Programa de intervención de Terapia Ocupacional en pacientes con fibromialgia.

El objeto de este trabajo consiste en realizar un programa de intervención desde terapia ocupacional en pacientes con fibromialgia. Se quiere determinar si la participación de estos pacientes, les produce mejoría en su calidad de vida y salud. Para ello se ha planteado como objetivo principal: valorar si un programa de intervención desde Terapia Ocupacional mejora la calidad de vida del paciente con fibromialgia, y los siguientes objetivos específicos: evaluar desde Terapia Ocupacional la calidad de vida y el grado de autonomía de los pacientes con fibromialgia y establecer un plan de intervención y desarrollar un sistema de evaluación de resultados. La metodología se ha basado en los siguientes instrumentos de evaluación: Fibromyalgia Impact Questionnaire (FIQ) versión español, Barthel-Lawton, Escala de Intensidad de la fatiga y el inventario de la depresión de Beck, posteriormente deben ser de nuevo utilizados para realizar una reevaluación y comprobar si se han cumplido los objetivos. Con este programa de intervención se podrá ver la eficacia del programa, de los resultados y los efectos en los usuarios con fibromialgia que reciben intervención desde Terapia ocupacional. Finalmente se pretende recoger la satisfacción de los pacientes con el programa. Esto se evaluará con un cuestionario ad hoc.<br /

Development and characterization of cell models harbouring mtDNA deletions for <i>in vitro</i> study of Pearson syndrome

Pearson syndrome is a rare multisystem disease caused by single large-scale mitochondrial DNA deletions (SLSMDs). The syndrome presents early in infancy and is mainly characterised by refractory sideroblastic anaemia. Prognosis is poor and treatment is supportive, thus the development of new models for the study of Pearson syndrome and new therapy strategies is essential. In this work, we report three different cell models carrying an SLMSD: fibroblasts, transmitochondrial cybrids and induced pluripotent stem cells (iPSCs). All studied models exhibited an aberrant mitochondrial ultrastructure and defective oxidative phosphorylation system function, showing a decrease in different parameters, such as mitochondrial ATP, respiratory complex IV activity and quantity or oxygen consumption. Despite this, iPSCs harbouring ‘common deletion’ were able to differentiate into three germ layers. Additionally, cybrid clones only showed mitochondrial dysfunction when heteroplasmy level reached 70%. Some differences observed among models may depend on their metabolic profile; therefore, we consider that these three models are useful for the in vitro study of Pearson syndrome, as well as for testing new specific therapies. This article has an associated First Person interview with the first author of the paper

Producción, Expresión And Purificación de Mutaciones en la Proteína Adar1 Involucrada en el Síndrome de Aicardi-Goutières Síndrome (AGS)

Peer reviewe

Diagnóstico molecular de enfermedades mitocondriales: síndrome de MELAS y síndrome de Leigh

El ADN mitocondrial tiene unas características propias y su localización en un orgánulo citoplasmático implica que la herencia de este genoma difiera de la del ADN nuclear en el modo de herencia, la poliplasmia, la segregación mitótica, la expresión umbral y la alta tasa de mutación. Las enfermedades causadas por mutaciones en el ADNmt tienen como característica común el estar producidas por una deficiente síntesis de ATP. Las mitocondrias son componentes fundamentales de todos los tejidos y órganos, por lo que estas enfermedades serán, en general, multisistémincas y darán lugar a amplio espectro de fenotipos. El síndrome de Leigh o encefalomiopatía mitocondrial necrotizante infantil subaguda es una enfermedad neurodegenerativa, genéticamente heterogénea causada por mutaciones tanto en el genoma nuclear como en el mitocondrial, por lo que puede presentar diversos tipos de herencia. El síndrome de MELAS es uno de los síndromes mitocondriales multisistémicos mejor definidos desde el punto de vista clínico. Los criterios invariables son episodios de accidentes cerebrovasculares antes de los 40 años, encefalopatía caracterizada por crisis epilépticas focales o generalizadas, acidosis láctica y/o fibras rojo rasgadas, además de dos de los siguientes síntomas: baja estatura, demencia, cefaleas recurrentes y vómitos. El objetivo de este estudio de este estudio es el análisis genetico-molecular del ADNmt de varios pacientes con síndrome de Leigh y síndrome de MELAS. Asimismo, se ha realizado la búsqueda de mutaciones en familiares relacionados por la vía materna. En este estudio se ha realizado el análisis genético mitocondrial de 22 personas, 20 pacientes y 2 familiares, principalmente por las técnicas de RFLP-PCR de los cuales se han obtenido 9 positivos

ITGB3-mediated uptake of small extracellular vesicles facilitates intercellular communication in breast cancer cells.

Metastasis, the spread of malignant cells from a primary tumour to distant sites, causes 90% of cancer-related deaths. The integrin ITGB3 has been previously described to play an essential role in breast cancer metastasis, but the precise mechanisms remain undefined. We have now uncovered essential and thus far unknown roles of ITGB3 in vesicle uptake. The functional requirement for ITGB3 derives from its interactions with heparan sulfate proteoglycans (HSPGs) and the process of integrin endocytosis, allowing the capture of extracellular vesicles and their endocytosis-mediated internalization. Key for the function of ITGB3 is the interaction and activation of focal adhesion kinase (FAK), which is required for endocytosis of these vesicles. Thus, ITGB3 has a central role in intracellular communication via extracellular vesicles, proposed to be critical for cancer metastasis.S

GDF15 is an Eribulin Response Biomarker also required for Survival of DTP Breast Cancer Cells.

Drug tolerant persister (DTP) cells enter into a reversible slow-cycling state after drug treatment. We performed proteomic characterization of the breast cancer (BC) DTP cell secretome after eribulin treatment. We showed that the growth differentiation factor 15 (GDF15) is a protein significantly over-secreted upon eribulin treatment. The biomarker potential of GDF15 was confirmed in 3D-cell culture models using BC cells lines and PDXs, as well as in a TNBC in vivo model. We also found that GDF15 is required for survival of DTP cells. Direct participation of GDF15 and its receptor GFRAL in eribulin-induction of DTPs was established by the enhanced cell killing of DTPs by eribulin seen under GDF15 and GFRAL loss of function assays. Finally, we showed that combination therapy of eribulin plus an anti-GDF15 antibody kills BC-DTP cells. Our results suggest that targeting GDF15 may help eradicate DTP cells and block the onset of acquired resistance

GDF15 is an eribulin response biomarker also required for survival of DTP breast cancer cells

Drug tolerant persister (DTP) cells enter into a reversible slow-cycling state after drug treatment. We performed proteomic characterization of the breast cancer (BC) DTP cell secretome after eribulin treatment. We showed that the growth differentiation factor 15 (GDF15) is a protein significantly over-secreted upon eribulin treatment. The biomarker potential of GDF15 was confirmed in 3D-cell culture models using BC cells lines and PDXs, as well as in a TNBC in vivo model. We also found that GDF15 is required for survival of DTP cells. Direct participation of GDF15 and its receptor GFRAL in eribulin-induction of DTPs was established by the enhanced cell killing of DTPs by eribulin seen under GDF15 and GFRAL loss of function assays. Finally, we showed that combination therapy of eribulin plus an anti-GDF15 antibody kills BC-DTP cells. Our results suggest that targeting GDF15 may help eradicate DTP cells and block the onset of acquired resistance

Laboratory-based molecular tests in SARS-CoV-2 infection diagnosed using RT-PCR

Background: Diagnosing patients with a SARS-CoV-2 infection played a critical role in managing the COVID-19 pandemic and remains a priority for the transition to long-term management of COVID-19. Initial shortages of extraction and RT-PCR reagents impaired the desired upscaling of testing in many countries, which led to the search for alternatives to extraction and RT-PCR testing. Reference standard methods for diagnosing the presence of SARS-CoV-2 infection rely primarily on real-time reverse transcription-polymerase chain reaction (RT-PCR). Alternatives to RT-PCR could, if sufficiently accurate, have a positive impact by expanding the range of diagnostic tools available for the timely identification of people infected by SARS-CoV-2 , access to testing and the use of resources.Objectives: To assess the diagnostic accuracy of alternative (to RT-PCR assays) laboratory-based molecular tests for diagnosing SARS-CoV-2 infection.Search methods: We searched the COVID‐19 Open Access Project living evidence database from the University of Bern until 30 September 2020 and the WHO COVID-19 Research Database until 31 October 2022. We did not apply language restrictions.Selection criteria: We included studies of people with suspected or known SARS‐CoV‐2 infection, or where tests were used to screen for infection, and studies evaluating commercially developed laboratory-based molecular tests for the diagnosis of SARS-CoV-2 infection considered as alternatives to RT-PCR testing. We also included all reference standards to define the presence or absence of SARS‐CoV‐2, including RT‐PCR tests and established clinical diagnostic criteria.Data collection and analysis: Two authors independently screened studies and resolved disagreements by discussing them with a third author. Two authors independently extracted data and assessed the risk of bias and applicability of the studies using the QUADAS‐2 tool. We presented sensitivity and specificity, with 95% confidence intervals (CIs), for each test using paired forest plots and summarised results using average sensitivity and specificity using a bivariate random‐effects meta‐analysis. We illustrated the findings per index test category and assay brand compared to the WHO's acceptable sensitivity and specificity threshold for diagnosing SARS-CoV-2 infection using nucleic acid tests.Main results: We included data from 64 studies reporting 94 cohorts of participants and 105 index test evaluations, with 74753 samples and 7517 confirmed SARS-CoV-2 cases. We did not identify any published or preprint reports of accuracy for a considerable number of commercially produced NAAT assays. Ninety-two cohorts were judged at unclear or high risk of bias in more than three QUADAS-2 domains. Around half of the cohorts were considered at high risk of selection bias because of recruitment based on COVID status. Seventy-five out of 94 cohorts were at high risk of bias in the reference standard domain because of reliance on a single RT-PCR result to determine the absence of SARS-CoV-2 infection. Seventy cohorts were at unclear risk of bias due to a lack of clarity about the time interval between the index test assessment and the reference standard, the number of missing results, or the absence of a participant flow diagram.For index tests categories with four or more evaluations and when summary estimations were possible, we found the following results: a) For RT-PCR assays omitting/adapting RNA extraction, the average sensitivity was 95.1% (95% CI 91.1% to 97.3%), and the average specificity was 99.7% (95% CI 98.5% to 99.9%; based on 27 evaluations, 2834 samples and 1178 SARS-CoV-2 cases); b) For RT-LAMP assays, the average sensitivity was 88.4% (95% CI 83.1% to 92.2%), and the average specificity was 99.7% (95% CI 98.7% to 99.9%; 24 evaluations, 29496 samples and 2255 SARS-CoV-2 cases); c) for TMA assays, the average sensitivity was 97.6% (95% CI 95.2% to 98.8%), and the average specificity was 99.4% (95% CI 94.9% to 99.9%; 14 evaluations, 2196 samples and 942 SARS-CoV-2 cases); d) for digital PCR assays, the average sensitivity was 98.5% (95% CI 95.2% to 99.5%), and the average specificity was 91.4% (95% CI 60.4% to 98.7%; five evaluations, 703 samples and 354SARS-CoV-2 cases); e) for RT-LAMP assays omitting/adapting RNA extraction, the average sensitivity was 73.1% (95% CI 58.4% to 84%), and the average specificity was 100% (95% CI 98% to 100%; 24 evaluations, 14342 samples and 1502 SARS-CoV-2 cases).Only two index test categories fulfil the WHO-acceptable sensitivity and specificity requirements for SARS-CoV-2 nucleic acid tests: RT-PCR assays omitting/adapting RNA extraction and TMA assays. In addition, WHO-acceptable performance criteria were met in a meta-analysis of data for two assays out of 35 when tests were used according to manufacturer instructions. At 5% prevalence using a cohort of 1000 people suspected of SARS-CoV-2 infection, the positive predictive value of RT-PCR assays omitting/adapting RNA extraction/purification will be 94%, with three in 51 positive results being false positives, and around 2 missed cases. For TMA assays, the positive predictive value of RT-PCR assays will be 89%, with 6 in 55 positive results being false positives, and around one missed case.Authors' conclusions: Alternative laboratory-based molecular tests aim to enhance testing capacity in different ways, such as reducing the time, steps and resources needed to obtain valid results. Several index test technologies with these potential advantages have not been evaluated or have been assessed by only a few studies of limited methodological quality, so the performance of these kits was undetermined.Only two index test categories with enough evaluations for meta-analysis fulfil the WHO set of acceptable accuracy standards for SARS-CoV-2nucleic acid tests: RT-PCR assays omitting/adapting RNA extraction and TMA assays. These assays might prove to be suitable alternatives to RT-PCR for identifying people infected by SARS-CoV-2, especially when the alternative would be not having access to testing. However, these findings need to be interpreted and used with caution because of several limitations in the evidence, including reliance on retrospective samples without information about the symptom status of participants and the timing of assessment. Although we used a comprehensive search and had broad eligibility criteria to include a wide range of tests that could be alternatives to RT-PCR methods, further research is needed to assess the performance of alternative COVID-19 tests and their role in pandemic management