Plasmodium falciparum parasite population structure and gene flow associated to anti-malarial drugs resistance in Cambodia

Background: Western Cambodia is recognized as the epicentre of emergence of Plasmodium falciparum multi-drug resistance. The emergence of artemisinin resistance has been observed in this area since 2008–2009 and molecular signatures associated to artemisinin resistance have been characterized in k13 gene. At present, one of the major threats faced, is the possible spread of Asian artemisinin resistant parasites over the world threatening millions of people and jeopardizing malaria elimination programme efforts. To anticipate the diffusion of artemisinin resistance, the identification of the P. falciparum population structure and the gene flow among the parasite population in Cambodia are essential. Methods: To this end, a mid-throughput PCR-LDR-FMA approach based on LUMINEX technology was developed to screen for genetic barcode in 533 blood samples collected in 2010–2011 from 16 health centres in malaria endemics areas in Cambodia. Results: Based on successful typing of 282 samples, subpopulations were characterized along the borders of the country. Each 11-loci barcode provides evidence supporting allele distribution gradient related to subpopulations and gene flow. The 11-loci barcode successfully identifies recently emerging parasite subpopulations in western Cambodia that are associated with the C580Y dominant allele for artemisinin resistance in k13 gene. A subpopulation was identified in northern Cambodia that was associated to artemisinin (R539T resistant allele of k13 gene) and mefloquine resistance. Conclusions: The gene flow between these subpopulations might have driven the spread of artemisinin resistance over Cambodia

PROJECTOS DE EXTENSÃO UNIVERSITÁRIA: FACTOR DA QUALIDADE NA FORMAÇÃO PROFISSIONAL E PROMOÇÃO DA IMAGEM DAS IES. ESTUDO REALIZADO NA ESCOLA SUPERIOR PEDAGÓGICA DO BENGO (ESPB)

A Higher Education Institution (HEI) is assessed on the basis of the quality of its students' exit profile, their efficiency in solving a particular personal and social problem. However, the question of quality in higher education was moderately motivated by the concern of governments with the relationship between economic investment at this level of education and school success. The university extension is an essential tool that universities have to guarantee quality in the professional training of students, thus providing an opportunity for more insertion in reality, which you will find when you become a professional, because it articulates teaching and research in solving social problems. The research is part of the qualitative approach model. A case study was carried out on ESPB's university extension project, entitled: Guidance to Families of Children with Special Educational Needs, with the aim of demonstrating the quality of student training as guarantee human capital and promote the image of HEI. The results show that the projects of university extension are characterized as one of the ways in which university extension is carried out, giving special importance to the construction of citizenship, seeking to overcome the social differences and contributing with their actions to the realization of inclusive policies. Its actions seek to contribute to the scientific and citizen education of students, teachers, employees and the community.Uma Instituição de Ensino Superior (IES) é avaliada em função da qualidade do perfil de saída dos seus estudantes e da sua eficiência na resolução de determinados problemas pessoais e sociais. Contudo, a questão da qualidade no ensino superior foi moderadamente motivada pela preocupação dos governos com a relação entre o investimento económico, neste nível de ensino, e o sucesso escolar. A extensão universitária figura-se como ferramenta imprescindível que as universidades dispõem para garantir a qualidade na formação profissional dos estudantes, proporcionando, deste modo, uma oportunidade a mais de inserção na realidade, que encontrará quando se tornar um profissional, pois, articula o ensino e a pesquisa na resolução de problemas sociais. A pesquisa enquadra-se no modelo de abordagem qualitativo, realizou-se um estudo de caso ao projecto de extensão universitária da ESPB, intitulado: Orientação às Famílias de Crianças com Necessidades Educativas Especiais, com o objectivo de demonstrar a qualidade da formação dos estudantes como garantia do capital humano e promoção da imagem da IES. Os resultados obtidos revelaram que os projectos de extensão universitária caracterizam-se como uma das vias pelas quais se realiza a extensão universitária, concedendo especial relevância à construção da cidadania, buscando superar as diferenças sociais e contribuindo com suas acções para se efectivarem políticas inclusivas. As suas acções buscam contribuir para a formação científica e cívicade estudantes, professores, funcionários e comunidade em geral, promovendo um diálogo de saberes: académico e popular

ENVEJECIMIENTO Y SOLEDAD

Objective: The aim of this study is to analyze whether elderly people in a state of fragility, visited by the Emergency Domiciliary Care service, have received a follow-up program by the nursing staff and whether they were labeled as “Elderly people in state of fragility.” Material and method: A two-phase observational study. Firstly, in phase I, a population comprising people older than 64 and visited by the emergency domiciliary service from the Raval Nord Primary Care Centre was selected. Whether they had received nursing follow-up and whether they had been diagnosed as elderly people in a frail state was analyzed. In phase II, a simple random sample from this population was chosen. Here, it was analyzed whether there had been changes in the nursing diagnosis and follow-up. Results: Of a total was seen that of 776 medical emergency domiciliary visits, 568 (73.19%) were people belonging to the over 64 group. Out of the total, 57 cases (10%) belonged to the age group between 75 and 85 years old (80%). Out of this group, 77.26% were women; 94% were diagnosed with a chronic condition (diabetes, hypertension, chronic obstructive pulmonary disease, etc); and 77.2% were polymedicated. In addition, 66.7% were not included in the Domiciliary Care Program and were not diagnosed as frail elderly people. In phase II, an increase in nursing care, as well as in the “frail elderly people” diagnosis were found. Conclusions: The use of nursing diagnosis for elderly people in a frail state is an indispensable tool for the monitoring and follow-up of that population.Objetivo: Analizar si en las personas mayores en situación de fragilidad que han sido atendidas por el servicio de urgencias domiciliarias, constaba el seguimiento por el personal de enfermería del centro y  si constaba  el diagnóstico de “Personas mayores en situación de fragilidad”. Material y método: Se realizó un estudio observacional en dos fases: 1ª se analizaron todas las personas mayores de 64 años que fueron atendidas en el servicio de urgencias domiciliarias del centro de Atención Primaría de Raval Nord, se analizó si en dicha población constaba el seguimiento de enfermería y el diagnóstico de personas mayores en situación de fragilidad ; 2ª fase, se realizó un muestreo simple  de la población analizada y se evaluó si se produjeron cambios en  los diagnósticos de enfermería y seguimiento de dicha población. Resultados: De un total de 776 visitas domiciliarias de urgencias médicas, se observó que 568 (73,19%) eran mayores de 64 años. Del total, se estudiaron 57 (10%) casos y se observó que el 80% pertenecían al grupo de edad comprendido entre 75 y 85 años; el 77,26% eran mujeres; el 94% con diagnóstico crónico (Diabetes, Hipertensión Epoc...); el 77,2% eran personas polimedicadas y el 66,7% del total no estaban incluidas en el programa de Atención Domiciliaria y no constaba ningún diagnóstico de “personas mayores frágiles”. En la segunda fase se observa un incremento  en la atención por parte de los profesionales de enfermería y un aumento en el diagnóstico de “Personas mayores frágiles”. Conclusiones: La utilización del diagnóstico de enfermería en las personas en situación de fragilidad es una herramienta imprescindible para el seguimiento y control de dicha población.  

Effects of sheep and horse manure and pine bark amendments on metal distribution and chemical properties of contaminated mine soils.

Mine soils usually contain large levels of heavy metals and poor fertility conditions which limit their reclamation and the application of phyto-remediation technologies. Two organic waste materials (pine bark compost and sheep and horse manure compost), with different pHs and varying degrees of humification and nutrient contents, were applied as amendments to assess their effects on copper (Cu) and zinc (Zn) bioavailability and on fertility conditions of mine soils. Soil samples collected from two abandoned mining areas near Madrid (Spain) were mixed with 0, 30 and 60 t ha?1 of the organic amendments. The concentrations of metals among the different mineral and organic fractions of soil were determined by several extraction procedures to study the metal distribution in the solid phase of the soil affected by the organic amendments. The results showed that the manure amendment increased the soil pH and the cation exchange capacity and enhanced the nutrient levels of these soils. The pine bark amendment decreased the soil pH and did not significantly change the nutrient status of soil. Soil pH, organic matter content and its degree of humification, which were altered by the amendments, were the main factors affecting Cu fractionation. Zn fractionation was mainly affected by soil pH. The addition of manure not only improved soil fertility, but also decreased metal bioavailability resulting in a reduction of metal toxicity. Conversely, pine bark amendment increased metal ioavailability. The use of sheep and horse manure could be a cost-effective practice for the restoration of contaminated mine soils

Using force spectroscopy analysis to improve the properties of the hairpin probe

The sensitivity of hairpin-probe-based fluorescence resonance energy transfer (FRET) analysis was sequence-dependent in detecting single base mismatches with different positions and identities. In this paper, the relationship between the sequence-dependent effect and the discrimination sensitivity of a single base mismatch was systematically investigated by fluorescence analysis and force spectroscopy analysis. The same hairpin probe was used. The uneven fluorescence analysis sensitivity was obviously influenced by the guanine-cytosine (GC) contents as well as the location of the mismatched base. However, we found that force spectroscopy analysis distinguished itself, displaying a high and even sensitivity in detecting differently mismatched targets. This could therefore be an alternative and novel way to minimize the sequence-dependent effect of the hairpin probe. The advantage offered by force spectroscopy analysis could mainly be attributed to the percentage of rupture force reduction, which could be directly and dramatically influenced by the percentage of secondary structure disruption contributed by each mismatched base pair, regardless of its location and identity. This yes-or-no detection mechanism should both contribute to a comprehensive understanding of the sensitivity source of different mutation analyses and extend the application range of hairpin probes

A

Abstract:  SARS-CoV-2 is causing COVID-19, a new respiratory virus from Wuhan, China, since December 2019 and devolved into the current pandemic. Viral circulation and infection would become endemic, making direct immunofluorescence (IF) a cost-effective diagnostic methodology for sustained sentinel surveillance. We proposed to develop an IF for SARS-CoV-2 in slides with respiratory samples from patients challenged with antibodies from the serum of convalescent patients as an intermediate reagent for subsequent labeling with fluorochrome conjugate. With Ethics Committee approvals, nasopharyngeal swabs and sera were obtained from patients with PCR-confirmed COVID-19. The slides were prepared following the standard method in a biological biosafety cabinet. The conditions for performing the IF technique were: 1-Sera were assayed with CovidAR, CMIA Arquitect-ABBOTT and Neutralization-titrated challenge sera. 2-Anti-FC IgG of human IgG obtained in mouse and goat conjugated with fluorescein. 3-Blocking with albumin and tween 20 at different stages of the technique. The results obtained so far indicate that the best combination of conditions involves the use of blocking with 1% albumin + 0.05% tween 20 on the slides for 20 minutes and a layer of human serum with specific antibody titer of 1/80 NT. In addition, the goat conjugate performed better. Validation and standardization with monoclonal antibodies are still pending. Descriptive analyses will be performed by means of tables and graphs, establishing absolute and relative frequencies (%).  Chi-square analysis will be applied estimating sensitivity, specificity, positive and negative predictive values. Interspecific and intraspecific validation assays and statistical analysis of DIF with respect to molecular biology will be performed. The R-Medic software will be used and in all cases the significance level will be 5%.  Although IF has lower sensitivity than molecular methods, it offers practicality in the initial screening task. The relevance of this work lies in being able to implement in the future, once the technique is standardized and validated, the detection of SARS-Cov-2 as differential diagnosis by IF, to distinguish it among the other viral agents of the respiratory panel, contributing to the diagnosis in clinical practice and epidemiological surveillance in Public Health.Resumen:  El SARS-CoV-2 es causante de COVID-19, un nuevo virus respiratorio de Wuhan, China, desde diciembre de 2019 y devino en la pandemia actual. La circulación e infección viral se volverían endémicas, por lo que la Inmunofluorescencia (IF) directa sería una metodología diagnóstica económica para la vigilancia centinela sostenida. Se propuso desarrollar una IF para SARS-CoV-2 en improntas con muestras respiratorias de pacientes enfrentadas con anticuerpos del suero de pacientes convalecientes como reactivo intermediario para el posterior marcaje con conjugado con fluorocromo. Con aprobaciones del Comités de Ética, se obtuvieron hisopados nasofaríngeos y sueros de pacientes con COVID-19 confirmado por PCR. Las improntas se elaboraron siguiendo el método estándar en cabina de bioseguridad biológica. Las condiciones para la realización de la técnica de IF fueron: 1-Se ensayó con sueros problemas titulados por CovidAR, CMIA Arquitect-ABBOTT y Neutralización. 2-IgG anti-FC de IgG humana obtenidas en ratón y cabra conjugada con fluoresceína. 3-Bloqueo con albúmina y tween 20 en diferentes etapas de la técnica. Los resultados obtenidos hasta ahora indican que la mejor combinación de condiciones implica el uso de bloqueo con albúmina al 1% + tween 20 al 0,05% sobre la impronta durante 20 minutos y una capa del suero humano con título de anticuerpos específicos de 1/80 NT.  Además, funcionó mejor el conjugado de cabra. Permanece a la espera la validación y estandarización con anticuerpos monoclonales. Se realizarán análisis descriptivos mediante tablas y gráficos, estableciendo frecuencias absolutas y relativas (%). Se aplicarán análisis Chi cuadrado estimando sensibilidad, especificidad, valores predictivos positivos y negativos. Se harán ensayos de validación interespecíficos e intraespecíficos y análisis estadísticos de IFD con respecto a biología molecular. Se utilizará el soft R-Medic y en todos los casos el nivel de significación será del 5%.  Si bien la IF posee menor sensibilidad que los métodos moleculares, ofrece practicidad en la tarea inicial de tamizaje. La relevancia del trabajo radica en poder implementar a futuro, una vez estandarizada y validada la técnica, la detección de SARS-Cov-2 como diagnóstico diferencial por IF, para distinguirlo entre los otros agentes virales del panel respiratorio, aportando al diagnóstico en la práctica clínica y a la vigilancia epidemiológica en Salud Pública.