Effects of deferoxamine on blood-brain barrier disruption after subarachnoid hemorrhage - Fig 3

<p><b>(A</b>) Occludin immunoreactivity and protein levels in cortex after sham or subarachnoid hemorrhage induction with deferoxamine (DFX) treatment or vehicle at day 3, scale bar = 20μm. Values are mean ± SD; n = 3 for each group, #p<0.01, *p<0.05 vs. SAH+vehicle group at day 3. <b>(B)</b> ZO-1 immunoreactivity and protein levels in cortex after sham or subarachnoid hemorrhage induction with deferoxamine (DFX) treatment or vehicle at day 3, scale bar = 20μm. Values are mean ± SD; n = 3 for each group, #p<0.01 vs. SAH+vehicle group at day 3. <b>(C)</b> Claudin-5 immunoreactivity and protein levels in cortex after sham or subarachnoid hemorrhage induction with deferoxamine (DFX) treatment or vehicle at day 3, scale bar = 20μm. Values are mean ± SD; n = 3 for each group, *p<0.05 vs. SAH+vehicle group at day 3.</p

Behavior and activity scores.

<p>Behavior and activity scores.</p

Ferritin immunoreactivity, and the ferritin heavy chain (FTH) and light chain (FTL) protein levels in cortex at day 3 after sham or subarachnoid hemorrhage induction with deferoxamine (DFX) treatment or vehicle, scale bar = 20μm.

<p>Values are mean ± SD; n = 3 for each group, #p<0.01 vs. SAH+vehicle group at day 3.</p

Effect of 15-PGJ₂ treatment on autophagic-related protein expression in ischemic cortex after cerebral I/R injury.

<p>The expression of LC3-II, Beclin 1, cathepisin-B, and LAMP1 expression significantly increased at 12 h after reperfusion. Treatment with 15-PGJ₂ at 1 to 50 pg significantly decreased LC3-II, Beclin 1, cathepisin-B, and LAMP1 expression after I/R injury. Optical density of respective protein bands were analyzed with Sigma Scan Pro 5 and normalized to the loading control (β-actin). ^#p<0.05 versus I/R group.</p

Neuroprotective effects of 15-PGJ₂ against cerebral I/R injury.

<p>15-PGJ₂ (1, 10, 50 pg) was administered icv immediately before reperfusion. (A) Five consecutive TTC-stained coronal brain slices arranged in cranial to caudal order 24 h after I/R. The white brain area represents infracted tissue. Infarct volume (B), neurological deficits (C) was measured 24 h after I/R. ^#p<0.05 versus I/R group.</p

Electron micrographs of morphological changes of cortical neurons after cerebral I/R injury.

<p>(A) N, nucleus; Broad arrows represent autophagosomes; Narrow arrows represent mitochondria. (B) Quantitative analysis of the nubmeber of autophagosomes. Three animals in each group and 10 fields for each animal were examined. *p<0.05 versus sham group.</p

PPAR-γ protein expression in cortex after cerebral I/R injury.

<p>(A) The level of PPAR-γ in ischemic cortex was higher than control in a time-dependent manner. (B) 15-PGJ₂ upregulated PPAR-γ expression in ischemic cortex in a concentration-dependent manner. *p<0.05 versus sham group, ^#p<0.05 versus I/R group.</p

Immunohistochemistry for LC3 and cathepsin B in neurons after cerebral I/R injury.

<p>(A) In sham-operated animals, cortical cells displayed diffuse and weak staining for LC3 in the cytosol. After I/R, intense LC3 staining appeared granular in the cytosol of cortical cells. Double staining for LC3 (green) and NeuN (red) showed that increase in LC3 punctate labeling occurred in cortical neurons. (B) In sham-operated animals, cortical cells displayed fine, granular, and perinuclear cathepsin-B staining. After I/R, cathepin-B granules became progressively larger and irregular, and the granular pattern was finally replaced with diffuse cytoplasmic staining. Double staining for cathepsin-B (green) and NeuN (red) showed that increased expression of cathepsin-B occurred mainly in neurons. Bar = 20 µm.</p

Nanosensor-Driven Detection of Neuron-Derived Exosomal Aβ₄₂ with Graphene Electrolyte-Gated Transistor for Alzheimer’s Disease Diagnosis

Blood-based tests have sparked tremendous attention in non-invasive early diagnosis of Alzheimer’s disease (AD), a most prevalent neurodegenerative malady worldwide. Despite significant progress in the methodologies for detecting AD core biomarkers such as Aβ42 from serum/plasma, there remains cautious optimism going forward due to its controversial diagnostic value and disease relevance. Here, a graphene electrolyte-gated transistor biosensor is reported for the detection of serum neuron-derived exosomal Aβ42 (NDE-Aβ42), which is an emerging, compelling trove of blood biomarker for AD. Assisted by the antifouling strategy with the dual-blocking process, the noise against complex biological background was considerably reduced, forging an impressive sensitivity gain with a limit of detection of 447 ag/mL. An accurate detection of SH-SY5Y-derived exosomal Aβ42 was also achieved with highly conformable enzyme-linked immunosorbent assay results. Importantly, the clinical analysis for 27 subjects revealed the immense diagnostic value of NDE-Aβ42, which can outclass that of serum Aβ42. The developed electronic assay demonstrates, for the first time, nanosensor-driven NDE-Aβ42 detection, which enables a reliable discrimination of AD patients from non-AD individuals and even the differential diagnosis between AD and vascular dementia patients, with an accuracy of 100% and a Youden index of 1. This NDE-Aβ42 biosensor defines a robust approach for blood-based confident AD ascertain