Development and validation of a novel approach based on ultrasonic-assisted enzymatic extraction and RP-UHPLC for simultaneous determination of astaxanthin isomers and alpha-tocopherol in Haematococcuspluvialis derived supplements.

A rapid, effective andgreenmethod was developed for simultaneous determination of total (free and esterified) astaxanthin (AX) isomers (all-E, 9Z and 13Z) and alpha-tocopherol(AT) in Haematococcuspluvialis derived supplements. The new method employed a highly efficient ultrasonic-assisted enzymatic extraction (UAEE) techniqueto perform deesterificationwith Cholesterol esterase from Pseudomonas fluorescenspermitting the concurrent detection. The subsequent RP-UHPLC method for separating and measuring was performed on a simple C18 column within 10.5 min by using methanol and ammonium acetate as mobile phase with a gradient elution. The proposed method was validated according to international guidelines and itproved to be highly accurate and robust. The optimized UHPLC method allowed easy transfer to HPLC, and allowed rapid analysis of active ingredients profiling in H. derived supplements.To our knowledge, this is the first quantification approach describing the rapid simultaneous analysis of the functional lipophilic substances including AX isomers in H. derived supplements using UAEE technique combined with RP-UHPLC.Moreover, this holistic approachcan be used to identify whether AX products are of natural origin or chemical synthesis, and may find more applications in new forms of H. derived productswith complexbiological matrix for more research on the bioavailability of AXisomersfrom natural source.

Inhibition of the JNK/MAPK signaling pathway by myogenesis-associated miRNAs is required for skeletal muscle development.

Skeletal muscle differentiation is controlled by multiple cell signaling pathways, however, the JNK/MAPK signaling pathway dominating this process has not been fully elucidated. Here, we report that the JNK/MAPK pathway was significantly downregulated in the late stages of myogenesis, and in contrast to P38/MAPK pathway, it negatively regulated skeletal muscle differentiation. Based on the PAR-CLIP-seq analysis, we identified six elevated miRNAs (miR-1a-3p, miR-133a-3p, miR-133b-3p, miR-206-3p, miR-128-3p, miR-351-5p), namely myogenesis-associated miRNAs (mamiRs), negatively controlled the JNK/MAPK pathway by repressing multiple factors for the phosphorylation of the JNK/MAPK pathway, including MEKK1, MEKK2, MKK7, and c-Jun but not JNK protein itself, and as a result, expression of transcriptional factor MyoD and mamiRs were further promoted. Our study revealed a novel double-negative feedback regulatory pattern of cell-specific miRNAs by targeting phosphorylation kinase signaling cascade responsible for skeletal muscle development.

(E)-1-(4-Chloro-phen-yl)-3-[4-(2,3,4,6-tetra-O-acetyl-ß-d-allopyranos-yloxy)phen-yl]prop-2-en-1-one.

The asymmetric unit of the title compound, C(29)H(29)ClO(11), contains two independent mol-ecules of similar geometry, both adopting an E conformation about the C=C double bond. The dihedral angles formed by benzene rings are 10.73 (16) and 13.79 (18)°. The pyran-oside rings adopt a chair conformation. Intra-molecular C-H⋯O close contacts occur. The crystal packing is stabilized by inter-molecular C-H⋯O hydrogen bonds.

1-[4-(ß-d-Allopyranos-yloxy)benzyl-idene]semicarbazide hemihydrate.

The mol-ecule of the title compound, C(14)H(19)N(3)O(7)·0.5H(2)O, exhibits an E conformation about the C=N double bond. The water mol-ecule possesses crystallographically imposed twofold symmetry. In the crystal structure, the mol-ecules are connected by inter-molecular O-H⋯O and N-H⋯O hydrogen bonds into a three-dimensional network.

4-[1-Acetyl-3-(4-methoxy-phen-yl)-2-pyrazolin-5-yl]phenol.

In the title compound, C(18)H(18)N(2)O(3), the dihedral angle formed by the benzene rings is 71.75 (4)°. In the crystal structure, centrosymmetrically related mol-ecules are linked into dimers by inter-molecular O-H⋯O hydrogen bonds and π-π stacking inter-actions with centroid-centroid distances of 3.5511 (6) Å.

4-Formyl-phenyl 2,3,4,6-tetra-O-acetyl-ß-d-allopyran-oside.

The title compound, C(21)H(24)O(11), crystallizes exclusively as the ß-anomer. The substituent of the protected sugar at position C-3 is in the axial position, while all other groups are in equatorial positions. The pyran-oside ring adopts a stable chair conformation.