DataSheet2_Neuroprotective effect of dexmedetomidine on autophagy in mice administered intracerebroventricular injections of Aβ25–35.PDF

Alzheimer’s disease (AD), one of the most prevalent neurodegenerative diseases is associated with pathological autophagy-lysosomal pathway dysfunction. Dexmedetomidine (Dex) has been suggested as an adjuvant to general anesthesia with advantages in reducing the incidence of postoperative cognitive dysfunction in Dex-treated patients with AD and older individuals. Several studies reported that Dex improved memory; however, evidence on the effects of Dex on neuronal autophagy dysfunction in the AD model is lacking. We hypothesized that Dex administration would have neuroprotective effects by improving pathological autophagy dysfunction in mice that received an intracerebroventricular (i.c.v.) injection of amyloid β-protein fragment 25–35 (Aβ25–35) and in an autophagy-deficient cellular model. In the Y-maze test, Dex reversed the decreased activity of Aβ25–35 mice. Additionally, it restored the levels of two memory-related proteins, phosphorylated Ca2+/calmodulin-dependent protein kinase II (p-CaMKII) and postsynaptic density-95 (PSD-95) in Aβ25–35 mice and organotypic hippocampal slice culture (OHSC) with Aβ25–35. Dex administration also resulted in decreased expression of the autophagy-related microtubule-associated proteins light chain 3-II (LC3-II), p62, lysosome-associated membrane protein2 (LAMP2), and cathepsin D in Aβ25–35 mice and OHSC with Aβ25–35. Increased numbers of co-localized puncta of LC3-LAMP2 or LC3-cathepsin D, along with dissociated LC3-p62 immunoreactivity following Dex treatment, were observed. These findings were consistent with the results of western blots and the transformation of double-membrane autophagosomes into single-membraned autolysosomes in ultrastructures. It was evident that Dex treatment alleviated impaired autolysosome formation in Aβ mice. Our study demonstrated the improvement of memory impairment caused by Dex and its neuroprotective mechanism by investigating the role of the autophagy-lysosomal pathway in a murine Aβ25–35 model. These findings suggest that Dex could be used as a potential neuroprotective adjuvant in general anesthesia to prevent cognitive decline.</p

DataSheet1_Neuroprotective effect of dexmedetomidine on autophagy in mice administered intracerebroventricular injections of Aβ25–35.PDF

Alzheimer’s disease (AD), one of the most prevalent neurodegenerative diseases is associated with pathological autophagy-lysosomal pathway dysfunction. Dexmedetomidine (Dex) has been suggested as an adjuvant to general anesthesia with advantages in reducing the incidence of postoperative cognitive dysfunction in Dex-treated patients with AD and older individuals. Several studies reported that Dex improved memory; however, evidence on the effects of Dex on neuronal autophagy dysfunction in the AD model is lacking. We hypothesized that Dex administration would have neuroprotective effects by improving pathological autophagy dysfunction in mice that received an intracerebroventricular (i.c.v.) injection of amyloid β-protein fragment 25–35 (Aβ25–35) and in an autophagy-deficient cellular model. In the Y-maze test, Dex reversed the decreased activity of Aβ25–35 mice. Additionally, it restored the levels of two memory-related proteins, phosphorylated Ca2+/calmodulin-dependent protein kinase II (p-CaMKII) and postsynaptic density-95 (PSD-95) in Aβ25–35 mice and organotypic hippocampal slice culture (OHSC) with Aβ25–35. Dex administration also resulted in decreased expression of the autophagy-related microtubule-associated proteins light chain 3-II (LC3-II), p62, lysosome-associated membrane protein2 (LAMP2), and cathepsin D in Aβ25–35 mice and OHSC with Aβ25–35. Increased numbers of co-localized puncta of LC3-LAMP2 or LC3-cathepsin D, along with dissociated LC3-p62 immunoreactivity following Dex treatment, were observed. These findings were consistent with the results of western blots and the transformation of double-membrane autophagosomes into single-membraned autolysosomes in ultrastructures. It was evident that Dex treatment alleviated impaired autolysosome formation in Aβ mice. Our study demonstrated the improvement of memory impairment caused by Dex and its neuroprotective mechanism by investigating the role of the autophagy-lysosomal pathway in a murine Aβ25–35 model. These findings suggest that Dex could be used as a potential neuroprotective adjuvant in general anesthesia to prevent cognitive decline.</p

Think Modular: A Simple Apoferritin-Based Platform for the Multifaceted Detection of Pancreatic Cancer

The generation of nanosized probes often requires time-intensive and application-specific optimization processes that involve conjugating a nanoconstruct to a targeting moiety. Herein, we genetically modify apoferritin and generate a universal interface system composed of protein G and 6×His-tag. The resulting construct, conferred with modularity and high targeting efficiency, is applied toward two distinct applications in the detection of a pancreatic cancer biomarker and used to demonstrate its potential in the facile exchange of nanoprobe components

Effect of over-expressed <i>TAS2R8</i> and <i>TAS2R10</i> on tumor incidence, final tumor volume, and tumor weight.

<p>Effect of over-expressed <i>TAS2R8</i> and <i>TAS2R10</i> on tumor incidence, final tumor volume, and tumor weight.</p

Over-expression of <i>TAS2R8</i> and <i>TAS2R10</i> suppressed HIF-1α-mediated regulation of <i>VEGF</i> and <i>GLUT1</i>.

<p>(A) After each TAS2Rs construct was over-expressed, each set of BE(2)C cells was treated with CoCl_2·6H₂O. After 24 h, protein levels of HIF-1α were compared with the levels detected in the cells that were transfected with an empty vector (EV) and maintained under hypoxic conditions. (B) The mRNA levels of two genes downstream of HIF-1α, <i>VEGF</i> and <i>GLUT1</i>, were compared with the control group. An unpaired two-tailed Student’s <i>t</i>-test was performed to compare each TAS2R data with the control cells. A <i>P</i>-value less than 0.05 was considered statistically significant. (* p < 0.05, ** p < 0.01, *** p < 0.001). N, normoxia; H, hypoxia.</p

The self-renewal capacity of BE(2)C cells was suppressed following the over-expression of TAS2Rs.

<p>BE(2)C cells were transfected with an empty vector (EV ctrl), or vectors expressing <i>TAS2R8</i>, or <i>TAS2R10</i>. (A) After 48 h, the transfected cells were plated in 6-well plates and cultured for additional 7–10 days. Colonies were then stained and the number of colonies with ≧ 50 cells was counted. (B) The number of spheres was counted. Magnification, x100. The percentages for colony and sphere formation were normalized to that of the empty vector group. An unpaired two-tailed Student’s <i>t</i>-test was performed to compare each TAS2R data with the control cells. A <i>P</i>-value less than 0.05 was considered statistically significant (* p < 0.05, ** p < 0.01, *** p < 0.001).</p

Over-expression of <i>TAS2R8</i> and <i>TAS2R10</i> suppressed <i>MMP-2</i> and <i>P-selectin</i> expression.

<p>The markers of adhesion, mRNA expressions of <i>MMP-2</i> and <i>P-selectin</i> were analyzed. An unpaired two-tailed Student’s <i>t</i>-test was performed to compare each TAS2Rs tumor group with the empty vector control (EV Ctrl) group. A <i>P</i>-value less than 0.05 was considered statistically significant. (* p < 0.05, ** p < 0.01, *** p < 0.001).</p

Bitter taste receptors were endogenously expressed and functional in neuroblastoma cell lines.

<p>(A) The mRNA levels of <i>TAS2R8</i> (a) and <i>TAS2R10</i> (b) were detected in mouse liver, tongue, and brain tissues and in BE(2)C and SY5Y cells (c). The bars represent the mean ± SEM (* p < 0.05). One-way analysis of variance (ANOVA) was applied to compare expressions in mouse tissues, and an unpaired two-tailed <i>t</i>-test was used to compare two cell lines. (B) mRNA expression of TAS2R8 (a) and TAS2R10 (b) over-expressed BE(2C) cells were analyzed. (C) Denatonium benzoate (0.01 μM) induced an increase in intracellular calcium [Ca²⁺] in HEK293 cells (a-b) and BE(2C) cells (c-e). HEK293 cells were transfected with TAS2R8 (a) and TAS2R10 (b). BE(2)C cells were transfected with an empty vector (EV ctrl) (c), or vectors expressing TAS2R8 (d), or TAS2R10 (e).</p

Migration and invasion were suppressed following the over-expression of <i>TAS2R8</i> and <i>TAS2R10</i> in the BE(2)C cells.

<p>BE(2)C cells were transfected with empty vector (EV ctrl), <i>TAS2R8</i>, and <i>TAS2R10</i>. (A) Cell migration was analyzed in wound-healing assays. Representative images of the migration assay results are shown (left panel). The fold increase in migration distance was compared with the empty vector group (right panel). (B) Cell invasion was evaluated in Transwell assays. Representative images of the invasion assays are shown (left panel). The number of invading cells was counted and the percentage of invading cells in each group was normalized to that of the empty vector group (right panel). (C-D) After <i>TAS2R8</i> and <i>TAS2R10</i> were over-expressed, MMP enzymatic activity and gene expressions were analyzed. (C) Enzyme activity of MMP-2 was detected in zymography assays. (D) The mRNA level of <i>MMP-2</i> was compared with the control group. An unpaired two-tailed Student‘s <i>t</i>-test was performed to compare each TAS2R data with the control cells. A <i>P</i>-value less than 0.05 was considered statistically significant (* p < 0.05, ** p < 0.01, *** p < 0.001).</p

Application of N‑Doped Three-Dimensional Reduced Graphene Oxide Aerogel to Thin Film Loudspeaker

We built a thermoacoustic loudspeaker employing N-doped three-dimensional reduced graphene oxide aerogel (N-rGOA) based on a simple template-free fabrication method. A two-step fabrication process, which includes freeze-drying and reduction/doping, was used to realize a three-dimensional, freestanding, and porous graphene-based loudspeaker, whose macroscopic structure can be easily modulated. The simplified fabrication process also allows the control of structural properties of the N-rGOAs, including density and area. Taking advantage of the facile fabrication process, we fabricated and analyzed thermoacoustic loudspeakers with different structural properties. The anlayses showed that a N-rGOA with lower density and larger area can produce a higher sound pressure level (SPL). Furthermore, the resistance of the proposed loudspeaker can be easily controlled through heteroatom doping, thereby helping to generate higher SPL per unit driving voltage. Our success in constructing an array of optimized N-rGOAs able to withstand input power as high as 40 W demonstrates that a practical thermoacoustic loudspeaker can be fabricated using the proposed mass-producible solution-based process