A5087018350- Extracellular vesicles in disease
- Renal Diseases and Glomerulopathies
- Chronic Kidney Disease and Diabetes
- Cardiovascular Disease and Adiposity
- Advanced Proteomics Techniques and Applications
- Adipokines, Inflammation, and Metabolic Diseases
- Mass Spectrometry Techniques and Applications
- Inflammatory Biomarkers in Disease Prognosis
- Clusterin in disease pathology
- Complement system in diseases
- MicroRNA in disease regulation
- Galectins and Cancer Biology
- Platelet Disorders and Treatments
- Glycosylation and Glycoproteins Research
- Dialysis and Renal Disease Management
- Protein Interaction Studies and Fluorescence Analysis
- Cell Adhesion Molecules Research
- nanoparticles nucleation surface interactions
- Biomedical Research and Pathophysiology
- Metabolomics and Mass Spectrometry Studies
- Ion Transport and Channel Regulation
- Atherosclerosis and Cardiovascular Diseases
- Cellular transport and secretion
- Coagulation, Bradykinin, Polyphosphates, and Angioedema
- Retinal Development and Disorders
University of Pennsylvania
2022-2025
University of Virginia
2018-2023
Center for Neuroscience and Regenerative Medicine
2023
Pennsylvania State University
2022
University of Virginia Health System
2017-2021
Charlottesville Medical Research
2018-2021
Dublin City University
2012-2019
Museu da Amazonia
2019
Istituto Giannina Gaslini
1999-2017
University of Helsinki
2014
Abstract A modified Neuhoff's colloidal Coomassie Blue G‐250 stain is reported, dubbed “blue silver” on account of its considerably higher sensitivity, approaching the one conventional silver staining. The main modifications, as compared to protocol, were: a 20% increment in dye concentration (from 0.1% up 0.12%) and much level phosphoric acid recipe 2% 10%). exhibits faster uptake (80% during first hour coloration, vs. none with commercial preparation from Sigma). Even at equilibrium (24 h...
Abstract Urine is commonly used for clinical diagnosis and biomedical research. The discovery of extracellular vesicles (EV) in urine opened a new fast‐growing scientific field. In the last decade urinary (uEVs) were shown to mirror molecular processes as well physiological pathological conditions kidney, urothelial prostate tissue. Therefore, several methods isolate characterize uEVs have been developed. However, methodological aspects EV separation analysis, including normalization...
The mechanisms underpinning the regenerative capabilities of mesenchymal stem cells (MSC) were originally thought to reside in their ability recognise damaged tissue and differentiate into specific cell types that would replace defective cells. However, recent work has shown molecules produced by MSCs (secretome), particularly those packaged extracellular vesicles (EVs), rather than themselves are responsible for repair.Here we have a secretome from adipose-derived (ADSC) is free exogenous...
Adaptive immune resistance in cancer describes the various mechanisms by which tumors adapt to evade anti-tumor responses. IFN-γ induction of programmed death-ligand 1 (PD-L1) was first defined and validated adaptive mechanism. The endoplasmic reticulum (ER) is central as modulatory secreted integral membrane proteins are dependent on ER. Sigma1 a unique ligand-regulated scaffolding protein enriched ER cells. PD-L1 an glycoprotein that translated into processed through cellular secretory...
Urinary extracellular vesicles provide a novel source for valuable biomarkers kidney and urogenital diseases: Current isolation protocols include laborious, sequential centrifugation steps which hampers their widespread research clinical use. Furthermore, large individual urine sample volumes or sizable target cohorts are to be processed (e.g. biobanking), the storage capacity is an additional problem. Thus, alternative methods necessary overcome such limitations. We have developed practical...
ABSTRACT. Podocin mutations (NPHS2 gene) are responsible for the autosomal recessive form of steroid-resistant nephrotic syndrome. As a result screening these gene alterations in cohort Italian patients with nonfamilial syndrome and histologic focal segmental glomerulosclerosis (FSGS), nine NPHS2 homozygous or composite heterozygous were found. In addition to previously described defects, two novel at exon 4 identified (frameshift, L169P); four single nucleotide polymorphisms (SNPs) one...
Even if nephrotic syndrome is characterized by massive urinary loss of major plasma proteins, a clear structural characterization based on proteomics has never been reported. Urine and 23 patients with different idiopathic syndromes (10 steroid-sensitive minimal-change nephropathy, seven steroid-resistant FSGS, six membranous glomerulonephritis) were analyzed two-dimensional electrophoresis in soft gel, Western blot, matrix-assisted laser desorption/ionization time flight mass spectrometry;...
Urinary exosomes represent a precious source of potential biomarkers for disease biology. Currently, the methods vesicle isolation are severely restricted by tendency entrapment, e.g. abundant Tamm-Horsfall protein (THP) polymers. Treatment reducing agents such as dithiothreitol (DTT) releases entrapped vesicles, thus increasing final yield. However, this harsh treatment can cause remodelling all those proteins which feature extra-vesicular domains stabilized internal disulfide bridges and...
Epithelial cells lining the urinary tract secrete exosomes (40–100 nm) that can be targeted to specific modulating their functionality. One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. This makes glycopeptide analysis of important. Exosomes reflect physiological state parent cells; therefore, they are a good source biomarkers for urological other diseases. Moreover, urine collection easy noninvasive give information about renal systemic...
This report summarises the presentations and activities of ISEV Workshop on extracellular vesicle biomarkers held in Birmingham, UK during December 2017. Among key messages was broad agreement about importance biospecimen science. Much greater attention needs to be paid towards provenance collected samples. The workshop also highlighted clear gaps our knowledge pre-analytical factors that alter vesicles (EVs). future utility certified standards for credentialing instruments software, analyse...
Abstract Urinary extracellular vesicles (uEVs) provide bio-markers for kidney and urogenital diseases. Centrifugation is the most common method used to enrich uEVs. However, a majority of studies date have focused on ultracentrifugation pellet, potentially losing novel source important biomarkers that could be obtained at lower centrifugation. Thus, aim this study rigorously characterize first time uEVs in low speed pellet determine minimal volume urine required proteomic analysis (≥9.0 mL...
The basic mechanism for idiopathic FSGS still is obscure. Indirect evidence in humans and generation of by oxidants experimental models suggest a role free radicals. In vitro studies demonstrate main plasma albumin as antioxidant, its modification representing chemical marker oxidative stress. With the use complementary liquid chromatography electron spray ionization tandem mass spectrometry (LC-ESI-MS/MS) biochemical methods, was characterized 34 patients with FSGS; 18 had received renal...
The secretome of human amniotic fluid stem cells (AFSCs) has great potential as a therapeutic agent in regenerative medicine. However, it must be produced clinically compliant manner before can used humans. In this study, we developed means producing biologically active from AFSCs that is free all exogenous molecules. We demonstrate the full capable promoting cell proliferation, migration, and protection against senescence. Furthermore, significant anti-inflammatory properties. Most...
ABSTRACT. A plasma factor displaying permeability activity in vitro and possibly determining proteinuria has been hypothesized idiopathic focal segmental glomerulosclerosis (FSGS). In (Palb) was determined sera of five patients with autosomal recessive steroid-resistant nephrotic syndrome (NPHS2), an inherited condition indistinguishable from FSGS on clinical grounds, but which is by homozygous mutations podocin, a key component the glomerular podocyte. All had presented intractable...
Urinary vesicles represent a newly established source of biological material, widely considered to faithfully pathological events in the kidneys and urogenital epithelium. The majority currently applied isolation protocols involve cumbersome centrifugation steps enrich from urine. To date, efficiency these approaches has not been investigated with respect performing quantitative qualitative analyses vesicle populations pellet supernatant (SN) fractions. After series differential...
Background Urinary extracellular vesicles (UEVs) represent an ideal platform for biomarker discovery. They carry different types of RNA species, and reported profile discrepancies related to the presence/absence 18s 28s rRNA remain controversial. Moreover, sufficient urinary yields respective quality profiles are still be fully established. Methods UEVs were enriched by hydrostatic filtration dialysis, content was extracted using 7 commercially available techniques. quantity assessed...
Extracellular vesicles (EVs) have been described as novel biomarkers and bioactivators in vascular dysfunction hypertension. However, the mechanism(s) by which EVs affect function is unknown. To examine effects of on endothelial-dependent vasodilation (acetylcholine), we isolated circulating from platelet-poor plasma using a low centrifugation speed (17 000 g ) mesenteric resistance arteries 12-week-old normotensive WKYs (Wistar-Kyoto rats) SHRs (spontaneously hypertensive rats). Arteries...
Abstract. Glomerular albumin permeability alterations can be induced in vitro by serum from patients with end-stage renal disease caused primary focal segmental glomerulosclerosis (FSGS). It was hypothesized that inhibitory substances may present normal serum, which prevent the isolated glomeruli, and study sought to isolate characterize these factors. Albumin determined change glomerular volume applying oncotic gradients across basement membrane of healthy rat glomeruli preincubated FSGS...
The initial events of vision at low light take place in vertebrate retinal rods. rod outer segment consists a stack flattened disks surrounded by the plasma membrane. A list proteins that reside has not been achieved yet. We present first comprehensive proteomic analysis purified disks, obtained combining results two-dimensional gel electrophoresis separation disk to MALDI-TOF or nLC-ESI-MS/MS mass spectrometry techniques. Intact were isolated from bovine segments method minimizes...
The airway surface is covered by a fluid, the liquid, interposed between mucous layer and epithelium. liquid contains proteins, secreted different cell types, that may have pro-/anti-inflammatory or bactericidal functions role in mucociliary clearance. We used proteomics approach to identify proteins an isolated vitro model of human epithelium, at resting under proinflammatory conditions, as strategy define factors involved epithelial barrier function. To this aim, we analyzed from bronchial...
Purpose Urinary extracellular vesicles (UEVs) are a novel source for disease biomarker discovery. However, Tamm–Horsfall protein (THP) is still challenge proteomic analysis since it can inhibit detection of low‐abundance proteins. Here, we introduce new approach that does not involve an ultracentrifugation step to enrich and reduces the amount THP manageable levels. Experimental design UEVs were dialyzed ultrafiltered after reduction alkylation. The retained fraction was digested with...
Urinary extracellular vesicles (UEVs) appear an ideal source of biomarkers for kidney and urogenital diseases. The majority protocols designed their isolation are based on differential centrifugation steps. However, little is still known the type amount left in supernatant. Here we used protocol UEVs which uses hydrostatic filtration dialysis as first pre-enrichment step, followed by centrifugation. Transmission electron microscopy (TEM), mass spectrometry (MS), western blot, ELISA assays...
CD40/CD40 ligand (CD40L) dyad, a co-stimulatory bi-molecular complex involved in the adaptive immune response, has also potent pro-inflammatory actions haematopoietic and non-haematopoietic cells. We describe here novel role for soluble CD40L (sCD40L) as modifier of glomerular permselectivity directly acting on epithelial cells (GECs). found that stimulation CD40, constitutively expressed GEC cell membrane, by sCD40L rapidly induced redistribution loss nephrin GECs, increased albumin...