Search

Seipin, a crucial protein for cellular lipid droplet (LD) assembly, oligomerizes at the interface between the endoplasmic reticulum and LDs to facilitate neutral lipid packaging. Using proximity labeling, we identified four proteins—Ldo45, Ldo16, Tgl4, and Pln1—that are recruited to the vicinity of yeast seipin, the Sei1-Ldb16 complex, exclusively when seipin function is intact, hence termed seipin accessory factors. Localization studies identified Tgl4 at the endoplasmic reticulum-LD contact site, in contrast to Ldo45, Ldo16, and Pln1 at the LD surface. Cells with compromised seipin function resulted in uneven distribution of these proteins with aberrant LDs, supporting a central role of seipin in orchestrating their association with the LD. Overexpression of any seipin accessory factor causes LD aggregation and affects a subset of LD protein distribution, highlighting the importance of their stoichiometry. Although single factor mutations show minor LD morphology changes, the combined mutations have additive effects. Lastly, we present evidence that seipin accessory factors assemble and interact with seipin in the absence of neutral lipids and undergo dynamical rearrangements during LD formation induction, with Ldo45 acting as a central hub recruiting other factors to interact with the seipin complex. [ABSTRACT FROM AUTHOR]

The CD1 family of antigen-presenting molecules adopt a major histocompatibility complex class I (MHC-I) fold. Whereas MHC molecules present peptides, the CD1 family has evolved to bind self- and foreign-lipids. The CD1 family of antigen-presenting molecules comprises four members—CD1a, CD1b, CD1c, and CD1d—that differ in their architecture around the lipid-binding cleft, thereby enabling diverse lipids to be accommodated. These CD1–lipid complexes are recognized by T cell receptors (TCRs) expressed on T cells, either through dual recognition of CD1 and lipid or in a new model whereby the TCR directly contacts CD1, thereby triggering an immune response. Chemical syntheses of lipid antigens, and analogs thereof, have been crucial in understanding the underlying specificity of T cell–mediated lipid immunity. This review will focus on our current understanding of how TCRs interact with CD1–lipid complexes, highlighting how it can be fundamentally different from TCR-MHC-peptide corecognition. [ABSTRACT FROM AUTHOR]

In this study, milk fat globule membrane (MFGM) ingredients enriched in polar lipids were prepared using membrane filtration, including microfiltration, diafiltration, and ultrafiltration from butter serum powder. Polar lipids (phospholipids, sterols, and gangliosides) in prepared MFGM ingredients were analyzed by 31P nuclear magnetic resonance spectroscopy, GC-MS, and ultra-performance liquid chromatography (UPLC)-MS/MS, respectively. The lipolysis degree and microstructure of MFGM ingredient and soybean lecithin (SL) emulsions during in vitro digestion were also analyzed. Microfiltration showed higher concentration efficiency than ultrafiltration, which increased by 2.16% and 2.73% in phospholipids, respectively. Moreover, diafiltration concentrated more polar lipids (6.39% of phospholipids) than microfiltration. Milk fat globule membrane ingredients had high levels of sphingomyelin (1.27%–1.36%) and ratio of GD 3 to GM 3 is 9.25- to 9.88-fold. The different lipolysis behaviors between MFGM ingredient emulsions and SL emulsions were correlated with their different polar lipid compositions. Phospholipids from both MFGM ingredients and SL could help maintain the initial structure during the gastric digestion. These results could provide a scientific basis for developing high-polar-lipids food, particularly infant formulas and special functional foods. [ABSTRACT FROM AUTHOR]

The cryo-EM resolution revolution has heralded a new era in our understanding of eukaryotic lipid flippases with a rapidly growing number of high-resolution structures. Flippases belong to the P4 family of ATPases (type IV P-type ATPases) that largely follow the reaction cycle proposed for the more extensively studied cation-transporting P-type ATPases. However, unlike the canonical P-type ATPases, no flippase cargos are transported in the phosphorylation half-reaction. Instead of being released into the intracellular or extracellular milieu, lipid cargos are transported to their destination at the inner leaflet of the membrane. Recent flippase structures have revealed multiple conformational states during the lipid transport cycle. Nonetheless, critical conformational states capturing the lipid cargo “in transit” are still missing. In this review, we highlight the amazing structural advances of these lipid transporters, discuss various perspectives on catalytic and regulatory mechanisms in the literature, and shed light on future directions in further deciphering the detailed molecular mechanisms of lipid flipping. [ABSTRACT FROM AUTHOR]

Human iPSC-derived airway organoids (iPSC-AOs) exhibit cellular heterogenicity resembling the human native airway. Bioinformatics analysis revealed a robust dysregulation in inflammation, lipid-metabolism pathway, and alternative splicing in SARS-CoV-2 infected airway organoids. Manipulating lipid homeostasis using cholesterol-lowering drugs (e.g. Statins) relocated the viral entry receptor angiotensin-converting enzyme-2 (ACE-2), leading to the reduction of SARS-CoV-2 entry and replication. [Display omitted] • Single-cell RNA sequencing showed that iPSC-AOs exhibition of the genetic profiles highly resembling the genotypes of human airways. • iPSC-AOs are susceptible to SARS-COV-2 infection and the expressions of S and N proteins after viral infection. • Bioinformatics analysis indicated the involvement of aberrant enrichment of lipid transport and lipid pathways in infected iPSC-AOs. • Statin preincubation relocated ACE2, reduced N protein, and limited SARS-CoV-2 infection in iPSC-AOs. • Lipid modifications consistently inhibited viral entry of various SARS-CoV-2 strains (wild-type, alpha, delta, omicron). Modifications of lipid metabolism were closely associated with the manifestations and prognosis of coronavirus disease of 2019 (COVID-19). Pre-existing metabolic conditions exacerbated the severity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection while modulations of aberrant lipid metabolisms alleviated the manifestations. To elucidate the underlying mechanisms, an experimental platform that reproduces human respiratory physiology is required. Here we generated induced pluripotent stem cell-derived airway organoids (iPSC-AOs) that resemble the human native airway. Single-cell sequencing (ScRNAseq) and microscopic examination verified the cellular heterogeneity and microstructures of iPSC-AOs, respectively. We subjected iPSC-AOs to SARS-CoV-2 infection and investigated the treatment effect of lipid modifiers statin drugs on viral pathogenesis, gene expression, and the intracellular trafficking of the SARS-CoV-2 entry receptor angiotensin-converting enzyme-2 (ACE-2). In SARS-CoV-2-infected iPSC-AOs, immunofluorescence staining detected the SARS-CoV-2 spike (S) and nucleocapsid (N) proteins and bioinformatics analysis further showed the aberrant enrichment of lipid-associated pathways. In addition, SARS-CoV-2 hijacked the host RNA replication machinery and generated the new isoforms of a high-density lipoprotein constituent apolipoprotein A1 (APOA1) and the virus-scavenging protein deleted in malignant brain tumors 1 (DMBT1). Manipulating lipid homeostasis using cholesterol-lowering drugs (e.g. Statins) relocated the viral entry receptor angiotensin-converting enzyme-2 (ACE-2) and decreased N protein expression, leading to the reduction of SARS-CoV-2 entry and replication. The same lipid modifications suppressed the entry of luciferase-expressing SARS-CoV-2 pseudoviruses containing the S proteins derived from different SARS-CoV-2 variants, i.e. wild-type, alpha, delta, and omicron. Together, our data demonstrated that modifications of lipid pathways restrict SARS-CoV-2 propagation in the iPSC-AOs, which the inhibition is speculated through the translocation of ACE2 from the cell membrane to the cytosol. Considering the highly frequent mutation and generation of SARS-CoV-2 variants, targeting host metabolisms of cholesterol or other lipids may represent an alternative approach against SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

Rumen-protected choline (RPC) promotes benefits in milk production, immunity, and health in dairy cows by optimizing lipid metabolism during transition period management and early lactation. However, the RPC success in dairy cows depends on choline bioavailability, which is affected by the type of protection used in rumen-protected choline. Therefore, our objectives were to determine the effects of a novel RPC on dry matter intake (DMI), identify markers of metabolism and immunity, and evaluate lactation performance. Dry Holstein (n = 48) cows at 245 ± 3 d of gestation were blocked by parity and assigned to control or RPC treatment within each block. Cows enrolled in the RPC treatment received 15 g/d of CholiGEM (Kemin Industries, Cavriago RE, Italy) from 21 d prepartum and 30 g/d of CholiGEM from calving to 21 d postpartum. During the transition period, DMI was measured daily, and blood was sampled weekly for energy-related metabolites such as β-hydroxybutyrate (BHB), glucose, and nonesterified fatty acids (NEFA), as well as immune function markers such as haptoglobin (Hp) and lipopolysaccharide-binding protein (LPB). Vaginal discharge samples were collected at the calving and 7 d postpartum and stored in microcentrifuge tubes at −80°C until 16S rRNA sequencing. The main responses of body condition score, body weight, DMI, milk yield, milk components, and immune function markers were analyzed using the GLIMMIX procedure of SAS with the effects of treatment, time, parity, and relevant covariates added to the models. The relative abundance of microbiome α-diversity was evaluated by 3 indexes (Chao1, Shannon, and Simpson) and β-diversity by principal coordinate analysis and permutational multivariate ANOVA. We found no differences in DMI in the pre- and postpartum periods. Cows fed RPC increased the yields of energy- and 3.5% fat-corrected milk and fat yield in primiparous and multiparous cows, with an interaction between treatment and parity for these lactation variables. However, we found no differences in milk protein and lactose up to 150 DIM between treatments. Glucose, NEFA, and BHB had no differences between the treatments. However, RPC decreased BHB numerically (control = 1.07 ± 0.13 vs. RPC = 0.63 ± 0.13) in multiparous on the third week postpartum and tended to reduce the incidence of subclinical ketosis (12.7% vs. 4.2%). No effects for Hp and LPB were found in cows fed RPC. Chao1, Shannon, and Simpson indexes were lower at calving in the RPC treatment than in the Control. However, no differences were found 7 d later for Chao1, Shannon, and Simpson indexes. The vaginal discharge microbiome was altered in cows fed RPC at 7 d postpartum. Fusobacterium , a common pathogen associated with metritis, was reduced in cows fed RPC. Rumen-protected choline enhanced lactation performance and health and altered the vaginal discharge microbiome which is a potential proxy for uterine healthy in dairy cows. The current study's findings corroborate that RPC is a tool to support adaptation to lactation and shed light on opportunities for further research in reproductive health. [ABSTRACT FROM AUTHOR]

The stability of ribosomal DNA (rDNA) is maintained through transcriptional silencing by the NAD+ -dependent histone deacetylase Sir2 in Saccharomyces cerevisiae. Alongside proteostasis, rDNA stability is a crucial factor regulating the replicative lifespan of S. cerevisiae. The unfolded protein response (UPR) is induced by misfolding of proteins or an imbalance of membrane lipid composition and is responsible for degrading misfolded proteins and restoring endoplasmic reticulum (ER) membrane homeostasis. Recent investigations have suggested that the UPR can extend the replicative lifespan of yeast by enhancing protein quality control mechanisms, but the relationship between the UPR and rDNA stability remains unknown. In this study, we found that the deletion of ARV1, which encodes an ER protein of unknown molecular function, activates the UPR by inducing lipid bilayer stress. In arv1Δ cells, the UPR and the cell wall integrity pathway are activated independently of each other, and the high osmolarity glycerol (HOG) pathway is activated in a manner dependent on Ire1, which mediates the UPR. Activated Hog1 translocates the stress response transcription factor Msn2 to the nucleus, where it promotes the expression of nicotinamidase Pnc1, a wellknown Sir2 activator. Following Sir2 activation, rDNA silencing and rDNA stability are promoted. Furthermore, the loss of other ER proteins, such as Pmt1 or Bst1, and ER stress induced by tunicamycin or inositol depletion also enhance rDNA stability in a Hog1-dependent manner. Collectively, these findings suggest that the induction of the UPR enhances rDNA stability in S. cerevisiae by promoting the Msn2-Pnc1- Sir2 pathway in a Hog1-dependent manner. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the lipid-lowering activity of LFBEP-C1 in high glucose-fed Caenorhabditis elegans (C. elegans). In this study, the fermented barley protein LFBEP-C1 was prepared and tested for its potential anti-obesity effects on C. elegans. The worms were fed Escherichia coli OP50 (E. coli OP50), glucose, and different concentrations of LFBEP-C1. Body size, lifespan, movement, triglyceride content, and gene expression were analyzed. The results were analyzed using ANOVA and Tukey's multiple comparison test. Compared with the model group, the head-swing frequency of C. elegans in the group of LFBEP-C1 at 20 μg/mL increased by 33.88%, and the body-bending frequency increased by 27.09%. This indicated that LFBEP-C1 improved the locomotive ability of C. elegans. The average lifespan of C. elegans reached 13.55 days, and the body length and width of the C. elegans decreased after LFBEP-C1 intake. Additionally, LFBEP-C1 reduced the content of lipid accumulation and triglyceride levels. The expression levels of sbp-1 , daf-2 , and mdt-15 significantly decreased, while those of daf-16 , tph-1 , mod-1 , and ser-4 significantly increased after LFBEP-C1 intake. Changes in these genes explain the signaling pathways that regulate lipid metabolism. LFBEP-C1 significantly reduced lipid deposition in C. elegans fed a high-glucose diet and alleviated the adverse effects of a high-glucose diet on the development, lifespan, and exercise behavior of C. elegans. In addition, LFBEP-C1 regulated lipid metabolism mainly by mediating the expression of genes in the sterol regulatory element-binding protein, insulin, and 5-hydroxytryptamine signaling pathways. [ABSTRACT FROM AUTHOR]

The oleaginous yeast, Lipomyces starkeyi can have diverse industrial applications due to its remarkable capacity to use various carbon sources for the biosynthesis intracellular triacylglycerides (TAGs). In L. starkeyi , TAG synthesis is enhanced through upregulation of genes involved in citrate-mediated acyl-CoA synthesis and Kennedy pathways through the transcriptional regulator LsSpt23p. High expression of LsSPT23 can considerably enhance TAG production. Altering the regulatory factors associated with lipid production can substantially augment lipid productivity. In this study, we identified and examined the L. starkeyi homolog sucrose nonfermenting 1 SNF1 (LsSNF1) of YlSNF1 , which encodes a negative regulator of lipid biosynthesis in the oleaginous yeast Yarrowia lipolytica. The deletion of LsSNF1 enhanced TAG productivity in L. starkeyi , suggesting that LsSnf1p is a negative regulator in TAG production. The enhancement of TAG production following deletion of LsSNF1 can primarily be attributed to the upregulation of genes in the citrate-mediated acyl-CoA synthesis and Kennedy pathways, pivotal routes in TAG biosynthesis. The overexpression of LsSPT23 enhanced lipid productivity; strain overexpressing LsSPT23 and without LsSNF1 exhibited increased TAG production capacity per cell. LsSnf1p also has a significant role in the utilization of carbon sources, including xylose or glycerol, in L. starkeyi. Our study results elucidated the role of LsSnf1p in the negative regulation of TAG synthesis in L. starkeyi , which has not previously been reported. • Lipomyces starkeyi SNF1 homolog (LsSNF1) was identified and the LsSNF1 deletion mutant was constructed. • LsSnf1p had important roles in lipid production and carbon-source utilization in L. starkeyi. • The overexpression of LsSpt23p and without LsSnf1p exhibited increased TAG production capacity per cell. [ABSTRACT FROM AUTHOR]

Lipids have been previously implicated in the lifecycle of neuroinvasive viruses. However, the role of lipids in programmed cell death and the relationship between programmed cell death and lipid droplets (LDs) in neuroinvasive virus infection remains unclear. Here, we found that the infection of neuroinvasive virus, such as rabies virus and encephalomyocarditis virus could enhance the LD formation in N2a cells, and decreasing LDs production by targeting diacylglycerol acyltransferase could suppress viral replication. The lipidomics analysis revealed that arachidonic acid (AA) was significantly increased after reducing LD formation by restricting diacylglycerol acyltransferase, and AA was further demonstrated to induce ferroptosis to inhibit neuroinvasive virus replication. Moreover, lipid peroxidation and viral replication inhibition could be significantly alleviated by a ferroptosis inhibitor, ferrostatin-1, indicating that AA affected neuroinvasive virus replication mainly through inducing ferroptosis. Furthermore, AA was demonstrated to activate the acyl-CoA synthetase longchain family member 4-lysophosphatidylcholine acyltransferase 3-cytochrome P450 oxidoreductase axis to induce ferroptosis. Our findings highlight novel cross-talks among viral infection, LDs, and ferroptosis for the first time, providing a potential target for antiviral drug development. [ABSTRACT FROM AUTHOR]

Messenger RNA (mRNA)-based therapy shows immense potential for broad biomedical applications. However, the development of safe and efficacious mRNA delivery vectors remains challenging due to delivery barriers and inefficient intracellular payload release. Herein, we presented a simple strategy to boost the mRNA intracellular release by incorporation of anionic poly(γ-glutamic acid) (PGA) into an ionizable lipid-based LNP/mRNA. We systematically investigated the impact of PGA incorporation on mRNA transfection both in vitro and in vivo. The molecular weights and formulation ratios of PGA greatly affected the transfection efficacy of LNP/mRNA. From in vitro study, the optimized LNP/mRNA/PGA was formulated by incorporation of PGA with the molecular weight of 80 kDa or 200 kDa and the charge ratio (N/P/C) of 25/1/1. The optimized formulation achieved around 3-fold mRNA expression in HeLa cells compared to the bare LNP/mRNA. The intracellular releasing study using specific DNA probe revealed that this enhancement of transfection efficacy was attributed to the elevated mRNA release into cytoplasm. Moreover, the optimized LNP/mRNA/PGA achieved up to 5-fold or 3-fold increase of luciferase mRNA expression in vivo after being injected into mice systematically or intramuscularly, respectively. In addition, the incorporation of PGA did not significantly alter the biodistribution profile of the complexes on both organ and cellular levels. Therefore, our work provides a simple strategy to boost mRNA delivery, which holds great promise to improve the efficacy of mRNA therapeutics for various biomedical applications. The process of designing and screening potent mRNA carriers is complicated and time-consuming, while the efficacy is not always satisfying due to the delivery barriers and inefficient mRNA release. This work presented an alternative strategy to boost the mRNA delivery efficacy by incorporating an anionic natural polymer poly(γ-glutamic acid) (PGA) into LNP/mRNA complexes. The optimized LNP/mRNA/PGA achieved up to 3-fold and 5-fold increase in transfection efficacy in vitro and in vivo , respectively. Intracellular releasing analysis revealed that the enhancement of transfection efficacy was mainly attributed to the elevated intracellular release of mRNA. In addition, the incorporation of PGA did not alter the biodistribution or the biosafety profile of the complexes. These findings indicate that PGA incorporation is a promising strategy to improve the efficacy of mRNA therapeutics. [Display omitted] [ABSTRACT FROM AUTHOR]

XK-related 8 (XKR8), in complex with the transmembrane glycoprotein basigin, functions as a phospholipid scramblase activated by the caspase-mediated cleavage or phosphorylation of its C-terminal tail. It carries a putative phospholipid translocation path of multiple hydrophobic and charged residues in the transmembrane region. It also has a crucial tryptophan at the exoplasmic end of the path that regulates its scrambling activity. We herein investigated the tertiary structure of the human XKR8-basigin complex embedded in lipid nanodiscs at an overall resolution of 3.66 Å. We found that the C-terminal tail engaged in intricate polar and van der Waals interactions with a groove at the cytoplasmic surface of XKR8. These interactions maintained the inactive state of XKR8. Point mutations to disrupt these interactions strongly enhanced the scrambling activity of XKR8, suggesting that the activation of XKR8 is mediated by releasing the C-terminal tail from the cytoplasmic groove. We speculate that the cytoplasmic tail region of XKR8 functions as a plug to prevent the scrambling of phospholipids. [ABSTRACT FROM AUTHOR]

Eukaryotic elongation factor 1A1 (EEF1A1) is canonically involved in protein synthesis but also has noncanonical functions in diverse cellular processes. Previously, we identified EEF1A1 as a mediator of lipotoxicity and demonstrated that chemical inhibition of EEF1A1 activity reduced mouse liver lipid accumulation. These findings suggested a link between EEF1A1 and metabolism. Therefore, we investigated its role in regulating metabolic substrate preference. EEF1A1-deficient Chinese hamster ovary (2E2) cells displayed reduced media lactate accumulation. These effects were also observed with EEF1A1 knockdown in human hepatocyte-like HepG2 cells and in WT Chinese hamster ovary and HepG2 cells treated with selective EEF1A inhibitors, didemnin B, or plitidepsin. Extracellular flux analyses revealed decreased glycolytic ATP production and increased mitochondrial-to-glycolytic ATP production ratio in 2E2 cells, suggesting a more oxidative metabolic phenotype. Correspondingly, fatty acid oxidation was increased in 2E2 cells. Both 2E2 cells and HepG2 cells treated with didemnin B exhibited increased neutral lipid content, which may be required to support elevated oxidative metabolism. RNA-seq revealed a >90-fold downregulation of a rate-limiting glycolytic enzyme, hexokinase 2, which we confirmed through immunoblotting and enzyme activity assays. Pathway enrichment analysis identified downregulations in TNFA signaling via NFKB and MYC targets. Correspondingly, nuclear abundances of RELB and MYC were reduced in 2E2 cells. Thus, EEF1A1 deficiency may perturb glycolysis by limiting NFKB- and MYC-mediated gene expression, leading to decreased hexokinase expression and activity. This is the first evidence of a role for a translation elongation factor, EEF1A1, in regulating metabolic substrate utilization in mammalian cells. [ABSTRACT FROM AUTHOR]

Background: For many years, United States' dietary policy recommended limiting egg intake to no more than 3/wk in the belief that restricting dietary cholesterol would lower plasma cholesterol levels and thereby reduce the risk of cardiovascular disease. The evidence supporting these recommendations is controversial. Objectives: To examine the impact of eggs, a major contributor to dietary cholesterol intake, on lipid levels and to determine whether these egg effects are modified by other healthy dietary factors in adults. Methods: Males and females aged 30-64 y with available 3-d diet record data, without cardiovascular disease and not taking lipid- or glucose-lowering medications in the prospective Framingham Offspring cohort were included (n = 1852). Analysis of covariance models were used to compare mean follow-up lipid levels adjusting for age, sex, BMI, and dietary factors. Cox proportional hazard's models were used to estimate risk for elevated lipid levels. Results: Consuming -5 eggs/wk was not adversely associated with lipid outcomes. Among men, consuming -5 (compared with <0.5) eggs/wk was associated with an 8.6 mg/dL lower total cholesterol level and a 5.9 mg/dL lower LDL cholesterol level, as well as lower triglycerides. Overall, higher egg intake combined with higher dietary fiber (compared with lower intakes of both) was associated with the lowest total cholesterol, LDL cholesterol, and LDL cholesterol-to-HDL cholesterol ratio. Finally, diets with higher (compared with lower) egg intakes in combination with higher total fish or fiber intakes, respectively, were associated with lower risks of developing elevated (>160 mg/dL) LDL cholesterol levels (hazard ratio: 0.61; 95% confidence interval: 0.44, 0.84; and HR: 0.70; 95% confidence interval: 0.49, 0.98, respectively). Conclusions: Higher egg intakes were beneficially associated with serum lipids among healthy adults, particularly those who consumed more fish and dietary fiber. [ABSTRACT FROM AUTHOR]

Many metabolic diseases are caused by disorders of lipid homeostasis. CIDEC, a lipid droplet (LD)-associated protein, plays a critical role in controlling LD fusion and lipid storage. However, regulators of CIDEC remain largely unknown. Here, we established a homogeneous time-resolved fluorescence (HTRF)-based high-throughput screening method and identified LPXN as a positive regulatory candidate for CIDEC. LPXN and Hic-5, the members of the Paxillin family, are focal adhesion adaptor proteins that contribute to the recruitment of specific kinases and phosphatases, cofactors, and structural proteins, participating in the transduction of extracellular signals into intracellular responses. Our data showed that Hic-5 and LPXN significantly increased the protein level of CIDEC and enhanced CIDEC stability not through triacylglycerol synthesis and FAK signaling pathways. Hic-5 and LPXN reduced the ubiquitination of CIDEC and inhibited its proteasome degradation pathway. Furthermore, Hic-5 and LPXN enlarged LDs and promoted lipid storage in adipocytes. Therefore, we identified Hic-5 and LPXN as novel regulators of CIDEC. Our current findings also suggest intervention with Hic-5 and LPXN might ameliorate ectopic fat storage by enhancing the lipid storage capacity of white adipose tissues. [ABSTRACT FROM AUTHOR]

Lipid nanovehicles are currently the most advanced vehicles used for RNA delivery, as demonstrated by the approval of patisiran for amyloidosis therapy in 2018. To illuminate the unique superiority of lipid nanovehicles in RNA delivery, in this review, we first introduce various RNA therapeutics, describe systemic delivery barriers, and explain the lipid components and methods used for lipid nanovehicle preparation. Then, we emphasize crucial advances in lipid nanovehicle design for overcoming barriers to systemic RNA delivery. Finally, the current status and challenges of lipid nanovehicle-based RNA therapeutics in clinical applications are also discussed. Our objective is to provide a comprehensive overview showing how to utilize lipid nanovehicles to overcome multiple barriers to systemic RNA delivery, inspiring the development of more high-performance RNA lipid nanovesicles in the future. This review provides a comprehensive overview of how to design lipid nanovehicles to overcome multiple extracellular and intracellular barriers to systemic RNA delivery. [Display omitted] [ABSTRACT FROM AUTHOR]

Polymeric prodrugs have been applied to control the delivery of various types of therapeutics. Similarly, conjugation of peptide therapeutics to lipids has been used to prolong systemic exposure. Here, we extend on these two approaches by conjugating brush polyethylene glycol (PEG) polymers with different lipid components including short-chain (1C2) or medium-chain (1C12) monoalkyl hydrocarbon tails, cholesterol (Cho), and diacylglycerols composed of two medium-chain (2C12) or long-chain (2C18) fatty acids. We uniquely evaluate the integration of these lipid-polymers into endogenous lipid trafficking pathways (albumin and lipoproteins) and the impact of lipid conjugation on plasma pharmacokinetics after intravenous (IV) and subcutaneous (SC) dosing to cannulated rats. The IV and SC elimination half-lives of Cho-PEG (13 and 22 h, respectively), 2C12-PEG (11 and 17 h, respectively) and 2C18-PEG (12 h for both) were prolonged compared to 1C2-PEG (3 h for both) and 1C12-PEG (4 h for both). Interestingly, 1C2-PEG and 1C12-PEG had higher SC bioavailability (40 % and 52 %, respectively) compared to Cho-PEG, 2C12-PEG and 2C18-PEG (25 %, 24 % and 23 %, respectively). These differences in pharmacokinetics may be explained by the different association patterns of the polymers with rat serum albumin (RSA), bovine serum albumin (BSA) and lipoproteins. For example, in pooled plasma (from IV pharmacokinetic studies), 2C18-PEG had the highest recovery in the high-density lipoprotein (HDL) fraction. In conclusion, the pharmacokinetics of brush PEG polymers can be tuned via conjugation with different lipids, which can be utilised to tune the elimination half-life, biodistribution and effect of therapeutics for a range of medical applications. Lipidation of therapeutics such as peptides has been employed to extend their plasma half-life by promoting binding to serum albumin, providing protection against rapid clearance. Here we design and evaluate innovative biomaterials consisting of brush polyethylene glycol polymers conjugated with different lipids. Importantly, we show for the first time that lipidated polymeric materials associate with endogenous lipoprotein trafficking pathways and this, in addition to albumin binding, controls their plasma pharmacokinetics. We find that conjugation to dialkyl lipids and cholesterol leads to higher association with lipid trafficking pathways, and more sustained plasma exposure, compared to conjugation to short and monoalkyl lipids. Our lipidated polymers can thus be utilised as delivery platforms to tune the plasma half-life of various pharmaceuticals. [Display omitted] [ABSTRACT FROM AUTHOR]

Fatty acid handling and complex lipid synthesis are altered in the kidney cortex of diabetic patients. We recently showed that inhibition of the renin-angiotensin system without changes in glycemia can reverse diabetic kidney disease (DKD) and restore the lipid metabolic network in the kidney cortex of diabetic (db/db) mice, raising the possibility that lipid remodeling may play a central role in DKD. However, the roles of specific enzymes involved in lipid remodeling in DKD have not been elucidated. In the present study, we used this diabetic mouse model and a proximal tubule epithelial cell line (HK2) to investigate the potential relationship between long-chain acyl-CoA synthetase 1 (ACSL1) and lipid metabolism in response to fatty acid exposure and inflammatory signals. We found ACSL1 expression was significantly increased in the kidney cortex of db/db mice, and exposure to palmitate or tumor necrosis factor-α significantly increased Acsl1 mRNA expression in HK-2 cells. In addition, palmitate treatment significantly increased the levels of long-chain acylcarnitines and fatty acyl CoAs in HK2 cells, and these increases were abolished in HK2 cell lines with specific deletion of Acsl1(Acsl1KO cells), suggesting a key role for ACSL1 in fatty acid β-oxidation. In contrast, tumor necrosis factor-α treatment significantly increased the levels of short-chain acylcarnitines and long-chain fatty acyl CoAs in HK2 cells but not in Acsl1KO cells, consistent with fatty acid channeling to complex lipids. Taken together, our data demonstrate a key role for ACSL1 in regulating lipid metabolism, fatty acid partitioning, and inflammation. [ABSTRACT FROM AUTHOR]

Microalgae have gained recognition as versatile candidates for the remediation of heavy metals (HMs). This study investigated the biosorption potential of Dunaliella sp. AL1 for copper (Cu(II)) and hexavalent chromium (Cr(VI)) in aqueous solutions. The marine microalga Dunaliella sp. AL1 was exposed to half-sublethal concentrations of both metals in single and bimetallic systems, and responses in algal growth, oxidative stress, photosynthetic pigment production, and photosynthetic performance were evaluated. Cu and/or Cr exposure increased the generation of reactive oxygen species (ROS) in microalgae cells but did not impact algal growth. In terms of photosynthesis, there was a decrease in chlorophylls and carotenoids production in the microalgae culture treated with Cr, either alone or in combination with Cu. The study recorded promising metal removal efficiencies: 26.67%–20.11% for Cu and 94.99%–95.51% for Cr, in single and bimetallic systems, respectively. FTIR analysis revealed an affinity of Cu and Cr ions towards aliphatic/aldehyde C–H, N–H bending, and phosphate groups, suggesting the formation of complex bonds. Biochemical analysis of microalgae biomass collected after the removal of Cr alone or in combination with Cu showed a significant decrease in total carbohydrate content and soluble protein levels. Meanwhile, higher lipid accumulation was recorded and evidenced by BODIPY 505/515 staining. Fatty acid composition analysis by GC revealed a modulation in lipid composition, with a decrease in the ratio of unsaturated fatty acids (UFA) to saturated fatty acids (SFA), in response to Cu, Cr, and Cu–Cr exposure, indicating the suitability of the biomass for sustainable biofuel production. [Display omitted] • Dunaliella sp. AL1 tolerates Cu and/or Cr, despite ROS accumulation. • Efficient removal of Cu and Cr unaffected by the bimetallic system. • Carbohydrates, proteins, and pigments decrease with Cu and/or Cr treatments. • Lipids accumulate, UFA:SFA ratio decreases with Cu and/or Cr treatments. • Feasibility of utilizing biomass after HMs removal for biofuel production. [ABSTRACT FROM AUTHOR]

Adsorption of surfactants to fluid interfaces occurs in numerous technological and daily-life contexts. The coverage at the interface and other properties of the formed adsorption layers determine the performance of a surfactant with regard to the desired application. Given the importance of these applications, there is a great demand for the comprehensive characterization and understanding of surfactant adsorption layers. In this review, we provide an overview of suitable experimental and simulation-based techniques and review the literature in which they were used for the investigation of surfactant adsorption layers. We come to the conclusion that, while these techniques have been successfully applied to investigate Langmuir monolayers of water-insoluble surfactants, their application to the study of Gibbs adsorption layers of water-soluble surfactants has not been fully exploited. Finally, we emphasize the great potential of these methods in providing a deeper understanding of the behavior of soluble surfactants at interfaces, which is crucial for optimizing their performance in various applications. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • LSA has excellent self-assembly capability. • LSA induces MERC via Vdac-Ip3r interaction, driving calcium ion influx and lipid metablism. • LSA enhances ferroptosis via augmented lipid peroxidation through MERC. • LSA has good in vivo antitumor capacity via ferroptosis. Mitochondrial-endoplasmic reticulum contacts (MERC), a crucial mediator in subcellular organelle communication, significantly influences the dynamics of tumor organelle. Despite the well-established correlation between MERC-associated enzymes and lipid peroxidation (LPO), the contribution of MERC to LPO-induced ferroptosis in tumor remains unclear. In this study, we unveil the role of MERC in amplifying ferroptosis within tumor cells by introducing a prodrug LSA (LA-SS-ART, Linoleic acid-disulfide bond-artesunate), which ingeniously utilizes fatty acids as MERC inducers and ferroptosis activators. LSA induces Vdac-Ip3r-mediated MERC, resulting in the influx of Ca2+ from ER into mitochondria. Consequently, mitochondrial dysfunction triggers lipid peroxidation, attributed to hindered β-oxidation. Furthermore, LSA upregulates the expression of Lpcat3 and Mfn2, promoting phospholipid synthesis and enhancing the production of polyunsaturated fatty acid-phospholipids, synergistically intensifying ferroptosis. In conclusion, this study leverages organelle interactions to finely modulate cellular metabolism and membrane function, ultimately amplifying the process of ferroptosis. These findings offer a perspective and direction for innovative cancer therapy. [ABSTRACT FROM AUTHOR]

Nucleic acid therapy is currently the most promising method for treating tumors and genetic diseases and for preventing infectious diseases. However, the biggest obstacle to this therapy is delivery of the nucleic acids to the target site, which requires overcoming problems such as capture by the immune system, the need to penetrate biofilms, and degradation of nucleic acid performance. Designing suitable delivery vectors is key to solving these problems. Lipids—which consist of a hydrophilic headgroup, a linker, and a hydrophobic tail—are crucial components for the construction of vectors. The headgroup is particularly important because it affects the drug encapsulation rate, the vector cytotoxicity, and the transfection efficiency. Herein, we focus on various headgroup structures (tertiary amines, quaternary ammonium salts, peptides, piperazines, dendrimers, and several others), and we summarize and classify important lipid-based carriers that have been developed in recent years. We also discuss applications of cationic lipids with various headgroups for delivery of nucleic acid drugs, and we analyze how headgroup structure affects transport efficiency and carrier toxicity. Finally, we briefly describe the challenges of developing novel lipid carriers, as well as their prospects. [ABSTRACT FROM AUTHOR]

[Display omitted] • LD-VP emits fluorescence at 520 nm in dioxane and 600 nm in viscous media without any interference from biological entities. • The LD-VP probe is pH-independent. • The quantum yield of probe LD-VP has changed from 0.5 % to 30% as the viscosity rises from 0.9 cP to 953 cP. • Probe LD-VP has provided green images in nonpolar intracellular milieu detection and red images for high viscosity. • Probe LD-VP concurrently labeled mitochondria, lysosomes, and lipid droplets in red and green channels based on intracellular viscosity and polarity. The viscosity and polarity of cells are inherent physical parameters, and changes to these attributes are frequently linked to malfunctioning conditions or different diseases. Cancer cells often harbor elevated nonpolar lipid droplets (LDs), acting as energy reserves. The rise in lipid droplets (LDs) and viscosity, distinctive traits of cancer cells, has motivated us to develop a fluorescent probe based on naphthalimide. The fluorescent marker LD-VP exhibits fluorescence signals at 520 nm in nonpolar dioxane and 600 nm in highly viscous glycerol. This implies that probe LD-VP can furnish details regarding cellular microviscosity and lipid droplets within live cells by providing dual-channel images. The probe LD-VP is chemically inert and harmless, enabling its safe utilization in live cell imaging. We conducted fluorescence imaging based on viscosity using probe LD-VP in the red channel, which is found to be co-localized in both mitochondria and lysosomes. Moreover, the probe effectively marks lipid droplets in the green channel. The probe LD-VP represents a distinctive fluorescent tool capable of monitoring both viscosity and lipid droplet expression in live cells. This tool holds the potential for visualizing physiological abnormalities or pathological conditions. [ABSTRACT FROM AUTHOR]

To treat colonic diseases more effectively, improved therapies are urgently needed. In this respect, delivering drugs locally to the colon is a key strategy to achieve higher local drug concentrations while minimizing systemic side effects. Understanding the luminal environment is crucial to efficiently develop such targeted therapies and to predict drug disposition in the colon. In this clinical study, we collected colonic contents from an undisturbed fasted proximal colon via colonoscopy and characterized their composition with regard to drug disposition. Colonic pH, osmolality, protein content, bile salts, lipids, phospholipids and short-chain fatty acids were investigated in 10 healthy volunteers (8 male and 2 female, age 19–25). The unique environment of the proximal colon was reflected in the composition of the sampled luminal fluids and the effect of the microbiota could be observed on the pH (median 6.55), the composition of bile salts (majority deconjugated and secondary), and the abundance of short-chain fatty acids. At the same time, an increase in phospholipid concentration, osmolality and total protein content compared to reported ileal values was seen, likely resulting from desiccation. Lipids could only be found in low quantities and mainly in the form of cholesterol and free fatty acids, showing almost complete digestion and absorption by the time luminal contents reach the colon. All characteristics also displayed the considerable intersubject variability found in different regions of the gastrointestinal tract. This study contributes to an improved understanding of the luminal conditions in the proximal colon and facilitates the development of new predictive tools to study colonic drug absorption. [Display omitted] [ABSTRACT FROM AUTHOR]

As an iron dependent regulatory cell death process driven by excessive lipid peroxides (LPO), ferroptosis is recognized as a powerful weapon for pancreatic cancer (PC) therapy. However, the tumor microenvironment (TME) with hypoxia and elevated glutathione (GSH) expression not only inhibits LPO production, but also induces glutathione peroxidase 4 (GPX4) mediated LPO clearance, which greatly compromise the therapeutic outcomes of ferroptosis. To address these issues, herein, a novel triple-enhanced ferroptosis amplifier (denoted as Zal@HM-PTBC) is rationally designed. After intravenous injection, the overexpressed H 2 O 2 /GSH in TME induces the collapse of Zal@HM-PTBC and triggers the production of oxygen and reactive oxygen species (ROS), which synergistically amplify the degree of lipid peroxidation (broaden sources). Concurrently, GSH consumption because of the degradation of the hollow manganese dioxide (HM) significantly weakens the activity of GPX4, resulting in a decrease in LPO clearance (reduce expenditure). Moreover, the loading and site-directed release of zalcitabine further promotes autophagy-dependent LPO accumulation (enhance effectiveness). Both in vitro and in vivo results validated that the ferroptosis amplifier demonstrated superior specificity and favorable therapeutic responses. Overall, this triple-enhanced LPO accumulation strategy demonstrates the ability to facilitate the efficacy of ferroptosis, injecting vigorous vitality into the treatment of PC. [Display omitted] • Zal@HM-PTBC was designed to facilitate ferroptosis by triple-enhanced LPO accumulation strategy. • TME induced the collapse of Zal@HM-PTBC and triggered the production of O 2 /ROS , which synergistically amplify LPO level. • The degradation of Zal@HM-PTBC consumed GSH in TME and attenuated "detoxification effect" on LPO by inactivating of GPX4. • The release of zalcitabine effectively led to autophagy dependent LPO accumulation. [ABSTRACT FROM AUTHOR]

Biodegradation of hydrophobic volatile organic compounds (VOCs) such as n -hexane is limited by their poor accessibility. Constructing fungal-bacterial degradation alliances is an effective approach, but the role of those fungi without the capability to degrade VOCs may have been overlooked. In this study, a non- n -hexane-degrading fungus, Fusarium keratoplasticum FK, was utilized to enhance n -hexane degradation by the bacterium Mycobacterium neworleansense WCJ. It was shown that strain WCJ removed 64.84% of n -hexane (at a concentration of 648.20 mg L−1) over 3 d, and 84.04% after introducing strain FK. Microbial growth kinetic studies revealed that the growth of strain WCJ was also promoted. Through a stepwise adsorption-degradation experiment combined with qPCR technology, it was found that the strain WCJ could utilize the n -hexane pre-adsorbed by strain FK, with an increase in copy number from 108.2662 to 108.7731. Therefore, the non-degrading fungi can improved the accessibility of n -hexane by providing n-hexane adsorbed by the mycelium to the degrading bacteria. In addition, the adsorption tests and characterization of the fungal samples before and after Soxhlet extraction indicated that the adsorption of n -hexane on strain FK conformed to Lagergren's pseudo-second-order kinetics and Freundlich adsorption isotherms, and was correlated with the presence of lipids and nonpolar groups. This study emphasizes the potential role of non-degrading fungi in bioremediation and proposes a viable strategy to enhance the bacterial degradation of hydrophobic VOCs. [Display omitted] • The introduction of strain FK increased the n -hexane removal to 84.04%. • The k value of strain WCJ was promoted by strain FK from 0.0395 to 0.1681 h−1. • Adsorption of VOCs by fungi through hyphae were confirmed by IFM and kinetics. • Bacterial degradation of the adsorbed n -hexane was indirectly proven by qPCR. • Lipids and nonpolar groups of fungi were found to contribute to the adsorption. [ABSTRACT FROM AUTHOR]

[Display omitted] • A solventless recovery of grease from sewage sludge was designed. • A very high yield of grease recovery was achieved (up to 70 wt%). • Final dewatered sludge resulted in 15 % less than the amount conventionally obtained. • A price of 800–900 € per ton of grease would guarantee the economic sustainability. For the first time, the recovery of lipids from urban sewage sludge was investigated through a solvent-free process based on elementary operations validated on a pilot scale (2 tons per hour). Sedimented sewage sludge was dewatered through centrifugation, acidified (and in the case treated with hydrogen peroxide), heated to 80 °C and centrifuged to obtain three different streams: i) raw grease, ii) exhausted aqueous phase, and iii) dewatered sludge. The recovery yield of grease from sludge could reach a very high value (70 wt%), depending essentially on the initial lipid content in sewage sludge, which was monitored and found to range between 1.1 and 6.6 wt% (13–25 wt% with respect to TS). The use of H 2 O 2 played a crucial role in the recoverability of lipids and the relevant pureness in free fatty acids content (>89 wt%). In addition, the total solids content in the exhausted aqueous phase and the final residual sludge were sensibly affected by using H 2 O 2. Final dewatered sludge had a very high content of solids (TS 31.0 ± 2.3 wt%) and resulted in 15 % less of the amount of sludge conventionally obtained from anaerobic digestion. According to these figures, for medium wastewater treatment plants, a selling price of 800–900 Euro per ton of grease would guarantee a return on investments within three years of operations. This simple procedure represents a quick and proficient way to treat sewage sludge, turning it into a valuable feedstock for fine chemicals, in agreement with Circular Bioeconomy principles. [ABSTRACT FROM AUTHOR]

Changes in protein and lipid levels may occur in the Alzheimer's disease brain, and DHA can have beneficial effects on it. To investigate the impact of DHA dietary intervention on brain protein and lipid profile in ApoE-/- mice and C57 mice. Three-month-old ApoE-/- mice and C57 mice were randomly divided into two groups respectively, and fed with control diet and DHA-fortified diet for five months. Cortical TC, HDL-C and LDL-C levels and cholesterol metabolism-related protein expression were measured by ELISA or immunohistochemistry methods. Hippocampus were collected for proteomic and lipidomics analysis by LC-MS/MS and differential proteins and lipid metabolites were screened and further analyzed by GO functional annotation and KEGG pathway enrichment analysis. DHA intervention decreased cortical TC level in both C57 and ApoE-/- mice (P < 0.05), but caused different change of cortical HDL-C, LDL-C level and LDL-C/HDL-C ratio in C57 and ApoE-/- mice (P < 0.05). Discrepant cortical and hippocampal LDLR, ABCG1, Lox1 and SORT1 protein expression was found between C57 and ApoE-/- mice (P < 0.05), and DHA treatment caused different changes of these proteins in C57 and ApoE-/- mice (P < 0.05). Differential hippocampal proteins and lipids profile were found in C57 and ApoE-/- mice before and after DHA treatment, which were mainly involved in vesicular transport and phospholipid metabolic pathways. ApoE genetic defect caused abnormal cholesterol metabolism, and affected protein and lipid profile, as well as discrepant response of hippocampal protein and lipids profile in the brain of mice given DHA fortified diet intervention. • C57 and ApoE-/- mice showed different protein and lipid profile in hippocampus • The hippocampal differential proteins in DHA-fed mice are mainly associated with vesicular transport. • DHA intervention caused change of hippocampal lipids profile, which mainly involved in the metabolism of glycerophospholipid. [ABSTRACT FROM AUTHOR]

[Display omitted] • Oilcane" represents a renewable feedstock for co-production of lipids, sugars, and lignin. • Two greener biomass deconstruction methods, i.e., NADES and HT pretreatments were investigated. • Treating with ChCl:LA enabled the removal of > 78.8 % lignin and 80.4 % of hemicellulose. • Enzymatic conversion yields 82.7 % of the available sugars using the recovered solids treatment with ChCl:LA. • High solid loading pretreatment (50 %) could be achieved using ChCl:LA NADES. • Lipids remained intact during bagasse processing and lipid content was enriched by 2.6-folds. Sugarcane is being enhanced as a bioenergy crop by engineering it to accumulate and store lipids along with polymeric sugars in vegetative tissues. However, there is no existing process that allows for processing this new crop to recover both lipid and cellulosic sugars from the oilcane bagasse. Therefore, a comprehensive investigation of two pretreatment methods—natural deep eutectic solvents (NADES) and chemical-free hydrothermal pretreatment (HT) was conducted to judge their suitability for recovering fermentable sugars, lipids, and lignin from bagasse. Two NADES, i.e., choline chloride: lactic acid (ChCl:LA) and betaine: lactic acid (BT:LA) were prepared using a 1:2 M ratio and were evaluated for pretreatment of oilcane bagasse at 10, 20, and 50 % (w/w) solids, followed by enzymatic hydrolysis at 10 % (w/w) solids. Notably, ChCl:LA NADES treatment at 10 % (w/w) solids at 140 °C for 2 h, solubilized 78.8 % of lignin and 80.4 % of hemicellulose and allowed 82.7 % enzymatic conversion of glucans to glucose. In contrast, HT pretreatment removed approximately 87.6 % of the hemicellulose and provided an enzymatic glucose yield of 69.7 %. Furthermore, ChCl:LA operated at 50 % solids loading the enriched lipids 2.6-fold (9.2 wt%) in recovered solids compared to HT (6.4 %) and BT:LA (5.1 %) pretreatment processes. NMR-HSQC and GPC analysis showed that ChCl:LA also cleaved the most lignin β–O–4 linkages and demonstrated lower molecular weight compared to HT. This study demonstrates that NADES pretreatment is an effective green processing method for recovering lipids, sugars, and lignin from bioenergy crops at high solid loading (50 % w/w) within the context of an integrated biorefinery. [ABSTRACT FROM AUTHOR]

Glucose and lipid metabolic disorders (GLMDs), such as diabetes, dyslipidemia, metabolic syndrome, nonalcoholic fatty liver disease, and obesity, are significant public health issues that negatively impact human health. The endoplasmic reticulum (ER) plays a crucial role at the cellular level for lipid and sterol biosynthesis, intracellular calcium storage, and protein post-translational modifications. Imbalance and dysfunction of the ER can affect glucose and lipid metabolism. As an essential trace element, selenium contributes to various human physiological functions mainly through 25 types of selenoproteins (SELENOs). At least 10 SELENOs, with experimental and/or computational evidence, are predominantly found on the ER membrane or within its lumen. Two iodothyronine deiodinases (DIOs), DIO1 and DIO2, regulate the thyroid hormone deiodination in the thyroid and some external thyroid tissues, influencing glucose and lipid metabolism. Most of the other eight members maintain redox homeostasis in the ER. Especially, SELENOF, SELENOM, and SELENOS are involved in unfolded protein responses; SELENOI catalyzes phosphatidylethanolamine synthesis; SELENOK, SELENON, and SELENOT participate in calcium homeostasis regulation; and the biological significance of thioredoxin reductase 3 in the ER remains unexplored despite its established function in the thioredoxin system. This review examines recent research advances regarding ER SELENOs in GLMDs and aims to provide insights on ER-related pathology through SELENOs regulation. [Display omitted] • Endoplasmic reticulum (ER) -resident selenoproteins are important molecules in regulation of ER-associated glucose and lipid metabolism. • ER-resident selenoproteins act as guardians to regulate ER function and homeostasis. • ER-resident selenoproteins may explain the effect of dietary selenium levels on glucose and lipid metabolic disorders. [ABSTRACT FROM AUTHOR]

In this study, the process conditions, physicochemical properties, structural composition and activity of polysaccharides isolated from leechee peel (LPP) by ultrasound-assisted extraction (UAE) with enzyme and alkali solution extraction (ASE) were compared. The results showed that the total sugar content of LPP extracted by UAE accounted for 75.65 %, which was significantly higher than that extracted by alkali solution. The optimum conditions were as follows: extraction temperature of 68.78 ℃, ultrasonic enzymolysis time of 39.68 min, pectinase dosage of 4.03 %, solid–liquid ratio of 1:30 g/mL, and ultrasonic power of 360 W. The antioxidant activities and structure of leechee peel polysaccharide (LPP) prepared under different conditions were compared. It was found that UAE-LPP was an α-type polysaccharide containing 15.83 % uronic acid. Moreover, LPP extracted by UAE showed strong activity in anti-lipid peroxidation and reducing ability. Ultrasound-assisted enzymatic method is an effective means to improve the content and activity of natural plant polysaccharides, and this method has the advantages of short time-consuming, simple process and easy operation, which can greatly improve the utilization rate of polysaccharides and lay a theoretical and scientific basis for the development and utilization of LPP. [ABSTRACT FROM AUTHOR]

In this study, a cyanobacteria-bacteria consortium containing native wastewater bacteria and immobilized Synechococcus sp. was constructed. The cyanobacterial cellular responses (including growth, biomass and lipid productivity) and contaminant removal ability (for TN, TP, COD and antibiotics) in the consortium were evaluated during the advanced treatment of wastewater containing 10–50 μg/L of mixed antibiotics (amoxicillin, tetracycline, erythromycin, sulfadiazine and ciprofloxacin) with the addition of a certain phytohormone (indole-3-acetic acid, gibberellin A3 or 6-benzylaminopurine) at trace level within a period of four days. Each phytohormone promoted the growth of Synechococcus sp. and increased the tolerance of Synechococcus sp. to mixed antibiotics. Indole-3-acetic acid coupled to moderate antibiotic stress could elevate lipid productivity and lipid content of Synechococcus sp. to 33.50 mg/L/day and 43.75%, respectively. Phytohormones increased the pollutant removal performance of the cyanobacteria-bacteria consortium through the stimulation of cyanobacterial growth and the regulation of cyanobacteria-bacteria interaction, which increased the abundances of microalgae-associated bacteria including Flavobacterium , Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium , Bosea , Sphingomonas and Emticicia. Up to 80.83%, 98.06%, 83.26%, 99.84%, 99.50%, 89.41%, 65.61% and 60.65% of TN, TP, COD, amoxicillin, tetracycline, erythromycin, sulfadiazine and ciprofloxacin were removed by the consortium with the addition of phytohormones. In general, indole-3-acetic acid was the optimal phytohormone for enhancing lipid production and contaminant removal performance of the cyanobacteria-bacteria consortium. [Display omitted] • Phytohormones increase the tolerance of cyanobacteria to mixed antibiotics. • Antibiotics and phytohormones regulate cyanobacteria-bacteria interaction. • Phytohormones elevate cyanobacterial lipid productivity. • Phytohormones elevate pollutant removal property of cyanobacteria-bacteria consortium. • Indole-3-acetic acid show optimal promotion performance. [ABSTRACT FROM AUTHOR]

The safe and effective delivery of messenger ribonucleic acid (mRNA) is crucial for its therapeutic effects in vivo. In this study, we developed a new type of ionizable lipid S-1, which contains an amino head, a cholesterol matrix, and a long hydrophobic carbon tail. We employed microfluidics to rapidly mix an ethanol phase containing S-1 lipid with an aqueous mRNA to form mRNA/S-1 lipid nanoparticles (LNPs, 100–200 nm). We observed low cytotoxicity and high transfection efficiency in RAW264.7 and HCT-116 cell lines for mRNA/S-1 LNPs, comparable to mRNA/SM-102 LNPs. Based on the obtained findings, mRNA/S-1 LNPs have good stability, low cytotoxicity, high transfection efficiency, and enhanced cellular uptake. The synthesized S-1 lipid ensures efficient assembly of lipid nanoparticles, protects mRNA from RNase degradation, and enables the delivery of mRNA into the cytoplasm for translation. • A novel ionizable lipid S-1 was synthesized. • Novel lipid nanoparticles S-1 LNPs were prepared by microfluidic equipment. • S-1 LNPs achieved successful mRNA delivery in vitro and vivo. [ABSTRACT FROM AUTHOR]

• We evaluated two grape pomace varieties as rearing media for black soldier fly larvae. • The growth rate was higher when the larvae were reared on Becuet than Moscato pomace. • The larvae fed on Moscato contained higher total lipids than the larvae fed on Becuet. • Both grape pomaces and black soldier fly larvae contained conjugated linoleic acids. • Grape pomace can be used to enrich larvae with potential health-promoting fatty acids. The black soldier fly (Hermetia illucens) is attracting increasing interest for its ability to convert low-value substrates into highly nutritious feed. This study aimed at evaluating grape pomace from two varieties (Becuet – B ; Moscato – M) as rearing substrates for black soldier fly larvae (BSFL), focusing on the related effects on larval growth performance, proximate composition, and fatty acid profile. A total of six replicates per treatment, and 1 000 BSFL per replica, were used. Larval development was assessed by larvae weight, which was recorded eight times during the trial: the day after the beginning of the trial, and then on days 5, 8, 13, 15, 20, 22, and 27 (day in which the 30% of BSFL reached the prepupal stage). Production and waste reduction efficiency parameters, namely the growth rate, substrate reduction and substrate reduction index, were calculated. The two grape pomace varieties were analysed for their proximate composition and fatty acid profile; the same analyses were conducted on BSFL (30 larvae per replica) that were collected at the end of the trial (day 27). The growth rate of BSFL showed a higher value when the larvae were reared on B substrate (4.4 and 3.2 mg/day for B and M, respectively; P < 0.01). The rearing substrate did not significantly affect the proximate composition of BSFL. The percentage of total lipids (TL) in M-fed BSFL was significantly higher than in B ones. Total saturated (P < 0.001) and monounsaturated fatty acids (P < 0.05) were significantly higher in M-fed BSFL, while an opposite trend was observed for total branched-chain (P < 0.001) and total polyunsaturated fatty acids (P < 0.001). Interestingly, some conjugated linoleic acid (CLA) isomers [i.e., C18:2 c 9 t 11(+ t 7 c 9+ t 8 c 10) and t 9 t 11] were detected in low amounts in both rearing substrates (total CLA equal to 0.085 and 0.16 g/100 g TL in B and M substrate, respectively). Some CLA isomers (i.e., C18:2 c 9 t 11, t 7 c 9, and t 10 c 12) were also found in BSFL, reaching a total CLA concentration equal to 2.95 and 0.052 g/100 g of TL in B-fed and M-fed BSFL, respectively. This study demonstrates that winery by-products from different grape varieties can significantly affect the development and lipid composition of BSFL. The CLA biosynthesis potential of BSFL opens newsworthy perspectives for a new valorisation of winery by-products to produce full-fat black soldier fly meal and black soldier fly oil enriched in specific fatty acids of potential health-promoting interest. [ABSTRACT FROM AUTHOR]

Thraustochytrids are marine microorganisms known for their fast growth and ability to store lipids, making them useful for producing polyunsaturated fatty acids (PUFAs), biodiesel, squalene, and carotenoids. However, the high cost of production, mainly due to expensive fermentation components, limits their wider use. A significant challenge in this context is the need to balance production costs with the value of the end products. This review focuses on integrating the efficient utilization of waste with Thraustochytrids fermentation, including the economic substitution of carbon sources, nitrogen sources, and fermentation water. This approach aligns with the 3Rs principles (reduction, recycling, and reuse). Furthermore, it emphasizes the role of Thraustochytrids in converting waste into lipid chemicals and promoting sustainable circular production models. The aim of this review is to emphasize the value of Thraustochytrids in converting waste into treasure, providing precise cost reduction strategies for future commercial production. • Excel in lipid chemical production and lipid accumulation (>60%). • Thraustochytrids have the ability to naturally utilize multiple waste types. • Waste has the ability to enhance the production capacity in Thraustochytrids. • Potential for circular economy via internal, external recycling loops. [ABSTRACT FROM AUTHOR]

• Urban stressors' impacts on freshwater biofilms were assessed in a 10-day experiment. • We tested 2 anthropogenic stressors: Artificial light at night (ALAN) and the biocide DDBAC. • ALAN affected DDBAC concentrations in water. • DDBAC exposure strongly inhibited photosynthetic activity. • DDBAC also impacted biofilm composition (taxonomy, pigments and lipids). Aquatic ecosystems and their communities are exposed to numerous stressors of various natures (chemical and physical), whose impacts are often poorly documented. In urban areas, the use of biocides such as dodecyldimethylbenzylammonium chloride (DDBAC) and their subsequent release in wastewater result in their transfer to urban aquatic ecosystems. DDBAC is known to be toxic to most aquatic organisms. Artificial light at night (ALAN) is another stressor that is increasing globally, especially in urban areas. ALAN may have a negative impact on photosynthetic cycles of periphytic biofilms, which in turn may result in changes in their metabolic functioning. Moreover, studies suggest that exposure to artificial light could increase the biocidal effect of DDBAC on biofilms. The present study investigates the individual and combined effects of DDBAC and/or ALAN on the functioning and structure of photosynthetic biofilms. We exposed biofilms in artificial channels to a nominal concentration of 30 mg.L–1 of DDBAC and/or ALAN for 10 days. ALAN modified DDBAC exposure, decreasing concentrations in the water but not accumulation in biofilms. DDBAC had negative impacts on biofilm functioning and structure. Photosynthetic activity was inhibited by > 90% after 2 days of exposure, compared to the controls, and did not recover over the duration of the experiment. Biofilm composition was also impacted, with a marked decrease in green algae and the disappearance of microfauna under DDBAC exposure. The integrity of algal cells was compromised where DDBAC exposure altered the chloroplasts and chlorophyll content. Impacts on autotrophs were also observed through a shift in lipid profiles, in particular a strong decrease in glycolipid content was noted. We found no significant interactive effect of ALAN and DDBAC on the studied endpoints. [Display omitted] [ABSTRACT FROM AUTHOR]

• Application of ultrasound technology to freeze storage. • UIF-175 W can effectively minimized the changes in MP structure. • UIF-175 W can protect the lipid during the freeze process. • A schematic illustration of protein and lipid oxidations was proposed. In this study, large yellow croaker (Larimichthys crocea) was frozen using multi-frequency ultrasound-assisted freezing (MUIF) with different powers (160 W, 175 W, and 190 W, respectively) and stored at −18 °C for ten months. The effect of different ultrasound powers on the myofibrillar protein (MP) structures and lipid oxidation of large yellow croaker was investigated. The results showed that MUIF significantly slowed down the oxidation of MP by inhibiting carbonyl formation and maintaining high sulfhydryl contents. These treatments also held a high activity of Ca2+-ATPase in the MP. MUIF maintained a higher ratio of α-helix to β-sheet during frozen storage, thereby protecting the secondary structure of the tissue and stabilizing the tertiary structure. In addition, MUIF inhibited the production of thiobarbituric acid reactive substances value and the loss of unsaturated fatty acid content, indicating that MUIF could better inhibit lipid oxidation of large yellow croaker during long-time frozen storage. [ABSTRACT FROM AUTHOR]

• The lipid profile of the larvae is influenced by the composition of the substrate. • The larvae synthesize nutraceutical fats by modulating them in different ways. • There is a different lipid synthesis strategy related to genetics of a population. • By changing the substrate, it is possible to adapt the larvae to different feed purposes. • Hermetia illucens can enhance different types of substrates. Black soldier fly larvae (Hermetia illucens ; BSFL) can transform organic wastes into nutritional biomass useful in animal feeding. The aim of this work was to study the effect of five diets (meat, fruit, vegetable substrates, a mix of them and control) on the profile of fatty acids (FA s) and sterols of BSFL. For a more exhaustive characterization of the nutritional properties, the profile of esterified FAs in the sn-2 position of the triglycerides, the most absorbed lipid component during animal digestion was evaluated. The dietary effect was estimated on two different Hermetia illucens populations (Greek − UTH and Italian − UNIPI). The diet affected all the lipid fractions examined. Regardless of diet, the fat was characterized mainly of lauric acid and other saturated FAs, which were found to be synthesized by the larvae, as it was not present in any of the five substrates. In general, UTH larvae contained a higher level of lipids (7.38 vs 2.48 g/100 g of larvae; P < 0.001) and saturated FAs (49.71 vs 36.10 g/100 g of Total Lipids; P < 0.001) and a lower percentage of monounsaturated FAs (14.74 vs 26.70 g/100 g of Total Lipids), C18:3n-3 (0.67 vs 1.13 g/100 g of Total Lipids; P < 0.001), and C18:2c9t11 (2.02 vs 2.80 g/100 g of Total Lipids; P < 0.001). Irrespective of the populations, BSFL reared on control and fruit substrates showed higher level of lipids (8.06 and 5.61 g/100 g of larvae, respectively), and saturated FA (38.99 and 71.19 g/100 g of Total Lipids, respectively), while the presence of meat increased the level of C20:4n-6, C20:5n-3 and C22:5n-3 (0.70, 0.13 and 0.45 g/100 g of Total Lipids, respectively). The results confirmed that BSFL accumulate phytosterols in their lipid fraction. The sterol profile was strongly influenced by the substrate on which the larvae were reared, with higher levels of cholesterol in the larvae of the meat group (38.55 mg/100 g of Total Lipids) and of stigmasterol and campesterol (9.04 and 15.23 mg/100 g of Total Lipids, respectively) in those of the vegetable group. The sterol content between the two populations was significantly different, with a higher percentage in UTH larvae (113.28 vs 34.03 mg/100 g of Total Lipids; P < 0.001). Finally, BSFLs showed a high plasticity of the lipid profile depending on both the substrate and the metabolism linked to the different populations. This variability allows the nutritional characteristics of the BSFL to be shaped by modifying the substrate, to adapt it to the technological and feeding needs to which the larvae are destined. [ABSTRACT FROM AUTHOR]

• Combining two CH 4 mitigation strategies can enhance their effectiveness for dairy cows. • 3-Nitrooxypropanol and cottonseeds were supplemented twice-daily at grazing. • 3-Nitrooxypropanol mildly decreased CH 4 with starch, but not with cottonseeds. • The additive efficacy was likely affected by supply mode and cottonseed composition. • Continuous additive delivery is needed to boost effectiveness in grazing systems. No single enteric CH 4 mitigating strategy has been consistently effective or is readily applicable to ruminants in grassland systems. When CH 4 mitigating strategies are effective under grazing conditions, mitigation is mild to moderate at best. A study was conducted to evaluate the potential of combining two CH 4 mitigation strategies deemed feasible to apply in grazing dairy cows, the methanogenesis inhibitor 3-nitrooxypropanol additive (3-NOP) and cottonseed supplementation (CTS), seeking to enhance their individual CH 4 mitigating potential. Forty-eight dairy cows were evaluated in a continuous grazing study and supplemented with either a starch-based concentrate (STA) or one that contained cottonseeds (1.75 kg DM/d; CTS), and with either 19 g/d of 10% 3-NOP (Bovaer®) or the additive's carrier (placebo), in a 2 × 2 factorial arrangement of treatments. Treatments were supplied mixed with a concentrate supplement (5 kg/d as fed) and offered in two equal rations at milking. Methane emissions were measured on weeks 4 and 8 using the sulphur hexafluoride tracer gas technique over a 5-d period. The 3-NOP and CTS treatments tended to interact on absolute CH 4 such that 3-NOP decreased CH 4 by 13.4% with STA, but there was no mitigation with 3-NOP and CTS. Treatment interactions were also obtained for CH 4 yield, where 3-NOP tended to decrease CH 4 when supplied with STA, and tended to increase it with CTS. The increase in CH 4 yield with the CTS diet was driven by a numerical decrease in DM intake. Methane intensity was not affected by the 3-NOP or CTS treatments. Total volatile fatty acids in ruminal fluid were not affected by 3-NOP supplementation, but a reduction in acetate and an increase in propionate proportion occurred, resulting in decreased acetate: propionate. The 3-NOP additive decreased grass intake; however, energy−corrected milk yield and milk composition were largely unaffected. Milk urea increased with 3-NOP supplementation. Combining twice daily supplementation of 3-NOP and CTS did not enhance their CH 4 mitigation potential when fed to grazing dairy cows. The relatively low inhibition of CH 4 production by 3-NOP compared to studies with total mixed rations may result from the mode of delivery (pulse dosed twice daily) and time gap caused by experimental handling and moving of animals to pasture after 3-NOP supplementation in the milking parlour, which could have impaired the synchrony between the additive presence in the rumen and grass intake in paddocks. [ABSTRACT FROM AUTHOR]

Microalgae are eukaryotic photosynthetic micro-organisms that convert CO 2 into carbohydrates, lipids, and other valuable metabolites. They are considered promising chassis for the production of various bioproducts, including fatty acid–derived biofuels. However, algae-based biofuels are not yet commercially available, mainly because of their low yields and high production cost. Optimizing strains to improve lipid productivity using the principles of synthetic biology should help move forward. This necessitates developments in the following areas: (1) identification of molecular bricks (enzymes, transcription factors, regulatory proteins etc.); (2) development of genetic tools; and (3) availability of high-throughput phenotyping methods. Here, we highlight the most recent developments in some of these areas and provide examples of the use of genome editing tools to improve oil content. [ABSTRACT FROM AUTHOR]

Ferroptosis is a programmed form of cell death regulated by iron and has been linked to the development of asthma. However, the precise mechanisms driving ferroptosis in asthma remain elusive. To gain deeper insights, we conducted an analysis of nasal epithelial and sputum samples from the GEO database using three machine learning methods. Our investigation identified a pivotal gene, Arachidonate 15-lipoxygenase (ALOX15), associated with ferroptosis in asthma. Through both in vitro and in vivo experiments, we further confirmed the significant role of ALOX15 in ferroptosis in asthma. Our results demonstrate that ferroptosis manifests in an HDM/LPS-induced allergic airway inflammation (AAI) mouse model, mimicking human asthma, and in HDM/LPS-stimulated 16HBE cells. Moreover, we observed an up-regulation of ALOX15 expression in HDM/LPS-induced mice and cells. Notably, silencing ALOX15 markedly decreased HDM/LPS-induced ferroptosis in 16HBE cells. These findings indicate that ferroptosis may be implicated in the onset and progression of asthma, with ALOX15 -induced lipid peroxidation raising the susceptibility to ferroptosis in asthmatic epithelial cells. • Ferroptosis may be implicated in the beginning and development of asthma. • ALOX15-induced lipid peroxidation raising the susceptibility to ferroptosis in asthmatic epithelial cells. • ALOX15 combines with PEBP1 to participate in ferroptosis of asthma epithelial cells. [ABSTRACT FROM AUTHOR]

The stratum corneum (SC) lipid matrix, composed primarily of ceramides (CERs), cholesterol and free fatty acids (FFA), has an important role for the skin barrier function. The presence of the long periodicity phase (LPP), a unique lamellar phase, is characteristic for the SC. Insight into the lipid molecular arrangement within the LPP unit cell is imperative for understanding the relationship between the lipid subclasses and the skin barrier function. In this study, the impact of the CER head group structure on the lipid arrangement and barrier functionality was investigated using lipid models forming the LPP. The results demonstrate that the positions of CER N -(tetracosanoyl)-sphingosine (CER NS) and CER N -(tetracosanoyl)-phytosphingosine (CER NP), two essentials CER subclasses, are not influenced by the addition of another CER subclass (N -(tetracosanoyl)-dihydrosphingosine (CER NdS), N -(2R-hydroxy-tetracosanoyl)-sphingosine (CER AS) or D-(2R-hydroxy-tetracosanoyl)-phytosphingosine (CER AP)). However, differences are observed in the lipid organization and the hydrogen bonding network of the three different models. A similar localization of CER NP and CER NS is also observed in a more complex lipid model, with the CER subclass composition mimicking that of human SC. These studies show the adaptability and insensitivity of the LPP unit cell structure to changes in the lipid head group structures of the CER subclasses. [Display omitted] • The LPP unit cell adapts to changes in the CER head group structure. • A change in CER headgroup structure leads to changes in hydrogen bonding and TEWL. • CER headgroup does not affect the acyl chain location of CERs in the LPP unit. • A similar lipid arrangement is observed for a model mimicking human CER composition. [ABSTRACT FROM AUTHOR]

Lipid-based formulations (LBFs) are an enabling-formulation approach for lipophilic poorly water-soluble compounds. In LBFs, drugs are commonly pre-dissolved in lipids, and/or surfactants/cosolvents, hereby avoiding the rate-limiting dissolution step. According to the Lipid formulation classification system, proposed by Pouton in 2006, in type II LBFs a surfactant with an HLB-value lower than 12 is added to the lipids. If high drug doses are required, e.g. for preclinical toxicity studies, supersaturated LBFs prepared at elevated temperatures may be a possibility to increase drug exposure. In the present study, the impact of digestion on drug absorption in rats was studied by pre-dosing of the lipase inhibitor orlistat. The lipid chain length of the type II LBFs was varied by administration of a medium-chain- (MC) and a long-chain (LC)-based formulation. Different drug doses, both non-supersaturated and supersaturated, were applied. Due to an inherent precipitation tendency of cinnarizine in supersaturated LBFs, the effect of the addition of the precipitation inhibitor Soluplus® was also investigated. The pharmacokinetic results were also evaluated by multiple linear regression. In most cases LC-based LBFs did not perform better in vivo, in terms of a higher area under the curve (AUC 0– 24 h) and maximal plasma concentration (C max), than MC-based LBFs. The administration of supersaturated LBFs resulted in increased AUC 0– 24 h (1.5 – 3.2-fold) and C max (1.1 – 2.6-fold)-values when compared to the non-supersaturated equivalents. Lipase inhibition led to a decreased drug exposure in most cases, especially for LC formulations (AUC 0– 24 h reduced to 47 – 67%, C max to 46 – 62%). The addition of Soluplus® showed a benefit to drug absorption from supersaturated type II LBFs (1.2 – 1.7-fold AUC 0– 24 h), due to an increased solubility of cinnarizine in the formulation. Upon dose-normalization of the pharmacokinetic parameters, no beneficial effect of Soluplus® could be demonstrated. [Display omitted] [ABSTRACT FROM AUTHOR]

Energy is an expensive component of diets with lipids providing a concentrated energy source to meet these needs; in addition, supplementary lipids affect milling efficiency and diet palatability. Because of the increased use of lipids as feedstocks in renewable energy production, typical fats and oils are becoming more limited and costly to the livestock industry necessitating the need to source and understand the caloric values of alternative lipids. The experiments reported herein determined the caloric value of typical and non-typical fats when supplemented to growing broilers and compared these empirical values to values predicted by commonly used equations. Thirteen sources of lipids consisting of an animal-vegetable fat blend, poultry fat, 3 soybean oils, 3 distillers corn oils, palm oil, acidulated canola-soybean oil soapstock, acidulated palm oil-soybean oil soapstock, flaxseed oil, and canola oil were evaluated. Differences in the determined nitrogen corrected apparent metabolizable energy (kcal/kg) were observed among the lipids tested (animal-vegetable fat blend, 7,671; poultry fat, 7,984; 3 soybean oils, 8,118, 7,535, and 8,767; 3 distillers corn oils, 8,205, 7,990, and 9,364; palm oil, 7,408; acidulated canola-soybean oil soapstock, 8,056;, acidulated palm oil-soybean oil soapstock, 7,685; flaxseed oil, 8,588; and canola oil, 7,854) and the use of a commonly used prediction equation did not adequately estimate the caloric value compared with the empirical values. Data presented also shows the difficulty of determining and predicting energy values of lipids when used at low inclusion levels. [ABSTRACT FROM AUTHOR]

PFOA is one of the most representative compounds in the family of perfluorinated organic compounds. Due to its varying toxicity, alternatives to PFOA are beginning to emerge. HFPO-TA is an alternative for PFOA. It is currently unclear whether HFPO-TA affects glucose and lipid metabolism. In this study, rats were used as an animal model to investigate the effects of HFPO-TA on liver glucose and lipid metabolism. We found that HFPO-TA can affect glucose tolerance. Through omics analysis and molecular detection, it was found that HFPO-TA mainly affects the PPAR signaling pathway in the liver of rats, inhibiting liver glycolysis while promoting glucose production. HFPO-TA not only promotes the synthesis of fatty acids in the liver, but also promotes the breakdown of fatty acids, which ultimately leads to the disruption of hepatic glucose and lipid metabolism. The effects of HFPO-TA on metabolism are discussed in this paper to provide a reference for the risk assessment of this PFOA substitute. [Display omitted] • HFPO-TA affects the genes, metabolites involved in glucose and lipid metabolism. • HFPO-TA inhibits glycolysis while promoting glucose synthesis. • HFPO-TA affects the PPAR signaling pathway. [ABSTRACT FROM AUTHOR]

[Display omitted] • Electrostatic interactions between PC and TAP phospholipid headgroups change bilayer packing at equilibrium. • At low DMTAP concentrations, lipid transfer takes place asymmetrically with delayed DMPC desorption. • At intermediate and high DMTAP concentrations, transfer is governed by time-dependent concentration changes in donor and acceptor vesicles. • Experimental results call for more complete models implementing time-dependent changes in desorption rates. The kinetics of spontaneous monomer transfer between vesicles consisting of zwitterionic phospholipids is dictated by the difference in desorption rate of lipid monomers from their donor vesicles and the concentration imbalance in the dispersion. In a system with two lipid species with the same headgroup, transfer is asymmetric, and takes place from the population of donor vesicles consisting of shorter chain lipids to acceptor ones of longer chain. Transfer typically proceeds until equilibrium is reached, resulting in populations of vesicles consisting of a binary mixture of both lipid species, whose concentration depends on the number of lipids in the precursor donor and acceptor vesicles before transfer. Upon the introduction of a second lipid species in the donor vesicle population, the desorption rate of monomers should change with time, since the composition of donor vesicles changes when monomers of a given lipid type desorb. To tackle this problem, we added a cationic lipid, 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), into donor zwitterionic lipid vesicles and assessed how the concentration of DMTAP affects the lipid transfer process. Lipid transfer is the result of the interplay between the initial concentration of DMTAP in the donor vesicles (and related probability of desorption at short transfer times) and their concomitant time-dependent concentration (and thus desorption rate) change due to the depletion of monomer species as the transfer process proceeds. [ABSTRACT FROM AUTHOR]

• mRNA-LNP suspensions were produced with a ceramic membrane as a new mixing device. • mRNA-LNPs were then treated by buffer exchange/concentration/ sterile filtration. • Particle size obtained was 100 nm, pdI 0.15, and encapsulation efficiency 95–96 % • Same characteristics were obtained using ethanol ratios of 1/4 and 1/6. • Membrane micromixing allows the use of low ethanol amount. Messenger RNA encapsulated in lipid nanoparticles (mRNA-LNPs) are successfully used worldwide for vaccination against COVID-19. mRNA-LNPs are manufactured by mixing an ethanolic phase (containing an ionizable lipid, a phospholipid, a PEGylated lipid and cholesterol) and an aqueous phase (citrate buffer at pH 4.5 containing mRNA). The characteristics of the mRNA-LNPs obtained (size, internal structure, mRNA functional delivery in vivo, etc) depend on several parameters and among them are the mixing conditions of the ethanolic and aqueous phases. While the impact of these parameters is well-recognized for small scale preparations (some microliters), their effect at a larger scale is less well-known. The purpose of this study is to investigate the production of some hundreds of millilitres of mRNA-LNP suspension using a ceramic membrane (SPG Technology) as a new mixing device. Summarily, in this technique, the ethanolic phase permeates through the membrane pores and mixes with the aqueous phase flowing in a small annular pipe between the tubular membrane and an inside rod. First, empty LNPs were prepared to determine the effect of membrane pore size, flow rate, and the ratio between the ethanolic and aqueous phase volumes. The experimental conditions tested did not change the characteristics of the LNPs obtained. Second, mRNA-LNPs were prepared and the suspensions obtained were then submitted to buffer exchange/concentration/ sterile filtration dialysis. The resulting mRNA-LNPs had a particle size of 103 ± 5 nm, a polydispersity index of 0.15 ± 0.005, and encapsulation efficiency of 96 ± 0.0 %. mRNA-LNPs with similar characteristics were obtained using ethanol ratios of 1/4 (20 % ethanol in the final preparation) and 1/6 (14 % ethanol). Overall, membrane micromixing using a ceramic membrane was shown to be a suitable technology for the preparation of mRNA-LNP suspensions. [ABSTRACT FROM AUTHOR]