Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study

Introduction: Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. Objective: To establish the concordance between the two tests as identifying methods for circulating species in Colombia. Materials and methods: A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen’s kappa coefficient and the 95% confidence interval (CI) were calculated. Results: The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed “very good” concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Conclusion: Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia

Capacidades do Talento Humano para a Pesquisa na Secretaria Distrital de Saúde e em Empresas Sociais do Estado

La generación de conocimiento aplicado a la resolución de problemas en salud es una necesidad inaplazable para impulsar los procesos de desarrollo competitivo en un país. Por eso, uno de los componentes esenciales dentro de los sistemas de ciencia, tecnología e innovación del mundo es el talento humano. En consecuencia, para fortalecer tales capacidades es necesario conocer suficientemente el estado actual. El objetivo del presente trabajo consiste en explorar las capacidades de investigación del talento humano de la Secretaría Distrital de Salud y la red adherida de empresas sociales del Estado. Esto se hizo a través de una metodología mixta donde se realizaron encuestas virtuales, revisiones del estado de los grupos de investigación en la plataforma ScienTI de Colciencias, grupos focales, y entrevistas para profundizar en una visión general y captar las ideas y estrategias probables de fortalecimiento. Los resultados mostraron un bajo porcentaje de servidores públicos y colaboradores con formación de alto nivel (< 6 %) y experiencia en investigación (27,9 %). Sin embargo, tienen un interés cada vez mayor por adherirse tanto al proceso de investigación (76,4 %) en cuanto a los grupos semillas de investigación (69,56 %). Igualmente, se identificaron ocho grupos de investigación dentro de esas instituciones, de los cuales cuatro fueron reconocidos y clasificados en la convocatoria 640-2013 de Colciencias. Estos hallazgos permitieron plantear algunas estrategias para el fortalecimiento de las capacidades para la investigación del talento humano desde la política, la gestión y la movilización del conocimiento institucional, siendo prioritaria la institucionalización de la investigación como eje de la misión en las organizaciones.Generating knowledge to solve health problems is a pressing need to boost the competitive development processes of a country. For this reason, human talent is one of the vital constituents of the world’s science, technology and innovation systems. Therefore, in order to strengthen such abilities, it is necessary to know the current state of the art. The goal of this study is exploring research competencies of the human talent at District Health Department and their networked State Social Enterprises. This was possible through a mixed methodology where we carried out virtual surveys, status checks of the research groups in Colciencias ScienTI platform, focus group, and interviews to deepen into a general overview and catching insights and probable strengthening strategies. Results showed a low percentage of public servants and collaborators with high-level training (< 6 %) and research experience (27,9 %). Nonetheless, there is an increasing interest to carry out both the research process (76,4 %) and research seedlings (69,56 %). In addition, we could identify eight research groups within these institutions, of which four recognized and classified at call 640-2013 by Colciencias. These findings allowed us to propose some strategies to strength research competencies by human talent taking into account policies, management and involvement where institutionalization of research activities may be a cornerstone for organizations

Human Macrophage Response to <em>L. (Viannia) panamensis</em>: Microarray Evidence for an Early Inflammatory Response

<div><h3>Background</h3><p>Previous findings indicate that susceptibility to <em>Leishmania (Viannia) panamensis</em> infection of monocyte-derived macrophages from patients and asymptomatically infected individuals were associated with the adaptive immune response and clinical outcome.</p> <h3>Methodology/Principal Findings</h3><p>To understand the basis for this difference we examined differential gene expression of human monocyte-derived macrophages following exposure to <em>L. (V.) panamensis</em>. Gene activation profiles were determined using macrophages from healthy volunteers cultured with or without stationary phase promastigotes of <em>L. (V.) panamensis</em>. Significant changes in expression (>1.5-fold change; p<0.05; up- or down-regulated) were identified at 0.5, 4 and 24 hours. mRNA abundance profiles varied over time, with the highest level of activation occurring at earlier time points (0.5 and 4 hrs). In contrast to observations for other <em>Leishmania</em> species, most significantly changed mRNAs were up- rather than down-regulated, especially at early time points. Up-regulated transcripts over the first 24 hours belonged to pathways involving eicosanoid metabolism, oxidative stress, activation of PKC through G protein coupled receptors, or mechanism of gene regulation by peroxisome proliferators via PPARα. Additionally, a marked activation of Toll-receptor mediated pathways was observed. Comparison with published microarray data from macrophages infected with <em>L. (Leishmania) chagasi</em> indicate differences in the regulation of genes involved in signaling, motility and the immune response.</p> <h3>Conclusions</h3><p>Results show that the early (0.5 to 24 hours) human monocyte-derived macrophage response to <em>L. (Viannia) panamensis</em> is not quiescent, in contrast to published reports examining later response times (48–96 hours). Early macrophage responses are important for the developing cellular response at the site of infection. The kinetics and the mRNA abundance profiles induced by <em>L. (Viannia) panamensis</em> illustrate the dynamics of these interactions and the distinct biologic responses to different <em>Leishmania</em> species from the outset of infection within their primary host cell.</p> </div

Variation of up- and down-regulated transcripts with time post-infection.

<p>A) The number of gene transcripts up- or down-regulated at 0.5, 4 and 24 hours post-infection/interaction with <i>Leishmania (Viannia) panamensis</i> promastigotes are schematically summarized. The decreasing response of the up-regulated gene transcripts with time is evident; the number of down-regulated genes is relatively consistent across the time points. B) The numbers of gene transcripts in common or differentially expressed at each time point. Gene transcript expression comparisons were made between infected and uninfected macrophages for each individual at the different time points. Gene transcripts with average intensity values less than 2× background intensity for both comparison groups, or fold change less than 1.5 fold (up or down-regulated) were excluded in further statistical analysis. A paired <i>t</i> test using p-value cutoff 0.05 was applied to determine if a gene was statistically differentially expressed.</p

Validation of transcript expression of <i>L. (V.) panamensis</i>-infected human monocyte-derived macrophages.

<p>Comparison of transcript expression in human macrophages infected with <i>L. (Viannia) panamensis</i> by quantitative real time PCR versus microarray analyses. Relative expression is expressed as a ratio of transcripts in infected cells over uninfected cells at 30 minutes or 4 hours after infection/interaction was determined. The values presented for the quantitative reverse transcriptase real time PCR analyses are the geometric mean of the ratios of macrophage transcripts from 6 to 7 different donors. The gene expression at each time point was normalized by the geometric mean of GAPDH and 39 S ribosomal protein L18 <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001866#pntd.0001866-Vandesompele1" target="_blank">[36]</a>. The genes listed are: IL1-β – interleukin-1β, TNFα – tumor necrosis factor-α, GMCSF2 –colony-stimulating factor 2 (granulocyte-macrophage), PTGS2 – prostaglandin-endoperoxide synthase 2, and IL6 – interleukin 6. The 95%CI are indicated between brackets.</p

Espirales de reflexividad crítica y propositiva para escribir la educación media de Bogotá

399 p. Libro digita