Cell Membrane Fluidity Research Articles

Fatty acid desaturases (FADs) modulate multiple lipid metabolism pathways to improve plant resistance.

Biological and abiotic stresses such as salt, extreme temperatures, and pests and diseases place major constraints on plant growth and crop yields. Fatty acids (FAs) and FA- derivatives are unique biologically active substance that show a wide range of functions in biological systems. They are not only participated in the regulation of energy storage substances and cell membrane plasm composition, but also extensively participate in the regulation of plant basic immunity, effector induced resistance and systemic resistance and other defense pathways, thereby improving plant resistance to adversity stress. Fatty acid desaturases (FADs) is involved in the desaturation of fatty acids, where desaturated fatty acids can be used as substrates for FA-derivatives. In this paper, the role of omega-FADs (ω-3 FADs and ω-6 FADs) in the prokaryotic and eukaryotic pathways of fatty acid biosynthesis in plant defense against stress (biological and abiotic stress) and the latest research progress were summarized. Moreover' the existing problems in related research and future research directions were also discussed. Fatty acid desaturases are involved in various responses of plants during biotic and abiotic stress. For example, it is involved in regulating the stability and fluidity of cell membranes, reactive oxygen species signaling pathways, etc. In this review, we have collected several experimental studies to represent the differential effects of fatty acid desaturases on biotic and abiotic species. Fatty acid desaturases play an important role in regulating biotic and abiotic stresses.

Microencapsulating role of whey protein isolate and sucrose in protecting the cell membrane and enhancing survival of probiotic lactobacilli strains during spray drying, storage, and simulated gastrointestinal passage

Limosilactobacillus reuteriKUB-AC5 andLactobacillus johnsoniiKUNN19-2 were spray-dried using whey protein isolate, sucrose, and whey protein isolate combined with sucrose at 130 °C and 150 °C. Differences in membrane fatty acid compositions of probiotics led to differences in cell membrane fluidity, cell injury, and cell survival during spray drying. Combination of whey protein isolate and sucrose improved survival of L. reuteriKUB-AC5, whose cell membrane were more fluid and more sensitive to stress, while whey protein isolate provided the best protection for L. johnsoniiKUNN19-2 against changes in protein secondary structures. Moreover, a relationship was found between membrane damage and the shelf-life ofmicrocapsules. Both microencapsulated cells prepared at the inlet temperature of 130 °C showed an enhanced survival upon simulated gastric digestion compared to the free cells. A thorough understanding of injury sites during spray drying will contribute to establishing optimal conditions and encapsulant formulations that maintain the cells’ viability during spray drying, storage, and gastrointestinal passage.

Antibacterial Activity and Components of the Methanol-Phase Extract from Rhizomes of Pharmacophagous Plant Alpinia officinarum Hance.

The rhizomes of Alpinia officinarum Hance (known as the smaller galangal) have been used as a traditional medicine for over 1000 years. Nevertheless, little research is available on the bacteriostatic activity of the herb rhizomes. In this study, we employed, for the first time, a chloroform and methanol extraction method to investigate the antibacterial activity and components of the rhizomes of A. officinarum Hance. The results showed that the growth of five species of pathogenic bacteria was significantly inhibited by the galangal methanol-phase extract (GMPE) (p < 0.05). The GMPE treatment changed the bacterial cell surface hydrophobicity, membrane fluidity and/or permeability. Comparative transcriptomic analyses revealed approximately eleven and ten significantly altered metabolic pathways in representative Gram-positive Staphylococcus aureus and Gram-negative Enterobacter sakazakii pathogens, respectively (p < 0.05), demonstrating different antibacterial action modes. The GMPE was separated further using a preparative high-performance liquid chromatography (Prep-HPLC) technique, and approximately 46 and 45 different compounds in two major component fractions (Fractions 1 and 4, respectively) were identified using ultra-HPLC combined with mass spectrometry (UHPLC-MS) techniques. o-Methoxy cinnamaldehyde (40.12%) and p-octopamine (62.64%) were the most abundant compounds in Fractions 1 and 4, respectively. The results of this study provide data for developing natural products from galangal rhizomes against common pathogenic bacteria.

How do humans and plants feel the heat?

The 2021 Nobel prize was awarded for the discovery of the animal thermosensory channel TRPV1. We highlight notable shared features with the higher plant thermosensory channel CNGC2/4. Both channels respond to temperature-induced changes in plasma membrane fluidity, leading to hyperphosphorylation of the HSF1 transcription factor via a specific heat-signaling cascade.

Saturated Fatty Acids in Cell Membrane Lipids Induce Resistance to 5-Fluorouracil in Colorectal Cancer Cells.

Resistance to chemotherapy is a major obstacle for patients with unresectable colorectal cancer (CRC); however, the factors that induce chemoresistance have not been elucidated. Lipid composition influences neoplastic behaviour. Therefore, this study examined whether lipid composition affects sensitivity to chemotherapeutic agents in CRC. We performed a lipidomic analysis of a CRC xenograft-derived spheroid model to identify potential relationships between the lipid profile and chemoresistance to 5-fluorouracil (5-FU). Genetic and pharmacological modulation of lipid synthesis were also used in the HCT-116 and DLD-1 CRC cell lines to further characterize resistance to 5-FU. Our lipidomic profiling revealed that phospholipids with saturated fatty acids (SFAs) were more abundant in 5-FU-resistant spheroids. The importance of phospholipids containing SFA in chemoresistance was confirmed by showing that in HCT-116 and DLD-1 cells, genetic or pharmacological inactivation of stearoyl-CoA desaturase-1, a key enzyme that converts SFAs to monounsaturated fatty acids, increased the proportion of SFAs in membranous phospholipids and reduced cell membrane fluidity, and this ultimately resulted in resistance to 5-FU. These data suggest that the saturated to monounsaturated fatty acid ratio in cellular membranous phospholipids affects sensitivity to chemotherapeutic agents.

Sphingolipid Metabolism and Signaling in Lung Cancer: A Potential Therapeutic Target.

Sphingolipids are important bioactive lipids that not only play an important role in maintaining the barrier function and fluidity of cell membranes but also regulate multiple processes in cancer development by controlling multiple signaling pathways in the signal transduction network. Dysregulation of sphingolipid metabolism is thought to be one of the most important dysregulated pathways in lung cancer, the most prevalent type of cancer in terms of incidence and mortality worldwide. This article focuses on lung cancer, reviewing the important lipids in sphingolipid metabolism and the related enzymes in relation to lung cancer progression and their effects on the tumor microenvironment and discussing their roles in the diagnosis and treatment of lung cancer.

Comparative transcriptomes and metabolomes reveal different tolerance mechanisms to cold stress in two different catfish species

Cold stress is detrimental to physiological fitness and can lower the survival rate, especially for the fishes with low ability of cold-tolerance. To further understand the coping mechanism on cold stress, we applied an integrated metabolomic and transcriptomic analyses to reveal the hepatic metabolic responses when cold-sensitive red-tail catfish and cold-tolerant longsnout catfish were exposed to lower temperature by a programmed cooling method. Differentially expressed genes (DEGs) and significantly different metabolites (DMs) related to lipid synthesis and metabolism were identified from comparative metabolomic and transcriptomic data, respectively. The results indicate that the two catfish species show significantly different responses to the cold stress. Synthesis and metabolism of related lipids can be mobilized quickly and efficiently to compensate for the integrity and maintain the fluidity of cell membranes in the early cooling stage in longsnout catfish, but not in red-tail catfish. The anti-oxidative and apoptosis system can be activated positively in the middle and late cooling stages in longsnout catfish, but it is inhibited in red-tail catfish, which may be the potential reason for red-tail catfish death. Therefore, this study provides key information for understanding low-temperature tolerance mechanisms in catfish.

Hydroxy-safflower yellow A composites: An effective strategy to enhance anti-myocardial ischemia by improving intestinal permeability

Hydroxy-safflower yellow A (HSYA) is the chief component of safflower against myocardial ischemia (MI), and belongs to biopharmaceutics classification system (BCS) III drugs. Its structure contains multiple hydroxyl groups, contributing to its high polarity and poor oral bioavailability. The main objective of this study was to probe the potential of oral penetration enhancer n-[8-(2-hydroxybenzoyl) amino] sodium octanoate (SNAC) and cationic copolymer Eudragit®EPO (EPO) to promote absorption of HSYA. HSYA composites (SNAC-HSYA-EPO) were formed by hydrogen bonding and van der Waals force. SNAC-HSYA-EPO has biocompatibility, and can improve the membrane fluidity, uptake, transport, and penetration of Caco-2 cells. The mechanism of promoting of SNAC-HSYA-EPO may be related to energy and P-glycoprotein (P-gp) when compared with the inhibitor NaN3 and verapamil group. In the pharmacokinetic (PK) results, SNAC-HSYA-EPO significantly improved oral bioavailability. Pharmacodynamics (PD) results determined that SNAC-HSYA-EPO could improve the symptoms of MI. The mechanism of the SNAC-HSYA-EPO anti-MI is related to alleviating inflammation and anti-apoptosis to protect the heart. In summary, SNAC-HSYA-EPO prepared in this study possessed a complete appearance, high recombination rate and excellent oral permeability promoting ability. SNAC-HSYA-EPO has the potential to improve oral bioavailability and further enhance the anti-MI effect of HSYA.

Abstract 2163: Gender and estrogens as key factors in the response to immunotherapy in non small cell lung cancer

Abstract Gender is an important stratification and prognostic factor in oncology in the response to treatments, particularly to the immunotherapy. The anti-PD-1/PD-L1 immune-checkpoint inhibitors (Pembrolizumab, Nivolumab, Atezolizumab) have improved the prognosis of non-small cell lung cancer (NSCLC) patients in 20-35% cases, but with a lower efficacy in females. The different steroid hormones - estrogens, progesterone or androgens - present in both gender have often contrasting effects on the activity of the host immune system, partly explaining the differences observed. It is not known if metabolic features of cancer cells of male and female patients may impact on the immune system response and immunotherapy efficacy. We set up an analytical pipeline based on patient-derived samples, cell lines and humanized male and female mice bearing gender-matched NSCLC xenografts. NanoString analysis in a cohort of 42 NSCLC patients of both genders treated with Pembrolizumab, identified the estrogen receptor α (ERα)/ESR1 gene as one of the most significantly associated with the response to Pembrolizumab, particularly in female patients. A panel of 30 human NSCLC cell lines of female and male origin, that reproduced the ERα/gender association observed in clinical samples, the amount of ERα and 17-β-estradiol was directly related to the expression of PD-L1, suggesting a potential direct linkage. ERα transcriptionally up-regulated CD274/PD-L1 gene, with higher effects in females. EGFR downstream effectors Akt and ERK1/2, increased the active form phospho(Ser118)ERα, upregulating PD-L1. Inhibiting the estrogen synthesis or switching off EGFR activity down-regulated PD-L1 levels. Consistently, the aromatase inhibitor letrozole increased the efficacy of Pembrolizumab in humanized NSCLC xenografts, by reducing tumor growth and tumor-associated PD-L1, increasing the infiltrating CD8+T-lymphocytes, NK cells and Vγ9Vδ2 anti-tumor T-lymphocytes. The benefit was maximal in 17-β-estradiol/ERα high female xenografts, intermediate in 17-β-estradiol/ERα medium female and male xenografts, minimal in 17-β-estradiol/ERα low male xenografts. Additionally, an untargeted lipidome analysis revealed that letrozole decreased the saturated fatty acids (SFAs)/mono- or poly-unsaturated fatty acids (MUFAs-PUFAs) ratio, and the cholesterol/cholesterol esters ratio. These changes increased plasma-membrane fluidity, an event that in turns reduced the affinity between PD-L1 and PD-1. As further proof of concept, ω6- or ω3-PUFA treated cells, displayed an increased membrane fluidity and a change in lipid metabolism that affect the response to anti-PD-1/PD-L1 as letrozole did. We propose a prototypical in vitro 3D spheroids culture gender-tailored platform that allows to investigate the metabolic circuitries causing such differences and to repurpose metabolic modifiers as novel immune-adjuvant agents in oncology. Citation Format: Sofia La Vecchia, Dario Pasquale Anobile, Fabrizio Tabbò, Iris C. Salaroglio, Joanna Kopecka, Luca Primo, Luisella Righi, Giorgio Vittorio Scagliotti, Silvia Novello, Cyril Corbet, Chiara Riganti. Gender and estrogens as key factors in the response to immunotherapy in non small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2163.

Effect of silica nanoparticles on cell membrane fluidity: The role of temperature and membrane composition

The increasing production and application of silica nanoparticles (SiO2 NPs) raise public concern regarding their environmental and health risks. The fluidity of the cell membrane is essential for supporting membrane proteins and regulating membrane transport. Changes in membrane fluidity inevitably influence the physiological activities of cells and even cause biological effects. In this study, the effect of SiO2 NPs on membrane fluidity was studied at 25 °C and 37 °C, and the role of membrane components in SiO2 NP-membrane interactions was investigated using giant plasma membrane vesicles (GPMVs) isolated from RBL-2H3 cells. SiO2 NPs cause a more serious membrane fluidity decrease at 37 °C than at 25 °C, which is revealed by the shift of Laurdan fluorescence emission and further quantified via forster resonance energy transfer (FRET) experiments. In addition, after the removal of 75 % cholesterol from the membrane, SiO2 NPs caused a greater extent of membrane gelation. These results indicate that SiO2 NPs prefer to interact with membranes that are more dynamic and less densely packed. Moreover, fluorescent experiments confirmed that the existence of phosphatidyl ethanolamine (PE) and phosphoinositide (PI) can mitigate NP-induced membrane gelation. Molecular dynamics simulation further demonstrated that SiO2 NPs form hydrogen bonds with the terminal of PE or PI but with the -PO4−- group in the middle of phosphatidylcholine (PC). The bonding that occurs in the terminal gives less restriction of phospholipid movement and a weaker effect on membrane fluidity. This research provides both evidence and mechanisms of SiO2 NP-induced membrane fluidity changes, which are helpful for understanding cell membrane damage and the biological effects of NPs.

Water-soluble improved NIR ratiometric fluorescent probe for specific imaging sulfur dioxide in cancer cell membranes and living mice

The comprehensive understanding of the mechanism between gas messenger molecules and cell membrane microenvironment is of great significance for basic biology research. Sulfur dioxide (SO2), as a vital gas molecule, is of great value in the permeability and fluidity of cell membranes. However, monitoring of sulfur dioxide in cell membranes with high temporal and spatial resolution remains a challenge. Herein, we designed and synthesized a ratiometric probe CMCB based on coumarin and indole sulfonate platform. The unsaturated CC is selected as the reaction site of SO2, utilizing sulfonic acid groups to increase CMCB hydrophilicity to achieve efficient targeting of cell membranes. The probe CMCB features a good cells membranes targeting and a NIR emission (720 nm). Interestingly, the probe showed low detection limit, high selectivity and specificity of targeting cell membranes. We expect that this probe will serve as an ideal tool in studying gas messenger molecule on cytomembrane in vivo.

A small molecule screen for paqr-2 suppressors identifies Tyloxapol as a membrane fluidizer for C. elegans and mammalian cells

Defects in cell membrane homeostasis are implicated in numerous disorders, including cancer, neurodegeneration and diabetes. There is therefore a need for a powerful model to study membrane homeostasis and to identify eventual therapeutic routes. The C. elegans gene paqr-2 encodes a homolog of the mammalian AdipoR1 and AdipoR2 proteins that, when mutated, causes a membrane homeostasis defect accompanied by multiple phenotypes such as intolerance to dietary saturated fatty acids, intolerance to cold and a characteristic tail tip morphology defect. We screened a compound library to identify molecules that can suppress the paqr-2 phenotypes. A single positive hit, Tyloxapol, was found that very effectively suppresses multiple paqr-2 phenotypes. Tyloxapol is a non-ionic detergent currently in use clinically as an expectorant. Importantly, we examined the potential of Tyloxapol as a fluidizer in human cells and found that it improves the viability and membrane fluidity of AdipoR2-deficient human cells challenged with palmitic acid, a membrane-rigidifying saturated fatty acid.

Seasonal changes in membrane structure and excitability in retinal neurons of goldfish (Carassius auratus) under constant environmental conditions.

Seasonal modifications in the structure of cellular membranes occur as an adaptive measure to withstand exposure to prolonged environmental change. Little is known about whether such changes occur independently of external cues, such as photoperiod or temperature, or how they may impact the central nervous system. We compared membrane properties of neurons isolated from the retina of goldfish (Carassius auratus), an organism well adapted to extreme environmental change, during the summer and winter months. Goldfish were maintained in a facility under constant environmental conditions throughout the year. Analysis of whole-retina phospholipid composition using mass spectrometry-based lipidomics revealed a twofold increase in phosphatidylethanolamine species during the winter, suggesting an increase in cell membrane fluidity. Atomic force microscopy was used to produce localized, nanoscale-force deformation of neuronal membranes. Measurement of Young's modulus indicated increased membrane-cortical stiffness (or decreased elasticity) in neurons isolated during the winter. Voltage-clamp electrophysiology was used to assess physiological changes in neurons between seasons. Winter neurons displayed a hyperpolarized reversal potential (Vrev) and a significantly lower input resistance (Rin) compared with summer neurons. This was indicative of a decrease in membrane excitability during the winter. Subsequent measurement of intracellular Ca2+ activity using Fura-2 microspectrofluorometry confirmed a reduction in action potential activity, including duration and action potential profile, in neurons isolated during the winter. These studies demonstrate chemical and biophysical changes that occur in retinal neurons of goldfish throughout the year without exposure to seasonal cues, and suggest a novel mechanism of seasonal regulation of retinal activity.

On the effects of diabetes mellitus on the mechanical properties of DRG sensory neurons and their possible relation with diabetic neuropathy

Diabetic neuropathy (DN) is one of the principal complications of diabetes mellitus (DM). Dorsal root ganglion (DRG) neurons are the primary sensory neurons that transduce mechanical, chemical, thermal, and pain stimuli. Diabetes-caused sensitivity alterations and presence of pain are due to cellular damage originated by persistent hyperglycemia, microvascular insufficiency, and oxidative and nitrosative stress. However, the underlying mechanisms have not been fully clarified. The present work addresses this problem by hypothesizing that sensitivity changes in DN result from mechanotransduction-system alterations in sensory neurons; especially, plasma membrane affectations. This hypothesis is tackled by means of elastic-deformation experiments performed on DGR neurons from a murine model for type-1 DM, as well a mathematical model of the cell mechanical structure. The obtained results suggest that the plasma-membrane fluidity of DRG sensory neurons is modified by the induction of DM, and that this alteration may correlate with changes in the cell calcium transient that results from mechanical stimuli.

Cell membrane fluidity and ROS resistance define DMSO tolerance of cryopreserved synovial MSCs and HUVECs

ObjectivesSynovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unclear. This study thus aimed to identify the biological and physical factors involved in the differences in freeze–thaw tolerance between MSCs and HUVECs.Materials and methodsFor biological analysis, MSC and HUVEC viability after freeze-thawing and alteration of gene expression in response to dimethyl sulfoxide (DMSO, a cryoprotectant) were quantitatively evaluated. For physical analysis, the cell membrane fluidity of MSCs and HUVECs before and after DMSO addition was assessed using a histogram for generalized polarization frequency.ResultsHUVECs showed lower live cell rates and higher gene expression alteration related to extracellular vesicles in response to DMSO than MSCs. Fluidity measurements revealed that the HUVEC membrane was highly fluidic and sensitive to DMSO compared to that of MSCs. Addition of CAY10566, an inhibitor of stearoyl-coA desaturase (SCD1) that produces highly fluidic desaturated fatty acids, decreased the fluidity of HUVECs and increased their tolerance to DMSO. The combination of CAY10566 and antioxidant glutathione (GSH) treatment improved HUVEC viability from 57 to 69%. Membrane fluidity alteration may thus contribute to pore-induced DMSO influx into the cytoplasm and reactive oxygen species production, leading to greater cytotoxicity in HUVECs, which have low antioxidant capacity.ConclusionsDifferences in freeze–thaw tolerance originate from differences in the cell membranes with respect to fluidity and antioxidant capacity. These findings provide a basis for analyzing cell biology and membrane-physics to establish appropriate long-term preservation methods aimed at promoting transplantation therapies.

N-acetylcysteine functionalized chitosan oligosaccharide-palmitic acid conjugate enhances ophthalmic delivery of flurbiprofen and its mechanisms

An N-acetylcysteine functionalized chitosan oligosaccharide-palmitic acid conjugate (NAC-COS-PA) with bioadhesive and permeation promoting properties was synthesized to enhance transocular drug delivery. Flurbiprofen (FB) loaded self-assembled NAC-COS-PA nanomicelles (NAC-COS-PA-FB) were prepared and the drug loading was 7.35 ± 0.32%. Human immortalized corneal epithelial (HCE-T) cell cytotoxicity and hen's egg test-chorioallantoic membrane assays confirmed that the conjugate had good biocompatibility. The transportation efficiency of coumarin-6 (C6) loaded nanomicelles in the HCE-T cell monolayer was approximately 1.97 times higher than that of free C6. Decreased intracellular Ca2+ concentration and cell membrane potential, increased cell membrane fluidity, and reversible changes in the F-actin cytoskeleton are presumed to be responsible for the enhanced drug permeation. NAC-COS-PA exhibited strong binding capacity with mucin and rabbit eyeball. In vivo pharmacokinetics indicated that the area under the curve (AUC0–6 h) and the maximum concentration (Cmax) of NAC-COS-PA-FB were approximately 1.92 and 2.44 times that of the FB solution, respectively. NAC-COS-PA-FB demonstrated the best in vivo anti-inflammatory efficacy compared to unfunctionalized nanomicelles (COS-PA-FB) and FB solution. Consequently, NAC-COS-PA appears to be a promising bioadhesive carrier for ophthalmic delivery.

Modulation of Cholesterol Pathways in Human Macrophages Infected by Clinical Isolates of Leishmania infantum.

To increase our understanding of factors contributing to therapeutic failure (TF) in leishmaniasis, we have studied some plasma membrane features of host THP-1 cells infected with clinical isolates of Leishmania infantum from patients with leishmaniasis and TF. The fluorescent probes DPH and TMA-DPH were used to measure changes in membrane fluidity at various depths of the plasma membranes. Steady-state fluorescence anisotropy of DPH embedded in the infected THP-1 membranes showed a significant increase, thereby suggesting a substantial decrease in plasma membrane fluidity relative to controls. Considering that cholesterol affects membrane fluidity and permeability, we determined the cholesterol content in plasma membrane fractions of human macrophages infected with these L. infantum lines and observed a significant increase in cholesterol content that correlates with the measured decrease in plasma membrane fluidity. In order to define the pathways that could explain the increase in cholesterol content, we studied the transcriptomics of the cholesterol-enriched pathways in host THP-1 cells infected with TF clinical isolates by RNA-seq. Specifically, we focused on four enriched Gene Ontology (GO) terms namely cholesterol efflux, cholesterol transport, cholesterol metabolic process and cholesterol storage. Additionally, we analyzed the genes involved in these pathways. Overall, this study shows that these clinical isolates are able to modulate the expression of specific genes in host cells, thereby modifying the cholesterol content in plasma membranes and inducing changes in plasma membrane fluidity that could be associated with the parasite’s ability to survive in the host macrophages, thereby possibly contributing to immune evasion and TF.

Wheat Gluten Peptides Enhance Ethanol Stress Tolerance by Regulating the Membrane Lipid Composition in Yeast.

Wheat gluten peptides (WGPs), identified as Leu-Leu (LL), Leu-Leu-Leu (LLL), and Leu-Met-Leu (LML), were tested for their impacts on cell growth, membrane lipid composition, and membrane homeostasis of yeast under ethanol stress. The results showed that WGP supplementation could strengthen cell growth and viability and enhance the ethanol stress tolerance of yeast. WGP supplementation increased the expressions of OLE1 and ERG1 and enhanced the levels of oleic acid (C18:1) and ergosterol in yeast cell membranes. Moreover, LLL and LML exhibited a better protective effect for yeast under ethanol stress compared to LL. LLL and LML supplementation led to 20.3 ± 1.5% and 18.9 ± 1.7% enhancement in cell membrane fluidity, 21.8 ± 1.6% and 30.5 ± 1.1% increase in membrane integrity, and 26.3 ± 4.8% and 27.6 ± 4.6% decrease in membrane permeability in yeast under ethanol stress, respectively. The results from scanning electron microscopy (SEM) elucidated that WGP supplementation is favorable for the maintenance of yeast cell morphology under ethanol stress. All of these results revealed that WGP is an efficient enhancer for improving the ethanol stress tolerance of yeast by regulating the membrane lipid composition.

Inactivation of Salmonella typhimurium SL1344 by Chlorogenic Acid and the Impairment of Cellular Integrity.

Chlorogenic acid (CGA) is an antibacterial agent that can be isolated from Eucommia ulmoides Oliver, a Chinese medicinal and edible plant food. The inhibitory effect of CGA on bacterial growth and stiffness of the outer membrane (OM) had been reported, while more evidence were required to elucidate its impairment of cell wall. In this study, the morphological and physiochemical changes of Salmonella cells under CGA treatment were investigated. Firstly, the minimum inhibitory concentration (MIC) of CGA against Salmonella was assayed. Later, the permeability of OM and activity of the proteins released were measured and observed to reveal the alteration of OM characteristic and cellular morphology. Finally, reactive oxygen species and cell membrane fluidity were analyzed, respectively, to elucidate how CGA damaged cell surface. The results showed that MIC of CGA against Salmonella was 6.25 mg/L. Under sub-lethal doses of CGA, the OM permeability and the release of soluble proteins were enhanced evidently, and Salmonella cells showed more deformed and shrunken, confirming the impairment of cellular integrity under CGA. Finally, the possible cause of cell surface damage was investigated. the fluidity of the membrane was increased upon CGA treatment, which may the possible cause of OM by CGA.

Endo- and Exometabolome Crosstalk in Mesenchymal Stem Cells Undergoing Osteogenic Differentiation.

This paper describes, for the first time to our knowledge, a lipidome and exometabolome characterization of osteogenic differentiation for human adipose tissue stem cells (hAMSCs) using nuclear magnetic resonance (NMR) spectroscopy. The holistic nature of NMR enabled the time-course evolution of cholesterol, mono- and polyunsaturated fatty acids (including ω-6 and ω-3 fatty acids), several phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelins, and plasmalogens), and mono- and triglycerides to be followed. Lipid changes occurred almost exclusively between days 1 and 7, followed by a tendency for lipidome stabilization after day 7. On average, phospholipids and longer and more unsaturated fatty acids increased up to day 7, probably related to plasma membrane fluidity. Articulation of lipidome changes with previously reported polar endometabolome profiling and with exometabolome changes reported here in the same cells, enabled important correlations to be established during hAMSC osteogenic differentiation. Our results supported hypotheses related to the dynamics of membrane remodelling, anti-oxidative mechanisms, protein synthesis, and energy metabolism. Importantly, the observation of specific up-taken or excreted metabolites paves the way for the identification of potential osteoinductive metabolites useful for optimized osteogenic protocols.