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Keywords = high-abundant protein depletion

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14 pages, 4605 KiB  
Article
Different Proteins as Biomarkers for Sac Shrinkage After Endovascular Aortic Repair of Abdominal Aortic Aneurysms
by Alexander Zimmermann, Daniela Reitnauer, Yankey Yundung, Anna-Leonie Menges, Lorenz Meuli, Jaroslav Pelisek and Benedikt Reutersberg
J. Cardiovasc. Dev. Dis. 2024, 11(11), 374; https://doi.org/10.3390/jcdd11110374 - 20 Nov 2024
Viewed by 1059
Abstract
Background: This study aims to identify circulating biomarkers by using proteomic analysis associated with sac shrinkage or expansion in patients undergoing endovascular aneurysm repair (EVAR) for abdominal aortic aneurysms (AAAs). Methods: Plasma samples were analysed from 32 patients treated with EVAR between 10/2009 [...] Read more.
Background: This study aims to identify circulating biomarkers by using proteomic analysis associated with sac shrinkage or expansion in patients undergoing endovascular aneurysm repair (EVAR) for abdominal aortic aneurysms (AAAs). Methods: Plasma samples were analysed from 32 patients treated with EVAR between 10/2009 and 10/2020. Patients were divided into two groups based on postoperative sac behaviour: sac shrinkage (≥5 mm reduction) and no shrinkage (stabilisation or expansion). Proteomic analysis was performed using high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS), with abundant protein depletion to enhance the detection of low-abundant proteins. Results: Of the 32 patients, 20 exhibited sac shrinkage, and 12 showed no shrinkage. Proteomic analysis identified 632 proteins, with significant differential abundance observed after adjusting for relevant clinical parameters. Notably, neurogranin (NRGN) levels were significantly associated with hypertension and smoking, while casein alpha S1 (CSN1S1) levels varied with statin use. Differentially abundant proteins related to aortic diameter included calpastatin, SCUBE3, and ubiquitin-conjugating enzyme E2, among others. Conclusions: Proteomic profiling revealed distinct biomarker patterns associated with sac behaviour in EVAR-treated AAA patients. These findings suggest potential therapeutic targets for enhancing EVAR outcomes and underscore the need for further investigation into the biological mechanisms underlying aneurysm sac shrinkage and stability. Full article
(This article belongs to the Special Issue Risk Factors and Outcomes in Cardiac Surgery)
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16 pages, 1926 KiB  
Article
TLR2/TLR5 Signaling and Gut Microbiota Mediate Soybean-Meal-Induced Enteritis and Declined Growth and Antioxidant Capabilities in Large Yellow Croaker (Larimichthys crocea)
by Lei Zheng, Chao Zeng, Wanqin Zhu, Jiaonan Zhang, Lei Wang, Jianchun Shao and Wei Zhao
J. Mar. Sci. Eng. 2024, 12(11), 2016; https://doi.org/10.3390/jmse12112016 - 8 Nov 2024
Cited by 4 | Viewed by 1179
Abstract
Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims [...] Read more.
Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims to elucidate the molecular basis of soybean-meal-induced enteritis and its impact on intestinal microbiota in L. crocea. To this end, four isonitrogenous and isolipidic diets with varying soybean meal levels (0% FM, 15% SBM15, 30% SBM30, and 45% SBM45) were administered to L. crocea for 8 weeks. The results indicated that the SBM30 and SBM45 treatments significantly hindered fish growth, digestive efficiency, and protein utilization. Furthermore, high soybean meal levels (SBM30 and SBM45) activated intestinal Toll-like receptors (TLR2A, TLR2B, TLR5, and TLR22), stimulating C-Rel and mTOR protein expression and elevating ERK phosphorylation. This led to increased pro-inflammatory cytokine production (IL-1β, IL-6, and TNF-α) and decreased anti-inflammatory cytokine expression (IL-4/13A, IL-4/13B, and TGF-β), suggesting a potential signaling pathway for soybean-meal-induced enteritis. Furthermore, enteritis induced by high soybean meal levels led to oxidative damage, evident from increased MDA levels and decreased antioxidant enzyme activities (SOD and CAT). The SBM30 and SBM45 treatments increased Firmicutes and Bacteroidetes abundance in fish gut microbiota, while Proteobacteria abundance decreased. This microbiota shift may enhance soybean meal nutrient utilization, yet high soybean meal concentrations still impair growth. A soybean-meal-rich diet promotes harmful bacteria like Rhodococcus and depletes probiotics like Ralstonia, increasing disease risks. L. crocea has limited tolerance for soybean meal, necessitating advanced processing to mitigate anti-nutritional factors. Ultimately, exploring alternative protein sources beyond soybean meal for fish meal replacement is optimal for L. crocea. Full article
(This article belongs to the Special Issue Nutrition and Physiology of Marine Fish)
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16 pages, 42416 KiB  
Article
FBXO11 Mediates Ubiquitination of ZEB1 and Modulates Epithelial-to-Mesenchymal Transition in Lung Cancer Cells
by Xinyue Zhao, Zhihui Han, Ruiying Liu, Zehao Li, Ling Mei and Yue Jin
Cancers 2024, 16(19), 3269; https://doi.org/10.3390/cancers16193269 - 26 Sep 2024
Cited by 2 | Viewed by 4273
Abstract
Epithelial-to-mesenchymal transition (EMT) affects the invasion and migration of cancer cells. Here, we show that FBXO11 recognizes and promotes ubiquitin-mediated degradation of ZEB1. There is a strong association between FBXO11 and ZEB1 in non-small cell lung cancer (NSLC) in a clinical database. FBXO11 [...] Read more.
Epithelial-to-mesenchymal transition (EMT) affects the invasion and migration of cancer cells. Here, we show that FBXO11 recognizes and promotes ubiquitin-mediated degradation of ZEB1. There is a strong association between FBXO11 and ZEB1 in non-small cell lung cancer (NSLC) in a clinical database. FBXO11 interacts with ZEB1, a core inducer of EMT. FBXO11 leads to increased ubiquitination and proteasomal degradation of ZEB1. Depletion of endogenous FBXO11 causes ZEB1 protein accumulation and EMT in A549 and H1299 cells, while overexpression of FBXO11 reduces ZEB1 protein abundance and cellular invasiveness. Importantly, the depletion of ZEB1 suppresses the increased migration and invasion of A549 and H1299 cells promoted by the depletion of FBXO11. The same results are shown in xenograft tumors. High FBXO11 expression is associated with a favorable prognosis in NSLC. Collectively, our study demonstrates that FBXO11 modulates EMT by mediating the stability of ZEB1 in lung cancer cells. Full article
(This article belongs to the Special Issue Molecular Mechanisms of Cancer Development and Metastasis)
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17 pages, 2636 KiB  
Article
The Low-Abundance Plasma Proteome Reveals Differentially Abundant Proteins Associated with Breast Implant Capsular Contracture: A Pilot Study
by Md. Arifur Rahman, Ardeshir Amirkhani, Maria Mempin, Seong Beom Ahn, Anand K. Deva, Mark S. Baker, Karen Vickery and Honghua Hu
Proteomes 2024, 12(3), 22; https://doi.org/10.3390/proteomes12030022 - 6 Aug 2024
Viewed by 2048
Abstract
Capsular contracture (CC) is one of the most common postoperative complications associated with breast implant-associated infections. The mechanisms that lead to CC remain poorly understood. Plasma is an ideal biospecimen for early proteomics biomarker discovery. However, as high-abundance proteins mask signals from low-abundance [...] Read more.
Capsular contracture (CC) is one of the most common postoperative complications associated with breast implant-associated infections. The mechanisms that lead to CC remain poorly understood. Plasma is an ideal biospecimen for early proteomics biomarker discovery. However, as high-abundance proteins mask signals from low-abundance proteins, identifying novel or specific proteins as biomarkers for a particular disease has been hampered. Here, we employed depletion of high-abundance plasma proteins followed by Tandem Mass Tag (TMT)-based quantitative proteomics to compare 10 healthy control patients against 10 breast implant CC patients. A total of 450 proteins were identified from these samples. Among them, 16 proteins were significantly differentially expressed in which 5 proteins were upregulated and 11 downregulated in breast implant CC patients compared to healthy controls. Gene Ontology enrichment analysis revealed that proteins related to cell, cellular processes and catalytic activity were highest in the cellular component, biological process, and molecular function categories, respectively. Further, pathway analysis revealed that inflammatory responses, focal adhesion, platelet activation, and complement and coagulation cascades were enriched pathways. The differentially abundant proteins from TMT-based quantitative proteomics have the potential to provide important information for future mechanistic studies and in the development of breast implant CC biomarkers. Full article
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20 pages, 7499 KiB  
Article
Bacteroides uniformis Ameliorates Carbohydrate and Lipid Metabolism Disorders in Diabetic Mice by Regulating Bile Acid Metabolism via the Gut–Liver Axis
by Xue-Xue Zhu, Chen-Yang Zhao, Xin-Yu Meng, Xiao-Yi Yu, Lin-Chun Ma, Tian-Xiao Chen, Chang Chang, Xin-Yu Chen, Yuan Zhang, Bao Hou, Wei-Wei Cai, Bin Du, Zhi-Jun Han, Li-Ying Qiu and Hai-Jian Sun
Pharmaceuticals 2024, 17(8), 1015; https://doi.org/10.3390/ph17081015 - 1 Aug 2024
Cited by 4 | Viewed by 2677
Abstract
Background: Type 2 diabetes mellitus (T2DM) is a metabolic syndrome characterized by chronic inflammation, insulin resistance, and islet cell damage. The prevention of T2DM and its associated complications is an urgent public health issue that affects hundreds of millions of people globally. Numerous [...] Read more.
Background: Type 2 diabetes mellitus (T2DM) is a metabolic syndrome characterized by chronic inflammation, insulin resistance, and islet cell damage. The prevention of T2DM and its associated complications is an urgent public health issue that affects hundreds of millions of people globally. Numerous studies suggest that disturbances in gut metabolites are important driving forces for the pathogenesis of diabetes. However, the functions and mechanisms of action of most commensal bacteria in T2DM remain largely unknown. Methods: The quantification of bile acids (BAs) in fecal samples was performed using ultra-performance liquid chromatography–tandem mass spectrometer (UPLC-MS/MS). The anti-diabetic effects of Bacteroides uniformis (B. uniformis) and its metabolites cholic acid (CA) and chenodeoxycholic acid (CDCA) were assessed in T2DM mice induced by streptozocin (STZ) plus high-fat diet (HFD). Results: We found that the abundance of B. uniformis in the feces and the contents of CA and CDCA were significantly downregulated in T2DM mice. B. uniformis was diminished in diabetic individuals and this bacterium was sufficient to promote the production of BAs. Colonization of B. uniformis and intragastric gavage of CA and CDCA effectively improved the disorder of glucose and lipid metabolism in T2DM mice by inhibiting gluconeogenesis and lipolysis in the liver. CA and CDCA improved hepatic glucose and lipid metabolism by acting on the Takeda G protein-coupled receptor 5 (TGR5)/adenosine monophosphate-activated protein kinase (AMPK) signaling pathway since knockdown of TGR5 minimized the benefit of CA and CDCA. Furthermore, we screened a natural product—vaccarin (VAC)—that exhibited anti-diabetic effects by promoting the growth of B. uniformis in vitro and in vivo. Gut microbiota pre-depletion abolished the favorable effects of VAC in diabetic mice. Conclusions: These data suggest that supplementation of B. uniformis may be a promising avenue to ameliorate T2DM by linking the gut and liver. Full article
(This article belongs to the Section Natural Products)
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12 pages, 2168 KiB  
Article
Impact of Serotonin Deficiency on Circadian Dopaminergic Rhythms
by Giacomo Maddaloni, Noemi Barsotti, Sara Migliarini, Martina Giordano, Serena Nazzi, Marta Picchi, Francesco Errico, Alessandro Usiello and Massimo Pasqualetti
Int. J. Mol. Sci. 2024, 25(12), 6475; https://doi.org/10.3390/ijms25126475 - 12 Jun 2024
Cited by 4 | Viewed by 2865
Abstract
Physiology and behavior are structured temporally to anticipate daily cycles of light and dark, ensuring fitness and survival. Neuromodulatory systems in the brain—including those involving serotonin and dopamine—exhibit daily oscillations in neural activity and help shape circadian rhythms. Disrupted neuromodulation can cause circadian [...] Read more.
Physiology and behavior are structured temporally to anticipate daily cycles of light and dark, ensuring fitness and survival. Neuromodulatory systems in the brain—including those involving serotonin and dopamine—exhibit daily oscillations in neural activity and help shape circadian rhythms. Disrupted neuromodulation can cause circadian abnormalities that are thought to underlie several neuropsychiatric disorders, including bipolar mania and schizophrenia, for which a mechanistic understanding is still lacking. Here, we show that genetically depleting serotonin in Tph2 knockout mice promotes manic-like behaviors and disrupts daily oscillations of the dopamine biosynthetic enzyme tyrosine hydroxylase (TH) in midbrain dopaminergic nuclei. Specifically, while TH mRNA and protein levels in the Substantia Nigra (SN) and Ventral Tegmental Area (VTA) of wild-type mice doubled between the light and dark phase, TH levels were high throughout the day in Tph2 knockout mice, suggesting a hyperdopaminergic state. Analysis of TH expression in striatal terminal fields also showed blunted rhythms. Additionally, we found low abundance and blunted rhythmicity of the neuropeptide cholecystokinin (Cck) in the VTA of knockout mice, a neuropeptide whose downregulation has been implicated in manic-like states in both rodents and humans. Altogether, our results point to a previously unappreciated serotonergic control of circadian dopamine signaling and propose serotonergic dysfunction as an upstream mechanism underlying dopaminergic deregulation and ultimately maladaptive behaviors. Full article
(This article belongs to the Special Issue Role of Serotonin in Brain Function)
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15 pages, 1476 KiB  
Article
Cyclosporin A-Based PROTACs Can Deplete Abundant Cellular Cyclophilin A without Suppressing T Cell Activation
by Katharina Hilbig, Russell Towers, Marc Schmitz, Martin Bornhäuser, Petra Lennig and Yixin Zhang
Molecules 2024, 29(12), 2779; https://doi.org/10.3390/molecules29122779 - 11 Jun 2024
Cited by 1 | Viewed by 2357
Abstract
Cyclophilin A (CypA), the cellular receptor of the immunosuppressant cyclosporin A (CsA), is an abundant cytosolic protein and is involved in a variety of diseases. For example, CypA supports cancer proliferation and mediates viral infections, such as the human immunodeficiency virus 1 (HIV-1). [...] Read more.
Cyclophilin A (CypA), the cellular receptor of the immunosuppressant cyclosporin A (CsA), is an abundant cytosolic protein and is involved in a variety of diseases. For example, CypA supports cancer proliferation and mediates viral infections, such as the human immunodeficiency virus 1 (HIV-1). Here, we present the design of PROTAC (proteolysis targeting chimera) compounds against CypA to induce its intracellular proteolysis and to investigate their effect on immune cells. Interestingly, upon connecting to E3 ligase ligands, both peptide-based low-affinity binders and CsA-based high-affinity binders can degrade CypA at nM concentration in HeLa cells and fibroblast cells. As the immunosuppressive effect of CsA is not directly associated with the binding of CsA to CypA but the inhibition of phosphatase calcineurin by the CypA:CsA complex, we investigated whether a CsA-based PROTAC compound could induce CypA degradation without affecting the activation of immune cells. P3, the most efficient PROTAC compound discovered from this study, could deplete CypA in lymphocytes without affecting cell proliferation and cytokine production. This work demonstrates the feasibility of the PROTAC approach in depleting the abundant cellular protein CypA at low drug dosage without affecting immune cells, allowing us to investigate the potential therapeutic effects associated with the endogenous protein in the future. Full article
(This article belongs to the Section Chemical Biology)
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19 pages, 9666 KiB  
Article
Plasma and Kidney Proteome Profiling Combined with Laser Capture Microdissection Reveal Large Increases in Immunoglobulins with Age
by Leanne J. G. Chan, Niclas Olsson, Magdalena Preciado López, Kayley Hake, Haruna Tomono, Matthew A. Veras and Fiona E. McAllister
Proteomes 2024, 12(2), 16; https://doi.org/10.3390/proteomes12020016 - 3 Jun 2024
Viewed by 5144
Abstract
One of the main hallmarks of aging is aging-associated inflammation, also known as inflammaging. In this study, by comparing plasma and kidney proteome profiling of young and old mice using LC–MS profiling, we discovered that immunoglobulins are the proteins that exhibit the highest [...] Read more.
One of the main hallmarks of aging is aging-associated inflammation, also known as inflammaging. In this study, by comparing plasma and kidney proteome profiling of young and old mice using LC–MS profiling, we discovered that immunoglobulins are the proteins that exhibit the highest increase with age. This observation seems to have been disregarded because conventional proteome profiling experiments typically overlook the expression of high-abundance proteins or employ depletion methods to remove them before LC–MS analysis. We show that proteome profiling of immunoglobulins will likely be a useful biomarker of aging. Spatial profiling using immunofluorescence staining of kidney sections indicates that the main increases in immunoglobulins with age are localized in the glomeruli of the kidney. Using laser capture microdissection coupled with LC–MS, we show an increase in multiple immune-related proteins in glomeruli from aged mice. Increased deposition of immunoglobulins, immune complexes, and complement proteins in the kidney glomeruli may be a factor leading to reduced filtering capacity of the kidney with age. Therapeutic strategies to reduce the deposition of immunoglobulins in the kidney may be an attractive strategy for healthy aging. Full article
(This article belongs to the Special Issue Quantitative Proteomics: Techniques and Applications)
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18 pages, 2619 KiB  
Article
Exploring the miRNAs Profile in Dark-Cutting Beef
by Laura González-Blanco, Luis J. Royo, Yolanda Diñeiro, Susana García-Torres, Ana Coto-Montes, Verónica Sierra and Mamen Oliván
Foods 2024, 13(6), 960; https://doi.org/10.3390/foods13060960 - 21 Mar 2024
Cited by 2 | Viewed by 2075
Abstract
In an animal production system, different stressors may cause the depletion of muscle glycogen stores, resulting in an elevated pH at 24 h post mortem (pH24), which leads to cell metabolism alterations that affect the conversion of muscle into meat, causing [...] Read more.
In an animal production system, different stressors may cause the depletion of muscle glycogen stores, resulting in an elevated pH at 24 h post mortem (pH24), which leads to cell metabolism alterations that affect the conversion of muscle into meat, causing meat quality defects, such as dark-cutting beef, also known as dark, firm, and dry (DFD) beef. This process may involve the alteration of small non-coding RNAs (miRNAs), which play critical regulatory roles in cellular processes. Here, we determined whether differential miRNA expression in the Longissimus thoracis et lumborum muscle from the Asturiana de los Valles breed at 24 h post mortem could serve as an early indicator of beef quality defects. Following total RNA extraction, complete miRNAome sequencing revealed 12 miRNAs that were significantly upregulated (p < 0.001) in DFD beef compared to the levels in CONTROL beef. These miRNAs are mainly involved in the cellular responses to redox imbalances and apoptosis. Among these, four miRNAs known to be related to oxidative stress (bta-miR-1246, bta-miR-2332, bta-miR-23b-5p, and bta-miR-2411-3p) were validated via quantitative RT-PCR. Some of their target proteins were also analyzed using Western blotting. High 70 kDa heat shock protein and low Caspase-9 expressions (p < 0.01) were found in DFD beef, suggesting the downregulation of apoptosis. These results suggest the importance of miRNAs in regulating stress in muscle cells during early post mortem, as differences in the abundance of some of these miRNAs are still observed at 24 h post mortem. These changes lead to an inadequate conversion of muscle into meat, resulting in meats with quality defects. Full article
(This article belongs to the Section Meat)
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22 pages, 8070 KiB  
Article
Preparation of Viable Human Neurites for Neurobiological and Neurodegeneration Studies
by Markus Brüll, Nils Geese, Ivana Celardo, Michael Laumann and Marcel Leist
Cells 2024, 13(3), 242; https://doi.org/10.3390/cells13030242 - 27 Jan 2024
Cited by 4 | Viewed by 2703
Abstract
Few models allow the study of neurite damage in the human central nervous system. We used here dopaminergic LUHMES neurons to establish a culture system that allows for (i) the observation of highly enriched neurites, (ii) the preparation of the neurite fraction for [...] Read more.
Few models allow the study of neurite damage in the human central nervous system. We used here dopaminergic LUHMES neurons to establish a culture system that allows for (i) the observation of highly enriched neurites, (ii) the preparation of the neurite fraction for biochemical studies, and (iii) the measurement of neurite markers and metabolites after axotomy. LUHMES-based spheroids, plated in culture dishes, extended neurites of several thousand µm length, while all somata remained aggregated. These cultures allowed an easy microscopic observation of live or fixed neurites. Neurite-only cultures (NOC) were produced by cutting out the still-aggregated somata. The potential application of such cultures was exemplified by determinations of their protein and RNA contents. For instance, the mitochondrial TOM20 protein was highly abundant, while nuclear histone H3 was absent. Similarly, mitochondrial-encoded RNAs were found at relatively high levels, while the mRNA for a histone or the neuronal nuclear marker NeuN (RBFOX3) were relatively depleted in NOC. Another potential use of NOC is the study of neurite degeneration. For this purpose, an algorithm to quantify neurite integrity was developed. Using this tool, we found that the addition of nicotinamide drastically reduced neurite degeneration. Also, the chelation of Ca2+ in NOC delayed the degeneration, while inhibitors of calpains had no effect. Thus, NOC proved to be suitable for biochemical analysis and for studying degeneration processes after a defined cut injury. Full article
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13 pages, 2340 KiB  
Article
Proteome Analysis of Serum Purified Using Solanum tuberosum and Lycopersicon esculentum Lectins
by Daisuke Nakajima, Ryo Konno, Yasuomi Miyashita, Masaki Ishikawa, Osamu Ohara and Yusuke Kawashima
Int. J. Mol. Sci. 2024, 25(2), 1315; https://doi.org/10.3390/ijms25021315 - 21 Jan 2024
Cited by 1 | Viewed by 2806
Abstract
Serum and plasma exhibit a broad dynamic range of protein concentrations, posing challenges for proteome analysis. Various technologies have been developed to reduce this complexity, including high-abundance depletion methods utilizing antibody columns, extracellular vesicle enrichment techniques, and trace protein enrichment using nanobead cocktails. [...] Read more.
Serum and plasma exhibit a broad dynamic range of protein concentrations, posing challenges for proteome analysis. Various technologies have been developed to reduce this complexity, including high-abundance depletion methods utilizing antibody columns, extracellular vesicle enrichment techniques, and trace protein enrichment using nanobead cocktails. Here, we employed lectins to address this, thereby extending the scope of biomarker discovery in serum or plasma using a novel approach. We enriched serum proteins using 37 different lectins and subjected them to LC–MS/MS analysis with data-independent acquisition. Solanum tuberosum lectin (STL) and Lycopersicon esculentum lectin (LEL) enabled the detection of more serum proteins than the other lectins. STL and LEL bind to N-acetylglucosamine oligomers, emphasizing the significance of capturing these oligomer-binding proteins when analyzing serum trace proteins. Combining STL and LEL proved more effective than using them separately, allowing us to identify over 3000 proteins from serum through single-shot proteome analysis. We applied the STL/LEL trace-protein enrichment method to the sera of systemic lupus erythematosus model mice. This revealed differences in >1300 proteins between the systemic lupus erythematosus model and control mouse sera, underscoring the utility of this method for biomarker discovery. Full article
(This article belongs to the Special Issue New Insights into Proteomics in Disease)
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21 pages, 1420 KiB  
Article
Identification of Plasma Biomarkers from Rheumatoid Arthritis Patients Using an Optimized Sequential Window Acquisition of All THeoretical Mass Spectra (SWATH) Proteomics Workflow
by Liang Jin, Fei Wang, Xue Wang, Bohdan P. Harvey, Yingtao Bi, Chenqi Hu, Baoliang Cui, Anhdao T. Darcy, John W. Maull, Ben R. Phillips, Youngjae Kim, Gary J. Jenkins, Thierry R. Sornasse and Yu Tian
Proteomes 2023, 11(4), 32; https://doi.org/10.3390/proteomes11040032 - 16 Oct 2023
Cited by 1 | Viewed by 4927
Abstract
Rheumatoid arthritis (RA) is a systemic autoimmune and inflammatory disease. Plasma biomarkers are critical for understanding disease mechanisms, treatment effects, and diagnosis. Mass spectrometry-based proteomics is a powerful tool for unbiased biomarker discovery. However, plasma proteomics is significantly hampered by signal interference from [...] Read more.
Rheumatoid arthritis (RA) is a systemic autoimmune and inflammatory disease. Plasma biomarkers are critical for understanding disease mechanisms, treatment effects, and diagnosis. Mass spectrometry-based proteomics is a powerful tool for unbiased biomarker discovery. However, plasma proteomics is significantly hampered by signal interference from high-abundance proteins, low overall protein coverage, and high levels of missing data from data-dependent acquisition (DDA). To achieve quantitative proteomics analysis for plasma samples with a balance of throughput, performance, and cost, we developed a workflow incorporating plate-based high abundance protein depletion and sample preparation, comprehensive peptide spectral library building, and data-independent acquisition (DIA) SWATH mass spectrometry-based methodology. In this study, we analyzed plasma samples from both RA patients and healthy donors. The results showed that the new workflow performance exceeded that of the current state-of-the-art depletion-based plasma proteomic platforms in terms of both data quality and proteome coverage. Proteins from biological processes related to the activation of systemic inflammation, suppression of platelet function, and loss of muscle mass were enriched and differentially expressed in RA. Some plasma proteins, particularly acute-phase reactant proteins, showed great power to distinguish between RA patients and healthy donors. Moreover, protein isoforms in the plasma were also analyzed, providing even deeper proteome coverage. This workflow can serve as a basis for further application in discovering plasma biomarkers of other diseases. Full article
(This article belongs to the Section Proteomics Technology and Methodology Development)
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21 pages, 9134 KiB  
Article
Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles
by Christian Freise, Andreas Zappe, Norbert Löwa, Jörg Schnorr, Kevin Pagel, Frank Wiekhorst and Matthias Taupitz
Int. J. Mol. Sci. 2023, 24(18), 14253; https://doi.org/10.3390/ijms241814253 - 18 Sep 2023
Cited by 4 | Viewed by 1987
Abstract
Uremic toxins exert pathophysiological effects on cells and tissues, such as the generation of a pro-calcifying subtype of exosome-like extracellular vesicles (EVs) in vascular cells. Little is known about the effects of the toxins on the surface structure of EVs. Thus, we studied [...] Read more.
Uremic toxins exert pathophysiological effects on cells and tissues, such as the generation of a pro-calcifying subtype of exosome-like extracellular vesicles (EVs) in vascular cells. Little is known about the effects of the toxins on the surface structure of EVs. Thus, we studied the effects of uremic toxins on the abundance of sulfated glycosaminoglycans (GAGs) in EVs, and the implications for binding of ligands such as very small superparamagnetic iron oxide particles (VSOPs) which could be of relevance for radiological EV-imaging. Vascular cells were treated with the uremic toxins NaH2PO4 and a mixture of urea and indoxyl sulfate. Uremia in rats was induced by adenine feeding. EVs were isolated from culture supernatants and plasma of rats. By proton T1-relaxometry, magnetic particle spectroscopy, and analysis of genes, proteins, and GAG-contents, we analyzed the roles of GAGs in the ligand binding of EVs. By influencing GAG-associated genes in host cells, uremic toxins induced higher GAG contents in EVs, particularly of sulfated chondroitin sulfate and heparan sulfate chains. EVs with high GAG content interacted stronger with VSOPs compared to control ones. This was confirmed by experiments with GAG-depleted EVs from genetically modified CHO cells and with uremic rat-derived EVs. Mechanistically, uremic toxin-induced PI3K/AKT-signaling and expression of the sulfate transporter SLC26A2 in host cells contributed to high GAG contents in EVs. In conclusion, uremic conditions induce enhanced GAG contents in EVs, which entails a stronger interaction with VSOPs. VSOPs might be suitable for radiological imaging of EVs rich in GAGs. Full article
(This article belongs to the Collection Feature Papers in “Molecular Biology”)
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19 pages, 1658 KiB  
Review
Evaluating Alternative and Sustainable Food Resources: A Review of the Nutritional Composition of Myctophid Fishes
by Bowen Zhang, Heidi Pethybridge, Patti Virtue, Peter D. Nichols, Kerrie Swadling, Alan Williams and Kim Lee-Chang
Sustainability 2023, 15(15), 12039; https://doi.org/10.3390/su151512039 - 6 Aug 2023
Cited by 3 | Viewed by 3178
Abstract
Additional and alternative sustainable food resources are needed as the global human population increases. Marine fishes have long provided essential nutrients, such as omega-3 long-chain (≥C20) polyunsaturated fatty acids (n-3 LC-PUFA), protein, and vitamins to meet human dietary requirements and feed for agricultural [...] Read more.
Additional and alternative sustainable food resources are needed as the global human population increases. Marine fishes have long provided essential nutrients, such as omega-3 long-chain (≥C20) polyunsaturated fatty acids (n-3 LC-PUFA), protein, and vitamins to meet human dietary requirements and feed for agricultural production. Many current commercial fish stocks are depleted or fully exploited, but oceanic mesopelagic fishes, particularly the myctophids (lanternfishes), represent a potentially very large and unfished resource. This review analysed the literature on nutritional and biochemical compositions of myctophids as a first step towards understanding the health benefits and risks of consuming them. We found that myctophids have high levels of protein (11–23% wet weight, WW) and variable lipid content (0.5–26% WW). In most species, desirable triacylglycerols or phospholipids dominated over less-desirable wax esters, and most have abundant amounts of health-promoting n-3 LC-PUFA, such as DHA and EPA. Myctophids have low levels of heavy metals and persistent organic pollutants. Most nutritional information is available for species from the Pacific and Southern Oceans and for the genera Benthosema, Electrona, and Diaphus. Myctophids generally possess favourable nutritional profiles, but major gaps in knowledge regarding their stock assessment, ecology and the economic viability for their harvest are barriers to developing sustainable fisheries. Full article
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6 pages, 242 KiB  
Proceeding Paper
Identification of Potential Protein Biomarkers in a Depressed Chinese Malaysian University Student Using Liquid Chromatography-Tandem Mass Spectrometry
by Sin Yee Yap, Chai Nien Foo, Yang Mooi Lim, Foong Leng Ng, Pek Yee Tang, Jagjit Kaur Najar Singh, Sherina Mohd-Sidik and Kai-Shuen Pheh
Med. Sci. Forum 2023, 21(1), 10; https://doi.org/10.3390/ECB2023-14089 - 9 Jun 2023
Viewed by 1426
Abstract
Depression is a serious psychological disorder with high prevalence rates, especially among university students. Serum proteins related to the immune system and oxygen and lipid transfer could have contributing roles in the development of depression and could act as biomarkers for depression. Currently, [...] Read more.
Depression is a serious psychological disorder with high prevalence rates, especially among university students. Serum proteins related to the immune system and oxygen and lipid transfer could have contributing roles in the development of depression and could act as biomarkers for depression. Currently, there is a lack of accurate biological methods that can be used to diagnose depression. Biomarkers could be an inexpensive and convenient way to predict depression and understand its pathophysiology. This study aimed to screen the serum proteome profile of a depressed student for the identification of potential depression biomarkers. A Malaysian private university student who was recruited from the pre-test study (n = 10) was further analyzed for serum proteome due to the fact that he was depressed, with scores of 15 out of 27 on the Patient Health Questionnaire (PHQ-9). After depleting the high-abundance proteins from the serum sample, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed to identify the expressed proteins. A total of 224 proteins were identified. Globins, globulins, apolipoproteins and glycoproteins were most commonly detected. Here, we show the potential biomarkers that can be used to identify depression vulnerable individuals. These findings may be relevant to the development of new diagnostic and treatment strategies. However, further studies with larger sample sizes and healthy controls are needed to confirm the role of these candidate biomarkers for the prediction and diagnosis of depression. Full article
(This article belongs to the Proceedings of The 2nd International Electronic Conference on Biomedicines)
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