ABSTRACT
Leigh syndrome (LS) associated with cytochrome c oxidase (COX) deficiency is an early onset, fatal mitochondrial encephalopathy, leading to multiple neurological failure and eventually death, usually in the first decade of life. Mutations in SURF1, a nuclear gene encoding a mitochondrial protein involved in COX assembly, are among the most common causes of LS. LSSURF1 patients display severe, isolated COX deficiency in all tissues, including cultured fibroblasts and skeletal muscle. Recombinant, constitutive SURF1-/- mice show diffuse COX deficiency, but fail to recapitulate the severity of the human clinical phenotype. Pigs are an attractive alternative model for human diseases, because of their size, as well as metabolic, physiological and genetic similarity to humans. Here, we determined the complete sequence of the swine SURF1 gene, disrupted it in pig primary fibroblast cell lines using both TALENs and CRISPR/Cas9 genome editing systems, before finally generating SURF1-/- and SURF1-/+ pigs by Somatic Cell Nuclear Transfer (SCNT). SURF1-/- pigs were characterized by failure to thrive, muscle weakness and highly reduced life span with elevated perinatal mortality, compared to heterozygous SURF1-/+ and wild type littermates. Surprisingly, no obvious COX deficiency was detected in SURF1-/- tissues, although histochemical analysis revealed the presence of COX deficiency in jejunum villi and total mRNA sequencing (RNAseq) showed that several COX subunit-encoding genes were significantly down-regulated in SURF1-/- skeletal muscles. In addition, neuropathological findings, indicated a delay in central nervous system development of newborn SURF1-/- piglets. Our results suggest a broader role of sSURF1 in mitochondrial bioenergetics.
Subject(s)
Central Nervous System/growth & development , Disease Models, Animal , Leigh Disease/genetics , Membrane Proteins/genetics , Mitochondrial Proteins/genetics , Sus scrofa/genetics , Animals , Animals, Genetically Modified , Animals, Newborn , Behavior, Animal , CRISPR-Cas Systems , Cells, Cultured , Down-Regulation , Electron Transport Complex IV/metabolism , Female , Fibroblasts , Gene Editing , Gene Knockout Techniques , Humans , Jejunum/pathology , Leigh Disease/pathology , Male , Mitochondria/pathology , Muscle, Skeletal/cytology , Muscle, Skeletal/pathology , Nuclear Transfer Techniques , Primary Cell CultureABSTRACT
since the beginning of 2020, a new virus named COVID-19 has rapidly spread to a global pandemic. The world economy has been strongly affected and early evidence suggests that its impact has involved all the sectors and business functions. In such context, logistics activities have been severely affected by the COVID-19 pandemic because of the introduction of new strict prevention rules; moreover, purchasing behaviours of customers have changed and systems had to rapidly adapt to unexpected events. This paper aims to investigate the impact of the sanitary emergency on logistics activities. A logistic provider specialized in the food and fast moving consumer goods sector has been involved in the research in order to analyse its activities and the main changes caused by the pandemic. A quantitative survey has been conducted, combining numerical data and qualitative answers. Results confirm strong effects on some sectors and an economic loss mainly due to the new procedures adopted to face the emergency.