Hydroxy Fatty Acids Research Articles

Serum metabolite profiles of thyroid autoimmunity patients in early pregnancy.

Research on serum metabolite profiles in thyroid autoimmunity (TAI) patients during early pregnancy is currently limited. The current study aimed to identify differential serum metabolites and assess the relationship between pregnancy outcomes and metabolic abnormalities in individuals with TAI. This research included 26 pregnant women with TAI and 30 healthy controls (HC). We employed a liquid chromatograph mass spectrometer (LC-MS) to analyze changes between the two groups. Newborns in the TAI patients had lower birth weights than those in the control group (P = 0.007). We identified 92 differential metabolites (including 50 upregulated and 42 downregulated) belonging to amino acids, fatty acyls, glycerophosphocholines, steroid and other categories and four significantly enrichment Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways including taurine and hypotaurine metabolism, citrate cycle (TCA cycle), glyoxylate and dicarboxylate metabolism and 2-oxocarboxylic acid metabolism. We further identified 15 characteristic metabolites (6-Methylquinoline, D-erythrose 4-phosphate, 4-Hydroxyisoleucine, phosphatidylcholine (PC)(16:2e/16:0), N3,N4-Dimethyl-L-arginine, N-desmethyltramadol, 3-Methoxybenzaldehyde, sphingomyelin (SM)(d14:3/28:2), gamma-Glutamylleucine, NSI-189, 3-(1-cyano-1,2-dihydroisoquinolin-2-yl)-3-oxopropyl propionate, lysophosphatidylinositol (LPI) 16:0, cis-Aconitic acid, polyamide (PA)(18:1/18:2) and fatty acyl esters of hydroxy fatty acid (FAHFA)(17:0/18:0)) using least absolute shrinkage and selection operator (LASSO) regression. Correlation analyses revealed that 6-Methylquinoline, D-erythrose 4-phosphate, gamma-Glutamylleucine, and LPI 16:0 exhibited a positive correlation with anemia before delivery, while 3-(1-cyano-1,2-dihydroisoquinolin-2-yl)-3-oxopropyl propionate had a negative correlation. LPI 16:0 displayed a positive correlation with uric acid (UA) during both middle and late pregnancy, whereas 3-Methoxybenzaldehyde exhibited a negative correlation with UA in late pregnancy. Cis-Aconitic acid showed a positive correlation with fasting blood glucose (FBG) in middle pregnancy. Conversely, 6-Methylquinoline and 4-Hydroxyisoleucine had a negative correlation with birth weight. Thyroid autoantibodies were found to be associated with 14 metabolites identified using LASSO, with the exception of PA (18:1/18:2). Our findings provide new evidence supporting the early screening of serum metabolites and their potential for predicting adverse pregnancy outcomes in women with TAI.

Discovery and Molecular Characterization of a Novel 9S-Lipoxygenase from Enhygromyxa salina for Fatty Acid Biotransformations.

Iron-dependent lipoxygenases (LOXs) are involved in the synthesis of oxylipins from polyunsaturated fatty acids. However, they are usually difficult to overexpress in functional form in microbial cell factories. Moreover, 9-LOXs, generating 9-hydroperoxy fatty acids from C18 polyunsaturated fatty acids, have rarely been found from microbial sources. Here, we discovered a novel 9S-LOX in the marine myxobacterium Enhygromyxa salina (Es-9S-LOX). The recombinant enzyme produced in Escherichia coli exhibited remarkable activity in the dioxygenation of linoleic acid (LA, 1), α-linolenic acid, γ-linolenic acid, and arachidonic acid specifically at the C9 position to yield the product with (S)-configuration at catalytic efficiency of 3.94, 1.42, 1.38, and 0.69 μM-1·s-1, respectively. The elucidated X-ray crystal structure of Es-9S-LOX reveals a long and narrow hydrophobic pocket that allows the substrate to be near the metal ion and the oxygen tunnel. The enzyme was successfully used in a chemoenzymatic reaction to generate a hydroxy fatty acid from LA. Our study thus contributes to the valorization of renewable polyunsaturated fatty acids into a variety of fatty acid derivatives, including hydroxy fatty acids.

Comprehensive overview of host plant resistance and its progress in wilt resistance in castor bean (Ricinus communis L.)

Castor bean, a non-edible monotypic species known for its unique hydroxy fatty acid, is susceptible to a range of pathogens. Wilt caused by Fusarium oxysporum f. sp. ricini is the most complex and destructive disease. Chemical control of wilt has proven ineffective due to the systemic nature of the disease and the seed and soil borne nature of the pathogen. Physical, chemical, biological, and integrated management methods have shown only limited success in controlling wilt. Host plant resistance, however, stands out as the most promising strategy, offering a viable pathway for the genetic improvement of castor beans for wilt resistance. Screening techniques are well established, and several resistant donors have already been identified. While significant progress has been made in understanding the inheritance of wilt resistance, a complete understanding of its genetic mechanisms still requires further research. Single Nucleotide Polymorphism (SNP) markers and genomic regions linked to wilt resistance have been successfully identified. Reniform nematode (Rotylenchulus reniformis) has been identified as a predisposing factor for wilt, and genomic regions linked to nematode resistance have also been identified. However, Marker-assisted selection has not yet been utilized to develop wilt-resistant castor varieties. Further research is required to explore the pathogen diversity, host-pathogen interactions, and mechanisms underlying wilt resistance including the metabolites responsible for preventing the emergence of new pathogenic races and ensuring long-term protection against wilt.

Metabolic abnormalities and reprogramming in cats with naturally occurring hypertrophic cardiomyopathy.

The heart is a metabolic organ rich in mitochondria. The failing heart reprograms to utilize different energy substrates, which increase its oxygen consumption. These adaptive changes contribute to increased oxidative stress. Hypertrophic cardiomyopathy (HCM) is a common heart condition, affecting approximately 15% of the general cat population. Feline HCM shares phenotypical and genotypical similarities with human HCM, but the disease mechanisms for both species are incompletely understood. Our goal was to characterize global changes in metabolome between healthy control cats and cats with different stages of HCM. Serum samples from 83 cats, the majority (70/83) of which were domestic shorthair and included 23 healthy control cats, 31 and 12 preclinical cats with American College of Veterinary Internal Medicine (ACVIM) stages B1 and B2, respectively, and 17 cats with history of clinical heart failure or arterial thromboembolism (ACVIM stage C), were collected for untargeted metabolomic analysis. Multiple linear regression adjusted for age, sex and body weight was applied to compare between control and across HCM groups. Our study identified 1253 metabolites, of which 983 metabolites had known identities. Statistical analysis identified 167 metabolites that were significantly different among groups (adjusted P<0.1). About half of the differentially identified metabolites were lipids, including glycerophospholipids, sphingolipids and cholesterol. Serum concentrations of free fatty acids, 3-hydroxy fatty acids and acylcarnitines were increased in HCM groups compared with control group. The levels of creatine phosphate and multiple Krebs cycle intermediates, including succinate, aconitate and α-ketoglutarate, also accumulated in the circulation of HCM cats. In addition, serum levels of nicotinamide and tryptophan, precursors for de novo NAD+ biosynthesis, were reduced in HCM groups versus control group. Glutathione metabolism was altered. Serum levels of cystine, the oxidized form of cysteine and cysteine-glutathione disulfide, were elevated in the HCM groups, indicative of heightened oxidative stress. Further, the level of ophthalmate, an endogenous glutathione analog and competitive inhibitor, was increased by more than twofold in HCM groups versus control group. Finally, several uremic toxins, including guanidino compounds and protein bound putrescine, accumulated in the circulation of HCM cats. Our study provided evidence of deranged energy metabolism, altered glutathione homeostasis and impaired renal uremic toxin excretion. Altered lipid metabolism suggested perturbed structure and function of cardiac sarcolemma membrane and lipid signalling.

Deducing formation routes of oxylipins by quantitative multiple heart-cutting achiral-chiral 2D-LC-MS

Several oxylipins are regulators of inflammation. They are formed by enzymes such as lipoxygenases or cyclooxygenases, but also stereorandomly by autoxidation. Reversed-phase liquid chromatography-tandem-mass-spectrometry (LC-MS/MS) methods for oxylipin quantification do not separate enantiomers. Here, we combine sensitive and selective oxylipin analysis with chiral separation using two-dimensional (2D)-LC-MS/MS. By multiple heart-cutting, the oxylipin peaks are transferred onto a chiral column. 45 enantiomeric pairs of (di-)hydroxy-fatty acids are separated with full gradient elution within 1.80min, yielding lower limits of quantification <1pg on column. Concentrations as well as enantiomeric fractions of oxylipins can be determined, even at low concentrations or at high enantiomeric excess of one isomer. The developed achiral-chiral multiple heart-cutting 2D-LC-MS/MS method offers unprecedented selectivity, enabling a better understanding of the formation route of these lipid mediators. This is demonstrated by distinguishing the formation of hydroxy-fatty acids by (acetylated) cyclooxygenase-2 and radical-mediated autoxidation. Applying the method to human M2-like-macrophages, we show that the so-called specialized pro-resolving mediators (SPM) 5,15-DiHEPE and 7,17-DiHDHA as well as 5,15-DiHETE were present as (S,S)-enantiomers, supporting their enzymatic formation. In contrast, at least eight isomers (including protectin DX but not neutroprotectin D1) of 10,17-DiHDHA are present in immune cells, indicating formation by autoxidation. In human plasma of healthy subjects, none of these dihydroxy-fatty acids are not present. However, we demonstrate that all four isomers quickly form via autoxidation if the samples are stored improperly. Thus, dihydroxy-FA should only be reported as SPM, such as resolvin D5 or resolvin E4, if an enantioselective analysis has been carried out.

Exploring the role of fatty acid esters of hydroxy fatty acids in metabolic dysfunction-associated steatotic liver disease in morbidly obese women

BackgroundFatty acid esters of hydroxy fatty acids (FAHFAs) present potential beneficial effects that could offer valuable insights into metabolic and inflammatory diseases. However, few FAHFAs have been studied, and their role is unclear. AimsTo assess FAHFA levels in individuals with metabolic dysfunction-associated steatotic liver disease (MASLD) associated with morbid obesity (MO) to explore the potential significance of FAHFAs under these conditions. MethodsUsing ultra-precise liquid chromatography, FAHFA serum levels were measured in 219 women, including 53 with normal weight (NW) and 166 with MO. The MO group was classified by histological diagnosis into 35 normal liver (NL), 38 simple steatosis (SS) and 93 metabolic dysfunction-associated steatohepatitis (MASH) groups. ResultsThirty-two FAHFA isoforms from 11 families were identified. Most FAHFAs presented low levels in MO, but tLAHOAs, LAHOA-1 and OAHOA-1 increased. In MASLD, elevated tLAHPO, LAHPO-2, PAHLA-3 and PAHLA-4 levels were observed. In MASH, increased POHLA-1, tLAHPOs, LAHPO-2 and LAHLA-3 and decreased PAHSA-1, tOAHOAs, OAHSA-2 and OAHSA-3 levels were reported. ConclusionThis study reveals novel insights into FAHFAs in a cohort of women with MO with MASLD. In MASLD, we reported only increased levels of certain FAHFAs. In MASH, we found a different profile that could be characteristic.

Domestication and engineering of pennycress (Thlaspi arvense L.): challenges and opportunities for sustainable bio-based feedstocks.

Pennycress, as an emerging oilseed crop with high oil content, faces challenges but offers potential for sustainable bioproducts; ongoing research aims to enhance its traits and quality. Pennycress (Thlaspi arvense L.) is an emerging oilseed crop with many advantages, such as high seed oil (27-39%) and monounsaturated fatty acid (55.6%) content, making it an attractive candidate to produce sustainable bioproducts. However, several challenges are associated with domesticating pennycress, including high silicle shatter, which reduces seed yield during harvest, non-uniformed germination rate and high contents of erucic acid and glucosinolates, which have adverse health effects on humans and animals. Pennycress, which can be easily and rapidly transformed using the floral dip method under vacuum, can achieve trait improvements. Ongoing research for pennycress domestication using mutation breeding, including ethylmethylsulfonate treatment and genome editing, aims to improve its quality. Pennycress can be used as an excellent platform for producing industrially important fatty acids such as hydroxy and epoxy fatty acids and docosahexaenoic acid. In conclusion, pennycress is a promising oilseed crop with multiple advantages and potential applications. Continuous improvements in quality and engineering for producing high-value bio-based feedstocks in pennycress will establish it as a sustainable and economically valuable crop.

Expanding the biosynthesis spectrum of hydroxy fatty acids: unleashing the potential of novel bacterial fatty acid hydratases

BackgroundHydroxy fatty acids represent an emerging class of compounds with promising applications in the chemical, medicinal and functional food sectors. The challenges associated with their chemical synthesis have spurred exploration of biological synthesis as an alternative route, particularly through the use of fatty acid hydratases. Fatty acid hydratases catalyse the regioselective addition of a hydrogen atom and a hydroxyl group from a water molecule to the carbon–carbon cis-double bond of unsaturated fatty acids to form hydroxy fatty acids. Despite having been discovered in the early 1960s, previous research has primarily focused on characterizing single fatty acid hydratase variants with a limited range of substrates. Comprehensive studies that systematically examine and compare the characteristics of multiple variants of fatty acid hydratases are still lacking.ResultsIn this study, we employed an integrated bioinformatics workflow to identify 23 fatty acid hydratases and characterized their activities against nine unsaturated fatty acid substrates using whole-cell biotransformation assays. Additionally, we tested a dual-protein system involving two fatty acid hydratases of distinct regioselectivity and demonstrated its suitability in enhancing the biosynthesis of di-hydroxy fatty acids.ConclusionsOur study demonstrates that fatty acid hydratases can be classified into three subtypes based on their regioselectivity and provides insights into their preferred substrate structures. These understandings pave ways for the design of optimal fatty acid hydratase variants and bioprocesses for the cost-efficient biosynthesis of hydroxy fatty acids.

Distinguishing Artifactual Fatty Acid Dimers from Fatty Acid Esters of Hydroxy Fatty Acids in Untargeted LC-MS Pipelines.

Untargeted metabolomics is a powerful profiling tool for the discovery of possible biomarkers of disease onset and progression. Analytical pipelines applying liquid chromatography (LC) and mass spectrometry (MS)-based methods are widely used to survey a broad range of metabolites within various metabolic pathways, including organic acids, amino acids, nucleosides, and lipids. Accurate and complete identification of putative metabolites is an ongoing challenge in untargeted metabolomics studies. Highly sensitive instrumentation can result in the detection of adduct and fragment ions that form reproducibly and contain identifiable ions that are difficult to distinguish from metabolic pathway intermediates, which may result in false-positive identification. At concentrations as low as 10μM, free fatty acids have been found to form homo- and heterodimers in untargeted metabolomics pipelines that resemble the lipid class fatty acid esters of hydroxy fatty acids (FAHFAs), resulting in misidentification. This chapter details a protocol for LC-MS-based untargeted metabolomics using hydrophilic interaction chromatography (HILIC) that specifically aids in distinguishing artifactual fatty acid dimers from endogenous FAHFAs.

ENSO modulates soil organic carbon retention and deposition in the East China Sea

Accurately quantifying the contribution of terrestrial soil organic carbon (SOC) in marine environment is still a challenge. Moreover, reassessing the effect of climate change on SOC variability is necessary, with the increasing variability of El Niño under greenhouse warming. Here, we present new insights from bacterial 3-hydroxy fatty acids (3-OH-FAs) by systematically analyzing the distributions of 3-OH-FA in soils, lake/river sediments and marine sediments, and propose a novel proxy (RIA, the ratio of odd carbon number iso to anteiso 3-OH-FAs) to quantificationally evaluate the contribution of OC from different components. The reliability of the RIA proxy is tested by using a three end-member model and Monte Carlo simulations along a transect from the Yangtze River estuary to the ECS shelf. Then the RIA proxy is applied to a core in the East China Sea covering the past 700 years, and the results reveal that El Niño (La Niña) induced more (less) precipitation and increased (decreased) SOC deposition. This study provides a novel approach for quantifying terrigenous OC inputs and burial in marginal seas and shows the impact of El Niño-Southern Oscillation (ENSO) modes on SOC loss and burial in marginal seas under global warming.

Non-Targeted Metabolomics of Serum Reveals Biomarkers Associated with Body Weight in Wumeng Black-Bone Chickens.

Growth performance is an important economic trait of broilers but the related serum metabolomics remains unclear. In this study, we utilized non-targeted metabolomics using ultra-high-performance liquid phase tandem mass spectrometry (UHPLC-MS/MS) to establish metabolite profiling in the serum of Chinese Wumeng black-bone chickens. The biomarker metabolites in serum associated with growth performance of chickens were identified by comparing the serum metabolome differences between chickens that significantly differed in their weights at 160 days of age when fed identical diets. A total of 766 metabolites were identified including 13 differential metabolite classes such as lipids and lipid-like molecules, organic acids and their derivatives, and organoheterocyclic compounds. The results of difference analysis using a partial least squares discriminant analysis (PLS-DA) model indicated that the low-body-weight group could be differentiated based on inflammatory markers including prostaglandin a2, kynurenic acid and fatty acid esters of hydroxy fatty acids (FAHFA), and inflammation-related metabolic pathways including tryptophan and arachidonic acid metabolism. In contrast, the sera of high-body-weight chickens were enriched for riboflavin and 2-isopropylmalic acid and for metabolic pathways including riboflavin metabolism, acetyl group transfer into mitochondria, and the tricarboxylic acid (TCA) cycle. These results provide new insights into the practical application of improving the growth performance of local chickens.

Seasonal Variability of Lipophilic Compounds in Oat (Avena sativa L.) Straw: A Comprehensive Chemical Study.

Oat straw, a residue of Avena sativa L., is recognized for its abundance in cellulose, hemicelluloses, and lignin. However, its potential as a source of lipophilic compounds within the framework of a biorefinery concept still remains unexplored. In this study, we conducted an extensive investigation into the content and chemical composition of the lipophilic compounds present in acetone extracts from oat straws of two distinct oat varieties, namely, Karen and Isaura. Furthermore, we examined their seasonal variability in content and composition in straw samples from oats planted in both spring and winter seasons. The extracted lipophilic compounds were predominantly composed of high molecular weight esters (26.0-38.1%), steroids (16.6-24.0%), n-fatty alcohols (10.9-20.7%), n-fatty acids (10.9-16.0%), and n-aldehydes (10.7-15.8%), with lower amounts of n-alkanes (1.1-3.0%), acylglycerides (2.3-3.8%), phytol and phytyl esters (0.6-2.9%), β-diketones (0.1-2.5%), triterpenoids (0.9-1.2%), tocopherols and tocopheryl esters (0.2-0.7%), 2-hydroxy fatty acids (0.1-0.2%), and n-alkylresorcinols (0.1%). Notably, these different classes of compounds exhibited variations in their contents depending on the oat variety and the specific planting season. Of particular interest was the Karen variety, which presented significant amounts of high molecular weight esters, free fatty acids, and acylglycerols, especially when it was cultivated during the winter season. These findings underline the potential of oat straw as a valuable resource for lipid extraction within a biorefinery context and emphasize the importance of selecting the appropriate variety and season for optimal lipid yield.

Lipidomics of herbal tea revealed their potential lipid nutrients including novel fatty acid esters of hydroxy fatty acids

Lipidomics of herbal tea revealed their potential lipid nutrients including novel fatty acid esters of hydroxy fatty acids

Association of Lipopolysaccharide-Type Endotoxins with Retinal Neurodegeneration: The Alienor Study

PurposeLipopolysaccharide (LPS) -type endotoxins are naturally found in the gut microbiota and there is emerging evidence linking gut microbiota and neuroinflammation leading to retinal neurodegeneration. Thinning of the retinal nerve fiber layer (RNFL) is a biomarker of retinal neurodegeneration, and a hallmark of glaucoma, the second leading cause of blindness worldwide. We assessed the association of a blood biomarker of LPS with peripapillary RNFL thickness (RNFLT) and its longitudinal evolution up to 11 years. DesignThe Alienor study is a single center prospective population-based cohort study. SubjectsThe studied sample of this study includes 1062 eyes of 548 participants receiving at least one gradable RNFL measurement. MethodsPlasma esterified 3-hydroxy fatty acids (3-OH FAs) were measured as a proxy of LPS burden. RNFLT was acquired using spectral domain optical coherence tomography imaging every two years from 2009 to 2020 (up to 5 visits). Main Outcome MeasureAssociations of plasma esterified 3-OH FAs with RNFLT were assessed using linear mixed models. ResultsMean age of the included 548 participants was 82.4 ± 4.3 years and 62.6 % were women. Higher plasma esterified 3-OH FAs was significantly associated with thinner RNFLT at baseline (Coefficient Beta (ß)= -1.42 microns for 1 SD-increase in 3-OH FAs, 95% confidence interval [-2.56; -0.28], p = 0.02). This association remained stable after multivariate adjustment for potential confounders. No statistically significant association was found between 3-OH FAs and longitudinal RNFLT change. ConclusionsHigher plasma esterified 3-OH FAs were associated with thinner RNFLT at baseline, indicating an involvement of LPS in the early processes of optic nerve neurodegeneration and highlighting the potential importance of the human microbiota in preserving retinal health.

Impact of Nonthermal Plasma Treatment on the Oxidation of Lipids with Different Unsaturation Degrees.

Nonthermal plasma (NTP) treatment of food presents a new technology for the industry but raises concerns about lipid oxidation due to the presence of reactive species. Considering the critical role of the degree of unsaturation in lipid oxidation, this study investigates NTP-induced oxidation across various unsaturated lipids. These lipids are six oil samples primarily containing one of the following methylesters: oleate, linoleate, linolenate, arachidonate, eicosapentaenoate, and docosahexaenoate. Samples were treated with a nonthermal surface dielectric barrier discharge. Plasma-induced effects were first examined by classical lipid oxidation indicators, such as the peroxide value and p-anisidine value. The specific volatile oxidation products, including hexanal, nonanal, trans-2-hexenal, and methyl 9-oxononanoate, were determined to further elucidate the impact of ozone-related oxidation. Monitoring the production of selected nonvolatile oxidation products, such as epoxy-, oxo-, and hydroxy fatty acid methylesters, confirmed that plasma treatment facilitated the decomposition of lipid hydroperoxide. Generally, the level of plasma-induced oxidation increased in parallel with the unsaturation degree of the studied samples, except for the quantity of individual volatile carbonyls. The long-term effect of NTP treatment was investigated by a stability test, revealing that the oxidative stability depended on the input gas of plasma treatment, the sensitivity of the treated sample, and the presence of antioxidants. Except for the focus on the NTP impact, this study offered a case study of a comprehensive investigation into lipid oxidation.

Testing sample selection criteria and loss of biomarkers during cleaning of archaeological unglazed pottery to maximize organic residue quantities

AbstractAs the number of studies in organic residue analysis (ORA) of ancient pottery—a sensitive but as of today also a destructive method—increases, archaeologists are interested in knowing which samples promise the biggest abundance of analytes in order to avoid unnecessary loss of artefacts. Another frequently asked question is whether the routinely performed cleaning procedure should be omitted for samples intended for ORA to preserve the availability of analytes. We tested several selection criteria commonly accessible to archaeologists (texture, position, shape) for lipid quantities in ancient pottery in order to determine the most productive sherds for analysis. Moreover, we monitored loss of lipids during the water‐and‐brush cleaning process. Beside the usually targeted straight‐chain fatty acids (FA), less abundant biomarkers such as α,ω‐dicarboxylic acids (DCA), ω‐(o‐alkylphenyl)alkanoic acids (APAA) and hydroxy fatty acids (HFA) were also screened. The highest concentrations of analytes were observed in rims of coarse‐textured plates and cooking pots, demonstrating the usefulness of the proposed criteria. The washing procedure applied here did not lead to a loss of bulk FA, although the effect on minor components was not uniform.

Tissue-specific sex-dependent difference in the metabolism of fatty acid esters of hydroxy fatty acids

Fatty acid esters of hydroxy fatty acids (FAHFAs) are endogenous bioactive lipids known for their anti-inflammatory and anti-diabetic properties. Despite their therapeutic potential, little is known about the sex-specific variations in FAHFA metabolism. This study investigated the role of sex and Androgen Dependent TFPI Regulating Protein (ADTRP), a FAHFA hydrolase. Additionally, tissue-specific differences in FAHFA levels, focusing on the perigonadal white adipose tissue (pgWAT), subcutaneous white adipose tissue (scWAT), brown adipose tissue (BAT), plasma, and liver, were evaluated using metabolomics and lipidomics. We found that female mice exhibited higher FAHFA levels in pgWAT, scWAT, and BAT compared to males. FAHFA levels were inversely related to testosterone and Adtrp mRNA, which showed significantly lower expression in females compared with males in pgWAT and scWAT. However, no significant differences between the sexes were observed in plasma and liver FAHFA levels. Adtrp deletion had minimal impact on both sexes' metabolome and lipidome of pgWAT. However, we discovered higher endogenous levels of triacylglycerol estolides containing FAHFAs, a FAHFA metabolic reservoir, in the pgWAT of female mice. These findings suggest that sex-dependent differences in FAHFA levels occur primarily in specific WAT depots and may modulate local insulin sensitivity in adipocytes, and the role of ADTRP is limited to adipose depots. However, further investigations are warranted to fully comprehend the underlying mechanisms and implications of sex-dependent regulation of human FAHFA metabolism.

Hydroxy Fatty Acid Synthesis-Related mRNA as the Biomarker for Detecting Mislabeling of Honey Entomological Origin.

The authentication of the entomological origin of honey is a widespread concern, necessitating the prompt establishment of an effective approach for distinguishing between Apis cerana cerana honey (ACH) and Apis mellifera ligustica honey (AMH). Hydroxy fatty acids (HFAs) found in honey are bee-derived components synthesized by the mandibular glands of worker bees. We previously discovered significant variations in the hydroxy fatty acid composition between ACH and AMH, suggesting their potential as indicators for identifying the authenticity of the entomological origin of honey. Herein, we identified differentially expressed genes associated with HFA synthesis by conducting transcriptome sequencing of the mandibular glands of AC and AM honeybees. Subsequently, we proposed a method for the relative quantitative analysis of bee-derived RNA components using real-time fluorescence quantitative polymerase chain reaction, which was supplemented by multivariate statistical analysis to further discern differences in HFA synthesis-related mRNA between ACH and AMH. The results showed that the mRNAs of FAXDC2 (fatty acid hydroxylase domain-containing protein 2) and FAS (fatty acid synthase) may serve as indicators to discern the entomological origin of honey. This study presents two novel biomarkers for detecting mislabeling of the entomological origin in ACH and AMH based on variations in bee-derived components.

Lipidomic Profiling of Flammulina velutipes (Curtis) Singer (Agaricomycetes) through Ultra-Performance Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry: Examining Lipid Dynamics Changes during Fruiting Body Formation and Development.

Flammulina velutipes (enokitake) is widely recognized for its nutritional and medicinal properties. Understanding the biochemical processes, such as lipid metabolism during fruiting body formation, is essential for enhancing mushroom cultivation and utilization. This study aimed at elucidating the dynamic lipidomic changes during seven growth stages of F. velutipes using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Our results revealed significant increases in ceramides along with the growth and a sharp decline in phosphatidylinositols from mycelial to primordial stages. Fatty acid esters of hydroxy fatty acids, recently discovered for their bioactivities, showed high intensities in the mycelial and primordial stages but decreased rapidly thereafter. These findings provide profound insights into the lipid profiles associated with mushroom morphology and development. This lipidomics study establishes a foundational understanding for future research in agricultural and food chemistry applications, potentially improving industrial production and quality control of F. velutipes.

Bufadienolide-Fatty Acid Conjugates from the Fertilized Eggs of Toad Bufo gargarizans: Isolation, Characterization, Toxicity, and Antiproliferative Evaluation.

Bufadienolides (BDs) are a class of naturally occurring toxins present in amphibian toads. Serving as the chemical weapons, they exist not only in the adult toads but also in toad eggs. Guided by mass spectrometry (MS)-based component analysis and feature-based molecular networking (FBMN), 30 bufadienolide-fatty acid conjugates (BDFs) were isolated from the fertilized eggs of toad Bufo gargrizans, including 25 previously undescribed compounds (1-25). Their chemical structures were elucidated by extensive spectroscopic analysis, chemical methods, and GC-MS. The toxicities of all BDFs and their corresponding free BDs were assessed using the zebrafish model. The structure-toxicity relationship analysis showed that the modification of BDs by hydroxy fatty acids can cause a significant increase of the toxicity. Furthermore, all the isolated compounds were evaluated for their antiproliferative activities in pancreatic cancer cell lines ASPC-1 and PANC10.05. The structure-activity relationship (SAR) analysis revealed that BDFs with hellebrigenin as the bufogenin moiety (6 and 7) exhibited the most potent antiproliferative effect. Further investigation into their functional mechanism demonstrated that 6 and 7 induced apoptosis in pancreatic cancer cells PANC10.05 and significantly suppressed the expression of the apoptosis-related gene c-MYC. In addition, 6 and 7 effectively inhibited the expression of the PI3K/Akt/mTOR pathway in PANC10.05. Moreover, we assessed the efficacy of 6 and 7 on cancer cells from various tissues and observed their broad-spectrum antiproliferative activity.