Browsing by MeSH term "Biological Assay"
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Publication Diagnostics to support mycetoma management-Development of two target product profiles(Wiley, 2022-12) Fongwen, Noah; Asiedu, Kingsley B; Bakhiet, Sahar; Bonifaz, Alexandro; Cruz, Israel; Argaw, Daniel; Estrada-Chavez, Guadalupe; Fahal, Ahmed H; Litvintseva, Ana; Marks, Michael; Salinas-Carmona, Mario C; Sow, Doudou; van de Sande, Wendy W JObjective: Mycetoma is a neglected tropical disease caused by more than 70 different microorganisms and identified by the WHO as one of the high-priority diseases for developing diagnostic tests. To ensure the production of diagnostic assays for use by clinical staff in endemic regions, target product profiles (TPPs) were designed. Methods: We describe the development of two TPPs: one for a diagnostic test able to identify the causative agent of mycetoma and another that would determine when treatment could be stopped. The TPPs were developed by considering product use, design, performance, product configuration and costs. Results: Version 1.0 TPPs for two uses were posted by WHO for a 1-month online public consultation on 25 October 2021, and the final TPP was posted online on 5 May 2022. Conclusion: A major difficulty encountered in developing both TPPs was the large number of agents able to cause mycetoma and the lack of specific biomarkers for most of them.Publication Identification of small molecules capable of enhancing viral membrane fusion(BioMed Central (BMC), 2023-05-24) García-Murria, Mª Jesús; Gadea-Salom, Laura; Moreno, Sandra; Rius-Salvador, Marina; Zaragoza, Oscar; Brun, Alejandro; Mingarro, Ismael; Martínez-Gil, Luis; Generalitat Valenciana (España); Ministerio de Ciencia e Innovación (España); Unión Europea. Comisión Europea. H2020; Agencia Estatal de Investigación (España)Several approaches have been developed to analyze the entry of highly pathogenic viruses. In this study, we report the implementation of a Bimolecular Multicellular Complementation (BiMuC) assay to safely and efficiently monitor SARS-CoV-2 S-mediated membrane fusion without the need for microscopy-based equipment. Using BiMuC, we screened a library of approved drugs and identified compounds that enhance S protein-mediated cell-cell membrane fusion. Among them, ethynylestradiol promotes the growth of SARS-CoV-2 and Influenza A virus in vitro. Our findings demonstrate the potential of BiMuC for identifying small molecules that modulate the life cycle of enveloped viruses, including SARS-CoV-2.Publication International network for comparison of HIV neutralization assays: the NeutNet report II(Public Library of Science (PLOS), 2012-05-09) Heyndrickx, Leo; Heath, Alan; Sheik-Khalil, Enas; Alcamí, José; Bongertz, Vera; Jansson, Marianne; Malnati, Mauro; Montefiori, David; Moog, Christiane; Morris, Lynn; Osmanov, Saladin; Polonis, Victoria; Ramaswamy, Meghna; Sattentau, Quentin; Tolazzi, Monica; Schuitemaker, Hanneke; Willems, Betty; Wrin, Terri; Fenyö, Eva Maria; Scarlatti, Gabriella; Unión Europea. Comisión Europea; World Health Organization (WHO/OMS); Europrise; Instituto de Salud Carlos III; Fundación para la Innovación y la Prospectiva en Salud en España; Fund for Scientific Research (Belgica); Istituto Superiore di Sanita (Italia); Swedish Research Council; Swedish International Development Cooperation Agency; Bill & Melinda Gates FoundationBACKGROUND: Neutralizing antibodies provide markers for vaccine-induced protective immunity in many viral infections. By analogy, HIV-1 neutralizing antibodies induced by immunization may well predict vaccine effectiveness. Assessment of neutralizing antibodies is therefore of primary importance, but is hampered by the fact that we do not know which assay(s) can provide measures of protective immunity. An international collaboration (NeutNet) involving 18 different laboratories previously compared different assays using monoclonal antibodies (mAbs) and soluble CD4 (Phase I study). METHODS: In the present study (Phase II), polyclonal reagents were evaluated by 13 laboratories. Each laboratory evaluated nine plasmas against an 8 virus panel representing different genetic subtypes and phenotypes. TriMab, a mixture of three mAbs, was used as a positive control allowing comparison of the results with Phase I in a total of nine different assays. The assays used either uncloned virus produced in peripheral blood mononuclear cells (PBMCs) (Virus Infectivity Assays, VIA), or Env (gp160)-pseudotyped viruses (pseudoviruses, PSV) produced in HEK293T cells from molecular clones or from uncloned virus. Target cells included PBMC and genetically engineered cell lines in either single- or multiple-cycle infection format. Infection was quantified by using a range of assay read-outs including extra- or intra-cellular p24 antigen detection, luciferase, beta-galactosidase or green fluorescent protein (GFP) reporter gene expression. FINDINGS: Using TriMab, results of Phase I and Phase II were generally in agreement for six of the eight viruses tested and confirmed that the PSV assay is more sensitive than PBMC (p = 0.014). Comparisons with the polyclonal reagents showed that sensitivities were dependent on both virus and plasma. CONCLUSIONS: Here we further demonstrate clear differences in assay sensitivities that were dependent on both the neutralizing reagent and the virus. Consistent with the Phase I study, we recommend parallel use of PSV and VIA for vaccine evaluation.Publication Longer intervals between SARS-CoV-2 infection and mRNA-1273 doses improve the neutralization of different variants of concern(Wiley, 2023-03) García-Pérez, Javier; Bermejo, Mercedes; Ramírez-García, Almudena; de la Torre-Tarazona, Humberto Erick; Cascajero Díaz, Almudena; Castillo de la Osa, María; Jiménez Santana, Paloma; Aparicio Gómez, Marta; Calonge, Esther; Sancho-López, Aránzazu; Payares-Herrera, Concepción; Layunta Acero, Rocio; Vicente-Izquierdo, Laura; Avendaño-Solá, Cristina; Alcamí, José; Perez-Olmeda, Mayte; Díez-Fuertes, Francisco; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)The humoral immune response against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern elicited by vaccination was evaluated in COVID-19 recovered individuals (Rec) separated 1-3 months (Rec2m) or 4-12 months (Rec9m) postinfection and compared to the response in naïve participants. Antibody-mediated immune responses were assessed in 66 participants by three commercial immunoassays and a SARS-CoV-2 lentiviral-based pseudovirus neutralization assay. Immunoglobulin (Ig) levels against SARS-CoV-2 spike were lower in naïve participants after two doses than in Rec after a single dose (p < 0.05). After two doses in Rec, levels of total Ig to receptor-binding domain were significantly increased in Rec9m compared to Rec2m (p < 0.001). The neutralizing potency observed in Rec9m was consistently higher than in Rec2m against variants of concern (VOCs) Alpha, Beta, Delta, and BA.1 sublineage of Omicron with 2.2-2.8-fold increases. Increasing the interval between SARS-CoV-2 infection and the vaccination with messenger RNA-based vaccines to more than 3 months generates a more efficient heterologous humoral immune response against VOCs by allowing enough time to mount a strong recall memory B cell response.Publication Monitoring Autophagy in Muscle Stem Cells.(Humana Press, 2017-03) García-Prat, Laura; Munoz-Canoves, Pura; Martínez-Vicente, Marta; Ministerio de Economía, Industria y Competitividad (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Instituto de Salud Carlos III; Fundación La Marató TV3; Unión EuropeaAutophagy is critical not only for the cell's adaptive response to starvation but also for cellular homeostasis, by acting as quality-control machinery for cytoplasmic components. This basal autophagic activity is particularly needed in postmitotic cells for survival maintenance. Recently, basal autophagic activity was reported in skeletal muscle stem cells (satellite cells) in their dormant quiescent state. Satellite cells are responsible for growth as well as for regeneration of muscle in response to stresses such as injury or disease. In the absence of stress, quiescence is the stem cell state of these cells throughout life, although which mechanisms maintain long-life quiescence remains largely unknown. Our recent findings showed that autophagy is necessary for quiescence maintenance in satellite cells and for retention of their regenerative functions. Importantly, damaged organelles and proteins accumulated in these cells with aging and this was connected to age-associated defective autophagy. Refueling of autophagy through genetic and pharmacological strategies restored aged satellite cell functions, and these finding have biomedical implications. In this chapter, we describe different experimental strategies to evaluate autophagic activity in satellite cells in resting muscle of mice. They should facilitate our competence to investigate stem cell functions both during tissue homeostasis as in pathological conditions.Publication Robust, universal biomarker assay to detect senescent cells in biological specimens(Wiley, 2017-02) Evangelou, Konstantinos; Lougiakis, Nikolaos; Rizou, Sophia V; Kotsinas, Athanassios; Kletsas, Dimitris; Muñoz-Espín, Daniel; Kastrinakis, Nikolaos G; Pouli, Nicole; Marakos, Panagiotis; Townsend, Paul; Serrano Marugan, Manuel; Bartek, Jiri; Gorgoulis, Vassilis G; Swedish Research Council for Health, Working Life and Welfare; Danish National Research Foundation; Novo Nordisk FoundationCellular senescence contributes to organismal development, aging, and diverse pathologies, yet available assays to detect senescent cells remain unsatisfactory. Here, we designed and synthesized a lipophilic, biotin-linked Sudan Black B (SBB) analogue suitable for sensitive and specific, antibody-enhanced detection of lipofuscin-containing senescent cells in any biological material. This new hybrid histo-/immunochemical method is easy to perform, reliable, and universally applicable to assess senescence in biomedicine, from cancer research to gerontology.