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1.
Anal Chem ; 95(41): 15384-15393, 2023 10 17.
Artículo en Inglés | MEDLINE | ID: mdl-37801728

RESUMEN

Glass is by far the most common substrate for biomolecular arrays, including high-throughput sequencing flow cells and microarrays. The native glass hydroxyl surface is modified by using silane chemistry to provide appropriate functional groups and reactivities for either in situ synthesis or surface immobilization of biologically or chemically synthesized biomolecules. These arrays, typically of oligonucleotides or peptides, are then subjected to long incubation times in warm aqueous buffers prior to fluorescence readout. Under these conditions, the siloxy bonds to the glass are susceptible to hydrolysis, resulting in significant loss of biomolecules and concomitant loss of signal from the assay. Here, we demonstrate that functionalization of glass surfaces with dipodal silanes results in greatly improved stability compared to equivalent functionalization with standard monopodal silanes. Using photolithographic in situ synthesis of DNA, we show that dipodal silanes are compatible with phosphoramidite chemistry and that hybridization performed on the resulting arrays provides greatly improved signal and signal-to-noise ratios compared with surfaces functionalized with monopodal silanes.


Asunto(s)
Ensayos Analíticos de Alto Rendimiento , Silanos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Silanos/química , Hibridación de Ácido Nucleico/métodos , ADN/química , Vidrio/química , Propiedades de Superficie
2.
Sci Rep ; 14(1): 15408, 2024 07 04.
Artículo en Inglés | MEDLINE | ID: mdl-38965271

RESUMEN

Chemosensory impairment is an outstanding symptom of SARS-CoV-2 infections. We hypothesized that measured sensory impairments are accompanied by transcriptomic changes in the foliate papillae area of the tongue. Hospital personnel with known SARS-CoV-2 immunoglobulin G (IgG) status completed questionnaires on sensory perception (n = 158). A subcohort of n = 141 participated in forced choice taste tests, and n = 43 participants consented to donate tongue swabs of the foliate papillae area for whole transcriptome analysis. The study included four groups of participants differing in IgG levels (≥ 10 AU/mL = IgG+; < 10 AU/mL = IgG-) and self-reported sensory impairment (SSI±). IgG+ subjects not detecting metallic taste had higher IgG+ levels than IgG+ participants detecting iron gluconate (p = 0.03). Smell perception was the most impaired biological process in the transcriptome data from IgG+/SSI+ participants subjected to gene ontology enrichment. IgG+/SSI+ subjects demonstrated lower expression levels of 166 olfactory receptors (OR) and 9 taste associated receptors (TAS) of which OR1A2, OR2J2, OR1A1, OR5K1 and OR1G1, as well as TAS2R7 are linked to metallic perception. The question raised by this study is whether odorant receptors on the tongue (i) might play a role in metal sensation, and (ii) are potential targets for virus-initiated sensory impairments, which needs to be investigated in future functional studies.


Asunto(s)
COVID-19 , SARS-CoV-2 , Lengua , Transcriptoma , Humanos , COVID-19/virología , COVID-19/genética , COVID-19/metabolismo , Masculino , Femenino , Adulto , Persona de Mediana Edad , Lengua/metabolismo , Lengua/virología , Lengua/patología , Inmunoglobulina G , Metales/metabolismo , Papilas Gustativas/metabolismo , Percepción del Gusto/genética , Gusto , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Percepción Olfatoria
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