The levels of IFN-γ in CSF and serum of ALS patients with bulbar or limb onset and their correlations with disease progression.

<p>Fig 2A shows the levels of IFN-γ in CSF of ALS patients with bulbar or limb onset. There are significant differences in the CSF levels of IFN-γ in ALS patients (both bulbar and limb onset) compared with non-ALS patients (P<0.01), but no difference is observed between the bulbar and limb onset subgroups (P>0.05). Fig 2B-C show the correlations between the CSF IFN-γ levels and the DPR. The CSF IFN-γ levels do not correlate with the DPR in bulbar onset patients (B) but positively correlate with the DPR in limb onset patients (C). Fig 2D shows the levels of IFN-γ in serum of ALS patients with bulbar or limb onset. There are significant differences in the serum levels of IFN-γ in ALS patients (both bulbar and limb onset) compared with non-ALS patients (P<0.01), but no difference is observed between the bulbar and limb onset subgroups (P>0.05). Fig 2E-F show the correlations between the serum IFN-γ levels and the DPR. The serum IFN-γ levels do not correlate with the DPR in bulbar onset patients (E) but positively correlate with the DPR in limb onset patients (F). * indicates significant differences (p<0.01) in the CSF or serum IFN-γ levels between the ALS patients (both bulbar and limb onset) and the non-ALS patients.</p

Summary of samples from ALS patients (ALS) and from non-ALS patients (Non-ALS).

<p>M: male; F: Female. B: bulbar onset; L: limb onset; MFO: interval from the observation of initial symptoms to diagnosis; DPR: disease progression rate; ALSFRS-r: Amyotrophic Lateral Sclerosis Functional Rating Scale revised.</p><p>* indicates a significant difference (p<0.01) in the IFN-γ levels between ALS and Non-ALS.</p><p># indicates a significant difference (p<0.01) in the CSF IFN-γ levels compared with the serum IFN-γ levels; mean±se.</p><p>Summary of samples from ALS patients (ALS) and from non-ALS patients (Non-ALS).</p

Correlations between the levels of IFN-γ and disease progression.

<p>Fig 1A-C show the correlations between the CSF IFN-γ levels and the DPR, ALSFRS-r, and MFO in the ALS patients. The IFN-γ levels in CSF positively correlate with the DPR (A) and the MFO (C) but not the ALSFRS-r score (B). Fig 1D-F show the correlations between the serum IFN-γ levels and the DPR, the ALSFRS-r score, and the MFO in the ALS patients. The IFN-γ level in serum significantly correlate with the DPR (D) but not the ALSFRS-r score (E) or the MFO (F).</p

Summary of samples from ALS patients (ALS) categorized according to MFO and from non-ALS patients (Non-ALS).

<p>ALS<12 m: the time from ALS onset to diagnosis was less than 12 months; AL≥12 m: the time from ALS onset to diagnosis was equal to or more than 12 months</p><p>* indicates a significant difference (p<0.01) in the IFN-γ levels between ALS and Non-ALS.</p><p># indicates a significant difference (p<0.01) in the CSF IFN-γ levels between the ALS≥12 m and ALS<12 m subgroups.</p><p>+ indicates a significant difference (p<0.01) in the CSF IFN-γ levels in the ALS≥12 m subgroup compared with either the serum IFN-γ level in the ALS<12 m subgroup or the serum IFN-γ levels in the ALS≥12 m subgroup; mean±se.</p><p>Summary of samples from ALS patients (ALS) categorized according to MFO and from non-ALS patients (Non-ALS).</p

The levels of IFN-γ in CSF and serum of ALS patients stratified according to MFO and their correlations with disease progression.

<p>Fig 3A shows the levels of IFN-γ in CSF of ALS patients stratified according to MFO. There are significant differences in both ALS subgroups (ALS<12 m and ALS≥12 m) compared with the non-ALS patients (P<0.01), and a difference is detected between the ALS<12 m and ALS≥12 m subgroups (P<0.01). Fig 3B-C show the correlations between the CSF IFN-γ levels and the DPR in the ALS<12 m and ALS≥12 m subgroups. The CSF IFN-γ levels positively correlate with the DPR in both subgroups. Fig 3D shows the levels of IFN-γ in serum of ALS patients stratified according to MFO. There are significant differences in both ALS subgroups (ALS<12 m and ALS≥12 m) compared with the non-ALS patients (P<0.01), but no difference is observed between the ALS<12 m and ALS≥12 m subgroups (P>0.05). Fig 3E-F show the correlations between the serum IFN-γ levels and the DPR in ALS<12 m and ALS≥12 m subgroups. The serum IFN-γ levels positively correlate with the DPR in the ALS<12 m subgroup but not in the ALS≥12 m subgroup. * indicates significant differences (p<0.01) in the CSF or serum IFN-γ levels between the ALS patients (both ALS<12 m and ALS≥12 m) and the non-ALS patients. # indicates a significant difference (p<0.01) in the CSF IFN-γ levels between the ALS≥12 m and ALS<12 m subgroups.</p

Supplementary document for Design method of imaging optical systems using confocal flat phase elements - 6158248.pdf

Detailed discussions of Eqs. (7) and (8) in the paper are demonstrate

Novel Strategy for Preparation of Graphene-Pd, Pt Composite, and Its Enhanced Electrocatalytic Activity for Alcohol Oxidation

As advanced electrodes for direct alcohol fuel cells, graphene-Pd and graphene-Pt composites with a trace of SnO₂ have been successfully synthesized by a modified electroless plating technique. The surface of graphene oxide is first sensitized by Sn²⁺ ions, and subsequently, Pd or Pt nanoparticles are deposited on the surface of graphene oxide. Finally, graphene oxide was reduced to graphene by further adding NaBH₄. Compared to other carbon-(e.g., Vulcan XC-72R) supported Pd and Pt, the resultant graphene-Pd and Pt composites exhibit better electrocatalytic activity and long-term stability toward alcohol electrooxidation. Additionally, a trace amount of SnO₂ formed around active catalysts may also be beneficial to the enhancement of electrochemical activity

Palmitoylation of the Cysteine Residue in the DHHC Motif of a Palmitoyl Transferase Mediates Ca²⁺ Homeostasis in <i>Aspergillus</i>

<div><p>Finely tuned changes in cytosolic free calcium ([Ca²⁺]_c) mediate numerous intracellular functions resulting in the activation or inactivation of a series of target proteins. Palmitoylation is a reversible post-translational modification involved in membrane protein trafficking between membranes and in their functional modulation. However, studies on the relationship between palmitoylation and calcium signaling have been limited. Here, we demonstrate that the yeast palmitoyl transferase <i>Sc</i>Akr1p homolog, AkrA in <i>Aspergillus nidulans</i>, regulates [Ca²⁺]_c homeostasis. Deletion of <i>akrA</i> showed marked defects in hyphal growth and conidiation under low calcium conditions which were similar to the effects of deleting components of the high-affinity calcium uptake system (HACS). The [Ca²⁺]_c dynamics in living cells expressing the calcium reporter aequorin in different <i>akrA</i> mutant backgrounds were defective in their [Ca²⁺]_c responses to high extracellular Ca²⁺ stress or drugs that cause ER or plasma membrane stress. All of these effects on the [Ca²⁺]_c responses mediated by AkrA were closely associated with the cysteine residue of the AkrA DHHC motif, which is required for palmitoylation by AkrA. Using the acyl-biotin exchange chemistry assay combined with proteomic mass spectrometry, we identified protein substrates palmitoylated by AkrA including two new putative P-type ATPases (Pmc1 and Spf1 homologs), a putative proton V-type proton ATPase (Vma5 homolog) and three putative proteins in <i>A</i>. <i>nidulans</i>, the transcripts of which have previously been shown to be induced by extracellular calcium stress in a CrzA-dependent manner. Thus, our findings provide strong evidence that the AkrA protein regulates [Ca²⁺]_c homeostasis by palmitoylating these protein candidates and give new insights the role of palmitoylation in the regulation of calcium-mediated responses to extracellular, ER or plasma membrane stress.</p></div

Phenotypic characterization of Golgi-localized AkrA.

<p>A. The phenotypic characterization of <i>akrA</i> under control of the <i>alcA(p)</i> conditional promoter. The colony images show corresponding strains grown on the non-inducing medium (<i>RE</i>::<i>akrA</i>), inducing medium (<i>EX</i>::<i>akrA</i>) and overexpressing medium (<i>OE</i>::<i>akrA</i>) at 37°C for 2.5 days. B. Western blot analysis indicated a fusion protein of GFP-AkrA was detected with a predicted size of approximately 100 kDa by using an anti-GFP antibody. GFP-AkrA non-inducing and GFP-AkrA inducing represent <i>alcA(p)</i>::GFP<i>-akrA</i> grown in liquid non-inducing medium and inducing medium, respectively. Anti-actin antibody against actin was used as an internal control of loading. C. Colocalization of GFP-AkrA and the GEs marker mRFP-PH^OSBP. A strain carrying transgenes expressing the two fluorescent reporters was imaged using GFP and mRFP specific filter sets. The yellow color in the merged image shows the co-localization. Bar, 5 μm.</p

The cysteine residue in the DHHC motif, is correspondingly required for AkrA palmitoylation.

<p>A. A schematic diagram of the acyl-biotin exchange (ABE) assay. ① Blocking the free sulfhydryls with N-ethylmaleimide (NEM); ② Cleavaging the thioester bonds with or without hydroxylamine (HA); ③ Biotinylating the palmitoylated proteins with HPDP-biotin; Lastly, ④ Enriching the biotinylated proteins bound to streptavidin agarose (SA). B. Flag-AkrA and Flag-AkrA^C487S were detected by Western blotting with anti-Flag antibodies using the ABE assay, treated or not with 100 μM 2-bromopalmitate (2-BP). Hydroxylamine (HA) was used to specifically cleave S-acyl groups revealing sulfhydryl groups, which were subsequently labeled with biotin. Samples were then bound to streptavidin beads. For the negative control HA was substituted by Tris. Anti-actin antibody was used as an internal control of loading. A band was detected in the +HA treated sample, indicating that it was bound to an acyl group via a thioester linkage confirming that it is auto-acylated. However, no signal was detected for Flag-AkrA^C487S and 2-BP treatment samples and therefore they are not auto-acylated. C. Western blot analysis indicated a fusion protein of GFP-AkrA^C487S was detected with a predicted size of approximately 100 kDa by using an anti-GFP antibody. D. GFP-AkrA and GFP-AkrA^C487S localization was assessed after culturing for 18 h in liquid induced medium supplemented with or without the indicated concentration of 2-BP. Localization within the Golgi was less distinct as punctate structures in the GFP-AkrA^C487S strain compared with that in the wild-type and its localization within the Golgi was completely abolished after 2-BP treatment. Bar, 2 μm. E. Total proteins from wild type and Δ<i>akrA</i> strains subjected to the ABE assay with (HA+) or without (HA-) hydroxylamine treatment. The samples were then electrophoresed by SDS-PAGE and detected by silver nitrate staining.</p