Phospholipid Content Research Articles

High Content Image Analysis of Cellular Responses of the Murine J774A.1 Cell Line and Primary Human Cells Alveolar Macrophages to an Extended Panel of Pharmaceutical Agents.

In vitro screening of macrophages for drug-induced effects, such as phospholipidosis, is useful for detecting potentially problematic compounds in the preclinical development of oral inhaled products. High-content image analysis (HCIA) is a multi-parameter approach for cytotoxicity screening. This study provides new insights into HCIA-derived response patterns of murine J774A.1 cells and primary human alveolar macrophages (hAM). Several compounds were compared with reference groups (cationic amphiphilic drugs and apoptosis inducers) at different concentrations (0.01 to 10µM). After incubation, cells were stained with fluorescence markers and HCIA was performed (Cytation™ 5 Cell Imaging System). Ten parameters were analysed: non-adherent cells, increased or reduced mitochondrial activity, membrane permeability, cell area, nuclear area, polynucleated cells, vacuole area, neutral and phospholipid content. A new system of response categorisation was developed for data analysis. Murine J774A.1 cells exhibited a drug-induced response pattern that was distinct to the corresponding pattern of hAM cells. Comparison with the literature revealed that primary cells (rat or human origin) have similar response patterns, while cell lines (mouse, rat or human) exhibited a different response pattern. Hierarchical clustering revealed toxicologically aligned clusters of compounds, suggesting potential use for understanding mechanisms of drug effects in cell lines and primary cells. Valuable information for selecting a suitable cell type for HCIA screening of macrophage responses to drug compounds is provided. All cell types were suitable for screening drug-induced phospholipidosis. Still, human primary alveolar macrophages responded differently to drug treatment compared to macrophage cell lines and may be required to evaluate broader response-patterns and mechanisms of toxicity.

Determination of the critical aggregation concentration of phospholipids widely used in nanoliposomal development from different experimental methodologies

Nanoliposomes are increasingly recognized for their potential in drug delivery due to their biocompatibility and versatility. Achieving optimal performance in these systems necessitates a comprehensive understanding of the physicochemical properties of the phospholipids used in their formulation. This study focuses on determining the critical aggregation concentration (CAC) of a phospholipid system, during liposomal development via the ethanol injection method. Key findings include the identification of CAC within the range of 10–11 mg/mL, where the formation of nanoliposomes is confirmed. The study systematically evaluated the inflection points from second derivative plots of surface tension, contact angle, work of adhesion, viscosity, electrical conductivity, density, and transmittance relative to phospholipid concentration. Additionally, dynamic light scattering-DLS and nanoparticle tracking analysis-NTA were employed to assess size distribution, polydispersity index-PDI, and zeta potential. The results revealed that at the CAC, nanoliposomes exhibited a particle size range of 188–232 nm, a PDI between 0.284 and 0.578, and a zeta potential between −6 mV and −14 mV. Finally, the stability assay of aqueous dispersions of the phospholipid system carried out at 4 °C and 25 °C for 4 weeks demonstrated that these aggregated systems tend to conserve their morphology and that a change occurs from a size monomodal distribution to a bimodal distribution depending on the stability test condition, which was evidenced by field emission scanning electron microscopy-SEM, as well as by DLS and NTA. These findings highlighted the critical concentration range for effective nanoliposome formation and provide insight into the optimal conditions for their development.

Associations between omega-3 fatty acid-derived lipid mediators and markers of inflammation in older subjects with low-grade chronic inflammation.

Cardiovascular disease (CVD), the leading cause of death in the United States and globally, is a chronic inflammatory disease likely caused by an impaired ability to resolve inflammation. Pre-clinical studies have provided strong evidence of the activating role of specialized pro-resolving lipid mediators (SPMs) derived from the omega-3 fatty acids eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosaheaxaenoic acid (DHA) on the resolution of inflammation. However, there is a dearth of information on the role of SPMs on inflammation in humans. Therefore, the aim of this study was to assess whether plasma concentrations of omega-3 fatty acids and their derived SPMs are associated with inflammatory markers in subjects with low-grade chronic inflammation (C-reactive protein >2µg/mL). The plasma phospholipid content of omega-3 fatty acids, a marker of dietary intake, plasma concentrations of SPMs, and serum concentrations of inflammatory markers were measured in 21 older men and postmenopausal women (age 53-73y) at the end of a four-week placebo phase (3g/day high oleic acid sunflower oil). The phospholipid DHA content was inversely related to interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and IL-10 concentrations. Moreover, MCP-1 was inversely associated with the DHA-derived 14-HDHA and 4-HDHA, and IL-10 was inversely associated with EPA-derived 18-HEPE, 12-HEPE and 5-HEPE, DPA-derived Rv5DPA, and DHA-derived 4-HDHA. These findings support the anti-inflammatory effect of dietary omega-3 fatty and suggest that lipid mediators derived from EPA, DPA, and DHA participate in the regulation of inflammation in subjects with chronic inflammation.

Anaerobic plasmalogen production in recombinant Escherichia coli carrying plasmalogen synthase gene from Selenomonas ruminantium.

Escherichia coli expressing SrPlsAR from Selenomonas ruminantium produces plasmalogen, comprising almost 60% of the total phospholipid content under anaerobic conditions. Both plasmenylethanolamine and plasmenylglycerol were detected, and the major acyl aldehyde derived from sn-1 vinyl ether was C16:1. Plasmalogen synthesis is affected by mutations in ATP-binding sites and Cys expected to be involved in the formation of the [4Fe-4S] cluster.

Different effects of low- and high-density lipoproteins in egg yolk on lipid metabolism of mouse: Role of phospholipids-to-cholesterol intake ratio.

Recent evidence increasingly indicates that moderate egg consumption (up to one egg per day) poses no significant risk of cardiovascular diseases in healthy individuals, leading researchers to speculate whether other lipids in eggs affect lipid metabolism. In this research, C57BL/6J mice were given chow diets containing egg yolk low-density lipoprotein (LDL) and high-density lipoprotein (HDL), two natural components with different proportions of phospholipids and cholesterol, to explore the effects of the phospholipids-to-cholesterol intake ratio on lipid metabolism. Increases in liver weight and liver index were observed following LDL intake, while body weight and epididymal fat decreased after HDL intake in a dose-dependent manner. The food efficiency of chow diets decreased with increasing phospholipids content. Lipid biomarkers indicated that LDL intake significantly increased plasma LDL cholesterol and liver cholesterol levels. At equivalent cholesterol doses, the cholesterol-raising effects of HDL were weaker than those of LDL. Higher protein expression of cytochrome P450 family 7 subfamily A member 1 in the HDL groups suggested increased bile acid excretion at higher phospholipids-to-cholesterol intake ratios. Fecal metabolite analysis revealed significant changes in lipid composition, primarily involving glycerophospholipids, sphingolipids, and sterol lipids, consistent with the lipid profiles of LDL and HDL. The upregulation of cholic acid and deoxycholic acid in feces further confirmed increased bile acid excretion. This study highlights the differences in lipid metabolism in mice under varying phospholipids-to-cholesterol intake ratios, which may be associated with changes in dietary energy supply and bile excretion following phospholipids involvement.

Protective Effect of Pentoxifylline on the Development of Acute Gastric Mucosal Injury in a Model of LPS-Induced Sepsis.

Alterations in the gastric mucosal barrier, one of whose fundamental components is phosphatidylcholine (PC), may play an important role in the pathophysiology of erosive gastritis secondary to sepsis. Pentoxifylline (PTX) has been shown to reduce tissue damage in various experimental models of sepsis. The aim of this study was to investigate the effect of PTX on gastric mucosa PC synthesis, leukocyte infiltration, arachidonic acid-related metabolites, inflammation, oxidative stress, NO, CO, and somatostatin in a rat model of LPS-induced sepsis. Rats were administered LPS (10 mg/kg b.w.) intraperitoneally. After 30 min (early treatment group) or 120 min (late treatment group) of LPS administration, they were randomly divided into two groups that were intraperitoneally administered saline (5 mL/kg; LPS + Saline group) or PTX (45 mg/kg; 5 mL/kg; LPS + PTX group). Control rats received only saline instead of LPS and/or PTX. Two hours after saline or PTX administration (total of 150 or 240 min of procedure), animals were anesthetized, and then gastric lavage, gastric mucosa and plasma samples were obtained and kept frozen until determination. LPS-induced sepsis changed the gastric mucosal barrier by reducing its phospholipid content, PGI2, and somatostatin levels, as well as increasing MPO, TXB2, LTB4, PLA2, and MDA. Alterations may be mediated, at least in part, by modifications in the production of NO, CO, and cGMP content. PTX had a beneficial effect on gastric lesions secondary to sepsis by restoring PC production.

Modulation of the Plasma Lipidomic Profile in Piglets Fed Polar Lipid-Rich Diets.

Background: Polar lipids from dairy are novel sources of energy that may replace other dietary lipids and impact plasma lipidomic profiles in piglets. This study evaluated the impact of feeding diets rich in polar lipids on the plasma lipidome of piglets during the weaning period. Material and Methods: Weaned male piglets (n = 240; 21 days of age; 6.3 ± 0.5 kg of BW) were blocked by initial weight and distributed into 48 pens of five animals each in a complete randomized block design with a 2 × 3 factorial arrangement of treatments as follows: a plant-based diet rich in neutral lipids from soybeans (24 pens; SD) or a polar lipid-rich diet by-product of cheese making (24 pens; PD) from weaning until the 21st day of the nursery phase. Within each diet group, animals received one of three milk replacers (MR; 0.5 L/d/animal) for the first 7 days after weaning: (1) commercial MR containing animal and coconut lipids (CO); (2) polar lipid-based MR (PO); or (3) soybean lipids-based MR (SO). Results: The PD diet group increased the plasma concentrations of sphingolipids, phospholipids, and cholesterol esters, but did not impact the concentrations of glycerolipids (GLs). Both the PO and CO milk replacers increased the plasma concentrations of ceramide, acyl-chain phosphatidyl choline, and cholesterol esters. The plasma concentrations of GLs containing 18-carbon fatty acids such as 18:0, 18:1, 18:2, and 18:3, were higher in SD, whereas GLs containing 16:0 and 20:3 were higher in PD. Conclusions: In summary, the diet lipid type significantly modulated the plasma lipid composition in piglets 7 days after weaning. The dietary inclusion of polar lipids in diets for growing pigs can modulate the plasma lipidomic profile, relative to plant-based diets rich in soybean lipids. Cost may be a major consideration when using these lipids in pig diets. Their health benefits need to be further characterized in other models of stress and inflammation.

Damage of the Phospholipid Bilayer by Aβ42 at Physiologically Relevant Peptide Concentrations.

Amyloid β (Aβ) aggregates are implicated in the pathology of several neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, and Parkinson's disease, and damage to membranes is considered one of the pathology-related effects of Aβ. Experiments in vitro indicate that Aβ can damage these membranes; however, such experiments were performed at Aβ concentrations in the micromolar range, several orders above the physiologically relevant conditions. Our studies with Aβ42 in the low nanomolar concentrations did not reveal any damage to the supported lipid bilayer, questioning this membrane damage mechanism of Aβ. However, the phospholipid composition can be a factor contributing to the interaction of Aβ with the membrane. Therefore, in this study, we investigated the interaction of 50 nM Aβ42 with supported lipid bilayers composed of equimolar ratios of POPS and POPC at phospholipid concentrations of 0.1 and 0.25 mg/mL. Using atomic force microscopy (AFM), we observed that Aβ42 induced damage to bilayers at 0.1 mg/mL, characterized by forming defects that grew in size and number over time. The defects penetrate only the upper leaflet of the bilayer, but no such defects were observed at 0.25 mg/mL phospholipid concentrations. We additionally determined Young's modulus of these bilayers as a measure of stiffness, and these values were 6.9 ± 3.6 MPa and 16.6 ± 5.3 MPa for the 0.1 mg/mL and the 0.25 mg/mL bilayers, respectively. These findings suggest that Aβ42's ability to induce bilayer damage depends on membrane stiffness, with softer bilayers (0.1 mg/mL) being more susceptible to Aβ42-induced damage. The results are discussed and compared with models in which Aβ42 oligomers create localized membrane damage. The implication of the results to the mechanisms of the Aβ42 oligomer pathology is discussed.

Abstract A011: Assessing pro-tumorigenicity of the aging microenvironment in a murine ovarian cancer model

Abstract Over 70% of high-grade serous ovarian cancer (HGSOC) patients are diagnosed after age 55 and the average patient who succumbs to HGSOC is 70 years old. Despite this, the majority of ovarian cancer studies, and cancer studies generally, utilize young mice that are more representative of human children. This approach overlooks age as a variable, despite its status as a top risk factor for cancer. Our preliminary studies reveal tumor burden and disease severity is significantly higher in aged mice compared to young animals. We have developed an epithelial cell line derived from C57BL/6 oviducts and used CRISPR-Cas9 genome editing to establish tumorigenic lines for syngeneic studies. One line harbors Trp53, Pten, and Brca2 deletions, is tumorigenic, and can be serially passaged in vitro. When injected intraperitoneally (IP) into 8-week and 56-week-old mice, equating to approximately 10- and 45-year-old humans respectively, a stark increase in tumor mass was evident in aged mice. Furthermore, the aged mice succumbed to disease at a faster rate after inoculation than the young group, where all mice succumbed to disease within 1-month of IP injection compared to some young mice surviving up to 3 months after injection. These findings, combined with epidemiologic data, suggest a metastatic driver in the aged microenvironment that facilitates worsened ovarian cancer progression. We are investigating two factors that change with age, (1) a suppressed immune system and (2) age-related changes to adipose tissue, as possible mechanisms driving this difference. Histologic analysis of tumors from each cohort showed decreased cleaved caspase-3 in aged tumors, indicating a reduction in tumor cell killing in older animals. Characterization of the pre-cancerous, tumor-naive microenvironment via flow cytometry and cytokine array have revealed that aged mice microenvironment is highly inflamed and features changes in immune populations and inflammatory states. Meanwhile, total RNA-sequencing of tumors harvested from aged and young mice showed minimal changes, but DEGs point to possible roles of fat transport and metabolism in these aged tumors, indicated by upregulated CD36 and lipoprotein lipase. Follow-up ex vivo experiments had demonstrated increased adhesion and proliferation of the previously described murine cancer cells to fat pads when cultured in vitro. When assessing tumor-naive fat depots, aged adipose tissue is comprised of significantly larger adipocytes and the omentum, a common secondary site of ovarian cancer and fatty organ, was found to have increased phospholipid content in older animals. Both findings point to a change in metabolic function in aged mice. Our preliminary findings indicate an important role of the aged microenvironment in driving enhanced diseases progression in older individuals and identifying the mechanisms behind this phenotype are critical; an improved understanding of how age affects HGSOC progression could enable the development of treatment strategies to prevent cancer cells from establishing in the peritoneum. Citation Format: Katherine A. Cummins, Ronny Drapkin. Assessing pro-tumorigenicity of the aging microenvironment in a murine ovarian cancer model [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A011.

Distribution of Dietary Phospholipids in Selected Agri-Foods: Versatile Nutraceutical Ingredients.

Phospholipids (PLs) play a crucial role in the nutraceutical field due to their various health benefits, including supporting acetylcholine production, enhancing cell membrane fluidity, and promoting cognitive functions. This study aimed to investigate the PL composition of selected agri-foods, including grains, vegetables, and fruits, and assess the effects of cooking methods. The major PLs identified in most agri-foods were phosphatidylethanolamine (PE) and phosphatidylcholine (PC). Additionally, lyso-phosphatidylethanolamine and lyso-phosphatidylcholine were found in rice, grains, and wheat, while N-acyl-phosphatidylethanolamine was detected in grains, wheat, and some vegetables. Phosphatidylinositol was present in fruits and vegetables, and phosphatidylserine was exclusively found in mushrooms. The PL composition was influenced by cooking methods, with boiling, steaming, blanching, and roasting increasing the PL content, while salting tended to decrease it. Although most agri-foods contained higher levels of PC than PE, citrus fruits under long-term low-temperature storage had significantly more PE than PC. This study established a PL database for the selected agri- and processed/cooked foods, providing insights into changes in PL composition and content based on cooking methods. Given the important health functions of each PL, consuming various agri-foods and incorporating different cooking methods for optimal health benefits is advisable.

From a metabolomics profiling perspective explaining the deterioration impact induced by chicken meat exudate

Exudate is the liquid released from meat during storage or processing. This study investigated the influence of exudate removal on the quality of pressed and freeze-thawed chicken meat during 36 h storage, focusing on exudate metabolites. Incorporating exudate increased lipid oxidation but only slightly affected TVB-N values in pressed and defrozen samples. This may be due to the high concentration of phospholipids and their derivatives in the exudate, which are prone to oxidation. For pressed samples, adding exudate raised total bacterial count (from 5.88 to 6.13 log cfu/g) and pH (from 6.14 to 6.17) at 36 h storage point, but no significant changes were observed in defrozen samples. While exudate removal had little effect on cooking loss and texture of cooked meat, it improved the meat's ability to maintain its structure and flavor during storage. These findings suggest removing exudate chicken meat processing can improve its quality and shelf life.

Integration of metabolomics and transcriptomics reveals the toxicological mechanism of deltamethrin exposure in Chinese mitten crab Eriocheir sinensis

This study investigated the toxicological mechanism of deltamethrin on Chinese mitten crab Eriocheir sinensis juveniles in fresh water. We first conducted an acute toxicity test, followed by laboratory methods to detect changes in immune-related indices in terms of antioxidant enzyme markers, lipid metabolism-related genes, and autophagy-related and apoptosis genes. The acute toxicity (96-h LC50) of deltamethrin to E. sinensis was 7.195 μg/L. After 48 h of exposure, serum showed elevated immune-related indices (P < 0.05) for alkaline phosphatase (AKP), acid phosphatase (ACP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), complement components C3 and C4, and the key pro-inflammatory cytokines interleukin-6, interleukin-1β, and tumor necrosis factor alpha (TNF-α). In hepatopancreas at 48 h, indicators related to the antioxidant system, namely superoxide dismutase (SOD) and glutathione (GSH), were significantly elevated, whereas nitric oxide and total antioxidant capacity (T-AOC) were decreased (P < 0.05). In contrast, lipid metabolism indices for triglyceride (TG), total cholesterol (TC), and malondialdehyde (MDA) were increased (P < 0.05). Transcriptomics and metabolomics revealed that exposure to deltamethrin disrupted the lipid metabolic process in the hepatopancreas mainly by altering fatty acid synthesis, amino acid metabolism, immune signaling, and autophagy activation, while the exposure increased the content of phospholipids and cholesterol but decreased the levels of amino acids and palmitoleic acid. Quantitative genetics revealed significantly aberrantly expressed (P < 0.05) lipid metabolism-related genes, including acc1, fasn, scd1, and pnpla2, all key genes involved in lipid accumulation. Deltamethrin exposure also significantly altered (P < 0.05) gene expression levels for Toll-like receptor (tlr), myeloid differentiation factor 88 (myd88), crustin1, anti-lipopolysaccharide factor isoform 3 (alf3), tumor necrosis factor alpha (tnf-α), and NF-κB transcription factor relish. Furthermore, deltamethrin activated the toll-like receptor/major myeloid differentiation response gene 88/nuclear factor kappa-light-chain-enhancer of activated B cells (TLR/MyD88/NF-kB) signaling pathway, which activates a nonspecific immune response in E. sinensis. Additionally, carnitine palmitoyltransferase 1 A (cpt1a), cytochrome c (cyt-c), adenosine 5′-monophosphate (amp)-activated protein kinase (ampk), the autophagosomal protein microtubule-associated protein 1 light chain 3c (lc3c), and the autophagy-related proteins beclin1, atg5, atg12 were significantly induced (P < 0.05) in the adenosine monophosphate-activated protein kinase/rapamycin (AMPK/mTOR) signaling pathway. These changes resulted in excess free radicals, causing oxidative stress in the mitochondrial membrane, promoting mitochondrial autophagy. The results confirm that deltamethrin exposure can induce hepatopancreatic injury by promoting mitochondrial autophagy, activating an immune response, and inhibiting lipid metabolism. Overall, this study provides multi-level information to reveal the toxic effects of deltamethrin on E. sinensis.

The assessment of myopenia and muscle biopsy in pediatric patients with liver disease awaiting liver transplantation - A cross-sectional analysis.

Little is known about the skeletal muscle characteristics (fibre type proportion and size, location of nuclei, presence of fat infiltration) in children with liver disease with radiologically determined myopenia (low muscle mass). During liver transplantation (LTx) surgery, biopsies from the rectus abdominis muscle were collected. Muscle fibre types (I, I/IIA, IIA, IIA/X, IIX) and CSA index (µm/m2) were determined using immunofluorescence staining. Triacylglycerol (TAG) and phospholipid content of muscle was determined using gas chromatography. Myopenia was defined using study-specific cut offs (skeletal muscle index <-2SD) from age-sex matched healthy control scans. Myopenia was prevalent in 41% of children. Children also had a high prevalence of high muscle adiposity (37%). Children with myopenia were older (8.4 vs. 0.7 yrs; p<0.001), had smaller total (median 595 vs. 844µm/m2; p=0.04) and hybrid IIA/X (612±143 vs. 993±341µm/m2; p=0.04) muscle fibre size index, lower prevalence of type I fibres (53 vs. 64%; p=0.01) and higher prevalence of type IIA/X hybrid fibres (median 7.5 vs. 0%; p=0.04). Children with myopenia also had a higher prevalence of elevated TAG content (>75 percentile) within the muscle compared to children without myopenia (36% vs 0%; p=0.009). Percent of muscle fibres with centralized nuclei were not different between groups. In conclusion, children with myopenia experience differences in skeletal muscle biological characteristics when compared to children without myopenia at LTx and these findings may have implications for dietary and exercise rehabilitation pre-and post-LTx.

Balancing act: How cholesterol and phospholipids influence juvenile mud crab Scylla paramamosain growth and lipid metabolism

A 2 × 3 two-factor experiment was designed to assess the effects of dietary cholesterol (CHO) and phospholipids (PLs) on growth and lipid metabolism in early juvenile mud crabs (0.01 g crab−1). The experimental diets were designed with two CHO levels: 0.40 % (LCH) and 0.80 % (HCH), and three PLs levels: 1.80 % (LPL), 2.50 % (MPL), and 3.20 % (HPL). The 58-day aquaculture trial demonstrated that the LCH-HPL group achieved the best growth performance in mud crabs, characterized by the highest final body weight, weight gain, and specific growth rate. Regarding whole-body composition, dietary PLs increased total cholesterol (T-CHO), PLs, triglyceride (TG), and crude lipid content in the LCH groups. Mud crabs in the HCH groups had a higher proportion of saturated fatty acids, a lower proportion of monounsaturated fatty acids, and increased gene expression of sterol regulatory element binding protein 1c (srebp-1c) and fatty acid synthase (fas) compared to those in the LCH groups. As the dietary PLs increased, mud crabs in the LCH groups exhibited up-regulation in the expression of genes, including srebp-1c, fas, elongation of very long-chain fatty acid protein 4 (elovl4), elovl6, and Δ9-fatty acid desaturase (Δ9-fad). In the HCH groups, elevated dietary PLs resulted in the down-regulation of fatty acid binding protein (fabp) gene expression. In addition, high levels of dietary CHO and PLs (HCH-HPL) inhibited the catalase activity of mud crabs, resulting in a significant increase in the malondialdehyde content. Correlation analysis revealed that dietary CHO was positively correlated with the expression of long-chain polyunsaturated fatty acids synthesizing genes, and dietary PLs were positively correlated with the whole-body lipid content of mud crabs. Additionally, a positive correlation was found between the growth performance and whole-body content of crude protein, crude lipid, and antioxidant capacity in mud crabs.

Interfacial behaviour of human bile and its substitution for in vitro lipolysis studies

This study examined the interfacial evolution of individual bile salts (BSs) and their blends with phosphatidylcholine (BS/PC) to simulate the complex behaviour of human bile (HB) during lipolysis at the triglyceride/water interface. Using adsorption and desorption cycles, mimicking exposure to small intestinal fluids, we demonstrate that the interfacial behaviour of real HB can be replicated using simple mixtures of BSs and PC. Interfacial tension (IFT) measurements after lipolysis and desorption showed no significant differences (P > 0.05) between HB samples and BS/PC mixtures across the total BS concentrations analysed (2.23–7.81 mM). However, individual BSs without PC yielded significantly different IFT results (P < 0.01) compared to HB, highlighting the importance of phospholipids. Dilatation rheology further emphasised the need for accurate phospholipid representation in bile models. Our results suggest that phospholipids in HB and in BS/PC systems enhance resistance to desorption, potentially affecting lipolysis. This is important, as current in vitro digestion models often replicate only intestinal BS concentrations to mimic the behaviour of HB in the intestinal lumen. Furthermore, the specific composition of BSs in HB appears less critical than the overall BS and phospholipid contents, suggesting that the kinetics of triglyceride digestion is influenced by the combined luminal concentrations of these components. These findings have significant implications for understanding the role of bile in digestion and offer insights for designing more accurate in vitro models to study the gastrointestinal behaviour of food emulsions and lipid-based delivery systems.

Lipid Compositions of Liquid-Ordered and Liquid-Disordered Phases in Ternary Membranes of Sphingomyelin, Cholesterol, and Dioleoylphosphatidylcholine Determined by 2H NMR: Stearoyl-Sphingomyelin Compared with Its Palmitoyl Counterpart.

Sphingomyelin (SM) and cholesterol are the major lipids in the signaling platforms of cell membranes, known as lipid rafts. In particular, SM with a stearoyl chain (C18-SM) is abundant in specific tissues such as the brain, the most cholesterol-rich organ, whereas the distribution of palmitoyl (C16)-SM is ubiquitous. Here, we reveal the differences between palmitoyl- and stearoyl-SM in lipid-lipid interactions based on the tie lines obtained from the 2H solid-state NMR spectra of bilayer systems composed of SM/dioleoylphosphatidylcholine/cholesterol 33:33:33 and 40:40:20. Lipid probes carrying position-selective deuterations, 10',10'-d2-SM, 24-d1-cholesterol, and 6″,6″-d2-dioleoyl-phosphatidylcholine, were incorporated into the membranes. 2H NMR peaks from these probes in the membranes directly provide the lipid compositions of the liquid-ordered (Lo) and liquid-disordered (Ld) regions. Without using bulky fluorescent groups, these probes allow us to obtain the end points of the tie lines in a ternary phase diagram based on the lever rule. Consequently, the tie lines of the stearoyl-SM membranes were steeper than those of the palmitoyl-SM membranes, indicating that cholesterol content was higher in the Lo domains of stearoyl-SM, regardless of the total concentration of unsaturated phospholipids. When comparing the content of unsaturated lipids in the Lo domain, the stearoyl-SM membranes had a higher content than palmitoyl-SM membranes. These results revealed that stearoyl-SM is suitable for stabilizing biologically functional microdomains in cholesterol-rich organs, whereas palmitoyl-SM may be better suited for stabilizing domains in tissue membranes with normal cholesterol content. The small but significant differences in the lipid interactions between stearoyl-SM and palmitoyl-SM may be related to the spatiotemporal formation of functional domains in biological environments.

Preparation of Paeonol Ethosomes by Microfluidic Technology Combined with Gaussians and Evaluation of Biological Activity by Zebrafish.

Paeonol, a monoterpene glycoside compound, has extensive pharmacological activities. However, its applications are restricted by poor water solubility and low bioavailability. In this study, paeonol ethosomes (PAE-ethosomes) were successfully prepared with a microfluidic method by optimizing the single factors and RSM test. The enhanced PAE-ethosomes were assessed using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), vesicle size (VS), zeta potential (ZP), and polydispersity index (PDI). Density functional theory analysis was employed to verify the molecular interaction. The optimized RSM conditions were a phospholipid concentration of 6 mg/mL, a cholesterol concentration of 1 mg/mL, and a total flow rate of 600 μL/min with a presumed value of 60.3% and confirmation results of 61.2 ± 0.3%. The prepared PAE-ethosomes showed better storage stability and a slow-release effect. The Q n of PAE-ethosomes rose from 167.0 ± 15.8 to 272.0 ± 16.4 μg/cm2 after 24 h, which was substantially greater than that from a 25% hydroethanolic solution of paeonol, according to in vitro skin retention and transdermal absorption. The Q s of PAE-ethosomes in the skin increased by 225% with 265.5 ± 15.4 vs 81.8 ± 8.2 μg/cm2, compared with 25% hydroethanolic solution of paeonol. Molecular interaction between paeonol and lecithin by Gaussians showed that the paeonol compound may have a higher probability of spreading in the hydrophilic phosphate group ("head") position for the PAE-ethosomes. The Tg (Lyz: EGFP) transgenic zebrafish results showed that PAE-ethosomes had better anti-inflammatory effects than paeonol. The microfluidic approach was efficient with good characteristics in physics and pharmacology with the potential in pharmaceutical use.

Berberine and Berberine Nanoparticles Mitigate Scopolamine-Induced Amnesia in Rats

Abstract Introduction/Objective Berberine (BER) has garnered attention in research for its multifaceted biochemical and pharmacological properties, positioning it as a promising natural remedy. Conversely, scopolamine (SCO) functions as a non-selective muscarinic receptor antagonist, disrupting central cholinergic neurotransmission and impairing learning and short-term memory. Methods/Case Report Forty-two male Wistar rats were procured from the National Research Center in Cairo, Egypt, and allocated randomly into seven groups, each comprising six rats. The groups were as follows: Sham Control (no treatment), BSA Control (received oral administration of 1 ml Bovine Serum Albumin-NP solution daily for 28 days), BRB-NP Control (received oral administration of BRB-NP nanoparticles at 5 mg/kg/day for 28 days), Induced Group (intraperitoneal injection of SCO at 2 mg/kg dissolved in saline once daily for 28 days), BER Treated Group (received BER at 50 mg/kg suspended in saline followed by intraperitoneal injection of SCO at 2 mg/kg after 40 minutes daily for 28 days), BSA Treated Group (received oral administration of 1 ml BSA-NP solution followed by intraperitoneal injection of SCO at 2 mg/kg after 40 minutes daily for 28 days), and BER-NP Treated Group (received oral administration of BER-NP at 5 mg/kg followed by intraperitoneal injection of SCO at 2 mg/kg after 40 minutes daily for 28 days). Whole blood was collected via heart puncture, and brain tissue was promptly extracted for brain homogenate preparation. Routine biochemical assays were conducted on serum and brain samples. Results (if a Case Study enter NA) Statistically significant differences were observed among the seven experimental rat groups concerning plasma AST, albumin, total protein, and triglyceride levels, while differences in ALT and total cholesterol were non-significant. Regarding brain parameters, significant differences were noted among groups in total protein, cholesterol, phospholipids, and uric acid levels. Conclusion Treatment with BRB NPs or free berberine led to a progressive increase in brain protein levels and a decrease in cholesterol content (p&amp;lt;0.05). Additionally, SCO administration resulted in a gradual reduction in brain phospholipid content, while treatment with BER NPs, regardless of the rat’s condition, exhibited the highest phospholipid levels (p&amp;lt;0.05). Furthermore, administration of BSA or free BRB improved brain uric acid levels (p&amp;lt;0.05).

Feeding Docosahexaenoic Acid and Arachidonic Acid during Suckling and Weaning Contributes to Oral Tolerance Development by Beneficially Modulating the Intestinal Cytokine and Immunoglobulin Levels in an Allergy-Prone Brown Norway Rat Model

BackgroundSuckling and weaning arachidonic acid (ARA) + docosahexaenoic acid (DHA) supplementation promoted oral tolerance (OT) development in pups, however, the effect of it on the intestine to promote OT development remains unknown. ObjectiveWe aimed to explore the impact of this supplementation on intestinal fatty acid composition, structure, and indicators that are supportive of OT development. MethodsAllergy-prone Brown Norway dams were randomly assigned to a control (0% ARA, 0% DHA) or ARA + DHA diet (0.45% ARA, 0.8% DHA) during suckling (0–3 wk). At weaning (3–8 wk), offspring were randomly assigned to a control (0% ARA, 0% DHA) or ARA + DHA diet (0.5% ARA, 0.5% DHA). At 3 wk, offspring in each group received an oral gavage of sucrose or ovalbumin (OVA) solution for five consecutive days. At 7 wk, all offspring received an intraperitoneal OVA injection. At 8 wk, offspring were terminated to evaluate jejunum morphology and measure mucosal food allergy-related secretory immunoglobulin A (sIgA) and cytokines, ileum phospholipid and triglyceride fatty acid compositions, and fecal calprotectin. ResultsWeaning ARA + DHA resulted in a higher percentage of DHA in ileum phospholipids and triglycerides (both P < 0.001), without affecting the percentage of ARA. Despite no lasting effect of suckling ARA + DHA on the DHA content in ileum phospholipids, a programming effect was found on the allergy-related intestinal immune profile [higher concentrations of mucosal IL-2 (P = 0.049) and sIgA (P = 0.033)]. OVA treatment resulted in a lower concentration of mucosal IL-6 (P = 0.026) regardless of dietary interventions. Offspring fed ARA + DHA during suckling and/or weaning had a higher concentration of mucosal transforming growth factor-beta (TGF-β) after OVA treatment but this was not observed in offspring fed control diets during suckling and weaning (P = 0.04). ConclusionsEarly life dietary ARA + DHA supplementation to allergy-prone rats enhanced the DHA concentration in intestinal phospholipids (weaning period) and increased the mucosal sIgA, IL-2, and TGF-β levels (suckling and weaning period), indicating its ability to create a tolerogenic intestinal environment to support OT development.

A Pair of Indicators for Characterizing Cerebral Microbleeds Based on Raman Spectrum and Two-Photon Imaging.

Cerebral microbleeds (CMBs) lead to cognitive decline, linked to the axonal structure composed of phospholipid bilayers. Current methods are difficult to obtain in situ changes of biochemical component concentration during CMB. In this study,by Raman spectrum and two-photon imaging, we achieve in situ changes in the information of biochemical components concentration during CMB. The overall concentration of phospholipids in the damaged tissue significantly decreases after CMB, forming a large region of low concentration, but the relative concentration of phosphatidylinositol (PI) increases, reflecting the inhibition role of the phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) pathway. Accordingly, two-photon images of neurons show a clear decrease in the number of axons, indicating a close correlation between phospholipid hydrolysis and axon damage, as well as cognitive impairment. Therefore, the decrease in phospholipid concentration and theincrease in the PI concentration might serve as a pair of indicators for characterizing CMB and its relationship with cognitive decline.