Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: cf-mtDNA content and deletion ratio in cross-sectional study cohort.

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques:

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: cf-mtDNA content and deletion ratio in longitudinal study cohort.

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques:

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: cf-mtDNA deletions in CSF and CD9-EVs immunocaptured from CSF in prodromal Lewy body disorders . mt64-ND1 quantifies the total number of cf-mtDNA molecules. mt96-ND5 quantifies only mtDNA molecules without major arc mtDNA deletions . a) cf-mtDNA deletions in CSF. b) cf-mtDNA deletions in CD9-EVs from CSF. c) Positive relationship between cf-mtDNA deletions and duration of IRBD (n = 51). The dotted lines represent 95% CI. Ctrl, control participants. IRBD, patients with idiopathic REM sleep behaviour disorder who did not convert (NC) or converted (C) to a Lewy body disorder at the time of the study. Values are mean with 95% CI. Numbers within squares indicate the number of participants per group. a) (NC-vs Ctrl) p = 0.054, (C vs Ctrl) p = 0.002. b) (NC vs Ctrl) p = 0.006, (C vs Ctrl) p < 0.0001, (NC vs C) p = 0.011. (ANOVA with Fisher's LSD test).

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques: Control

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: cf-mtDNA copy number in CSF and CD9-EVs immunocaptured from CSF in prodromal Lewy body disorders . Quantification of cf-mtDNA copies by multiplex dPCR assay using two different primer pairs (mt64-ND1 and mt96-ND5). a) cf-mtDNA copy number in CSF. b) cf-mtDNA copy number in CD9-EVs from CSF. Ctrl, control participants. IRBD, patients with idiopathic REM sleep behaviour disorder who did not convert (NC) or converted (C) to a Lewy body disorder at the time of the study. NTC, non-template control. Values are mean with 95% CI. Numbers within squares indicate the number of participants per group. b) mt64-ND1 copies/μl (NC vs Ctrl) p = 0.005, (C vs Ctrl) p = 0.001; mt96-ND5 copies/μl (NC vs Ctrl) p = 0.001, (C vs Ctrl) p < 0.0001. (ANOVA with Fisher's LSD test).

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques: Multiplex Assay, Control

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: Longitudinal study of cf-mtDNA deletions in CSF and CD9-EVs immunocaptured from the CSF of patients with IRBD before and after conversion to a Lewy body disorder . mt64-ND1 quantifies cf-mtDNA molecules with and without deletions. mt96-ND5 quantifies molecules without major arc mtDNA deletions. a) cf-mtDNA deletions in CSF. b) cf-mtDNA deletions in CD9-EVs from CSF. Ctrl, control participants. Pre-C, patients with idiopathic REM sleep behaviour disorder (IRBD) before conversion. Post-C, the same patients after conversion to a Lewy body disorder. Dotted lines indicate measurements performed in samples obtained from the same patient at IRBD diagnosis (Pre-C) and after conversion to a Lewy body disorder (Post-C). Values are mean with 95% CI. Numbers within squares indicate the number of participants per group. a) (Pre-C vs. Ctrl) p = 0.043, (Post-C vs. Ctrl) p = 0.035. b) (Pre-C vs. Ctrl) p = 0.003, (Post-C vs. Ctrl) p = 0.001. (ANOVA with Brown-Forsythe (a) and Fisher's LSD test (b)).

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques: Control, Biomarker Discovery

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: Longitudinal study of cf-mtDNA copy number in CSF and CD9-EVs immunocaptured from the CSF of patients with IRBD before and after conversion to a Lewy body disorder . cf-mtDNA copies were quantified with two different primer pairs (mt64-ND1 and mt96-ND5) directly in CSF (a) and in CD9-EVs from CSF (b) . Ctrl, control participants. Pre-C, patients with idiopathic REM sleep behaviour disorder (IRBD) before conversion. Post-C, the same patients after conversion to a Lewy body disorder. Dotted lines indicate measurements performed in samples obtained from the same patient at IRBD diagnosis (Pre-C) and after conversion to a Lewy body disorder (Post-C). Values are mean with 95% CI. Numbers within squares indicate the number of participants per group. b) mt64-ND1 copies/μl (Pre-C vs. Ctrl) p = 0.043 (Post-C vs. Ctrl) p = 0.047; mt96-ND5 copies/μl (Pre-C vs. Ctrl) p = 0.012 (Post-C vs. Ctrl) p = 0.012. (ANOVA with Fisher's LSD test).

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques: Control, Biomarker Discovery

Journal: eBioMedicine

Article Title: Mitochondrial DNA deletions in the cerebrospinal fluid of patients with idiopathic REM sleep behaviour disorder

doi: 10.1016/j.ebiom.2024.105065

Figure Lengend Snippet: cf-mtDNA copy number in serum and CD9-EVs immunocaptured from serum in prodromal Lewy body disorders . Quantification of circulating cf-mtDNA copies by multiplex dPCR with two different primer pairs (mt64-ND1 and mt96-ND5) directly in serum (a) and in CD9-EVs from serum (b) . Ctrl, control participants. IRBD, patients with idiopathic REM sleep behaviour disorder who did not convert (NC) or converted (C) to a Lewy body disorder at the time of the study. NTC, non-template control. Values are mean with 95% CI. Numbers within squares indicate the number of participants per group. a) mt64-ND1 copies/μl (C vs Ctrl) p = 0.003; mt96-ND5 copies/μl (C vs Ctrl) p = 0.007. b) mt64-ND1 copies/μl (C vs Ctrl) p = 0.036; mt96-ND5 copies/μl (C vs Ctrl) p = 0.034. (ANOVA with Fisher's LSD test).

Article Snippet: CD9-EVs were immunocaptured from CSF and serum samples using plates which contained covalently attached antibodies against the tetraspanin CD9 (HBM-POS-CC/T1, Hansa-BioMed).

Techniques: Multiplex Assay, Control

Journal: bioRxiv

Article Title: Salivary exosomal miR-1307-5p predicts disease aggressiveness and poor prognosis in oral squamous cell carcinoma patients

doi: 10.1101/2022.07.13.499918

Figure Lengend Snippet: (a) Representative TEM images of exosomes from OSCC patients, (Scale: 200nm); (b) Estimation of size and concentration of salivary exosomes from OSCC patients using Nanoparticle Tracking Analysis (NTA). The x-axis indicates the size distribution of particles and the y-axis shows the signal intensity in NTA; (c-e) Characterization of exosomes by flow cytometry. We used a membrane-binding dye vFRed to stain exosomes and examined the violet side scatter v/s vFRed staining of these exosomes. This profile was similar to the profile of Lipo-50 liposomes used as a reference standard. Salivary exosomes were stained with antibodies against tetraspanins (CD63, CD81 and CD9) labelled with PE, and APC labelled CD47 antibody. We identified a distinct CD47+/vFRed+ population in the exosome population that was absent in the unstained control. This population was then examined for tetraspanin expression and showed detectable levels of tetraspanins. Since these particles stain with vFRed, have a violet side scatter similar to 50 nm liposomes, express CD47 and tetraspanins, we concluded that these are exosomes.

Article Snippet: The samples (at optimal dilution) were then incubated with PE tagged antibodies against tetraspanins (TS; CD9, CD63, and CD81) (Cellarcus Biosciences, CA, USA) and APC tagged CD47 antibody (Thermo Scientific, Waltham, MA, USA) for one hour at room temperature.

Techniques: Concentration Assay, Flow Cytometry, Membrane, Binding Assay, Staining, Liposomes, Control, Expressing

Journal: Scientific Reports

Article Title: Digital Detection of Exosomes by Interferometric Imaging

doi: 10.1038/srep37246

Figure Lengend Snippet: The expression of the typical exosomal biomarkers CD81 CD63 as well as the neural adhesion protein CD171 is significantly different than in the artificial CSF, negative control. An additional negative control, a non correlated IgG, shows a low level of non-specific binding. Each multiplexed chip was run in duplicate independent replicate tests.

Article Snippet: Monoclonal antibodies against the human tetraspanins CD63, CD81, CD9, and CD171 were purchased from Nexgenarrays LLC (Boston, MA).

Techniques: Expressing, Negative Control, Binding Assay