The effects of NAC on the decreased number of myotube formations caused by arecoline.

C2C12 cells were cultured in differentiation medium with 0–5 mM NAC and 0.04 mM (A, B) or 0.08 mM (C, D) arecoline for 7 days. The myotubes were photographed by phase contrast microscopy (A, C). Scale bar, 100 μm. The numbers of myotubes from 30 random fields were counted (B, D). The values of ANOVA with 0.04 and 0.08 mM were (F5,12 = 241.90, P = 1.30E-11) and (F5,12 = 379.27, P = 8.99E-13) (mean ± SD; **p < 0.01, as compared to cells that were treated only with arecoline, Tukey HSD test).</p

Effect of NAC on the arecoline-inhibited multinucleated myotube formation.

C2C12 cells were cultured in differentiation medium with 0–5 mM NAC and 0.04 mM (A, B) or 0.08 mM (C, D) arecoline for 7 days and H&E staining. The myotubes were photographed by light microscopy (A, C). Arrows indicate the nuclei in the multinucleated myotubes. Scale bar, 100 μm. The numbers of nuclei per myotube from 30 random fields were shown (B, D). The values of ANOVA with 0.04 and 0.08 mM were (F5,12 = 14.51, P = 9.87E-05) and (F5,12 = 154.72, P = 1.83E-10). (mean ± SD; *p p < 0.01, as compared to cells treated with arecoline only, Tukey HSD test).</p

Effect of NAC on decreased MYH expression caused by arecoline in C2C12 cells.

The C2C12 cells were cultured in differentiation medium with 0–2 mM NAC and 0.04 mM (A, C) or 0.08 mM (B, D) arecoline for 7 days. The distribution of MYH was examined by immunofluorescence under a fluorescence microscope (A and B, left, upper panel). Scale bar represents 100 μm at 100x magnification. Nuclei in the same field were counterstained with DAPI (A and B, left, middle panel). The images of MYH and DAPI were merged (A and B, left, lower panel). The intensities of MYH were quantified and normalized to the no-treatment control from random images of three independent experiments (Fig 5A and 5B, right panels). The values of ANOVA with 0.04 and 0.08 mM were (F4,10 = 2.73, P = 0.09) and (F4,10 = 14.49, P = 3.63E-04). The expression of MYH protein was detected by Western blotting (C and D, upper panel). β-actin served as a loading control. The band intensities of MYH from three independent experiments were quantified and normalized to no-treatment control (C and D, lower panel). The values of ANOVA with 0.04 and 0.08 mM were (F4,10 = 0.63, P = 0.65) and (F4,10 = 5.15, P = 0.02) (mean ± SD; *p p < 0.01, as compared to cells treated only with arecoline, Tukey HSD test).</p

Effect of NAC on cell proliferation in C2C12 myoblasts.

Cells were exposed to 0–20 mM arecoline for 24 h (solid bars) or 48 h (open bars) (A) and 7 days (B). The viability of cells was measured with CellTiter 96 Aqueous One Solution Reagent (Promega) in duplicate. The means of duplicate from three independent experiments were used for further analysis. The values of ANOVA at 24 h, 48 h and 7 days were (F7,16 = 56.44, P = 4.28E-10), (F7,16 = 26.97, P = 1.03E-07) and (F5,12 = 7.93, P = 1.66E-3), respectively (mean ± SD; **p < 0.01, as compared to a no-treatment control, Tukey HSD test).</p

The effect of NAC on decreased ERK1/2 Tyr-204 phosphorylation caused by arecoline during myogenic differentiation in C2C12 cells.

(A) The expression of Tyr-204 phosphorylated ERK1/2, ERK1/2 and β-actin proteins were detected by Western blotting at 0, 6, 12, 24 h, 4 and 7 days of myogenic differentiation in C2C12 cells treated with 0.08 mM arecoline and 0–2 mM NAC. p44:ERK1, p42:ERK2. β-actin served as the loading control. (B) The relative expression of p-ERK1/2 protein to β-actin were quantified and normalized to the no-treatment control. The values of ANOVA at 12 and 24h were (F4,10 = 5.03, P = 0.02) and (F4,10 = 5.07, P = 0.02). (C) The relative expression of ERK1/2 protein to β-actin were quantified and normalized to the no-treatment control. (D) The ratio of p-ERK1/2 to ERK1/2 from figure B and C were relative to β-actin and normalized to the no-treatment control. The values of ANOVA at 12 and 24h were (F4,10 = 5.54, P = 0.01) and (F4,10 = 7.94, P = 3.78E-03). Data from three independent experiments were presented (mean ± SD; *p p < 0.01, as compared to cells treated with arecoline, Tukey HSD test).</p

Effect of NAC on decreased myogenin expression caused by arecoline in C2C12 cells.

The C2C12 cells were cultured in differentiation medium with 0–2 mM NAC and 0.08 mM arecoline for 7 days. (A) The expression of myogenin protein was detected by Western blotting. β-actin served as a loading control. (B) The band intensities of myogenin were quantified and normalized to the no-treatment control. Data from three independent experiments were used. The values of ANOVA were (F4,10 = 22.02, P = 6.05E-05)(mean ± SD; **p < 0.01 as compared to cells treated with arecoline only, Tukey HSD test).</p

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Tuning Dimensionality of Benzimidazole Aggregates by Using Tetraoctylammonium Bromide: Enhanced Electrochemiluminescence Studies

Exploring the depolymerization strategy of liposoluble luminophores in the aqueous phase is vital for the development of electrochemiluminescence (ECL). In this work, tetraoctylammonium bromide (TOAB) with four long hydrophobic chains and short hydrophilic ends is used as a template to limit the aggregation of benzimidazole (BIM). By adjusting the loading of BIM on the hydrophobic chains of TOAB, a two-dimensional lamellar BIM/TOAB is formed, the ECL intensity of which is 6.4 times higher than that of the aggregated BIM (H2O2 as the coreactant). In terms of ECL spectroscopies, cyclic voltammetry , ECL transients, and the adjustment of the scanning potential range, the ECL mechanism is thoroughly studied. This work provides a new way to depolymerize organic luminophores and reveals a possible pathway in the annihilation ECL mechanism

Table_1_A new species of the genus Catillopecten (Bivalvia: Pectinoidea: Propeamussiidae): morphology, mitochondrial genome, and phylogenetic relationship.xls

Catillopecten is a small genus of deep-sea glass scallops, but its diversity is poorly known in many parts of the world ocean. We described C. margaritatus n. sp. (Pectinoidea: Propeamussiidae), and performed morphological analyses and DNA sequencing, and estimated the divergence time of scallops based on samples collected from Haima cold seep in the South China Sea. Morphologically, the new species can be distinguished from congeneric species by its large shell size, relatively small auricle length, absence of without monocrystal aerials, presence of longitudinal radial ridges on the left valve, and the alternated rounded striae and distal and proximal growth lines of prisms on the right valve. Anatomically, this new species can be distinguished from C. vulcani by its anteriorly located auriculate gills, compared to the centrally located lamellar gills of the latter, and the different locations of the pericardium. Sequence comparison and phylogenetic analysis based on the 18S rRNA fragments supported the placement of the new species in Catillopecten. We also report the mitogenome of C. margaritatus n. sp. as the only reported mitogenome of the family Propeamussiidae, which differs from those of other scallops substantially in gene order arrangement. Divergence time estimation revealed that Propeamussiidae and Pectinidae diverged in the early Carboniferous, while Catillopecten and Parvamussium diverged during the late Cretaceous to early Eocene. Finally, we presented a key to the species of Catillopecten.</p

Micelle-Assisted Confined Coordination Spaces for Benzimidazole: Enhanced Electrochemiluminescence for Nitrite Determination

Selective and sensitive detection of nitrite has important medical and biological implications. In the present work, to obtain an enhanced electrochemiluminescence (ECL) determination of nitrite, a novel nano-ECL emitter CoBIM/cetyltrimethylammonium bromide (CTAB) was prepared via a micelle–assisted, energy-saving, and ecofriendly method based on benzimidazole (BIM) and CTAB. Unlike conventional micelle assistance, the deprotonated BIM (BIM–) preferential placement was in the palisade layer of cationic CTAB-based micelles. Enriching the original CTAB micelle with BIM– disrupted its stability and resulted in the formation of considerably smaller BIM/CTAB-based micelles, providing a confined coordination environment for BIM– and Co2+. As a result, the growth of CoBIM/CTAB was also limited. Owing to the unusual nitration reaction between BIM and nitrite, the prepared CoBIM/CTAB was successfully applied as a novel ECL probe for the detection of nitrite with a wide linear range of 1–1500 μM and a low detection limit of 0.67 μM. This work also provides a promising ECL platform for ultrasensitive monitoring of nitrite and it was applied with sausages and pickled vegetables