Publications by authors named "Spickett C"

Endothelial dysfunction is implicated in the development and aggravation of cardiovascular complications. Among the endothelium-released vasoactive factors, hydrogen sulfide (HS) has been investigated for its beneficial effects on the vasculature through anti-inflammatory and redox-modulating regulatory mechanisms. Reduced HS bioavailability is reported in chronic diseases such as cardiovascular disease, diabetes, atherosclerosis and preeclampsia, suggesting the value of investigating mechanisms, by which HS acts as a vasoprotective gasotransmitter.

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Saccharomyces cerevisiae is an important unicellular yeast species within the biotechnological and the food and beverage industries. A significant application of this species is the production of ethanol, where concentrations are limited by cellular toxicity, often at the level of the cell membrane. Here, we characterize 61 S.

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Protein lipoxidation is a non-enzymatic post-translational modification that consists of the covalent addition of reactive lipid species to proteins. This occurs under basal conditions but increases in situations associated with oxidative stress. Protein targets for lipoxidation include metabolic and signalling enzymes, cytoskeletal proteins, and transcription factors, among others.

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Ischemic stroke is one of the leading causes of death and disability worldwide. This acute vascular event interferes with blood supply to the brain and induces a burst of free radicals such as nitric oxide and superoxide, producing peroxynitrite, a precursor of strong nitrating agents. Fibrinogen is one of the most abundant plasma proteins; it plays a role in the hemostatic system, mediating clot formation, which can be affected by nitrotyrosine formation.

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While often regarded as a subset of metabolomics, lipidomics can better be considered as a field in its own right. While the total number of lipid species in biology may not exceed the number of metabolites, they can be modified chemically and biochemically leading to an enormous diversity of derivatives, many of which retain the lipophilic properties of lipids and thus expand the lipidome greatly. Oxidative modification by radical oxygen species, either enzymatically or chemically, is one of the major mechanisms involved, although attack by non-radical oxidants also occurs.

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  • PTEN is a crucial tumor suppressor that regulates the PI3K-AKT signaling pathway, impacting cell survival and metabolism, and is often mutated in cancer and associated with diseases like diabetes.
  • The review explores the interactions of PTEN with various proteins, highlighting its roles beyond just signaling regulation and emphasizing its importance in cell biology.
  • It compares different research methods used to study PTEN's interactome, from hypothesis-driven techniques like co-immunoprecipitation to exploratory high-data approaches, ultimately showcasing the diverse cellular functions of PTEN revealed by recent studies.
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  • Wilson disease is a genetic disorder linked to dysfunctional copper metabolism, leading to liver and neurological issues due to mutations in a specific gene.
  • The variant NM_000053.3:c.1934T > G (Met645Arg) is common among patients of Spanish descent and is classified as pathogenic, but its actual impact on protein function was previously debated.
  • Recent studies using a minigene system and gene-edited cells found that this variant causes significant exon skipping, confirming its pathogenicity and paving the way for potential genetic therapies.
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The adenosine 2A receptor (AR), a G-protein-coupled receptor (GPCR), was solubilised and purified encapsulated in styrene maleic acid lipid particles (SMALPs). The purified AR-SMALP was associated with phospholipids characteristic of the plasma membrane of Pichia pastoris, the host used for its expression, confirming that the AR-SMALP encapsulated native lipids. The fluorescence spectrum of the AR-SMALP showed a characteristic broad emission peak at 330 nm, produced by endogenous Trp residues.

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  • * These modifications can cause proteins to change shape, potentially losing their activity, although some proteins might be activated through this process, enhancing cell signaling.
  • * ALEs impact various cellular functions, including metabolism and protein interactions, and their formation has been linked to both healthy and disease states, suggesting that reducing ALEs could be beneficial.
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BRG1, an active subunit of the SWI/SNF chromatin-remodeling complex, enables the EP300-dependent transcription of proliferation and DNA repair genes from their E2F/CpG-driven promoters in breast cancer cells. In the current study, we show that BRG1-EP300 complexes are accompanied by poly-ADP-ribose polymerase 1 (PARP1), which emerges as the functional component of the promoter-bound multiprotein units that are capable of controlling gene expression. This enzyme is co-distributed with BRG1 at highly acetylated promoters of genes such as CDK4, LIG1, or NEIL3, which are responsible for cancer cell growth and the removal of DNA damage.

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Oxidized LDL (oxLDL) has been shown to play a crucial role in the onset and development of cardiovascular disorders. The study of oxLDL, as an initiator of inflammatory cascades, led to the discovery of a variety of oxidized phospholipids (oxPLs) responsible for pro-inflammatory actions. Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (PAPC) is frequently used by the scientific community as a representative oxPL mixture to study the biological effects of oxidized lipids, due to the high abundance of PAPC in human tissues and the biological activities of oxidized arachidonic acids derivatives.

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Pyruvate kinase catalyses the last step in glycolysis and has been suggested to contribute to the regulation of aerobic glycolysis in cancer cells. It can be inhibited by oxidation of cysteine residues in vitro and in vivo, which is relevant to the more pro-oxidant state in cancer and proliferating tissues. These conditions also favour lipid peroxidation and the formation of electrophilic fragmentation products, including short-chain aldehydes that can covalently modify proteins.

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Most membrane proteins function through interactions with other proteins in the phospholipid bilayer, the cytosol or the extracellular milieu. Understanding the molecular basis of these interactions is key to understanding membrane protein function and dysfunction. Here we demonstrate for the first time how a nano-encapsulation method based on styrene maleic acid lipid particles (SMALPs) can be used in combination with native gel electrophoresis to separate membrane protein complexes in their native state.

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Silver is widely used for its antimicrobial properties, but microbial resistance to heavy metals is increasing. Silver(II) compounds are more oxidizing and therefore have the potential to overcome resistance via extensive attack on cellular components, but have traditionally been hard to stabilize for biological applications. Here, the high oxidation state cation was stabilised using pyridinecarboxylate ligands, of which the 2,6-dicarboxypyridine Ag(II) complex (Ag2,6P) was found to have the best tractability.

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  • * Researchers used HNE-treated human serum albumin to create sheep antiserum and employed peptide arrays to compare binding patterns between a commercial antibody and an in-house developed antibody against HNE.
  • * Both antibodies identified common peptide bindings on HNE-treated human serum albumin, but the commercial antibody was more specific, recognizing more adducts than the in-house antibody, while the latter showed unique recognition of specific peptides.
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So far, the investigation in cancer cell lines of the modulation of cancer growth and progression by oxysterols, in particular 27-hydroxycholesterol (27HC), has yielded controversial results. The primary aim of this study was the quantitative evaluation of possible changes in 27HC levels during the different steps of colorectal cancer (CRC) progression in humans. A consistent increase in this oxysterol in CRC mass compared to the tumor-adjacent tissue was indeed observed, but only in advanced stages of progression (TNM stage III), a phase in which cancer has spread to nearby sites.

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  • Biological characterisation of membrane proteins is challenging due to traditional methods losing essential lipids during extraction, which affects protein structure and function.
  • Styrene maleic acid (SMA) copolymers provide a detergent-free alternative for isolating membrane proteins while preserving their surrounding lipids, creating SMA-lipid particles (SMALPs).
  • A new LC-MS/MS method was developed to analyze bacterial phospholipids, revealing distinct lipid profiles associated with different membrane proteins, indicating specific lipid-protein interactions.
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Lipoproteins are essential systemic lipid transport particles, composed of apolipoproteins embedded in a phospholipid and cholesterol monolayer surrounding a cargo of diverse lipid species. Many of the lipids present are susceptible to oxidative damage by lipid peroxidation, giving rise to the formation of reactive lipid peroxidation products (rLPPs). In view of the close proximity of the protein and lipid moieties within lipoproteins, the probability of adduct formation between rLPPs and amino acid residues of the proteins, a process called lipoxidation, is high.

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Although electrophiles are considered as detrimental to cells, accumulating recent evidence indicates that proliferating non-cancerous and particularly cancerous cells utilize these agents for pro-survival and cell cycle promoting signaling. Hence, the redox shift to mild oxidant release must be balanced by multiple defense mechanisms. Our latest findings demonstrate that cell cycle progression, which dictates oxidant level in stress-free conditions, determines PARP1 transcription.

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Unlabelled: Cardiovascular diseases (CVDs) represent the most important cause of mortality in women and in men. Contrary to the long-standing notion that the effects of the major risk factors on CVD outcomes are the same in both sexes, recent evidence recognizes new, potentially independent, sex/gender-related risk factors for CVDs, and sex/gender-differences in the clinical presentation of CVDs have been demonstrated. Furthermore, some therapeutic options may not be equally effective and safe in men and women.

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  • * Researchers used model proteins, lysozyme and human serum albumin (HSA), to explore the effects of two aldehydes, acrolein and pentanal, on protein modifications, employing mass spectrometry (MS) for analysis.
  • * The findings reveal specific amino acid modifications and provide diagnostic ions for identifying these adducts, showcasing a refined approach combining intact protein analysis with LC-MS/MS techniques for studying complex biological systems.
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  • The generation of 3-nitrotyrosine in proteins is a result of oxidative or nitrative stress, potentially serving as a biomarker for inflammatory diseases.
  • A new highly sensitive electrochemiluminescence-based ELISA for measuring nitrotyrosine has been developed, offering 50 times greater sensitivity than some existing tests and showing accuracy in quantifying levels in serum samples.
  • The ELISA was validated against mass spectrometry, and in a clinical study of surgical patients, it detected a significant increase in nitrotyrosine levels post-surgery, indicating it can effectively measure inflammatory responses.
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  • Oxidation from reactive species, like hypochlorous acid (HOCl), can alter the function of proteins such as PTEN, a regulator of the PI3K/AKT signaling pathway, leading to downstream cellular effects.
  • Experiments revealed that PTEN's activity significantly decreased with higher HOCl:protein ratios (60:1 and above), resulting in protein instability and aggregation, as shown by SDS-PAGE analysis.
  • A total of 20 oxidative modifications were identified in PTEN, with methionine and cysteine residues showing the most oxidation, influencing protein aggregation, structure, and overall function.
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Unlabelled: Cardiovascular diseases (CVDs) represent the most important cause of mortality in women and in men. Contrary to the long-standing notion that the effects of the major risk factors on CVD outcomes are the same in both sexes, recent evidence recognizes new, potentially independent, sex/gender-related risk factors for CVDs, and sex/gender-differences in the clinical presentation of CVDs have been demonstrated. Furthermore, some therapeutic options may not be equally effective and safe in men and women.

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