Sex-Specific Differences in Extracellular Vesicle Protein Cargo in Synovial Fluid of Patients with Osteoarthritis

Women are at a significantly higher risk of developing osteoarthritis (OA) compared to males. The pathogenesis of osteoarthritis (OA) in women is poorly understood. Extracellular vesicles (EVs) have been shown to play an essential role in numerous signaling processes during the pathogenesis of age-related diseases via paracrine signaling. Molecular profiling of the synovial fluid-derived EVs cargo in women may help in the discovery of novel biomarkers and therapeutics for the treatment of OA in women. Previously, we reported that synovial fluid-derived EV miRNA cargo differs in a sex-specific manner. This study aims to characterize synovial fluid-derived EV protein cargo in OA patients. Our data showed sex-specific EVs protein content in OA. We found haptoglobin, orosomucoid, and ceruloplasmin significantly up-regulated, whereas apolipoprotein down-regulated in female OA EVs. In males, we discovered β-2-glycoprotein, and complement component 5 proteins significantly up-regulated and Spt-Ada-Gcn5 acetyltransferase (SAGA)-associated factor 29 down-regulated in male OA EVs. Database for Annotation, Visualization, and Integrated Discovery (DAVID) and QuickGO analysis revealed OA-specific protein involvement in several biological, molecular, and cellular pathways, specifically in inflammatory processes. In conclusion, synovial fluid EV protein content is altered in a sex-specific manner with OA, explaining the increased prevalence and severity of OA in women.

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Looking to the horizon: the role of bilirubin in the development and prevention of age-related chronic diseases

Clinical Science ◽

10.1042/cs20140566 ◽

2015 ◽

Vol 129 (1) ◽

pp. 1-25 ◽

Cited By ~ 59

Author(s):

Karl-Heinz Wagner ◽

Marlies Wallner ◽

Christine Mölzer ◽

Silvia Gazzin ◽

Andrew Cameron Bulmer ◽

...

Keyword(s):

Disease Resistance ◽

Chronic Diseases ◽

Elevated Serum ◽

Future Research ◽

Bile Pigment ◽

Therapeutic Modalities ◽

Age Related ◽

All Cause Mortality ◽

Novel Biomarkers ◽

Future Research Directions

Bilirubin, the principal tetrapyrrole, bile pigment and catabolite of haem, is an emerging biomarker of disease resistance, which may be related to several recently documented biological functions. Initially believed to be toxic in infants, the perception of bilirubin has undergone a transformation: it is now considered to be a molecule that may promote health in adults. Data from the last decade demonstrate that mildly elevated serum bilirubin levels are strongly associated with reduced prevalence of chronic diseases, particularly cardiovascular diseases (CVDs), as well as CVD-related mortality and risk factors. Recent data also link bilirubin to other chronic diseases, including cancer and Type 2 diabetes mellitus, and to all-cause mortality. Therefore, there is evidence to suggest that bilirubin is a biomarker for reduced chronic disease prevalence and a predictor of all-cause mortality, which is of important clinical significance. In the present review, detailed information on the association between bilirubin and all-cause mortality, as well as the pathological conditions of CVD, cancer, diabetes and neurodegenerative diseases, is provided. The mechanistic background concerning how bilirubin and its metabolism may influence disease prevention and its clinical relevance is also discussed. Given that the search for novel biomarkers of these diseases, as well as for novel therapeutic modalities, is a key research objective for the near future, bilirubin represents a promising candidate, meeting the criteria of a biomarker, and should be considered more carefully in clinical practice as a molecule that might provide insights into disease resistance. Clearly, however, greater molecular insight is warranted to support and strengthen the conclusion that bilirubin can prevent disease, with future research directions also proposed.

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Mass Spectrometry-Based Proteomic Characterization of Cutaneous Melanoma Ectosomes Reveals the Presence of Cancer-Related Molecules

International Journal of Molecular Sciences ◽

10.3390/ijms21082934 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2934 ◽

Cited By ~ 1

Author(s):

Magdalena Surman ◽

Sylwia Kędracka-Krok ◽

Dorota Hoja-Łukowicz ◽

Urszula Jankowska ◽

Anna Drożdż ◽

...

Keyword(s):

Cell Proliferation ◽

Protein Content ◽

Cell Lines ◽

Cutaneous Melanoma ◽

Epithelial Mesenchymal Transition ◽

Mesenchymal Transition ◽

Starting Point ◽

Novel Biomarkers

Cutaneous melanoma (CM) is an aggressive type of skin cancer for which effective biomarkers are still needed. Recently, the protein content of extracellular vesicles (ectosomes and exosomes) became increasingly investigated in terms of its functional role in CM and as a source of novel biomarkers; however, the data concerning the proteome of CM-derived ectosomes is very limited. We used the shotgun nanoLC–MS/MS approach to the profile protein content of ectosomes from primary (WM115, WM793) and metastatic (WM266-4, WM1205Lu) CM cell lines. Additionally, the effect exerted by CM ectosomes on recipient cells was assessed in terms of cell proliferation (Alamar Blue assay) and migratory properties (wound healing assay). All cell lines secreted heterogeneous populations of ectosomes enriched in the common set of proteins. A total of 1507 unique proteins were identified, with many of them involved in cancer cell proliferation, migration, escape from apoptosis, epithelial–mesenchymal transition and angiogenesis. Isolated ectosomes increased proliferation and motility of recipient cells, likely due to the ectosomal transfer of different cancer-promoting molecules. Taken together, these results confirm the significant role of ectosomes in several biological processes leading to CM development and progression, and might be used as a starting point for further studies exploring their diagnostic and prognostic potential.

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Short-term increases in intraluminal pressure reverse age-related decrements in endothelium-dependent dilation in soleus muscle feed arteries

Journal of Applied Physiology ◽

10.1152/japplphysiol.00416.2007 ◽

2007 ◽

Vol 103 (4) ◽

pp. 1172-1179 ◽

Cited By ~ 17

Author(s):

Christopher R. Woodman ◽

Daniel W. Trott ◽

M. Harold Laughlin

Keyword(s):

Protein Content ◽

Soleus Muscle ◽

Intraluminal Pressure ◽

Separate Experiment ◽

Enos Expression ◽

Short Term ◽

Age Related ◽

Enos Mrna ◽

Feed Arteries ◽

Increased Pressure

We tested the hypothesis that short-term increases in intraluminal pressure improve endothelium-dependent dilation and increase endothelial nitric oxide (NO) synthase (eNOS) expression in senescent soleus muscle feed arteries (SFA). SFA isolated from young (4 mo) and old (24 mo) Fischer 344 rats were cannulated and pressurized at 90 (p90) or 130 (p130) cmH2O for 4 h. At the end of the 4-h protocol, pressure in p130 SFA was lowered to 90 cmH2O for examination of endothelium-dependent (flow- or ACh-induced) vasodilation. Flow- and ACh-induced dilations were blunted in old p90 SFA relative to young p90 SFA. Pretreatment with increased pressure (p130) improved flow- and ACh-induced dilations in old SFA, such that vasodilator responses were similar to those in young SFA. In the presence of Nω-nitro-l-arginine (l-NNA) or l-NNA + indomethacin (Indo), flow-induced dilation was inhibited in old p130 SFA, such that the response was not greater than the response in old p90 SFA. In old p130 SFA, ACh-induced dilation was inhibited by l-NNA + Indo (not l-NNA alone). In a separate experiment, SFA were pressurized at 70, 90, 110, or 130 cmH2O for 4 h, and eNOS mRNA and protein content were assessed. Increased pressure induced eNOS mRNA expression in young (not old) SFA. eNOS protein content was not altered in young or old SFA. These results indicate that short-term increases in intraluminal pressure improve endothelium-dependent dilation in senescent SFA, in part by enhancing NO bioavailability; however, the beneficial effect was not associated with increased eNOS expression.

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STRAIN IMPROVEMENT OF A FUNGUS PRODUCING CHITINASE BY A CHEMICAL MUTAGEN

INDIAN DRUGS ◽

10.53879/id.50.11.p0025 ◽

2013 ◽

Vol 50 (11) ◽

pp. 25-28

Author(s):

K Narayanan ◽

◽

N.D. Chopade ◽

V.M Subrahmanyam ◽

J. Venkata Rao

Keyword(s):

Protein Content ◽

Ethidium Bromide ◽

Yeast Extract ◽

Wild Strain ◽

Strain Improvement ◽

Cost Effective ◽

Specific Protein ◽

Chemical Mutagen ◽

Yeast Extract Medium ◽

Extract Medium

Microbial chitinases are commercially exploited for their biocontrol properties and generation of useful products from chitinous waste. Availability of highly active chitinolytic enzymes is a major problem. The present study was carried out to improve chitinase production by Aspergillus terreus using a chemical mutagen, ethidium bromide. The organism was cultivated on lactose- yeast extract medium. The production medium consisting of chitin- yeast extract medium was seeded at 10% level. The wild strains were exposed to ethidium bromide in the concentration range 1.5- 6.0 µg/mL. Generally, all the mutated strains showed an improved chitinase yield compared to the control. Highest yield was observed with the strain exposed to 6 µg/mL of ethidium bromide. The yield was 25.03 % higher compared to the wild strain. The mutated strain was slimy in nature. Protein content of the mutated strain decreased by 11%. Ethidium bromide at a concentration of 1.5 µg/mL was considered optional, at which the strain was stable with increase of 21.80 % in enzyme activity and 4.41% increase in protein content. Increased enzyme yield with decreased non-specific protein could be useful in producing cost effective enzyme.

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Proinflammatory Soluble Interleukin-15 Receptor Alpha Is Increased in Rheumatoid Arthritis

Arthritis ◽

10.1155/2012/943156 ◽

2012 ◽

Vol 2012 ◽

pp. 1-7 ◽

Cited By ~ 6

Author(s):

Ana Cecilia Machado Diaz ◽

Araceli Chico Capote ◽

Celia Aurora Arrieta Aguero ◽

Yunier Rodríguez Alvarez ◽

Diana García del Barco Herrera ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Synovial Fluid ◽

Enzyme Linked Immunosorbent Assay ◽

Reverse Signaling ◽

Synovial Fluids ◽

Inflammatory Processes ◽

Membrane Bound ◽

Interleukin 15 ◽

Interleukin 15 Receptor Alpha ◽

Pro Inflammatory Cytokines

Rheumatoid arthritis (RA) is an autoimmune and inflammatory disease in which many cytokines have been implicated. In particular, IL-15 is a cytokine involved in the inflammatory processes and bone loss. The aim of this study was to investigate the existence in synovial fluid of soluble IL-15Rα, a private receptor subunit for IL-15 which may act as an enhancer of IL-15-induced proinflammatory cytokines. Soluble IL-15Rα was quantified by a newly developed enzyme-linked immunosorbent assay (ELISA) in samples of synovial fluid from patients with RA and osteoarthritis (OA). The levels of IL-15Rα were significantly increased in RA patients compared to OA patients. Also, we studied the presence of membrane-bound IL-15 in cells from synovial fluids, another element necessary to induce pro-inflammatory cytokines through reverse signaling. Interestingly, we found high levels of IL-6 related to high levels of IL-15Rα in RA but not in OA. Thus, our results evidenced presence of IL-15Rα in synovial fluids and suggested that its pro-inflammatory effect could be related to induction of IL-6.

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EVQuant; high-throughput quantification and characterization of extracellular vesicle (sub)populations

10.1101/2020.10.21.348375 ◽

2020 ◽

Author(s):

T.A. Hartjes ◽

J.A. Slotman ◽

M.S. Vredenbregt ◽

N. Dits ◽

R. Van der Meel ◽

...

Keyword(s):

Protein Content ◽

Size Distribution ◽

High Throughput ◽

Liquid Biopsy ◽

Ideal Liquid ◽

Extracellular Vesicle ◽

Cell Of Origin ◽

Clinical Implementation ◽

High Throughput Assay

AbstractExtracellular vesicles (EVs) reflect the cell of origin in terms of nucleic acids and protein content. They are found in biofluids and represent an ideal liquid biopsy biomarker source for many diseases. Unfortunately, clinical implementation is limited by available technologies for EV analysis. We have developed a simple, robust and sensitive microscopy-based high-throughput assay (EVQuant) to overcome these limitations and allow widespread use in the EV community. The EVQuant assay can detect individual immobilized EVs as small as 35 nm and determine their concentration in biofluids without extensive EV isolation or purification procedures. It can also identify specific EV subpopulations based on combinations of biomarkers and is used here to identify prostate-derived urinary EVs as CD9-/CD63+. Moreover, characterization of individual EVs allows analysis of their size distribution. The ability to identify, quantify and characterize EV (sub-)populations in high-throughput substantially extents the applicability of the EVQuant assay over most current EV quantification assays.

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Peptide location fingerprinting reveals modification-associated biomarkers of ageing in human tissue proteomes

10.1101/2020.09.14.296020 ◽

2020 ◽

Author(s):

Matiss Ozols ◽

Alexander Eckersley ◽

Kieran T Mellody ◽

Venkatesh Mallikarjun ◽

Stacey Warwood ◽

...

Keyword(s):

Structural Modification ◽

Protein Structures ◽

Cytoskeletal Proteins ◽

Structural Modifications ◽

Location Fingerprinting ◽

Age Related ◽

Analysis Technique ◽

Key Factor ◽

Aged Skin ◽

Novel Biomarkers

AbstractAlthough dysfunctional protein homeostasis (proteostasis) is a key factor in many age-related diseases, the untargeted identification of structural modifications in proteins remains challenging. Peptide location fingerprinting is a proteomic analysis technique capable of identifying structural modification-associated differences in mass spectrometry (MS) datasets of complex biological samples. A new webtool (Manchester Peptide Location Fingerprinter), applied to photoaged and intrinsically aged skin proteomes, can relatively quantify peptides (spectral counting) and map statistically significant differences to regions within protein structures. New photoageing biomarkers were identified in multiple proteins including matrix components (collagens and proteoglycans), oxidation and protease modulators (peroxiredoxins and SERPINs) and cytoskeletal proteins (keratins). Crucially, for many extracellular biomarkers, structural modification-associated differences were not correlated with relative abundance (by ion intensity). By applying peptide location fingerprinting to published MS datasets, (identifying biomarkers including collagen V and versican in ageing tendon) we demonstrate the potential of the MPLF webtool to discover novel biomarkers.

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Complement Activation and Inflammatory Processes in Drusen Formation and Age Related Macular Degeneration

Experimental Eye Research ◽

10.1006/exer.2001.1094 ◽

2001 ◽

Vol 73 (6) ◽

pp. 887-896 ◽

Cited By ~ 344

Author(s):

Lincoln V Johnson ◽

William P Leitner ◽

Michelle K Staples ◽

Don H Anderson

Keyword(s):

Macular Degeneration ◽

Complement Activation ◽

Age Related Macular Degeneration ◽

Age Related ◽

Inflammatory Processes

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Advances in microglia cellular models: focus on extracellular vesicle production

Biochemical Society Transactions ◽

10.1042/bst20210203 ◽

2021 ◽

Author(s):

Lorenzo Ceccarelli ◽

Laura Marchetti ◽

Chiara Giacomelli ◽

Claudia Martini

Keyword(s):

Cell Communication ◽

Extracellular Vesicle ◽

Cellular Models ◽

Advantages And Disadvantages ◽

Inflammatory Processes ◽

The Central Nervous System ◽

Harmful Effects ◽

Brain Homeostasis

Microglia are the major component of the innate immune system in the central nervous system. They promote the maintenance of brain homeostasis as well as support inflammatory processes that are often related to pathological conditions such as neurodegenerative diseases. Depending on the stimulus received, microglia cells dynamically change their phenotype releasing specific soluble factors and largely modify the cargo of their secreted extracellular vesicles (EVs). Despite the mechanisms at the basis of microglia actions have not been completely clarified, the recognized functions exerted by their EVs in patho-physiological conditions represent the proof of the crucial role of these organelles in tuning cell-to-cell communication, promoting either protective or harmful effects. Consistently, in vitro cell models to better elucidate microglia EV production and mechanisms of their release have been increased in the last years. In this review, the main microglial cellular models that have been developed and validated will be described and discussed, with particular focus on those used to produce and derive EVs. The advantages and disadvantages of their use will be evidenced too. Finally, given the wide interest in applying EVs in diagnosis and therapy too, the heterogeneity of available models for producing microglia EVs is here underlined, to prompt a cross-check or comparison among them.

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