Distribution and metabolism of daidzein and its benzene sulfonates in vivo (in mice) based on MALDI-TOF MSI.

Daidzein (D1) has been proved to be of great benefit to human health. More and more attention was paid to the metabolic process of D1. Most studies focused on the metabolites of D1 and analogs were determined through the excretion of animals and humans by traditional HPLC-MS, while their in situ distribution and metabolism in organs in vivo has not been reported. In our group, novel daidzein sulfonate derivatives were synthesized and confirmed to have excellent pharmaceutical properties. They exhibited good anti-inflammatory, inhibitory activities on human vascular smooth muscle cell proliferation and other bioactivities. Compared with traditional analytical methods, matrix-assisted laser desorption ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI) can directly analyze the distribution of compounds in tissues and organs. In this study, we investigate the in situ distribution and metabolism of D1 and its derivatives (DD2, DD3) in the organs of mice based on MALDI-TOF MSI for the first time. Trace prototype compounds were detected in the plasma 4 h after the intravenous injection of D1, DD2, and DD3. Seven phase I metabolites and seven phase II metabolites were detected. D1 sulfates were found in the plasma and in organs except the heart. The presence of D1 and DD3 monosulfates in the brain indicated that they could penetrate the blood-brain barrier. DD2 and DD3 could be hydrolyzed into D1 and their metabolic pathways were similar to those of D1. In addition, a ligand-receptor docking of D1 and DD2 with mitogen-activated protein kinase 8 (JNK1) was performed because of their significant anti-inflammatory activities through the JNK signaling pathway. It showed that the binding energy of DD2 with JNK1 was obviously lower than that of D1 which was consistent with their anti-inflammatory activities. It provided a theoretical basis for further validation of their anti-inflammatory mechanism at the protein level. In summary, the research will provide beneficial guidance for further pharmacological, toxicological studies and the clinical-use research of these compounds.

Comparison of Biochemical Composition and Non-Volatile Taste Active Compounds of Back and Abdominal Muscles in Three Marine Perciform Fishes, Chromileptes altivelis, Epinephelus akaara and Acanthopagrus schlegelii.

Humpback grouper Chromileptes altivelis (HG), red-spotted grouper Epinephelus akaara (RG) and black seabream Acanthopagrus schlegelii (BS) are three popular perciform fishes with an increasingly important farming industry. The prices of BS are much lower than other grouper species; however, the differences in the nutritive values of these three perciform fishes with commercial specifications have not been reported. In this study, the biochemical composition and non-volatile taste active compounds of adult HG, RG and BS were investigated. Moisture contents in BS were significantly higher than in HG and RG (p < 0.05), and relatively lower crude protein contents in BS were observed. Lipid contents of back muscle were lower than that of abdomen muscle in the three fish species. C22:6n-3 (DHA) was the major poly-unsaturated fatty acid (PUFA) in HG and BS, while the main PUFA in RG was C18:2n-6. The total healthy omega-3 fatty acid (Σn-3) profiles in HG were the highest (24.08−24.59%), followed by RG (18.24−19.06%) and BS (13.63−15.91%) (p < 0.05). Glycine was the most abundant free amino acid (FAA) in HG and RG, while lysine was the major FAA in BS. Equivalent umami concentration (EUC) values in BS were the highest, followed by HG and RG (p < 0.05). Lactic acid and PO43− were the major organic acids and inorganic ions, respectively. In conclusion, HG and RG provided more protein and healthy omega-3 fatty acids than BS, while BS had a stronger umami taste according to the EUC values.

CC chemokine 1 protein from Cromileptes altivelis (CaCC1) promotes antimicrobial immune defense.

Chemokines are a family of small signaling proteins that are secreted by various cells. In addition to their roles in immune surveillance, localization of antigen, and lymphocyte trafficking for the maintenance of homeostasis, chemokines also function in induce immune cell migration under pathological conditions. In the present study, a novel CC chemokine gene (CaCC1) from humpback grouper (Cromileptes altivelis) was cloned and characterized. CaCC1 comprised a 435 bp open reading frame encoding 144 amino acid residues. The putative molecular weight of CaCC1 protein was 15 kDa CaCC1 contains four characteristic cysteines that are conserved in other known CC chemokines. CaCC1 also shares 11.64%-90.28% identity with other teleost and mammal CC chemokines. Phylogenetic analysis revealed that CaCC1 is most closely related to Epinephelus coioides EcCC1, both of which are in a fish-specific CC chemokine clade. CaCC1 was constitutively expressed in all examined C. altivelis tissues, with high expression levels in skin, heart, liver, and intestine. Vibrio harveyi stimulation up-regulated CaCC1 expression levels in liver, spleen, and head-kidney. Functional analyses revealed that the recombinant protein (rCaCC1) could induce the migration of head-kidney lymphocytes from C. altivelis. Moreover, rCaCC1 significantly enhanced phagocytosis in head-kidney macrophages from C. altivelis. In addition, rCaCC1 exhibited antimicrobial activities against Staphylococcus aureus, Edwardsiella tarda, and V. harveyi. In vivo, CaCC1 overexpression improved bacterial clearance in V. harveyi infected fish. Conversely, CaCC1 knockdown resulted in a significant decrease of bacterial clearance. These results demonstrate the important roles that CaCC1 plays in homeostasis and in inflammatory response to bacterial infection.