Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Outline of study time-course and sample processing for EV isolation. Antemortem plasma samples with postmortem pathological confirmation of neurological diagnoses were processed to isolate cell-specific EVs using our mTENPO microfluidic platform, alongside plasma protein biomarkers using commercial digital ELISA, for patients with LBD ( n = 30), AD ( n = 31), AD/LBD ( n = 30), AD/ALB ( n = 19), and controls ( n = 27). B The mTENPO platform, illustrating the external magnet, inlet reservoir, outlet ports, and tubing connections to syringe pumps. Syringes are connected to the waste outlet for blocking, washing, and sample addition steps, and then replaced and switched to the lysate outlet before captured EVs are lysed on-chip. The inset shows a photo of the mTENPO chip with a quarter for scale. C Schematic of operation of the mTENPO platform for cell-specific EV isolation using antibody-labeled magnetic nanoparticles (MNPs) for GluR2+ (top) and GLAST+ (bottom) EV pulldowns. D Scanning electron microscopy (SEM) images of GluR2+ EVs immobilized on the edges of pores of the mTENPO device’s surface. E SEM images of GLAST+ EVs immobilized on the edges of pores of the mTENPO device’s surface. F Representative cropped western blot images showing protein expression of GluR2, GLAST, and EV-associated marker TSG101 using mTENPO-isolated GluR2+ or GLAST + EV lysates from n = 2 human plasma samples. Full-length western blot images are shown in Supplementary Fig. .

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Isolation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Blocking Assay, Labeling, Electron Microscopy, Western Blot, Expressing, Marker

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for biomarkers with Benjamini-Hochberg FDR-corrected P value < 0.1. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending fold-change. B Volcano plot demonstrating differential expression of GluR2+ EV miRNAs, GLAST + EV miRNAs, and plasma proteins. C Venn diagram showing overlap in FDR P value significant miRNAs ( P value < 0.1) between GluR2+ EVs and GLAST+ EVs. D Top 30 biomarkers in all compartments ranked by descending area under the curve (AUC). Error bars represent standard error from bootstrapping 10x.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Quantitative Proteomics, Clinical Proteomics

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: GO and KEGG pathway analyses were performed on differentially expressed miRNAs using DIANA miRPath v4.0 using the TarBase v8.0 database. FDR P values for identified GO terms and KEGG pathways were calculated using a one-sided Fisher’s exact test and considered significant at P value < 0.05. The top 10 (ranked by number of target genes) terms within each of the three GO categories (BP, CC, MF) and top 10 (ranked by number of target genes) KEGG pathways were identified for each pulldown. A Top 10 terms within each GO category for GluR2+ EV miRNAs. B Top 10 KEGG pathways for GluR2+ EV miRNAs. C Top 10 terms within each GO category for GLAST + EV miRNAs. D Top 10 KEGG pathways for GLAST + EV miRNAs. In all panels, each bar is labeled to the right with the number of differentially expressed miRNAs associated with the given GO term or KEGG pathway.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Labeling

Journal: Npj Biosensing

Article Title: Microfluidic nanomagnetically isolated neuron- and astrocyte-derived extracellular vesicles to differentiate Lewy body and Alzheimer’s disease

doi: 10.1038/s44328-026-00086-x

Figure Lengend Snippet: A Heatmap of z-score of log 2 (expression) for LASSO-selected biomarkers. Subjects (columns) are hierarchically clustered within cohort and biomarkers within each compartment (rows) are sorted by descending AUC. B Kendall correlation staircase plots identifying the extent to which biomarker information was correlated between the LASSO-selected GluR2+ EV, GLAST + EV, and protein biomarkers. Biomarkers are sorted within compartments by AUC. The inset shows the correlation distribution of Kendall’s τ, where the dotted line represents the median count. C LASSO panel accuracy versus panel size for classifying LBD versus AD, shown in blue; accuracy is assessed through tenfold cross-validation, with error bars representing standard error from 5 repeats of panel training on the LBD vs AD patient groups. Average accuracy and standard error for control experiments performed by scrambling patient labels 10x are shown in orange. D LASSO panel AUC versus panel size for classifying LBD versus AD, shown in blue with error bars as described in ( C ). Average AUC and standard error for the same control experiments described in ( C ) are shown in orange. E AUCs for the 15-marker LASSO panel and individual LASSO biomarkers, sorted by descending AUC. Error bars represent 95% confidence intervals, calculated from 5x repeats of tenfold cross-validation for the 15-marker panel or from bootstrapping 10x for individual markers.

Article Snippet: Briefly, 500 μL of patient plasma was incubated for 20 min at a concentration of 1 μg/mL with either biotinylated GluR2 capture antibody (GluR1 + GluR2 polyclonal antibody, Bioss bs-10042R-Biotin) for neuron-derived EVs per our previous work or biotinylated GLAST capture antibody [GLAST (ACSA-1) antibody, anti-human/mouse/rat Biotin, Miltenyi Biotec, 130-118-984] for astrocyte-derived EVs, where the use of GLAST as a protein target for astrocyte EV isolation has been previously reported , – , .

Techniques: Expressing, Biomarker Discovery, Control, Marker

Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA can inhibit the activation of microglia in the pPD stage mediated by TLR2. ( A ) Western blot detection of TLR2 protein expression in the substantia nigra. ( B ) Western blot detection of MyD88 protein expression in the substantia nigra. ( C ) Western blot detection of p-NF-κB-p65 expression in the substantia nigra. ( D ) Western blot detection of NLRP3 protein expression in the substantia nigra. ( E ) Western blot detection of Caspase-1 protein expression in the substantia nigra. ( F ) Western blot detection of GSDMD protein expression in the substantia nigra. ( G ) Western blot detection of IL-1β protein expression in the substantia nigra. (n=6; compared with the control group, *** P <0.001; compared with the model group, ### P <0.001, ## P <0.01, # P <0.05).

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Activation Assay, Western Blot, Expressing, Control

Journal: Journal of Inflammation Research

Article Title: Electroacupuncture Inhibits the Early Neuroinflammatory Cascade Triggered by TLR2 in the Prodromal Period of PD

doi: 10.2147/JIR.S585729

Figure Lengend Snippet: EA intervention affects the expression of TLR2 in different regions of pPD, inhibits microglial activation, regulates the TLR2/NF-κB/NLRP3 pathway, and suppresses pyroptosis of microglia.

Article Snippet: The membrane was incubated overnight at 4°C with primary antibodies specific to TLR2 (#bs-1019R, Bioss, 1:1000), MyD88 (#bs-1047R, Bioss, 1:1000), p-NF-κB-p65 (#bs-7343R, Bioss, 1:1000), NLRP3 (#bs-41293R, Bioss, 1:1000), caspase-1 (#bsm-52441R, Bioss, 1:1000), IL-1β (#bs-0812R, Bioss, 1:1000), and GSDMD (#bs-14287R, Bioss, 1:1000), followed by incubation with an IgG secondary antibody (#bs-10900R, Bioss, 1:5000) for 1 hour in the dark.

Techniques: Expressing, Activation Assay

Journal: Nucleic Acids Research

Article Title: Functional transitions of the Aspergillus fumigatus iron regulator HapX are governed by conserved domains cooperatively binding [2Fe-2S] clusters

doi: 10.1093/nar/gkaf796

Figure Lengend Snippet: In liquid media, growth inhibition by combinatorial mutation of at least CRR-B and -C is more severe in +Fe compared to −Fe and accelerates with increased expression of the corresponding hapX alleles. For each strain, ( A ) pxylP:hapX , ( B ) pxylP:hapX A/B/C/D , ( C ) pxylP:hapX B/C , and ( D ) pxylP:hapX A/B/C/D,464 , 10 4 conidia were inoculated in triplicate into 96-well microplates in 0.1 mL minimal medium reflecting iron limitation (−Fe; in green) and iron sufficiency (+Fe; 10 μM FeSO 4 ; in red) supplemented with 0.1% xylose for moderate and 1% xylose for high expression of the respective hapX allele. Using the oCelloScope (BioSense Solutions, Denmark) and its UniExplorer software (version 14.0), growth was scored hourly at 37 °C from 2 h post-inoculation (graphs start at the 5 h time point as there was very limited growth before that) until 24 h incubation. In the software, the “ fungi filamentous growth module ” was used to assess the hyphal area up to 2.75 × 10 5 μm 2 when the growth measurement reached saturation. The values in the growth curves represent the mean ± standard deviation of biological triplicates. Below the growth curves are examples of microscopic images of the fungal cultures taken automatically by the oCelloScope at two time points, 12 h and 24 h, underlining the scored growth data.

Article Snippet: Therefore, we applied automated microbial live cell imaging and analysis using the oCelloScope (BioSense Solutions ApS, Denmark) measuring growth by determining hyphal area (Fig. ).

Techniques: Inhibition, Mutagenesis, Expressing, Software, Incubation, Standard Deviation