Lipid Content Research Articles

Algal Biofuels: A Comprehensive Review and Analysis

The depletion of fossil fuels and increased atmospheric CO2 levels have precipitated a global energy and environmental crisis. The cultivation of microalgae offers a viable remedy by utilizing solar energy and assimilating CO2 for biofuel production via photosynthesis. Microalgae demonstrate rapid growth in diverse environments, rendering them a dependable biomass source for large-scale biofuel generation. Furthermore, their biofuel production can be tailored through the manipulation of growth conditions or genetic engineering. This review aims to highlight the variety of biofuels produced from microalgae and the methodologies to optimize their yield. This review also explores the economic implications of algal biofuel production in both open ponds and closed photobioreactors. Various strategies for the photo-production of biohydrogen utilizing the hydrogenase enzyme from green algae are investigated. Additionally, certain microalgae species are recognized for their potential as biodiesel sources due to their high lipid content. The lipid profiles of leading oil-producing algal strains under optimal conditions are assessed. This review further examines the potential of microalgae in synthesizing petroleum-based chemicals and in generating bioethanol and biogas from algal biomass.

Transcriptome analysis of lipid biosynthesis during kernel development in two walnut (Juglans regia L.) varieties of ‘Xilin 3’ and ‘Xiangling’

BackgroundWalnut is an oilseed tree species and an ecologically important woody tree species that is rich in oil and nutrients. In light of differences in the lipid content, fatty acid composition and key genes expression patterns in different walnut varieties, the key gene regulatory networks for lipid biosynthesis in different varieties of walnuts were intensively investigated.ResultsThe kernels of two walnut varieties, ‘Xilin 3’ (X3) and ‘Xiangling’ (XL) were sampled at 60, 90, and 120 days post-anthesis (DPA) to construct 18 cDNA libraries, and the candidate genes related to oil synthesis were identified via sequencing and expression analysis. A total of 106 differentially expressed genes associated with fatty acid biosynthesis, fatty acid elongation, unsaturated fatty acid biosynthesis, triglyceride assembly, and oil body storage were selected from the transcriptomes. Weighted gene co-expression network analysis (WGCNA), correlation analysis and quantitative validation confirmed the key role of the FAD3 (109002248) gene in lipid synthesis in different varieties.ConclusionsThese results provide valuable resources for future investigations and new insights into genes related to oil accumulation and lipid metabolism in walnut seed kernels. The findings will also aid future molecular studies and ongoing efforts to genetically improve walnut.

Organosulfur Compounds, S-Allyl-L-Cysteine and S-Ethyl-L-Cysteine, Target PCSK-9/LDL-R-Axis to Ameliorate Cardiovascular, Hepatic, and Metabolic Changes in High Carbohydrate and High Fat Diet-Induced Metabolic Syndrome in Rats.

Metabolic syndrome (MetS) is an ever-evolving set of diseases that poses a serious health risk in many countries worldwide. Existing evidence illustrates that individuals with MetS have a 30%-40% higher chance of acquiring type 2 diabetes mellitus (T2DM), cardiovascular disease (CVD), or both. This study was undertaken to uncover the regulatory role of natural organosulfur compounds (OSCs), S-allyl-L-cysteine (SAC), and S-ethyl-L-cysteine (SEC), in targeting high carbohydrate high fat (HCHF)-diet-induced MetS-associated risk management. Our findings suggested that SAC and SEC ameliorated HCHF-diet-induced diabetic profiles, plasma lipid and lipoprotein level, liver function, oxidative-stress, inflammatory cytokines, and chemokines including monocyte chemoattractant protein-1 (MCP-1), lipid peroxidation, plasma proprotein convertase subtilisin/kexin type-9 (PCSK-9), and high-sensitivity C-reactive protein (hs-CRP). Moreover, the assessment of the hepatic mRNA expression of the key genes involved in cholesterol homeostasis depicted that SAC and SEC downregulated the PCSK-9 mRNA expression via targeting the expression of HNF-1α, a transcriptional activator of PCSK-9. On the other hand, the LDL-receptor (LDL-R) expression was upregulated through the activation of its transcriptional regulator sterol regulatory element binding protein-2 (SREBP-2). In addition, the activity and the mRNA expression of 3-hydroxy-3-methylglutaryl coenzyme-A reductases (HMG-R) and peroxisome proliferator-activated receptors (PPARs) were also improved by the treatment of SAC and SEC. We concluded that SAC and SEC can protect against MetS via improving the lipid and lipoprotein content, glycemic indices, hepatic function, targeting the inflammatory cascades, and oxidative imbalance, regulation of the mRNA expression of PCSK-9, LDL-R, SREBP-2, HNF-1α, PPARs, and inflammatory biomarkers.

Physicochemical and stability analysis of mung bean protein hydrolysates with lipid peroxidation inhibition

This study investigated mung bean protein hydrolysates (MBPH) produced using neutral protease, examining their physicochemical properties, stability, and lipid peroxidation inhibition capabilities. The research revealed that MBPH molecular weight ranged from 17 to 26 kDa and perform various functions, including catalytic, nutrient storage, and binding. Stability assessments showed that MBPH are stable at 45 °C and pH of 7.5 but are light-sensitive and unstable in solution or when combined with sugars. Additionally, increased concentrations of digestive enzymes reduce MBPH stability. Antioxidant tests in vitro and in Caenorhabditis elegans confirmed MBPH's ability to neutralizing radicals, enhance antioxidant enzyme activities, and reduce lipid peroxidation, thereby protecting against oxidative damage. Furthermore, in vivo experiments showed that MBPH extend the lifespan of worms and reduced their body lipid content, indicating potential benefits in mitigating cholesterol-related damage. This research demonstrates the potential of MBPH in inhibiting lipid peroxidation.

Characteristics of Malassezia furfur at various pH and effects of Malassezia lipids on skin cells

Malassezia species are commensal and opportunistic fungi found in human skin. All Malassezia species lack fatty acid synthesis genes and survive by utilizing several lipases to degrade and absorb fatty acids from external lipid sources, but little research has been done on their optimal active pH and temperature. Our skin protects itself from external stimuli and maintains homeostasis, involving bacteria and fungi such as Malassezia species that inhabit our skin. Hence, dysbiosis in the skin microbiome can lead to various skin diseases. The skin’s pH is slightly acidic compared to neutral, and changes in pH can affect the metabolism of Malassezia species. We used keratinocyte cell lines to determine the effect of lipids bio-converted by Malassezia furfur, Malassezia japonica, and Malassezia yamatoensis under pH conditions similar to those of healthy skin. Lipids bio-converted from Malassezia species were associated with the regulation of transcripts related to inflammation, moisturizing, and promoting elasticity. Therefore, to determine the effect of pH on lipid metabolism in M. furfur, which is associated with seborrheic dermatitis, changes in biomass, lipid content, and fatty acid composition were determined. The results showed that pH 7 resulted in low growth and reduced lipid content, which had a negative impact on skin health. Given that bio-converted Malassezia-derived lipids show positive effects at the slightly acidic pH typical of healthy skin, it is important to study their effects on skin cells under various pH conditions.Key points• pH 6, Malassezia spp. bio-converted lipid have a positive effect on skin cells• Malassezia spp. have different lipid, fatty acid, and growth depending on pH• Malassezia spp. can play a beneficial role by secreting lipids to the outside

The Effect of Acrylonitrile Butadiene Styrene (ABS) and Polypropylene (PP) Microplastics on Ulva lactuca L. and Ceramium diaphanum R. Algal Growth

The goal of this study was to determine the effects of acrylonitrile butadiene styrene and polypropylene microplastics as two typical classes for microplastic pollution on algal growth, lipids, carbohydrates, chlorophyll-a and total carotenoid content of Ulva lactuca L. and Ceramium diaphanum R. collected from the Gulf coast of Izmir province. The results demonstrate that ABS-MPs had no statistical effect (P>0.05) on the various physiological parameters of U. lactuca macroalgae after 5 days of exposure, except for the total carbohydrate and lipid concentration. The chl-a value of C. diaphanum was considerably increased under the low concentration of ABS (25 mg L-1), increasing by 81.56% compared with the control while the amount of chl-a decreases (32.05%) in parallel with the increasing concentration of PP-MPs. Laboratory incubation experiments showed that MPs affect relative growth rate, pigments efficiency or carbohydrate content of C. diaphanum until reaching an extremely high concentration (100 mg L-1), indicating a high tolerance to MPs. The results showed that two macroalgae species, especially U. lactuca, were not highly affected at low MPs concentrations under laboratory conditions which were much higher than the levels of environmentally relevant concentrations.

Discovery of natural anthraquinones as potent inhibitors against pancreatic lipase: structure-activity relationships and inhibitory mechanism

Obesity is acknowledged as a significant risk factor for various metabolic diseases, and the inhibition of human pancreatic lipase (hPL) can impede lipid digestion and absorption, thereby offering potential benefits for obesity treatment. Anthraquinones is a kind of natural and synthetic compounds with wide application. In this study, the inhibitory effects of 31 anthraquinones on hPL were evaluated. The data shows that AQ7, AQ26, and AQ27 demonstrated significant inhibitory activity against hPL, and exhibited selectivity towards other known serine hydrolases. Then the structure-activity relationship between anthraquinones and hPL was further analysed. AQ7 was found to be a mixed inhibition of hPL through inhibition kinetics, while AQ26 and AQ27 were effective non-competitive inhibition of hPL. Molecular docking data revealed that AQ7, AQ26, and AQ27 all could associate with the site of hPL. Developing hPL inhibitors for obesity prevention and treatment could be simplified with this novel and promising lead compound.

Physiological response and carbon dioxide sequestration mechanisms during photosynthesis in Mychonastes rotundus

Carbon sequestration by microalgae is a simple and easy-to-operate process, without secondary pollution that has broad application prospects due to rapid growth, efficient photosynthesis, and high lipid content produced. In this study, the potential of Mychonastes rotundus for CO2 fixation, and its growth pattern, enzyme activity, lipid production, and cellular microstructure were investigated with changes in CO2 input. The results showed that when the inlet CO2 concentration was 20% (v/v), stable carbon sequestration ability was obtained after a 590 min reaction with an outlet CO2 concentration of 0.32%. The algal cellular structure was intact when the lowest contents of malondialdehyde and reactive oxygen species (ROS) were 7.08 and 17.79 nmol/mg, respectively, indicating the highest cell activity. In addition, 20% CO2 effectively promoted the synthesis of C18:2n6c and C20:0 in M. rotundus. When the inlet CO2 concentration was increased to 30%, the average pH dropped to 5.7, resulting in the death of M. rotundus. 1O2 was identified as the major ROS damaging microalgal cells during carbon sequestration. This study determined the optimal CO2 concentration for carbon capture by M. rotundus as a raw material for biodiesel production. The results indicate that M. rotundus is a promising microalgae for carbon fixation and CO2 capture of power plant flue gas and biodiesel production for low-cost bioenergy production.

Can We Simplify Liposome Manufacturing Using a Complex DoE Approach?

Microfluidic liposome production presents a streamlined pathway for expediting the translation of liposomal formulations from the laboratory setting to clinical applications. Using this production method, resultant liposome characteristics can be tuned through the control of both the formulation parameters (including the lipids and solvents used) and production parameters (including the production speed and mixing ratio). Therefore, the aim of this study was to investigate the relationship between not only total flow rate (TFR), the fraction of the aqueous flow rate over the organic flow rate (flow rate ratio (FRR)), and the lipid concentration, but also the solvent selection, aqueous buffer, and production temperature. To achieve this, we used temperature, applying a design of experiment (DoE) combined with machine learning. This study demonstrated that liposome size and polydispersity were influenced by manipulation of not only the total flow rate and flow rate ratio but also through the lipids, lipid concentration, and solvent selection, such that liposome attributes can be in-process controlled, and all factors should be considered within a manufacturing process as impacting on liposome critical quality attributes.

Taxonomic Revision of Genus Ephedra Tourn. ex L. in Egypt with Intra-Gender Diversity in Morphometric Traits and Fatty Acid Composition.

The genus Ephedra Tourn. ex L. (Ephedraceae) still exhibits taxonomic complexity that has not yet been resolved. This study aimed to determine the taxonomic identity of the Ephedra species in Egypt and identify the fatty acid profile and its diversity at the gender level as a taxonomic tool for specimens lacking reproductive cones. The current study provides a pioneering approach that distinguishes Ephedra species at the gender level. A total of 120 fresh individuals were collected from 20 locations representing different habitats where Ephedra plants grow in Egypt. In addition, herbarium specimens were deposited in Egyptian herbaria. The studied morphological traits included 30 vegetative characteristics and 72 traits of the reproductive organs of both genders. The fatty acid content was measured using gas-mass chromatography (GC-Mass). The taxonomic revision revealed that the Genus Ephedra was represented in the Egyptian flora by five species, Ephedra alata in section Alatae and E. aphylla, E. ciliata, E. foemina, and E. pachyclada in section Ephedra. South Sinai hosts these five species and represents the center of diversity for this genus in Egypt. The vegetative characteristics were subjected to principal component analysis (PCA), which revealed a distinct separation of the five studied species. Similarly, the cone traits treated by hierarchical clustering revealed intra-gender variations. The taxonomic key was developed based on the morphological traits to distinguish the studied species at the gender level. In total, 51 fatty acids were identified from the studied species and grouped as 18 saturated, 16 monounsaturated, and 17 polyunsaturated fatty acids. In the absence of reproductive cones, the lipid content and fatty acid composition of the vegetative parts displayed significant interspecific and intra-gender variations. Therefore, fatty acids can be used to efficiently identify the studied species when they lack reproductive cones. This study proved the efficacy of a multidisciplinary approach to identify Ephedra species at the gender level and recommends this trend for future studies of this genus.

Hyperthermal stress potentiates enhanced lipid utilisation in genetically improved farmed Tilapia, Oreochromis niloticus juveniles

The present experiment evaluated whether dietary protein (P) or lipid (L) is preferred as an energy source by genetically improved farmed tilapia (GIFT) reared at high temperatures. A 60-day feeding trial was conducted at 28.3 °C and 33.3 °C, testing five diets with varying protein (34,36,38 %) and lipid (8,10,12 %) levels, viz., P38L8, P36L8, P34L8, P34L10, P34L12. Parameters assessed included growth, body composition, serum lipids, enzyme activities, fatty acid profiles, and PPAR-α mRNA expression. Results indicated that the fish fed optimum protein and highest lipid level (P34L12) showed significantly higher (P < 0.05) weight gain percent and thermal growth coefficient. Increasing dietary lipid content reduced whole-body lipid deposition and mobilised serum triglycerides and cholesterol at higher temperatures (HT). Hepatic malic enzyme activity decreased with rising temperature and lipid content, while lipoprotein lipase activity in muscle increased. The fatty acid composition altered substantially with the changes in rearing temperature and diets. Unsaturated fats were preferred as direct fuels for β-oxidation, wherein the P34L12 groups preserved body (area %) EPA, DHA, and linolenic acid, especially at HT. The expression of PPAR-α, a lipolytic marker, was upregulated with increasing temperature and high dietary lipid content, peaking in P34L12 groups. The study concludes that high-lipid diets (12 %) are metabolically superior to high-protein diets for GIFT tilapia at elevated temperatures, optimising growth, enhancing metabolic efficiency, and maintaining essential fatty acid profiles under hyperthermal stress.

Lysyl oxidase expression in smooth muscle cells determines the level of intima calcification in hypercholesterolemia-induced atherosclerosis

IntroductionCardiovascular calcification is an important public health issue with an unmeet therapeutic need. We had previously shown that lysyl oxidase (LOX) activity critically influences vascular wall smooth muscle cells (VSMCs) and valvular interstitial cells (VICs) calcification by affecting extracellular matrix remodeling. We have delved into the participation of LOX in atherosclerosis and vascular calcification, as well as in the mineralization of the aortic valve. MethodsImmunohistochemical and expression studies were carried out in human atherosclerotic lesions and experimental models, valves from patients with aortic stenosis, VICs, and in a genetically modified mouse model that overexpresses LOX in CMLV (TgLOXCMLV). Hyperlipemia and atherosclerosis was induced in mice through the administration of adeno-associated viruses encoding a PCSK9 mutated form (AAV-PCSK9D374Y) combined with an atherogenic diet. ResultsLOX expression is increased in the neointimal layer of atherosclerotic lesions from human coronary arteries and in VSMC-rich regions of atheromas developed both in the brachiocephalic artery of control (C57BL/6 J) animals transduced with PCSK9D374Y and in the aortic root of ApoE−/− mice. In TgLOXCMLV mice, PCSK9D374Y transduction did not significantly alter the enhanced aortic expression of genes involved in matrix remodeling, inflammation, oxidative stress and osteoblastic differentiation. Likewise, LOX transgenesis did not alter the size or lipid content of atherosclerotic lesions in the aortic arch, brachiocephalic artery and aortic root, but exacerbated calcification. Among lysyl oxidase isoenzymes, LOX is the most expressed member of this family in highly calcified human valves, colocalizing with RUNX2 in VICs. The lower calcium deposition and decreased RUNX2 levels triggered by the overexpression of the nuclear receptor NOR-1 in VICs was associated with a reduction in LOX. ConclusionsOur results show that LOX expression is increased in atherosclerotic lesions, and that overexpression of this enzyme in VSMC does not affect the size of the atheroma or its lipid content, but it does affect its degree of calcification. Further, these data suggest that the decrease in calcification driven by NOR-1 in VICs would involve a reduction in LOX. These evidences support the interest of LOX as a therapeutic target in cardiovascular calcification.

Full-fat black soldier fly larvae meal and yellow mealworm meal: Impact on feed protein quality, growth and nutrient utilization of Atlantic salmon (Salmo salar) post smolts

The shift towards insect-based alternatives in aquafeeds stems from the need to reduce the carbon footprint associated with the ingredients used in salmon feeds. In the present study, we describe the effects of full-fat insect meal incorporation on amino acid derivatives in feeds and the growth performance and nutrient utilization of Atlantic salmon post-smolts fed insect meals. A 74-day growth trial was conducted employing 520 fish (143 ± 12.8 g); these fish were fed either a control diet (CO) that contained 20 % fishmeal, 20 % soy protein concentrate (SPC), 14 % wheat gluten, 19 % plant raw materials, 13 % fish oil and 6 % rapeseed oil, or one of four insect meal diets, prepared with either 5 % and 10 % black soldier fly larvae meal (BS5, BS10) or 15 % and 30 % mealworm meal (MW15, MW30), by substituting mainly SPC, wheat gluten and rapeseed oil. The inclusion levels of BS and MW were determined based on their distinct chemical compositions, specifically their lipid content. As regards the results, HPLC-based high-throughput data on amino acid derivatives revealed the following: carboxymethyllysine was significantly higher in the MW30 feed than in BS5, BS10 and MW15 feeds; carboxyethyllysine was higher in the MW15 and MW30 feeds compared to the others; furosine was lower in the MW15 feed compared to the CO, BS5 and MW30 feeds. Regarding the effect of the insect meals on Atlantic salmon post-smolts, their inclusion had no significant effect on feed conversion ratio, health indicators and the retention efficiency of macronutrients. Apparent digestibility coefficients (ADC) of dry matter, lipid, ash and energy were not significantly different. However, protein digestibility in the 30 % MW group (80.4 %) was significantly lower (P ≤ 0.05) compared to the control and BS5 group (84.5 %). Furthermore, the ADC of protein was found to decrease linearly with increasing levels of insect meals, with MW showing a significant relationship (adjusted r2 = 0.77, p = 0.014). Overall, our study highlights the efficacy of 5 and 10 % BS and 15 % MW as ingredients in the feeds of Atlantic salmon. Other focal points included the undesirable amino acid derivatives and a reduction in protein digestibility associated with 30 % of MW in the diet of Atlantic salmon. Hence, future investigations should focus on the effects of feed processing conditions on the protein quality of mealworm before suggesting higher inclusion levels to the feed industry.

The Omega-6 Lipid pathway shift is associated with neutrophil influx and structural lung damage in early cysticfibrosis lung disease.

In cystic fibrosis (CF), an imbalanced lipid metabolism is associated with lung inflammation. Little is known about the role that specific lipid mediators (LMs) exert in CF lung inflammation, and whether their levels change during early disease progression. Therefore, we measured airway LM profiles of young CF patients, correlating these with disease-associated parameters. Levels of omega (ω)-3/6 PUFAs and their LM derivatives were determined in bronchoalveolar lavage fluid (BALF) of children with CF ages 1-5 using a targeted high-performance liquid chromatography-tandem mass spectrometry approach. Hierarchical clustering analysis was performed on relative LM levels. Individual relative LM levels were correlated with neutrophilic inflammation (BALF %Neu) and structural lung damage (PRAGMA-CF %Disease). Significant correlations were included in a backward multivariate linear regression model to identify the LMs that are best related to disease progression. A total of 65 BALF samples were analysed for ω-3/6 lipid content. LM profiles clustered into an arachidonic acid (AA)-enriched and a linoleic acid (LA)-enriched sample cluster. AA derivatives like 17-OH-DH-HETE, 5-HETE, 5,15-diHETE, 15-HETE, 15-KETE, LTB4 and 6-trans-LTB4 positively correlated with BALF %Neu and/or PRAGMA %Dis. Contrastingly, 9-HoTrE and the LA derivatives 9-HoDE, 9(10)-EpOME, 9(10)-DiHOME, 13-HoDE, 13-oxoODE and 12(13)-EpOME negatively correlated with BALF %Neu and/or PRAGMA %Dis. 6-trans-LTB4 was the strongest predictor for BALF %Neu. 5-HETE and 15-KETE contributed most to PRAGMA %Dis prediction. Our data provide more insight into the lung lipidome of infants with CF, and show that a shift from LA derivatives to AA derivatives in BALF associates with early CF lung disease progression.

Model development and process evaluation for algal growth and lipid production

Microalgae grabbed the attention worldwide because of their application in renewable energy with a number of environmental benefits such as carbon dioxide assimilation to produce biofuel. In this study, a kinetic model aiming to analyze algae growth and lipid production was developed. Biomass was divided into two parts, algae residual cell and lipid, to analyze their kinetics distinctively and to find out the inside process mechanism. The model was calibrated and validated with different experimental datasets, varying carbon sources, and phosphate concentrations. The capability of the model to predict the dynamics of algal culture over a broad range of growth conditions was investigated. The presence of acetate reduced the bicarbonate uptake for algal growth and growth on organic carbon was higher compared to that of carbon dioxide. The presence of ammonium showed a very strong inhibition effect on algae and lipid production rate but enhanced lipid content. Phosphate caused both limitation and inhibition effects on algae growth and lipid production rate. The maximum growth was found at 12 mg/L PO43- concentration and 3.10 mg/L NO3--N concentration. Lipid content was enhanced by limiting phosphate. Overall, the developed model allows optimizing of nutrient concentrations and operating conditions specifically to enhance lipid productivity.

Hepatocyte MMP14 mediates liver and inter-organ inflammatory responses to diet-induced liver injury.

The matrix metalloproteinase MMP14 is a ubiquitously expressed, membrane-bound, secreted endopeptidase that proteolyzes substrates to regulate development, signaling, and metabolism. However, the spatial and contextual events inciting MMP14 activation and its metabolic sequelae are not fully understood. Here, we introduce an inducible, hepatocyte-specific MMP14-deficient model (MMP14LKO mice) to elucidate cell-intrinsic and systemic MMP14 function. We show that hepatocyte MMP14 mediates diet-induced body weight gain, peripheral adiposity, and impaired glucose homeostasis and drives diet-induced liver triglyceride accumulation and induction of hepatic inflammatory and fibrotic gene expression. Single-nucleus RNA sequencing revealed that hepatocyte MMP14 mediates Kupffer cell and T-cell accumulation and promotes diet-induced hepatocellular subpopulation shifts toward protection against lipid absorption. MMP14 co-immunoprecipitation and proteomic analyses revealed MMP14 substrate binding across both inflammatory and cytokine signaling, as well as metabolic pathways. Strikingly, hepatocyte MMP14 loss-of-function suppressed skeletal muscle and adipose inflammation in vivo, and in a reductionist adipose-hepatocyte co-culture model. Finally, we reveal that trehalose-type glucose transporter inhibitors decrease hepatocyte MMP14 gene expression and nominate these inhibitors as translatable therapeutic metabolic agents. We conclude that hepatocyte MMP14 drives liver and inter-organ inflammatory and metabolic sequelae of obesogenic dietary insult. Modulating MMP14 activation and blockade thus represents a targetable node in the pathogenesis of hepatic inflammation.

Buttermilk treatment strategies to obtain MFGM-enriched isolates and comprehensive characterization of their lipid and protein components

The extraction of Milk Fat Globule Membrane (MFGM) from dairy by-products, particularly buttermilk, involves a highly intricate procedures to obtain a bioactive isolate with multiple health benefits. This study delves into a laboratory ultrafiltration-based method enabling the acquisition of a concentrated MFGM isolate. Pre-ultrafiltration treatments, designed to clarify the sample, such as casein coagulation and the addition of sodium citrate, were examined to evaluate their impact on the lipid and protein components of the MFGM. Treatments combining rennet coagulation with centrifugation steps, as well as those combining rennet with sodium citrate, yielded highly pure MFGM isolates exhibiting a substantial concentration of polar lipids and a rich content of MFGM proteins. This laboratory investigation offers valuable insights into the potential industrial-scale applications of MFGM isolation and purification.

P11-07 In vitro effects of polystyrene microplastics, alone or in combination with environmental pollutants, on viability and lipid content of a human hepatocarcinoma cell line

P11-07 In vitro effects of polystyrene microplastics, alone or in combination with environmental pollutants, on viability and lipid content of a human hepatocarcinoma cell line

Integrated ultrasensitive metabolomics and single-cell transcriptomics identify crucial regulators of sheep oocyte maturation and early embryo development in vitro

IntroductionDevelopmental competence of oocytes matured in vitro is limited due to a lack of complete understanding of metabolism and metabolic gene expression during oocyte maturation and embryo development. Conventional metabolic analysis requires a large number of samples and is not efficiently applicable in oocytes and early embryos, thereby posing challenges in identifying key metabolites and regulating their in vitro culture system. ObjectivesTo enhance the developmental competence of sheep oocytes, this study aimed to identify and supplement essential metabolites that were deficient in the culture systems. MethodsThe metabolic characteristics of oocytes and embryos were determined using ultrasensitive metabolomics analysis on trace samples and single-cell RNA-seq. By conducting integrated analyses of metabolites in cells (oocytes and embryos) and their developmental microenvironment (follicular fluid, oviductal fluid, and in vitro culture systems), we identified key missing metabolites in the in vitro culture systems. In order to assess the impact of these key missing metabolites on oocyte development competence, we performed in vitro culture experiments. Furthermore, omics analyses were employed to elucidate the underlying mechanisms. ResultsOur findings demonstrated that betaine, carnitine and creatine were the key missing metabolites in vitro culture systems and supplementation of betaine and L-carnitine significantly improved the blastocyst formation rate (67.48% and 48.61%). Through in vitro culture experiments and omics analyses, we have discovered that L-carnitine had the potential to promote fatty acid oxidation, reduce lipid content and lipid peroxidation level, and regulate spindle morphological grade through fatty acid degradation pathway. Additionally, betaine may participate in methylation modification and osmotic pressure regulation, thereby potentially improving oocyte maturation and early embryo development in sheep. ConclusionTogether, these analyses identified key metabolites that promote ovine oocyte maturation and early embryo development, while also providing a new viewpoint to improve clinical applications such as oocyte maturation or embryo culture.

Analysis of phthalate esters from vegetable oils by gas chromatography-mass spectrometry coupled with headspace solid-phase microextraction using cyclodextrin-based hypercrosslinked polymer coated fiber

Phthalate esters (PAEs) are used as additives to enhance the pliability and malleability of plastics. These substances frequently migrate from packaging materials to vegetable oils because of the absence of covalent bonds. Over time, this migration could result in the accumulation of PAEs in the human body through ingestion, contributing to various diseases. Therefore, accurate qualitative and quantitative analyses of PAEs in vegetable oils are imperative to assess the origins of contamination and investigate their toxicity, degradation, migration, and transformation patterns. However, the concentration of PAEs in most samples is low, and the composition of vegetable oils is complex. Thus, PAEs must be enriched and purified using appropriate sample pretreatment procedures before analysis. Common methods for pretreating PAEs in oil include solid-phase extraction (SPE), dispersive SPE, and magnetic SPE. These techniques require time-consuming and labor-intensive procedures such as oil dissolution, solvent extraction, and degreasing. These approaches also require numerous solvents and containers, increasing the risk of sample cross-contamination. Solid-phase microextraction (SPME) integrates sampling, extraction, purification, concentration, and injection into a single process, significantly accelerating analytical testing and reducing the potential for sample cross-contamination. In headspace (HS) mode, the analytes achieve equilibrium on the coating and are extracted in the gas phase. The fibers are shielded from nonvolatile and high-relative molecular mass substances in the sample matrix. Thus, SPME is an ideal method for extracting volatile compounds in vegetable oils. When HS-SPME coupled with gas chromatography-mass spectrometry (GC-MS), it can achieve the rapid screening of PAEs in vegetable oil. In this study, an SPME with cyclodextrin-based hypercrosslinked polymers (BnCD-HCP) coated on stainless steel fibers was employed to extract PAEs from vegetable oil. The structure and morphology of the polymers were characterized using Fourier-transform infrared spectroscopy, nuclear magnetic spectroscopy, and scanning electron microscopy. BnCD-HCP exhibited high stability and diverse interactions, including π-π, hydrophobic, and host-guest interactions. The oil samples were incubated with methanol, and the PAEs were extracted from the headspace using the probe. The optimal extraction parameters included an extraction time of 20 min, extraction temperature of 50 ℃, desorption time of 4 min, and desorption temperature of 275 ℃. The BnCD-HCP/HS-SPME method was evaluated under optimized experimental conditions. The limits of detection (LODs) and quantification (LOQs) were determined by applying signal-to-noise ratios (S/N) of 3 and 10, respectively. Method accuracy was evaluated using relative standard deviations (RSDs). Single-needle precision was evaluated by conducting three consecutive analyses at 3 h intervals within a day. Inter-needle precision was assessed by conducting the same analyses (three replicates) with differently coated fibers. The 12 PAE compounds exhibited good linearity with correlation coefficients (R2) of at least 0.99. The LODs and LOQs ranged from 0.21 to 3.74 μg/kg and from 0.69 to 12.34 μg/kg, respectively. The RSDs were in the range of 1.8%-11.4% and 5.1%-13.9% for the single-needle and needle-to-needle methods, respectively. The proposed method was applied to soybean, peanut, and sunflower oils, and two PAEs were found in all three oils. Moreover, the method demonstrated good precision (RSD=1.17%-11.73%) and recoveries (72.49%-124.43%). Compared with other methods, the developed method was able to extract many target analytes and had a low or comparable LOD and high recovery. More importantly, this method does not require tedious operations such as solvent extraction and purification. Consequently, the developed method can be used to extract not only PAEs in oils but also other substances with a high lipid content.