Synthesis, structure, magnetism and antibacterial properties of a 2-D nickel(II) metal–organic framework based on 3-nitrophthalic acid and 4,4′-bipyridine

<div><p>A 2-D nickel(II) mixed-ligand metal–organic framework [Ni(NPTA)(4,4′-bipy)(H₂O)]_n (<b>1</b>) was synthesized by reaction of 3-nitrophthalic acid (H₂NPTA) and 4,4′-bipyridine (4,4′-bipy) with Ni(II) under hydrothermal condition and characterized by elemental analysis, infrared spectroscopy, and single-crystal X-ray diffraction analysis. Compound <b>1</b> possesses a 2-D layer structure constructed from dinuclear nickel(II) building blocks in which two crystallographically equivalent Ni ions are bridged by two NPTA ligands. Furthermore, the layers are connected into 3-D supramolecular network by hydrogen bonds. The magnetism and antibacterial activity of <b>1</b> were investigated.</p></div

DataSheet1_The serum acylcarnitines profile in epileptic children treated with valproic acid and the protective roles of peroxisome proliferator-activated receptor a activation in valproic acid-induced liver injury.pdf

Valproic acid (VPA) is widely used as a major drug in the treatment of epilepsy. Despite the undisputed pharmacological importance and effectiveness of VPA, its potential hepatotoxicity is still a major concern. Being a simple fatty acid, the hepatotoxicity induced by VPA has long been considered to be due primarily to its interference with fatty acid β-oxidation (β-FAO). The aim of this study was to investigate the biomarkers for VPA-induced abnormal liver function in epileptic children and to determine potential mechanisms of its liver injury. Targeted metabolomics analysis of acylcarnitines (ACs) was performed in children’s serum. Metabolomic analysis revealed that VPA -induced abnormal liver function resulted in the accumulation of serum long-chain acylcarnitines (LCACs), and the reduced expression of β-FAO relevant genes (Carnitine palmitoyltrans-ferase (CPT)1, CPT2 and Long-chain acyl-CoA dehydrogenase (LCAD)), indicating the disruption of β-FAO. As direct peroxisome proliferator-activated receptor a (PPARα)- regulated genes, CPT1A, CPT2 and LCAD were up-regulated after treatment with PPARα agonist, fenofibrate (Feno), indicating the improvement of β-FAO. Feno significantly ameliorated the accumulation of various lipids in the plasma of VPA-induced hepatotoxic mice by activating PPARα, significantly reduced the plasma ACs concentration, and attenuated VPA-induced hepatic steatosis. Enhanced oxidative stress and induced by VPA exposure were significantly recovered using Feno treatment. In conclusion, this study indicates VPA-induced β-FAO disruption might lead to liver injury, and a significant Feno protective effect against VPA -induced hepatotoxicity through reversing fatty acid metabolism.</p

Complement-mediated hemolysis was inhibited by r<i>T</i>s-Pmy.

<p>Normal human serum (NHS) was pre-incubated with r<i>T</i>s-Pmy (0, 1, 2, or 4 μg) for 1 h prior to incubation with EAs. Hemolysis was measured at 412 nm for the supernatant of the reactions compared to complete lysis in water. The results shown are means ± SD and are representative of three independent experiments. BSA, C1q D and C3 D alone were added as controls. *<i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001. ns, no significant difference.</p

Classical complement activation was inhibited by r<i>Ts</i>-Pmy.

<p>Two μg of human C1q was pre-incubated with increasing amounts of r<i>T</i>s-Pmy (0, 2, 4 μg) or BSA (4 μg), then added to human IgM-coated plates. After washing with PBST for three times, a total of 2% C1q-deficient serum (C1q D) was added as a source of rest complement components. The deposition of C3 was detected with anti-C3 polyclonal antibody. The NHS alone and BSA added to the activation were used as controls. The results are shown as the means ± SD for three independent experiments.* <i>p</i><0.05, **<i>p</i><0.01, ***<i>p</i><0.001. ns, no significant difference.</p

r<i>Ts</i>-Pmy bound to A chain of human C1q.

<p><b>(A)</b> 96-well plates were coated with different amounts of C1q (a) or 2 μg BSA (b). Increasing amounts of r<i>Ts</i>-Pmy were added and anti-<i>Ts</i>-Pmy antibody 9G3 was used to detect the binding. ***<i>p</i><0.001. <b>(B)</b> Human C1q (5 μg) was subjected to SDS-PAGE under reducing condition and stained with Coomassie blue (a). C1q and BSA (each 5 μg) under reducing condition were transferred onto a NC membrane, incubated with r<i>Ts</i>-Pmy (5 μg/ml) and detected with anti-His mAb (1:5,000) (b). The same amount of r<i>Ts</i>-Pmy, r<i>Ts</i>87 or BSA (each 5 μg) were transferred to a NC membrane and then incubated with C1q (5 μg/ml). The proteins bound on the membrane were detected with anti-C1q antibody (1:1,000) (c). The r<i>Ts</i>-Pmy on the membrane was detected with anti-His mAb (d). The consistent results are repeated for three times. M, standard protein marker.</p

Native <i>T</i>s-Pmy from <i>T</i>. <i>spiralis</i> worm bound to human C1q.

<p>Protein G Micro Beads were pre-incubated with anti-<i>T</i>s-Pmy mAb 9G3 and <i>T</i>. <i>spiralis</i> adult worm crude extracts. Then, human complement C1q was added and the incubation was continued for 2h. The bound proteins were subjected to SDS-PAGE and transferred to a NC membrane. The membrane was probed with an anti-C1q mAb. The consistent results are repeated for three times. M, standard protein marker; Lane 1, pulled down immune complex (worm extracts + anti-<i>Ts</i>-Pmy + C1q); Lane 2, human C1q alone (2 ug); Lane 3, human C1q with the anti-<i>T</i>s-Pmy mAb 9G3 alone without the addition of <i>T</i>. <i>spiralis</i> adult extracts.</p

C1q-induced chemotaxis of THP-1-derived M2 macrophages was inhibited by r<i>Ts</i>-Pmy.

<p>Transwell-24-well plates were used for the migration assay. THP-1 cells were induced into M2 type macrophages in the upper chamber. LPS (100 ng/ml), C1q (10 nM) and C1q with different amounts of r<i>T</i>s-Pmy (0, 3, 6, or 12 μg) were added into the lower chamber. The cells that migrated though the membrane were counted under a microscope, and the cells from 8 randomly selected fields were counted. The results shown are means ± SD and are representative of three independent experiments. ***<i>p</i><0.001. ns, no significant difference.</p

The binding of C1q to THP-1-derived macrophages was inhibited by r<i>T</i>s-Pmy.

<p><b>(A)</b> After fixing with 4% paraformaldehyde and blocking with goat serum, M2 type macrophage cells were incubated with C1q, C1q plus r<i>T</i>s-Pmy or PBS alone for 1 h at 37°C, and then detected with an anti-C1q mAb or anti-<i>Ts</i>-Pmy mAb 9G3. Dylight 488 was used as a secondary antibody (green). The nuclei were stained with DAPI (blue). The imagine magnitude is 400 X with one cell 1000 X at the bottom right corner. <b>(B)</b> The fluorescence intensity was measured with high content analysis. The results shown are means ± SD and are representative of three independent experiments. ***<i>p</i><0.001.</p

r<i>Ts</i>-Pmy inhibited the binding of human C1q to IgM.

<p>One μg of C1q was pre-incubated with different amounts of r<i>Ts</i>-Pmy or BSA (0, 2, 3, 4 μg), then added to the IgM coated plates. After washing three times with PBST, the binding of C1q to IgM was determined with anti-C1q polyclonal antibody. The results shown are means ± SD and are representative of three independent experiments. ***<i>p</i><0.001.</p

r<i>Ts</i>-Hsp70 binds to DCs through TLR2 and TLR4 <i>in vitro</i> detected by flow cytometry.

<p>(A) Representative dot plots for the gating strategy: (I) gating on viable cells, (II) selection of non-adherent cells, (III) gating on CD11c⁺ cells, and (IV) selection of TLR2⁺ and Hsp70⁺ from gated CD11c⁺ cells (upper panel) and TLR4⁺ and Hsp70⁺ from gated CD11c⁺ cells (lower panel), respectively. (B) The binding of r<i>Ts</i>-Hsp70 to DCs derived from WT, TLR2^-/- or TLR4^-/- mice <i>in vitro</i>. DCs derived from WT, TLR2^-/- or TLR4^-/- mice were stained with anti-mouse CD11c APC and r<i>Ts</i>-Hsp70-PE. Representative dot plots for the gating strategy: (I) gating on viable cells, (II) gating on CD11c⁺ cells, (III) selection of Hsp70⁺ and CD11c⁺ from gated R1 cells, and (IV) selection of r<i>Ts</i>-Hsp70⁺ from gated CD11c⁺ cells. The percentage of CD11c+ cells binding to r<i>Ts</i>-Hsp70 shown on the right. All experiments were performed three times and data are shown with mean ± SD. n = 3, * <i>p</i> < 0.05, ** <i>p</i> < 0.01, *** <i>p</i> < 0.001.</p