Triacylglycerol Concentrations Research Articles

Propanol and 1, 3-propanediol enhance fatty acid accumulation synergistically in Schizochytrium ATCC 20888.

The effects of propanol and 1, 3-propanediol on fatty acid and biomass accumulation in Schizochytrium ATCC 20888 were explored. Propanol increased the contents of saturated fatty acids and total fatty acids by 55.4 and15.3%, while 1, 3-propanediol elevated the polyunsaturated fatty acids, total fatty acids and biomass contents by 30.7, 17.0, and 6.89%. Although both of them quench ROS to increase fatty acids biosynthesis, the mechanisms are different. The effect of propanol did not reflect on metabolic level while 1, 3-propanediol elevated osmoregulators contents and activated triacylglycerol biosynthetic pathway. The triacylglycerol content and the ratio of polyunsaturated fatty acids to saturated fatty acids were significantly increased by 2.53-fold, which explained the higher PUFA accumulation in Schizochytrium after adding 1, 3- propanediol. At last, the combination of propanol and 1, 3-propanediol further elevated total fatty acids by approximately 1.2-fold without compromising cell growth. These findings are valuable for scale-up production of designed Schizochytrium oil for various application purposes.

Applications of Diacylglycerol Acyltransferase for Triacylglycerol Production in Mortierella alpina.

Triacylglycerol (TG) with high-value long-chain polyunsaturated fatty acids is beneficial to human health; consequently, there is an urgent need to broaden its sources due to the current growing demand. Mortierella alpina, one of the most representative oleaginous fungi, is the only certificated source of dietary arachidonic acid-rich oil supplied in infant formula. This study was conducted to improve TG production in M. alpina by homologous overexpression of diacylglycerol acyltransferase (DGAT) and linseed oil (LSO) supplementation. Our results showed that the homologous overexpression of MaDGAT1B and MaDGAT2A strengthened TG biosynthesis and significantly increased the TG content compared to the wild-type by 12.24% and 14.63%, respectively. The supplementation with an LSO concentration of 0.5 g/L elevated the TG content to 83.74% and total lipid yield to 4.26 ± 0.38 g/L in the M. alpina-MaDGAT2A overexpression strain. Our findings provide an effective strategy for enhancing TG production and highlight the role of DGAT in TG biosynthesis in M. alpina.

Accurate quantification of TAGs to identify adulteration of edible oils by ultra-high performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry

Edible oils play important roles in biological functions, and triacylglycerols (TAGs) in edible oils are complex mixtures. This makes accurate TAGs quantitation quite difficult that bring economically motivated food adulteration. Herein, we demonstrated a strategy for accurate quantification of TAGs in edible oils, which could be applied in identification of olive oil adulteration. The results showed that the proposed strategy could significantly improve the accuracy of TAG content determination, reduce the relative error of fatty acids (FAs) content determination, and present a wider accurate quantitative range than that of gas chromatography-flame ionization detection. Most important, this strategy coupled with principal component analysis could be used to identify adulteration of high-priced olive oil with cheaper soybean oils, rapeseed oils or camellia oils at a lower concentration of 2%. These findings indicated that the proposed strategy could be regarded as a potential method for edible oils quality and authenticity analysis.

Changes in the content of pollen total lipid and TAG in Arabidopsis thaliana DGAT1 mutant as11.

In mature pollen grains, lipids are primarily stored in the form of lipid droplets that provide energy and act as a carbon source for normal pollen development and germination. Triacylglycerol (TAG) is the major form of stored plant lipids. Diacylglycerol transferase, which is encoded by DGAT1 in Arabidopsis thaliana, is an important enzyme regulating triacylglycerol synthesis. Within the seeds of the DGAT1 mutant as11, the content of TAG is significantly decreased and the fatty acid composition also differs from the wild type. Transcriptome data of mature anthers showed that the genes involved in the TAG synthesis pathway were downregulated in as11. Analysis of gene expression patterns via transcriptome data also revealed that the expression of PDAT1, which functions in a manner complementary to the DGAT1 gene, was significantly decreased in as11, whereas the amylopectin synthase genes SS1 and SS2 were upregulated in mutant as11. We also detected lower total lipid, TAG and fatty acid contents in mature as11 pollen, with palmitic acid (C16:0) and linolenic acid (C18:3) being the major fatty acids in mature pollen. The cytological results showed that the lipid droplet content was reduced in mature as11 pollen. In the binuclear pollen grain II stage, WT pollen contained lipid droplets that were primarily accumulated around the generative nucleus, whereas the pollen in the mutant as11 was rich in starch grains that were primarily distributed around the vegetative nucleus. Ultrastructural analysis indicated that during pollen development in as11, the amount of endoplasmic reticulum in tapetal cells and pollen grains decreased, whereas the Golgi body content increased, which directly or indirectly led to a decrease in the levels of lipidosomes and an increase in the starch content in as11. Changes in the lipid content and fatty acid composition of the mutant as11 differ from those in the wild type during pollen development.

Severe gestational diabetes mellitus in lean dams is associated with low IL-1α levels and affects the growth of the juvenile mouse offspring

We investigated how maternal gestational diabetes (GDM) impacts the metabolic status of offspring. GDM was induced in CD1 mice consuming a fast-food diet (FFD) by repeated low-dose streptozotocin injections before mating. Offspring of normoglycemic standard chow or the FFD consuming dams served as controls. In 4-week-old offspring weaned to standard chow, plasma concentrations of extracellular DNA, inflammatory markers, and parameters of the cardiometabolic status (glycemia, liver lipid content; body, organ, and fat weight) were determined. Two-factor analysis of variance indicated that the male offspring of GDM dams manifest postnatal growth retardation and lower relative kidney weight. Regardless of sex, GDM offspring manifest the lowest IL-1α levels, and other inflammatory markers showed mild and inconsistent alterations. Offspring of dams consuming the FFD displayed higher liver triacylglycerols content. The three groups of offspring showed no significant differences in glycemia and extracellular DNA. Partial least squares-discriminant analysis indicated that male GDM offspring present lower kidney, body, and brown adipose tissue weights; lower IL-1α levels, and higher concentrations of GM-CSF and IL-10 compared with their FFD counterparts. The model failed to select discriminative variables in females. In conclusion, in mice, maternal GDM in the absence of obesity adversely affects the early growth of juvenile male offspring.

Enriched functional milk fat ameliorates glucose intolerance and triacylglycerol accumulation in skeletal muscle of rats fed high-fat diets.

We examined the effect of a functional milk fat (FMF) on the glucose metabolism and its association with the intramuscular triacylglycerol (TAG) content in rats fed high-fat diets. Male Wistar rats were fed for 60days with S7 (soybean oil 7%), S30 (soybean oil 30%), MF30 (soybean oil 3% + milk fat 27%), or FMF30 (soybean oil 3% + FMF 27%) diets. An oral glucose tolerance test was performed. The levels of key metabolites in gastrocnemius muscle and mRNA levels of genes involved in glucose and lipid metabolism in muscle, epididymal white adipose tissue (EWAT), and serum were assessed. The S30 diet induced glucose intolerance and led to TAG, citrate, and glucose accumulation in muscle. Moreover, we observed a downregulation of uncoupling proteins (Ucp2 and Ucp3) and insulin receptor substrate-1 (Irs1) genes, lower carnitine palmitoyl transferase-1b (CPT-1b), and phosphofructokinase-1 (PFK1) activities in muscle and lower expression of adiponectin (Adipoq) in EWAT. The FMF30 diet ameliorated the glucose intolerance and normalized the glucose and TAG levels in muscle, preventing the accumulation of citrate and enhancing glucose utilization by the PFK1. The beneficial effects might also be related to the higher expression of Adipoq in EWAT, its receptor in muscle (Adipor1), and the expression of Ucp2, Ucp3, and Irs1 in muscle, restoring the alterations observed with the S30 diet. FMF30 modulated key genes involved in glucose and lipid metabolism in skeletal muscle, improving the glucose utilization and preventing TAG, glucose, and citrate accumulation.

Expression profile of genes associated with the accumulation of triacylglycerol (TAG) in Auxenochlorella protothecoides KP7 under alkaline pH and nitrogen starvation conditions

Identification of target genes related to lipid accumulation in microalgae can increase triacylglycerol production without affecting biomass production through genetic manipulation. This study is part of our previous study and aimed to determine the effects of nitrogen starvation, alkaline pH (pH 10), and a combination of these factors on the transcriptional profiles of eight genes potentially related to triacylglycerol accumulation in Auxenochlorella protothecoides KP7. As shown previously, triacylglycerol content in Auxenochlorella protothecoides KP7 reached the maximum level under combined stress conditions. In this study, the highest upregulation for the gene encoding phosphoenolpyruvate carboxylase (PEPC) was observed under both nitrogen starvation and combined stress. Nitrogen starvation alone resulted in a significant increase in the expression level of all genes except NADP-malic enzyme (MEchl), which is localized in the chloroplast. Compared with nitrogen starvation alone, the upregulation of pyruvate dehydrogenase complex E2 component (PDHC-E2), glycerol-3-phosphate dehydrogenase (GPDH), diacylglycerol acyltransferase 1 (DGAT1), and MEchl genes further increased under the combined stress. These results suggest that genes encoding the enzymes DGAT1, GPDH, ME, PEPC, and PDHC-E2 are associated with triacylglycerol accumulation and could be used as target genes for genetic modification of Auxenochlorella protothecoides and possibly other microalgal species.

Effect of starvation and pesticide exposure on neutral lipid composition of the digestive gland of males of the apple snails Pomacea canaliculata

Pollutants as well as starvation usually modify homeostasis of neutral lipids in aquatic organisms. However, studies on the simultaneous effects of both stressors are scarce. The aim of this study was to evaluate the effect of toxicant exposure under starvation conditions on neutral lipids of the freshwater snail Pomacea canaliculata, selected as the model organism. Starved adult male snails were exposed to sublethal concentration of the pesticide cypermethrin (100 µg/L) during 4 and 10 days. Fed snails were sacrificed at the onset of the experiment (T0), along with starved snails exposed to the pesticide vehicle (ethanol) and another group without solvent served as controls. Total lipid content, neutral lipid classes, fatty acid composition, and pesticide accumulation were determined in the digestive gland of snails. The ethanol concentration used was not an additional stressful agent. As expected, starvation caused a decrease in neutral lipid content in the digestive gland of snails with respect to T0 snails. Pesticide exposure caused, on the other hand, an increase in triacylglycerol content compared to ethanol exposure at day 10 of the bioassay. This increment correlated with the bioconcentration of cypermethrin, which was 47% higher by day 10 than by day 4. The fatty acid profile of triacylglycerols in the digestive gland was significantly altered under starvation and pesticide exposure. Stressed male snails showed the ability to preserve polyunsaturated fatty acids, as evidenced by their significant increase with respect to T0 snails. These results suggest that the alteration of lipid homeostasis could be involved in an adaptive mechanism of aquatic organisms to lipophilic and obesogenic pollutants.

Preparation of γ‐linolenic acid rich triacylglycerol from borage oil via a two‐step lipase‐catalyzed reaction

Abstractγ‐Linolenic acid (GLA) rich triacylglycerol (TAG) was successfully synthesized from glyceride, instead of glycerol, and fatty acid (FA) via Lipozyme TL IM‐catalyzed esterification as a novel strategy. In the first step, GLA was enriched into glyceride fraction from borage oil by Candida rugosa lipase‐catalyzed hydrolysis. The glyceride was separated from the reaction mixture by molecular distillation. GLA was enriched from 20.64% in borage oil to 45.94% in the glyceride fraction under optimum conditions. In the second step, the Lipozyme TL IM‐catalyzed synthesis of TAG was carried out with the glyceride, and the FA obtained by saponification of a portion of the glyceride. The optimum conditions were the temperature of 50°C, the enzyme loading of 10%, and the vacuum level of 20 mmHg, respectively. The maximum TAG content of approximately 92% was achieved after 12 h under the optimum conditions.

Goat FADS2 controlling fatty acid metabolism is directly regulated by SREBP1 in mammary epithelial cells.

Goat milk provides benefits to human health due to its richness in bioactive components, such as polyunsaturated fatty acids (PUFAs). The fatty acid desaturase 2 (FADS2) is the first rate-limiting enzyme in PUFAs biosynthesis. However, its role and transcriptional regulation mechanisms in fatty acid metabolism in dairy goat remain unclear. Here, our study revealed that the FADS2 gene was highly expressed during the peak lactation compared with the dry period, early lactation, and late lactation. The content of triacylglycerol (TAG) was enhanced with the increasing mRNA expression of TAG synthesis genes (diacylglycerol acyltransferase 1/2, DGAT1/2) in FADS2-overexpressed goat mammary epithelial cells (GMECs). Overexpression of FADS2 was positively correlated with the elevated concentrations of dihomo-gamma-linolenic acid (DGLA) and docosahexaenoic acid (DHA) in GMECs. BODIPY staining showed that FADS2 promoted lipid droplet accumulation in GMECs. To clarify the transcriptional regulatory mechanisms of FADS2, 2,226 bp length of FADS2 promoter was obtained. Deletion mutation assays revealed that the core region of FADS2 promoter was located between the -375 and -26 region, which contained SRE1 (-361/-351) and SRE2 (-191/-181) cis-acting elements of transcription factor sterol regulatory element-binding protein 1 (SREBP1). Overexpression of SREBP1 enhanced relative luciferase activity of the single mutant of SRE1 or SRE2, vice versa, and failed to alter the relative luciferase activity of the joint mutant of SRE1 and SRE2. Chromatin immunoprecipitation (ChIP) and site-directed mutation assays further demonstrated that SREBP1 regulated the transcription of the FADS2 gene by binding to SRE sites in vivo and in vitro. In addition, the mRNA levels of FADS2 were significantly decreased by targeting SRE1 and SRE2 sites in the genome via the CRISPR interference (CRISPRi) system. These findings establish a direct role for FADS2 regulating TAG and fatty acid synthesis by SREBP1 transcriptional regulation in dairy goat, providing new insights into fatty acid metabolism in mammary gland of ruminants.

FEATURES OF CARBOHYDRATE-LIPID METABOLISM IN DOGS WITH DIFFERENT TYPES OF HIGHER NERVOUS ACTIVITY

The relevance of the study is due to the lack of data on the cortical mechanisms of regulation of carbohydrate and lipid metabolism in the body of dogs. In this regard, this article is aimed at revealing the issue of lipid and carbohydrate metabolism in dogs with various types of higher nervous activity under the influence of short-term food deprivation. It is shown that the effect of short-term food deprivation is characterized by changes in metabolism in the body of dogs, which are limited by the state of the nervous system of these animals. In the intact state, the glucose content in the blood of dogs with different types of higher nervous activity does not reliably differ, while the lactate content in the blood of dogs with a weak type of higher nervous activity is higher by 16.2% (P < 0.01), and pyruvate is lower by 6.3% (P < 0.001) from the indicators of dogs with a strong balanced mobile type. Within a day after the beginning of food deprivation, the glucose content in the blood of dogs decreases by 4.6–8.2% depending on the type of nervous activity (P < 0.05–0.01). During the day after the beginning of deprivation, the ratio of lactate to pyruvate in the blood of dogs, depending on the type of higher nervous activity, increases by 19.1–36.0% (Р < 0.01), the content of total cholesterol and triacylglycerols in the blood plasma decreases by 3, 5–12.9% and the ratio of lipids of different densities changes significantly. In particular, in the blood plasma of dogs of a strong balanced mobile type of higher nervous activity, the cholesterol content of high-density lipoproteins decreased during the day by 7.1% (Р < 0.05). Thanks to the conducted research, it was possible to obtain fundamental knowledge of the cortical mechanisms of regulation of carbohydrates and lipids in the body of dogs, which will form the basis of the development of new, modern methods of correction of metabolism, taking into account the type of higher nervous activity. Prospects for further research consist in the development of new methods of correction of metabolism based on the use of nanoaquachelates of biogenic metals, taking into account the individual characteristics of the body of dogs.

Serum Cholesterol Differences Between Statin Users Who Take Dietary Supplements and Those Who Do Not: NHANES 2013–2018

BackgroundCardiovascular disease (CVD) is the leading cause of mortality in the United States and statins are the most commonly prescribed medication. It is important to understand the potential impact supplements may have when taken in combination with statins on serum lipid outcomes. ObjectivesTo evaluate the differences in the concentrations of cholesterol, triacylglycerol (TAG), and HbA1c between adults who use statins alone and those who combine statins and dietary supplements. MethodsA cross-sectional analysis using data from US adults aged ≥20 years who participated in the NHANES (2013–2018). The serum concentrations of lipids and the HbA1c levels were compared using independent sample t-tests. All analyses were adjusted for the complex survey design and used appropriate sample weights. ResultsOf 16,327 participants included in this analysis, 13% reported the use of statins alone, and 8.8% used statins and dietary supplements. Statin users who used dietary supplements tended to be women (50.5%), aged 65.8 ± 0.4 years, and were more likely to be White (77.4%). Participants who used statins in combination with dietary supplements were less likely to have higher levels of total cholesterol (5.1% ± 1.4% vs. 15.6% ± 2.7%, P < 0.001), HbA1c (6.0% ± 0.1% vs. 6.3% ± 0.1%, P < 0.05), and HDL cholesterol (50 ± 1.3 vs. 47 ± 0.8 mg/dL, P < 0.05) than those who used statins alone. No significant differences were identified between the two groups for LDL cholesterol and TAG concentrations. ConclusionsStatin users who coingested dietary supplements were less likely to have high levels of total cholesterol and HbA1c and greater HDL levels than statin users who did not take dietary supplements. Dietary intake, lifestyle choices, and other confounders may have influenced the observed outcome differences for those who took dietary supplements with statins and those who did not.

C/EBPα promotes triacylglycerol synthesis via regulating PPARG promoter activity in goat mammary epithelial cells.

CCAAT/enhancer binding protein α (C/EBPα) is the key transcription factor involved in lipid metabolism, however, the role of C/EBPα in milk fat synthesis of dairy goats remains unknown. The objective of the present research was to clarify the function of C/EBPα in goat mammary epithelial cells (GMECs) and its impact on peroxisome proliferator-activated receptor gamma (PPARG) promoter activity. In this study, C/EBPα overexpression increased its mRNA and protein levels by 42-fold and 6-fold, respectively. In contrast, transfecting siRNA targeting C/EBPα decreased its mRNA level to 20% and protein abundance to 80% of the basal level. The contents of lipid droplets, triacylglycerol (TAG), and cholesterol were increased (P < 0.05) in C/EBPα-overexpressing GMECs, and knockdown of C/EBPα led to the opposite results. Overexpression of C/EBPα significantly increased the expression levels of genes involved in TAG synthesis (AGPAT6, DGAT2, P < 0.01), lipid droplet formation (PLIN2, P < 0.01), and fatty acid synthesis (FADS2, P < 0.05; ELOVL6, P < 0.01). Knockdown of C/EBPα decreased (P < 0.05) the expression levels of AGPAT6, DGAT1, DGAT2, PLIN2, FADS2, and ELOVL6. C/EBPα upregulated the expression level of PPARG (P < 0.05), and four C/EBPα binding regions were identified in the PPARG promoter at -1,112 to -1,102 bp, -734 to -724 bp, -248 to -238 bp, and -119 to -109 bp. Knockdown of C/EBPα reduced (P < 0.05) the PPARG promoter activity when the C/EBPα binding regions were mutated at -1,112 to -1,102 bp, -734 to -724 bp, and -248 to -238 bp locations of the promoter. However, the promoter activity did not change when the mutation was located at -119 bp. In conclusion, our results suggest that C/EBPα can promote TAG synthesis in GMECs through its effects on mRNA abundance of genes related to lipid metabolism and regulation of the PPARG promoter activity via C/EBPα binding regions.

Functional and miRNA regulatory characteristics of INSIG genes highlight the key role of lipid synthesis in the liver of chicken (Gallus gallus)

The insulin-induced genes (INSIG1 and INSIG2) have been demonstrated to play a vital role in regulating lipid metabolism in mammals, however the function and regulation mechanism of them remains unknown in poultry. In this study, firstly the phylogenetic trees of INSIGs among various species were constructed and their subcellular locations were mapped in chicken LMH. Then the spatiotemporal expression profiles, over-expression and knockdown assays of chicken INSIGs were conducted. Furthermore, conservation of potential miRNA binding sites in INSIGs among species were analyzed, and the miRNA biological function and regulatory role were verified. The results showed that chicken INSIGs located in cellular endoplasmic reticulum, and were originated from the common ancestors of their mammalian counterparts. The INSIGs were widely expressed in all detected tissues, and their expression levels in the liver of chicken at 30 wk were significantly higher than that at 20 wk (P < 0.01). Over-expression of INSIGs led no significant increase in mRNA abundance of lipid metabolism-related genes and the contents of triacylglycerol (TG) and cholesterol (TC) in LMH cells. Knockdown of INSIG1 led to the decreased expressions of ACSL1, MTTP-L, ApoB, ApoVLDLII genes and TG, TC contents (P < 0.05). Knockdown of INSIG2 could significantly decrease the contents of TG and TC, and expressions of key genes related to the lipid metabolism (P < 0.05). Moreover, INSIG1 was directly targeted by both miR-130b-3p and miR-218-5p, and INSIG2 was directly targeted by miR-130b-3p. MiR-130b-3p mimic and miR-218-5p mimic treatment could significant decrease the mRNA and protein levels of INSIGs, mRNA levels of genes related to lipid metabolism, and the contents of TG and TC in LMH cells. The inhibition of miR-130b-3p and miR-218-5p on TG and TC contents could be restored by the overexpression of INSIGs, respectively. No significant alteration in expressions of sterol regulatory element binding protein (SREBPs) and SREBP cleavage-activating protein (SCAP) were observed when INSIGs were over-expressed. SCAP was down-regulated when INSIG1 was knocked down, while SREBP1 was down-regulated when INSIG2 was knocked down. Taken together, these results highlight the role of INSIG1 and INSIG2 in lipid metabolism and their regulatory mechanism in chicken.

Effect of Acute Walking on Endothelial Function and Postprandial Lipemia in South Asians and White Europeans.

South Asians (SAs) have an elevated risk of cardiovascular disease (CVD) compared with White Europeans (WEs). Postprandial endothelial function (flow-mediated dilatation (FMD%)) in SA women and SA men with central obesity has not been investigated. Research in other populations has highlighted that a 1% higher FMD% is associated with a ~13% lower risk of future CVD events. We investigated whether FMD% and lipemia, two markers for CVD risk, were higher in SAs versus WEs, whether walking improved FMD% and lipemia, and if there were ethnic differences in the response. Lean premenopausal women (study 1; 12 SA, 12 WE) and men with central obesity (study 2; 15 SA, 15 WE) completed two 2-d trials. On day 1, participants walked for 60 min at 60% of their peak oxygen uptake or rested. On day 2, participants rested and consumed two high-fat meals over 8 h. Repeated ultrasound assessments of endothelial function and venous blood samples for CVD risk markers were taken. Compared with WEs, SAs had lower postprandial FMD% (study 1, -1.32%; study 2, -0.54%) and higher postprandial triacylglycerol concentrations (study 1, 0.31 mmol·L -1 ·h -1 ; study 2, 0.55 mmol·L -1 ·h -1 ). Walking improved postprandial FMD% (study 1, 1.12%; study 2, 0.94%) and resulted in no significant change or small reductions in postprandial triacylglycerol concentrations (study 1, -0.01 mmol·L -1 ·h -1 ; study 2, -0.25 mmol·L -1 ·h -1 ). Exercise-induced changes in FMD% and triacylglycerol were consistent between ethnic groups. Walking mitigated the adverse postprandial effect of a high-fat diet on FMD% to a similar extent in SA and WE women and men, even with no/small improvements in triacylglycerol. This study highlights the importance of exercise to clinically improve FMD% in SAs and WEs.

Fatty acid exporter 1 enhances seed oil content in Brassica napus.

The online version contains supplementary material available at 10.1007/s11032-022-01346-0.

Distinct subcellular localisation of intramyocellular lipids and reduced PKCε/PKCθ activity preserve muscle insulin sensitivity in exercise-trained mice.

Athletes exhibit increased muscle insulin sensitivity, despite increased intramuscular triacylglycerol content. This phenomenon has been coined the 'athlete's paradox' and is poorly understood. Recent findings suggest that the subcellular distribution of sn-1,2-diacylglycerols (DAGs) in the plasma membrane leading to activation of novel protein kinase Cs (PKCs) is a crucial pathway to inducing insulin resistance. Here, we hypothesised that regular aerobic exercise would preserve muscle insulin sensitivity by preventing increases in plasma membrane sn-1,2-DAGs and activation of PKCε and PKCθ despite promoting increases in muscle triacylglycerol content. C57BL/6J mice were allocated to three groups (regular chow feeding [RC]; high-fat diet feeding [HFD]; RC feeding and running wheel exercise [RC-EXE]). We used a novel LC-MS/MS/cellular fractionation method to assess DAG stereoisomers in five subcellular compartments (plasma membrane [PM], endoplasmic reticulum, mitochondria, lipid droplets and cytosol) in the skeletal muscle. We found that the HFD group had a greater content of sn-DAGs and ceramides in multiple subcellular compartments compared with the RC mice, which was associated with an increase in PKCε and PKCθ translocation. However, the RC-EXE mice showed, of particular note, a reduction in PM sn-1,2-DAG and ceramide content when compared with HFD mice. Consistent with the PM sn-1,2-DAG-novel PKC hypothesis, we observed an increase in phosphorylation of threonine1150 on the insulin receptor kinase (IRKT1150), and reductions in insulin-stimulated IRKY1162 phosphorylation and IRS-1-associated phosphoinositide 3-kinase activity in HFD compared with RC and RC-EXE mice, which are sites of PKCε and PKCθ action, respectively. These results demonstrate that lower PKCθ/PKCε activity and sn-1,2-DAG content, especially in the PM compartment, can explain the preserved muscle insulin sensitivity in RC-EXE mice.

Metabolomic characterization of congenital microtia: a possible analysis for early diagnosis.

Although metabolic abnormalities have been deemed one of the essential risk factors for growth and development, the relationship between metabolic abnormalities and microtia is still unclear. In this study, we aimed to establish a cell model of microtia and the changes of serum metabolites in patients with microtia. After constructing a cell model of microtia with low expression of BMP5, we performed integrative metabolomics analysis. For the altered metabolites, the content of glycerophosphocholine (PC), triacylglycerol (TG), and choline in the serum of 28 patients (15 patients with microtia and 13 controls) with microtia was verified by enzyme-linked immunosorbent assay (ELISA). Detailed metabolomic evaluation showed distinct clusters of metabolites between BMP5-low expressing cells and normal control (NC) cells. The cell model of microtia had significantly higher levels of TG, PC, glycerophosphoethanolamine (PE), sphingomyelin, sulfatide, glycerophosphoglycerol, diacylglycerol, and glycosphingolipid. The main abnormal metabolites were mainly concentrated in the glycerophospholipid metabolism pathway, and PC and choline were closely related. In the serum of patients with microtia, the contents of PC, TG, and choline were significantly increased. The individual serum samples confirmed the different metabolites between patients with microtia and controls. In particular, we showed that a newly developed metabolic biomarker panel has a high sensitivity and specificity for separating patients with microtia from controls.

Predictive value of the complement factors B and H for women with gestational diabetes mellitus who are at risk of preeclampsia

We aimed to define the relationships between the serum concentrations of complement factors B and H in mid-pregnancy and the risk of preeclampsia (PE) in patients with gestational diabetes mellitus (GDM). We performed a prospective nested case-control study of 503 patients with GDM who attended Peking University Third Hospital between March 2018 and December 2018. 456 patients were followed until delivery and blood samples were collected between gestational weeks 24 and 28. Thirty patients developed PE, 12 developed gestational hypertension (GH), and 42 matched cases were selected as a control group. The incidence of PE was 5.96%. The serum concentrations of triacylglycerol (TG), FB, and FH of women with GDM who developed PE (GDM-PE group) in mid-pregnancy were significantly higher than those of the GDM group [TG: 3.60 (2.94-4.63) vs 2.54 (2.14-3.01) mmol/L, p<0.001; FB: 346 (314-378) vs 284 (263-323) mg/L, p<0.001; FH: 417±45 vs 379±47mg/L, p=0.003]. Multivariate regression analysis showed that high serum concentrations of TG and FB in mid-pregnancy were related to the risk of patients with GDM developing PE [TG: odds ratio (OR) 2.035 (95% confidence interval (CI) 1.032-4.013); FB: OR 1.018 (95% CI 1.001-1.035)]. The area under the curve (AUC) of FB for the prediction of PE was 0.821 (95% CI 0.722-0.921) and that for a combination of TG, FB, and FH was 0.857 (95% CI 0.770-0.944). High serum FB concentration during mid-pregnancy is a potential predictor of the risk of PE in patients with GDM, and the combination of FB, FH, and TG increased this predictive value.

Intervalos de referência bioquímica para Crocodylus moreletii e Crocodylus acutus criados em cativeiro na Lagoa Alcuzahue, Colima México

ABSTRACT Crocodylus moreletii and Crocodylus acutus are two endangered crocodile species endemic to Mexico. In this descriptive cross-sectional study, a total of 58 crocodiles (30 females and 28 males) were examined to determine and evaluate 24 blood biochemical indicators associated with energy, lipid, protein, mineral and enzymatic metabolic processes. Most of the serum biochemistry parameters were similar among sexes by species. However, male crocodiles showed higher triacylglycerol concentration and alkaline phosphatase activity, and lower globulin concentration than females. There were some significant differences between species. Total bilirubin, triacylglycerol, carbon dioxide, and hydrogen carbonate were higher in Crocodylus moreletii, and urea nitrogen, creatinine, alanine aminotransferase, and aspartate aminotransferase were higher in Crocodylus acutus. These reference values are very important for the protection of crocodiles. The calculated confidence intervals could be used to detect alert situations when at least 5% of the sampled crocodiles would fall outside of the calculated reference interval for a given parameter.