Spinal Cord Injury Affects Gene Expression of Transmembrane Proteins in Tissue and Release of Extracellular Vesicle in Blood: In Silico and In Vivo Analysis

Objective: Spinal cord injury (SCI) can disrupt membrane transmission by affecting transmembrane channels orneurotransmitter release. This study aimed to explore gene expression changes of transmembrane proteins underlyingSCI through bioinformatics approaches and confirming in SCI model in rats.Materials and Methods: In this experimental study, the differentially expressed genes (DEGs) in acute and subacuteSCI were obtained based on microarray data downloaded from the gene expression omnibus (GEO). Transmembraneproteins of DEGs were recognized by using the UniProt annotation and transmembrane helices prediction (TMHMM)methods. The model of SCI was established through a weight-dropping procedure in rats. To confirm the SCI model,hematoxylin and eosin (H&E) staining was performed. Total mRNA was extracted from spinal cord tissues, and the RNAexpression profile of some of the significantly changed genes in the previous part that has been confirmed by real-timepolymerase chain reaction (PCR). Blood was collected from rats before sacrificing. Extracellular vesicles (EVs) wereisolated by high-speed centrifugation from plasma. For the assessment of protein expression, western blotting wasused.Results: Based on bioinformatics analysis, we candidated a set of membrane proteins in SCI’s acute and sub-acutephases, and confirmed significant upregulation in Grm1, Nrg1, CD63, Enpp3, and Cxcr4 between the acute and controlgroups and downregulation in Enpp3 between acute and subacute groups at the RNA level. Considering CD63 as anEV marker, we examined the protein expression of CD9 and CD63 in the plasma-derived EVs, and CD9 has significantexpression between acute and control groups. We also demonstrate no significant CD63 and Cxcr4 expressionsbetween groups.Conclusion: Our results provide new insight into the relationship between candidate transmembrane protein expressionand different stages of SCI using in-silico approaches. Also, results show the release of EVs in blood in each group afterSCI helping enlarge strategies to enhance recovery following SCI

Healthy Male Individuals Possess Higher Plasma HER-2 Level than Females

Considering HER2 as one of the well-known biomarkers in the cancer field, and published articles regarding serumlevels of HER2, in this paper we tried to highlight the issue that most studies don’t stratify the HER-2 concentrationof individuals in terms of gender. In this brief survey, healthy individuals with no prior non-communicable diseaseswere categorized as males (n=34) and females (n=43), and all samples were evaluated for plasma HER-2 levelsat once. Surprisingly, the plasma level of HER-2 of healthy male individuals (mean= 2.28 ± 0.21 ng/mL) wassignificantly (P<0.0001) higher than the plasma level of HER-2 of healthy females (mean: 0.06 ± 0.09 ng/mL),with no overlap. Therefore, we suggest that more studies are required to re-check the cutoff values for HER-2plasma levels based on gender since the clinical implications of a unique HER-2 cutoff for both genders may beseriously concerning

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

© 2024 The Authors. Journal of Extracellular Vesicles, published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.Peer reviewe

Minimal information for studies of extracellular vesicles 2018 (MISEV2018):a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points

Isolation and Characterization of Human Blood Extracellular Vesicles: a Promising Method of Fluid Biopsy

Aims: Extracellular vesicles (EV) are nanoscale vesicles that were previously thought to be secreted into the bloodstream by apoptotic cells. Today, EVs have been demonstrated to be secreted by almost all cells in the body, which contain valuable biomarkers for early diagnosis of a wide variety of diseases, particularly cancers. Efficient separation of EVs encounter challenges due to the presence of proteins and lipoproteins. Hence, the need for a time-saving and non-invasive diagnostic method with the ability to quantify isolated EVs, which is also applicable to resource-limited situations, can be the major necessities to advance EV-based diagnostic studies. The aim of the present study was to provide a relatively rapid and efficient method for isolating EV from blood as an important biological fluid in humans. Materials & Methods: In this case study, with the combination of size exclusion chromatography and ultracentrifuge, EVs were successfully isolated from human blood plasma samples within an hour. To confirm the isolation process according to MISEV guidelines, three tetraspanin membrane proteins CD63, CD81, CD9, and one luminal protein TG101, were evaluated as positive protein markers and connexin protein as a negative marker by Western blot. Findings: The maximum presence of positive markers and the absence of negative markers in F7 to F9 fractions were confirmed. Additionally, their size distribution evaluation was done using the DLS technique with an average diameter in the range of 100 nm. Quantification of the protein content of EVs was also performed by BCA assay. Conclusion: The present study shows that the combination of high-speed centrifugation and size-based chromatography methods is very effective in isolating EV from the blood plasma of individual

Data for whole and mitochondrial proteome of human embryonic stem cells

The data presented here pertain to the research article entitled “Proteome Analysis of Human Embryonic Stem Cell Organelles” (Shekariet al., 2017 [1]). In the present article we endeavour to locate new proteins and pathways in human embryonic stem cells (hESCs) by mass spectrometry and bioinformatics analysis. We have analyzed total and mitochondrial proteins extracted from three biological replicates of the hESC H9 cell line according to mass spectrometry proteomics and bioinformatics investigations

Editorial: Mesenchymal stem cell-derived extracellular vesicles: considerations and therapeutic applications

Editorial on the research topic: Mesenchymal stem cell-derived extracellular vesicles: considerations and therapeutic applications.</p

Correction: Karimi, P., et al. Crocetin Prevents RPE Cells from Oxidative Stress through Protection of Cellular Metabolic Function and Activation of ERK1/2. Int. J. Mol. Sci. 2020, 21, 2949

The authors wish to make the following correction to Figure 6B of this article [...

Earlier Detection of Alzheimer’s Disease Based on a Novel Biomarker <em>cis</em> P-tau by a Label-Free Electrochemical Immunosensor

Early detection of cis phosphorylated tau (cis P-tau) may help as an effective treatment to control the progression of Alzheimer’s disease (AD). Recently, we introduced for the first time a monoclonal antibody (mAb) with high affinity against cis P-tau. In this study, the cis P-tau mAb was utilized to develop a label-free immunosensor. The antibody was immobilized onto a gold electrode and the electrochemical responses to the analyte were acquired by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), and differential pulse voltammetry (DPV). The immunosensor was capable of selective detection of cis P-tau among non-specific targets like trans P-tau and major plasma proteins. A wide concentration range (10 × 10−14 M–3.0 × 10−9 M) of cis P-tau was measured in PBS and human serum matrices with a limit of detection of 0.02 and 0.05 pM, respectively. Clinical applicability of the immunosensor was suggested by its long-term storage stability and successful detection of cis P-tau in real samples of cerebrospinal fluid (CSF) and blood serum collected from human patients at different stages of AD. These results suggest that this simple immunosensor may find great application in clinical settings for early detection of AD which is an unmet urgent need in today’s healthcare services