Comparison of reproducibility, accuracy, sensitivity, and specificity of miRNA quantification platforms

Given the increasing interest in their use as disease biomarkers, the establishment of reproducible, accurate, sensitive, and specific platforms for microRNA (miRNA) quantification in biofluids is of high priority. We compare four platforms for these characteristics: small RNA sequencing (RNA-seq), FirePlex, EdgeSeq, and nCounter. For a pool of synthetic miRNAs, coefficients of variation for technical replicates are lower for EdgeSeq (6.9%) and RNA-seq (8.2%) than for FirePlex (22.4%); nCounter replicates are not performed. Receiver operating characteristic analysis for distinguishing present versus absent miRNAs shows small RNA-seq (area under curve 0.99) is superior to EdgeSeq (0.97), nCounter (0.94), and FirePlex (0.81). Expected differences in expression of placenta-associated miRNAs in plasma from pregnant and non-pregnant women are observed with RNA-seq and EdgeSeq, but not FirePlex or nCounter. These results indicate that differences in performance among miRNA profiling platforms impact ability to detect biological differences among samples and thus their relative utility for research and clinical use

Large Differences in Small RNA Composition Between Human Biofluids.

Extracellular microRNAs (miRNAs) and other small RNAs are implicated in cellular communication and may be useful as disease biomarkers. We systematically compared small RNAs in 12 human biofluid types using RNA sequencing (RNA-seq). miRNAs and tRNA-derived RNAs (tDRs) accounted for the majority of mapped reads in all biofluids, but the ratio of miRNA to tDR reads varied from 72 in plasma to 0.004 in bile. miRNA levels were highly correlated across all biofluids, but levels of some miRNAs differed markedly between biofluids. tDR populations differed extensively between biofluids. Y RNA fragments were seen in all biofluids and accounted for >10% of reads in blood plasma, serum, and cerebrospinal fluid (CSF). Reads mapping exclusively to Piwi-interacting RNAs (piRNAs) were very rare, except in seminal plasma. These results demonstrate extensive differences in small RNAs between human biofluids and provide a useful resource for investigating extracellular RNA biology and developing biomarkers

Mitogen-activated protein kinase kinase 5 regulates proliferation and biosynthetic processes in procyclic forms of Trypanosoma brucei

The pathogenic protozoan T. brucei alternates into distinct developmental stages in the mammalian and insect hosts. The mitogen-activated protein kinase (MAPK) signaling pathways transduce extracellular stimuli into a range of cellular responses, which ultimately lead to the adaptation to the external environment. Here, we combined a loss of function approach with stable isotope labeling with amino acids in cell culture (SILAC)-based mass spectrometry (MS) to investigate the role of the mitogen-activated protein kinase kinase 5 (MKK5) in T. brucei. The silencing of MKK5 significantly decreased the proliferation of procyclic forms of T. brucei. To shed light on the molecular alterations associated with this phenotype, we measured the total proteome and phosphoproteome of cells silenced for MKK5. In the total proteome, we observed a general decrease in proteins related to ribosome and translation as well as down-regulation of several components of the fatty acids biosynthesis pathway. In addition, we observed alterations in the protein levels and phosphorylation of key metabolic enzymes, which point toward a suppression of the oxidative metabolism. Taken together, our findings show that the silencing of MKK5 alters cell growth, energy metabolism, protein and fatty acids biosynthesis in procyclic T. brucei

Functional analysis of recurrent CDC20 promoter variants in human melanoma

Small nucleotide variants in non-coding regions of the genome can alter transcriptional regulation, leading to changes in gene expression which can activate oncogenic gene regulatory networks. Melanoma is heavily burdened by non-coding variants, representing over 99% of total genetic variation, including the well-characterized TERT promoter mutation. However, the compendium of regulatory non-coding variants is likely still functionally under-characterized. We developed a pipeline to identify hotspots, i.e. recurrently mutated regions, in melanoma containing putatively functional non-coding somatic variants that are located within predicted melanoma-specific regulatory regions. We identified hundreds of statistically significant hotspots, including the hotspot containing the TERT promoter variants, and focused on a hotspot in the promoter of CDC20. We found that variants in the promoter of CDC20, which putatively disrupt an ETS motif, lead to lower transcriptional activity in reporter assays. Using CRISPR/Cas9, we generated an indel in the CDC20 promoter in human A375 melanoma cell lines and observed decreased expression of CDC20, changes in migration capabilities, increased growth of xenografts, and an altered transcriptional state previously associated with a more proliferative and less migratory state. Overall, our analysis prioritized several recurrent functional non-coding variants that, through downregulation of CDC20, led to perturbation of key melanoma phenotypes

Historia del cultivo de la judía: su evolución más allá de las áreas de origen y domesticación

The common bean (Phaseolus vulgaris L.) is the most important grain legume for direct human consumption on a global scale. Current bean germplasm collections show a wide variation of phenotypes, although genetic erosion is gradually affecting this species as in many countries local traditional varieties are being replaced by elite cultivars. This crop has spread to every continent over the past few centuries, which has resulted in a complex genetic structure of bean germplasm outside its areas of origin and domestication (South and Central America). Some evidence indicates that this germplasm is more complex than previously thought and contains additional, as yet unexplored, diversity. This is especially the case in southern Europe, particularly in the Iberian Peninsula, where it was introduced in the early sixteenth century and has been documented as a secondary focus of domestication of the species. The integration of omic data into bean germplasm documentation databases and its combination with genotypic, phenotypic and agro-ecological data is opening a new era for the enhancement and efficient use of common bean genetic resources as the main grain legume in Europe and worldwide.La judía común (Phaseolus vulgaris L.) es la leguminosa de grano más relevante para el consumo humano directo en escala global. Las colecciones de germoplasma de judía actuales muestran una amplia variación de fenotipos, aunque en muchos países las variedades locales están siendo reemplazados por cultivares de élite, concentrando la producción agraria en un número cada vez más reducido de cultivares con la consecuente erosión genética o pérdida de biodiversidad. Este cultivo se ha extendido por todos los continentes durante los últimos siglos, lo que ha dado lugar a una compleja estructura genética fuera de sus áreas de origen y domesticación (Mesoamérica y Sudamérica). Diversas evidencias indican que el germoplasma europeo contiene una diversidad adicional mayor de la esperada especialmente en el Sur de Europa, y particularmente en la Península Ibérica, dónde fue introducida a comienzos del siglo XVI, y que ha sido documentada como un centro de domesticación secundaria de la especie. La integración de datos ómicos en las bases de datos de documentación del germoplasma de judía y su combinación con datos genéticos, fenotípicos y agro-ecológicos está abriendo una nueva era para la valorización y el uso eficiente de los recursos genéticos de la judía común como la principal leguminosa de grano para consumo humano en Europa y globalmente

Displaced Voices: A Journal of Migration, Archives and Cultural Heritage, Volume 3 Issue 2 (Autumn 2023)

Twentieth Century Histories of Civic Society’s Responses to Crises of Displacement: A Special Issue to mark the 70th Anniversary of Refugee Council Displaced Voices is a biannual digital magazine produced twice a year by the Living Refugee Archive team at the University of East London. Displaced Voices aims to provide a digital platform for activists, archivists, researchers, practitioners and academics to contribute to issues pertaining to refugee and migration history; refugee and migrant rights; social justice; cultural heritage and archives. We welcome a range of contributions to the magazine including articles of between 1000-2000 words; reports on fieldwork in archival collections; book recommendations and reviews; and more creative pieces including (but not limited too) cartoons; photography; and poetry. We would also welcome news on activities; publication of reports, projects; letters and news from your own networks. We welcome submissions from all writers whether you are a student, practitioner, activist or established academic. The Displaced Voices online magazine is born out of the collaborative and intersectional work that we have been undertaking through our work with the refugee and migration archives housed at the University of East London. Our work to date has explored the intersections of refugee and migration studies with narrative and life history research linked to oral history methods and archival approaches to the preservation, documentation and accessibility of archival resources recording the refugee experience. This magazine is a collaborative project between the Living Refugee Archive at the University of East London; the Oral History Society Migration Special Interest Group and the International Association for the Study of Forced Migration Working Group on the History of Forced Migration and Refugees. Thematically we are looking to engage with articles that explore the intersection of refugee and forced migration studies; history and cultural heritage studies; narrative research; oral history and archival science

Phenotyping Superagers Using Resting-State fMRI

BACKGROUND AND PURPOSE: Superagers are defined as older adults with episodic memory performance similar or superior to that in middle-aged adults. This study aimed to investigate the key differences in discriminative networks and their main nodes between superagers and cognitively average elderly controls. In addition, we sought to explore differences in sensitivity in detecting these functional activities across the networks at 3T and 7T MR imaging fields. MATERIALS AND METHODS: Fifty-five subjects 80 years of age or older were screened using a detailed neuropsychological protocol, and 31 participants, comprising 14 superagers and 17 cognitively average elderly controls, were included for analysis. Participants underwent resting-state-fMRI at 3T and 7T MR imaging. A prediction classification algorithm using a penalized regression model on the measurements of the network was used to calculate the probabilities of a healthy older adult being a superager. Additionally, ORs quantified the influence of each node across preselected networks. RESULTS: The key networks that differentiated superagers and elderly controls were the default mode, salience, and language networks. The most discriminative nodes (ORs > 1) in superagers encompassed areas in the precuneus posterior cingulate cortex, prefrontal cortex, temporoparietal junction, temporal pole, extrastriate superior cortex, and insula. The prediction classification model for being a superager showed better performance using the 7T compared with 3T resting-state-fMRI data set. CONCLUSIONS: Our findings suggest that the functional connectivity in the default mode, salience, and language networks can provide potential imaging biomarkers for predicting superagers. The 7T field holds promise for the most appropriate study setting to accurately detect the functional connectivity patterns in superagers