Unexpected omega-3 activities in intracellular lipolysis and macrophage foaming revealed by fluorescence lifetime imaging.

Omega-3 polyunsaturated fatty acids (PUFA) found primarily in fish oil have been a popular supplement for cardiovascular health because they can substantially reduce circulating triglyceride levels in the bloodstream to prevent atherosclerosis. Beyond this established extracellular activity, here, we report a mode of action of PUFA, regulating intracellular triglyceride metabolism and lipid droplet (LD) dynamics. Real-time imaging of the subtle and highly dynamic changes of intracellular lipid metabolism was enabled by a fluorescence lifetime probe that addressed the limitations of intensity-based fluorescence quantifications. Surprisingly, we found that among omega-3 PUFA, only docosahexaenoic acid (DHA) promoted the lipolysis in LDs and reduced the overall fat content by approximately 50%, and consequently helped suppress macrophage differentiation into foam cells, one of the early steps responsible for atherosclerosis. Eicosapentaenoic acid, another omega-3 FA in fish oil, however, counteracted the beneficial effects of DHA on lipolysis promotion and cell foaming prevention. These in vitro findings warrant future validation in vivo.

Metabolic syndrome prediction model using Bayesian optimization and XGBoost based on traditional Chinese medicine features.

Metabolic syndrome (MetS) has a high prevalence and is prone to many complications. However, current MetS diagnostic methods require blood tests that are not conducive to self-testing, so a user-friendly and accurate method for predicting MetS is needed to facilitate early detection and treatment. In this study, a MetS prediction model based on a simple, small number of Traditional Chinese Medicine (TCM) clinical indicators and biological indicators combined with machine learning algorithms is investigated. Electronic medical record data from 2040 patients who visited outpatient clinics at Guangdong Chinese medicine hospitals from 2020 to 2021 were used to investigate the fusion of Bayesian optimization (BO) and eXtreme gradient boosting (XGBoost) in order to create a BO-XGBoost model for screening nineteen key features in three categories: individual bio-information, TCM indicators, and TCM habits that influence MetS prediction. Subsequently, the predictive diagnostic model for MetS was developed. The experimental results revealed that the model proposed in this paper achieved values of 93.35 %, 90.67 %, 80.40 %, and 0.920 for the F1, sensitivity, FRS, and AUC metrics, respectively. These values outperformed those of the seven other tested machine learning models. Finally, this study developed an intelligent prediction application for MetS based on the proposed model, which can be utilized by ordinary users to perform self-diagnosis through a web-based questionnaire, thereby accomplishing the objective of early detection and intervention for MetS.

Identification of Bioactive Components of Stephania epigaea Lo and Their Potential Therapeutic Targets by UPLC-MS/MS and Network Pharmacology.

Stephania epigaea, an important traditional folk medicinal plant, elucidating its bioactive compound profiles and their molecular mechanisms of action on human health, would better understand its traditional therapies and guide their use in preclinical and clinical. This study aims to detect the critical therapeutic compounds, predict their targets, and explore potential therapeutic molecular mechanisms. This work first determined metabolites from roots, stems, and flowering twigs of S. epigaea by a widely targeted metabolomic analysis assay. Then, the drug likeness of the compounds and their pharmacokinetic profiles were screened by the ADMETlab server. The target proteins of active compounds were further analyzed by PPI combing with GO and KEGG cluster enrichment analysis. Finally, the interaction networks between essential compounds, targets, and disease-associated pathways were constructed, and the essential compounds binding to their possible target proteins were verified by molecular docking. Five key target proteins (EGFR, HSP90AA1, SRC, TNF, and CASP3) and twelve correlated metabolites, including aknadinine, cephakicine, homostephanoline, and N-methylliriodendronine associated with medical applications of S. epigaea, were identified, and the compounds and protein interactions were verified. The key active ingredients are mainly accumulated in the root, which indicates that the root is the main medicinal tissue. This study demonstrated that S. epigaea might exert the desired disease efficacy mainly through twelve components interacting via five essential target proteins. EGFR is the most critical one, which deserves further verification by biological studies.

RNA interference of GhPEPC2 enhanced seed oil accumulation and salt tolerance in Upland cotton.

Phosphoenolpyruvate carboxylase (PEPCase) mainly produces oxaloacetic acid for tricarboxylic acid (TCA) cycle. Here we reported that GhPEPC2 silencing with PEPC2-RNAi vector could regulate oil and protein accumulation in cottonseeds. In GhPEPC2 transgenic plants, PEPCase activities in immature embryos were significantly reduced, and the oil content in seed kernel was increased 7.3 percentages, whereas total proteins decreased 5.65 percentages. Compared to wild type, agronomical traits of transgenic plant were obviously unaffected. Furthermore, gene expression profile of GhPEPC2 transgenic seeds were investigated using RNA-seq, most lipid synthesis related genes were up-regulated, but amino acid metabolic related genes were down-regulated. In addition, the GhPEPC2 transgenic cotton seedlings were stressed using sodium salts at seedling stage, and the salt tolerance was significantly enhanced. Our observations of GhPEPC2 in cotton would shade light on understanding the regulation of oil content, protein accumulation and salt tolerance enhancement in other plants.

Overexpression of GhDof1 improved salt and cold tolerance and seed oil content in Gossypium hirsutum.

A homologous GhDof1, which belongs to a large family of plant-specific transcription factor DOF, was isolated from Upland cotton (Gossypium hirsutum L.). GhDof1 protein was located in the nucleus of onion epidermal cells, the core domain of transcriptional activity existed in the C-terminal, and the activity elements of GhDof1 promoter existed in the regions of -645∼ -469bp and -286∼ -132bp of transcriptional start codon. GhDof1 constitutively expressed in leaves, roots and stems, accumulated highest in leaves. The salinity and cold treatments induced GhDof1 transcript accumulation. The GhDof1-overexpressed cotton showed significantly higher salt and cold tolerance over the wild-type plants. Under salt stress, the root growth of overexpressed GhDof1 lines was promoted. The expression levels of stress-responsive genes, GhP5CS, GhSOD and GhMYB, were differently up-regulated in transgenic lines. Oil contents increased in some transgenic plants, and protein contents reduced compared with transformed receptor. These results suggested that GhDof1 was a functional transcription factor for improving the abiotic tolerance and seed oil content in Upland cotton.

Fractionation of grape seed extract and identification of gallic acid as one of the major active constituents causing growth inhibition and apoptotic death of DU145 human prostate carcinoma cells.

The anti-cancer efficacy of grape seed extract (GSE) against prostate cancer (PCA) via its anti-proliferative, pro-apoptotic and anti-angiogenic activities in both cell culture and animal models have recently been described by us. GSE is a complex mixture containing gallic acid (GA), catechin (C), epicatechin (EC) and several oligomers (procyanidins) of C and/or EC, some of which are esterified to GA. To determine which components are most active against PCA, an ethyl acetate extract of GSE was separated by reverse-phase high-performance liquid chromatography (HPLC) into three fractions. Fraction 1 was far more effective than others in causing growth inhibition and apoptotic death of human PCA DU145 cells. Of the components in this fraction, GA showed a very strong dose- and time-dependent growth inhibition and apoptotic death of DU145 cells, but C and procyanidins B1 (EC-C dimer), B2 (EC-EC dimer) and B3 (C-C dimer) were nearly ineffective. Mechanistic studies demonstrated a strong caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP) cleavages by GA in DU145 cells. Procyanidin oligomers eluting in HPLC Fractions 2 and 3 were obtained in larger quantities by separating GSE into eight fractions (I-VIII) on a gel filtration column. All fractions were analyzed by HPLC-UV and negative-ion electrospray mass spectrometry. Fractions I-III contained the active compound GA and inactive components C, EC, B1 and B2. Fraction IV contained other dimers and a dimer-GA ester and was also less active than GSE in DU145 cells. Fractions V-VIII, however, caused significant growth inhibition and apoptosis with the highest activity present in the later fractions that contained procyanidin trimers and GA esters of dimers and trimers. Together, these observations identify GA as one of the major active constituents in GSE. Several procyanidins, however, and especially the gallate esters of dimers and trimers also may be efficacious against PCA and merit further investigation.

Isolation and structures of avicins D and G: in vitro tumor-inhibitory saponins derived from Acacia victoriae.

Two new saponins named avicins D (1) and G (2) were isolated from the seed pods of the desert legume plant Acacia victoriae. The structures, elucidated by 1D and 2D NMR studies and by chemical means, were characterized as acacic acid-bearing oligosaccharides at C-3 and C-28 with a side chain linked to C-21 comprised of two monoterpene carboxylic acids and a quinovose moiety. Both compounds exhibited potent cytotoxicity (apoptosis) against human T-cell leukemia (Jurkat cells) in vitro.