of wine. Therefore, in the context of viticulture management, it is extremely important to assess the physical and chemical parameters of vineyards soils. In this study, the soils of two vineyards were analysed by near-infrared (NIR) spectroscopy and established analytical reference procedures. The main objective of this study was to verify if NIR spectroscopy is a potential tool to discriminate the soils of both vineyards as well as to quantify differences of soil's parameters. For that, a total of eight sampling spots were selected at each vineyard taking into consideration the soil type and sampled at different depths. The data analysis was performed using analysis of variance (ANOVA), principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) and partial least squares (PLS) regression. The ANOVA results revealed that 12 out of the 18 parameters analysed through the reference procedures can be considered statistically different (p < 0.05). Regarding PCA, the obtained results revealed a clear separation between the scores of both vineyards either considering NIR spectra or the chemical parameters. The PLS-DA model was able to obtain 100 % of correct predictions for the discrimination of both vineyards. PLS regression analysis using NIR spectra revealed R 2 P and RER values higher than 0.85 and 10, respectively, for 8 (pH (H 2 O), N, Ca 2+ , Mg 2+ , SB, CEC, ECEC and GSB) of the 18 chemical parameters evaluated. Concluding, these results demonstrate that it is possible to discriminate the soils of the different vineyards through NIR spectroscopy as well as to quantify several chemical parameters through soils NIR spectra in a rapid, accurate, cost-effective, simple and environmentally friendly way when compared to the reference procedures., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)

Indoxyl sulfate (INDS) and p-cresol sulfate (pCS) are two of the most relevant uremic toxins that are recognized to have an essential role in chronic kidney disease (CKD) progression and associated cardiovascular risk. Thus, it is crucial to accurately assess their circulating levels in the body. Aiming at establishing an analytical strategy for quantification of INDS and pCS in human plasma, an automatic on-line micro-solid-phase extraction (μSPE) procedure hyphenated to tandem mass spectrometry (MS/MS) detection without previous chromatographic separation was herein developed. The bead injection (BI) concept was used to implement the μSPE procedure in the lab-on-valve (LOV) format. After studying the extraction conditions, the anion-exchange OASIS WAX sorbent beads (10 mg) and 99% ACN-H 2 O (15:85, v/v)-1% (v/v) NH 4 OH were chosen as sorbent and eluent, respectively, as they provided the highest analyte recoveries. Subsequently, the μSPE-BI-LOV system was hyphenated on-line to a MS/MS detector and the full analytical cycle, comprising sample preparation and analytes detection, was completed in <20 min. The developed μSPE-BI-LOV-MS methodology presented good linearity (r 2  > 0.999) for quantification of the target analytes at concentrations ranging from 18 to 360 μg mL -1 in plasma. LOQ values were 2 μg mL -1 for INDS and 7 μg mL -1 for pCS in plasma. Human plasma samples from healthy subjects and individuals with CKD were successfully analyzed using the developed approach. The proposed automatic methodology can be described as an eco-friendly strategy, with a favorable score of 0.64 after greenness evaluation using the AGREE metric., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Nanoparticles used for medical applications commonly possess coatings or surface functionalities intended to provide specific behavior in vivo , for example, the use of PEG to provide stealth properties. Direct, quantitative measurement of the surface chemistry and composition of such systems in a hydrated environment has thus far not been demonstrated, yet such measurements are of great importance for the development of nanomedicine systems. Here we demonstrate the first use of cryo-XPS for the measurement of two PEG-functionalized nanomedicines: a polymeric drug delivery system and a lipid nanoparticle mRNA carrier. The observed differences between cryo-XPS and standard XPS measurements indicate the potential of cryo-XPS for providing quantitative measurements of such nanoparticle systems in hydrated conditions.

The quantification of the drug associated to nanoparticle carriers, often expressed in terms of encapsulation efficiency, is a regulatory requirement. The establishment of independent methods to evaluate this parameter provides a means for measurement validation, which is critical in providing confidence in the methods and enabling the robust characterization of nanomedicines. Chromatography is traditionally used to measure drug encapsulation into nanoparticles. Here, we describe an additional independent strategy based on analytical centrifugation. The encapsulation of diclofenac into nanocarriers was quantified based on the mass difference between placebo (i.e. unloaded) and loaded nanoparticles. This difference was estimated using particle densities measured by differential centrifugal sedimentation (DCS) and size and concentration values measured by particle tracking analysis (PTA). The proposed strategy was applied to two types of formulations, namely poly(lactic-co-glycolic acid) (PLGA) nanoparticles and nanostructured lipid carriers, which were analysed by DCS operated in sedimentation and flotation modes, respectively. The results were compared to those from high performance liquid chromatography (HPLC) measurements. Additionally, X-ray photoelectron spectroscopy analysis was used to elucidate the surface chemical composition of the placebo and loaded nanoparticles. The proposed approach enables the monitoring of batch-to-batch consistency and the quantification of diclofenac association to PLGA nanoparticles from 0.7 ng to 5 ng of drug per 1 μg of PLGA, with good linear correlation between DCS and HPLC results (R 2  = 0.975). Using the same approach, similar quantification in lipid nanocarriers was possible for a loading of diclofenac ≥1.1 ng per 1 μg of lipids, with results in agreement with the HPLC method (R 2  = 0.971). Hence, the strategy proposed here expands the analytical tools available for evaluating nanoparticles encapsulation efficiency, being thus significant for increasing the robustness of drug-delivery nanocarriers characterization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Nanoparticles (NPs) concentration directly impacts the dose delivered to target tissues by nanocarriers. The evaluation of this parameter is required during NPs developmental and quality control stages, for setting dose-response correlations and for evaluating the reproducibility of the manufacturing process. Still, faster and simpler procedures, dismissing skilled operators and post-analysis conversions are needed to quantify NPs for research and quality control operations, and to support result validation. Herein, a miniaturized automated ensemble method to measure NPs concentration was established under the lab-on-valve (LOV) mesofluidic platform. Automatic NPs sampling and delivery to the LOV detection unit were set by flow programming. NPs concentration measurements were based on the decrease in the light transmitted to the detector due to the light scattered by NPs when passing through the optical path. Each analysis was accomplished in 2 min, rendering a determination throughput of 30 h -1 (6 samples h -1 for n = 5) and only requiring 30 μL (≈0.03 g) of NPs suspension. Measurements were performed on polymeric NPs, as these represent one of the major classes of NPs under development for drug-delivery aims. Determinations for polystyrene NPs (of 100, 200, and 500 nm) and for NPs made of PEGylated poly-d,l-lactide- co -glycolide (PEG-PLGA, a biocompatible FDA-approved polymer) were accomplished within 10 8 -10 12 particles mL -1 range, depending on the NPs size and composition. NPs size and concentration were maintained during analysis, as verified for NPs eluted from the LOV by particle tracking analysis (PTA). Moreover, concentration measurements for PEG-PLGA NPs loaded with an anti-inflammatory drug, methotrexate (MTX), after their incubation in simulated gastric and intestinal fluids were successfully achieved (recovery values of 102-115%, as confirmed by PTA), showing the suitability of the proposed method to support the development of polymeric NPs targeting intestinal delivery.

The analysis and interpretation of data retrieved from Oxygen Radical Absorbance Capacity (ORAC) assays represent a challenging task. ORAC indexes originate from different mathematical approaches often lacking correct elucidation of kinetic features concerning radical scavenging reactions by antioxidant compounds. In this work, the expression of ORAC values as area under fluorescein (FL) decay curves (AUC) and lag time are critically compared. This multi-parametric analysis showed the extension of radical scavenging reactions beyond the lag time period for caffeic acid, gallic acid, reduced glutathione and quercetin, extending their antioxidant protection of FL. Ethanol delayed the reaction of both FL and antioxidant compounds with free radical species generated from 2,2'-azobis(2-amidinopropane) dihydrochloride thermolysis. Trolox equivalent values, commonly used to express ORAC values, were more affected by the differences in radical scavenging kinetics between the reference and the tested antioxidant compounds when calculated from AUC than from lag time. These findings stressed the importance of choosing calibrator compounds presenting ORAC kinetics similar to samples to prevent biased estimation of the antioxidant capacity. Additionally, the framework proposed here provides a sustainable analytical method for the evaluation of antioxidant capacity, with an AGREE score of 0.73.

Protein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multianalyte methods allowing the establishment of a uremic metabolite profile are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fluorescence detection for the simultaneous quantification of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12 min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate buffer containing an ion-pairing reagent, at a flow rate of 2 mL min -1 . Standards were prepared in plasma and quantification was performed using the background subtraction approach. LOQ values were ≤ 0.2 µg mL -1 for all analytes except for pCS (LOQ of 2 µg mL -1 ). The method proved to be accurate (93.5-112%) and precise (CV ≤ 14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting significantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression., (© 2022. Springer-Verlag GmbH Germany, part of Springer Nature.)

Protein-bound uremic retention solutes, such as indole-3-acetic acid, indoxyl sulfate, p-cresol and p-cresol sulfate, are associated with the development of several pathologies, namely renal, cardiovascular, and bone toxicities, due to their potential accumulation in the human body, thus requiring analytical methods for monitoring and evaluation. The present review addresses conventional and advanced sample treatment procedures for sample handling and the chromatographic analytical methods developed for quantification of these compounds in different biological fluids, with particular focus on plasma, serum, and urine. The sample preparation and chromatographic methods coupled to different detection systems are critically discussed, focusing on the different steps involved for sample treatment, namely elimination of interfering compounds present in the sample matrix, and the evaluation of their environmental impact through the AGREEprep tool. There is a clear trend for the application of liquid-chromatography coupled to tandem mass spectrometry, which requires protein precipitation, solid-phase extraction and/or dilution prior to analysis of biological samples. Furthermore, from a sustainability point of view, miniaturized methods resorting to microplate devices are highly recommended., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Vascular calcification (VC) is a frequent condition in chronic kidney disease (CKD) and a well-established risk factor for the development of cardiovascular disease (CVD). Gut dysbiosis may contribute to CVD and inflammation in CKD patients. Nonetheless, the role of gut and blood microbiomes in CKD-associated VC remains unknown. Therefore, this pilot study aimed to explore the link between gut and blood microbiomes and VC in CKD patients on peritoneal dialysis (CKD-PD). Our results showed relative changes in specific taxa between CKD-PD patients with and without VC, namely Coprobacter, Coprococcus 3, Lactobacillus, and Eubacterium eligens group in the gut, and Cutibacterium, Pajaroellobacter, Devosia, Hyphomicrobium, and Pelomonas in the blood. An association between VC and all-cause mortality risk in CKD-PD patients was also observed, and patients with higher mortality risk corroborate the changes of Eubacterium eligens in the gut and Devosia genus in the blood. Although we did not find differences in uremic toxins, intestinal translocation markers, and inflammatory parameters among CKD-PD patients with and without VC, soluble CD14 (sCD14), a nonspecific marker of monocyte activation, positively correlated with VC severity. Therefore, gut Eubacterium eligens group, blood Devosia, and circulating sCD14 should be further explored as biomarkers for VC, CVD, and mortality risk in CKD.

An automatic micro-solid-phase extraction (μSPE) method using on-line renewable sorbent beads followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was established for the determination of tranexamic acid (TXA) in urine. The μSPE method was based on the bead injection (BI) concept combined with the mesofluidic lab-on-valve (LOV) platform. All steps of the μSPE-BI-LOV were implemented by computer programming, rendering enhanced precision on time and flow events. Several parameters, including the type of sorbent, volume and composition of the conditioning solution, washing solution, and eluent composition, were evaluated to improve the extraction efficiency. The best results were obtained with a hydrophilic-lipophilic balanced mixed-mode sorbent, decorated with sulfonic acid groups (Oasis MCX), and 99% acetonitrile-water (50:50, v/v)-1% ammonium hydroxide as eluent. Chromatographic separation was performed using a BEH amide column coupled to MS/MS detection in positive ionization mode. Good linearity was achieved (R 2  > 0.998) for TXA concentrations in urine ranging from 300 to 3000 ng mL -1 , with LOD and LOQ of 30 and 65 ng mL -1 , respectively. Dilution integrity was observed for dilution factors up to 20,000 times, providing the extension of the upper limit of quantification to 12 mg mL -1 . The method was validated according to international guidelines and successfully applied to urine samples collected during scoliosis surgery of pediatric patients treated with TXA., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

The quantitative analysis of pharmaceuticals in biomatrices by liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is often hampered by adduct formation. The use of the molecular ion resulting from solvent adducts for quantification is uncommon, even if formed in high abundance. In this work, we propose the use of a protonated acetonitrile adduct for the quantitative analysis of tranexamic acid (TXA) by LC-MS/MS. The high abundance of the protonated acetonitrile adduct [M + ACN + H] + was found to be independent of source-dependent parameters and mobile phase composition. The results obtained for TXA analysis in clinical samples were comparable for both [M + ACN + H] + and [M + H] + , and no statistically significant differences were observed. The relative stability and structure of the [M + ACN + H] + ions were also studied by analyzing probable structures from an energetic point of view and by quantum chemical calculations. These findings, and the studied fragmentation pathways, allowed the definition of an acetimidium structure as the best ion to describe the observed acetonitrile protonated adduct of TXA.

Supplemental zinc from organic sources has been suggested to be more bioavailable than inorganic ones for dog foods. However, the bioavailability of zinc might be affected by dietary constituents such as phytates. The present study aimed to evaluate the effects of two zinc sources (zinc sulfate and zinc proteinate) and the addition of a multi-enzymatic complex from the solid-state fermentation of Aspergillus niger on end-products of fecal fermentation and fecal microbiota of adult Beagles fed a high-phytate diet. The experimental design consisted of three 4 × 4 Latin Squares with a 2 × 2 factorial arrangement of treatments ( n = 12 Beagles), with four periods and four diets: zinc sulfate without (IZ) or with (IZ +) enzyme addition, and zinc proteinate without (OZ) or with (OZ +) enzyme addition. Enzyme addition significantly affected Faith's phylogenetic diversity index, whereas zinc source did not affect either beta or alpha diversity measures. Linear discriminant analysis effect size detected nine taxa as markers for organic zinc, 18 for inorganic source, and none for enzyme addition. However, with the use of a negative binomial generalized linear model, further effects were observed. Organic zinc was associated with a significantly higher abundance of Firmicutes and lower Proteobacteria and Bacteroidetes, although at a genus level, the response varied. The DNA abundance of Clostridium cluster I, Clostridium cluster XIV, Campylobacter spp., Ruminococcaceae, Turicibacter , and Blautia was significantly higher in dogs fed IZ and IZ + diets. Higher abundance of genus Lactobacillus was observed in dogs fed enzyme-supplemented diets. End-products of fecal fermentation were not affected by zinc source or enzymes. An increase in some taxa of the phyla Actinobacteria and Firmicutes was observed in feces of dogs fed organic zinc with enzyme addition but not with inorganic zinc. This study fills a gap in knowledge regarding the effect of zinc source and enzyme addition on the fecal microbiota of dogs. An association of zinc bioavailability and bacteria abundance is suggested, but the implications for the host (dog) are not clear. Further studies are required to unveil the effects of the interaction between zinc sources and enzyme addition on the fecal microbial community., Competing Interests: AP’s Ph.D. scholarship was co-funded by Alltech Portugal. Alltech Portugal approved the publication of the manuscript without interfering in any stage of the study, including experimental design, analysis, and report of the results. EM was employed by the company Sorgal, which produced the diets according to the design of the study. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Pereira, Maia, Pinna, Biagi, Matos, Segundo, Fonseca and Cabrita.)

The determination of contaminants of emerging concern (CECs) in environmental samples has become a challenging and critical issue. The present work focuses on miniaturized analytical strategies reported in the literature for the determination of CECs. The first part of the review provides brief overview of CECs whose monitoring in environmental samples is of particular significance, namely personal care products, pharmaceuticals, endocrine disruptors, UV-filters, newly registered pesticides, illicit drugs, disinfection by-products, surfactants, high technology rare earth elements, and engineered nanomaterials. Besides, an overview of downsized sample preparation approaches reported in the literature for the determination of CECs in environmental samples is provided. Particularly, analytical methodologies involving microextraction approaches used for the enrichment of CECs are discussed. Both solid phase- and liquid phase-based microextraction techniques are highlighted devoting special attention to recently reported approaches. Special emphasis is placed on newly developed materials used for extraction purposes in microextraction techniques. In addition, recent contributions involving miniaturized analytical flow techniques for the determination of CECs are discussed. Besides, the strengths, weaknesses, opportunities and threats of point of need and portable devices have been identified and critically compared with chromatographic methods coupled to mass chromatography. Finally, challenging aspects regarding miniaturized analytical methods for determination of CECs are critically discussed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)