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1.
J Equine Vet Sci ; 128: 104881, 2023 09.
Article in English | MEDLINE | ID: mdl-37422140

ABSTRACT

Microminerals are necessary for all bodily functions. In animal species, selenium (Se), copper (Cu) and zinc (Zn) all form part of antioxidant enzymes. Micromineral deficiencies, particularly Se, are well recognized in large animal species in Chile. Glutathion peroxidase (GPx) is a widely used biomarker for Se nutritional status and to diagnose Se deficiency in horses. Superoxide dismutase (SOD) is a Cu and Zn-dependant antioxidant enzyme, although it is not commonly used as a proxy for the nutritional status of these minerals. Ceruloplasmin (CP) is used as a biomarker of Cu nutritional status. This study´s objective was to explore correlation between said minerals and biomarkers in adult horses from southern Chile. Se, Cu, Zn, GPx, SOD, and CP were measured in whole blood of a group of 32 adult horses (5-15 years old). Further, a second group of 14 adult horses (5-15 years old) underwent gluteal muscle biopsies to determine Cu, Zn, GPx, and SOD. Correlations were determined by means of Pearson´s r coefficient. Significant correlations were found for blood GPx and Se (r = 0.79), blood GPx and SOD (r = -0.6), muscular GPx and SOD (r = 0.78), and Cu and CP (r = 0.48). These results confirm previously described strong association between blood GPx and Se in horses, validating the former´s use as a diagnostic proxy of Se deficiency in Chilean horses, and suggest significant interactions between GPx and SOD in blood and muscle tissue.


Subject(s)
Selenium , Horses , Animals , Copper , Zinc , Antioxidants , Chile , Superoxide Dismutase , Muscle, Skeletal , Biomarkers
2.
Sci Rep ; 11(1): 3318, 2021 02 08.
Article in English | MEDLINE | ID: mdl-33558635

ABSTRACT

Despite unprecedented global efforts to rapidly develop SARS-CoV-2 treatments, in order to reduce the burden placed on health systems, the situation remains critical. Effective diagnosis, treatment, and prophylactic measures are urgently required to meet global demand: recombinant antibodies fulfill these requirements and have marked clinical potential. Here, we describe the fast-tracked development of an alpaca Nanobody specific for the receptor-binding-domain (RBD) of the SARS-CoV-2 Spike protein with potential therapeutic applicability. We present a rapid method for nanobody isolation that includes an optimized immunization regimen coupled with VHH library E. coli surface display, which allows single-step selection of Nanobodies using a simple density gradient centrifugation of the bacterial library. The selected single and monomeric Nanobody, W25, binds to the SARS-CoV-2 S RBD with sub-nanomolar affinity and efficiently competes with ACE-2 receptor binding. Furthermore, W25 potently neutralizes SARS-CoV-2 wild type and the D614G variant with IC50 values in the nanomolar range, demonstrating its potential as antiviral agent.


Subject(s)
Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Antibody Affinity/genetics , COVID-19/immunology , SARS-CoV-2/immunology , Single-Domain Antibodies/immunology , Spike Glycoprotein, Coronavirus/immunology , Angiotensin-Converting Enzyme 2/immunology , Animals , COVID-19/virology , Camelids, New World/immunology , Escherichia coli/genetics , Escherichia coli/metabolism , Green Fluorescent Proteins/genetics , HeLa Cells , Humans , Immunization , Male , Neutralization Tests , Peptide Library , Protein Binding/genetics , SARS-CoV-2/chemistry , SARS-CoV-2/isolation & purification , Spike Glycoprotein, Coronavirus/genetics , Transfection
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