Cell Membrane Fluidity Research Articles

Mechanism analysis of methionine inhibiting growth of Bifidobacterium bifidum

Bifidobacterium bifidum rarely utilises yeast extract, which causes significant challenges in its industrial production. This study characterised the utilisation of amino acids by B. bifidum to improve the viability of the strain. In this study, the proliferation effect of B. bifidum in different nitrogen sources was determined, and the amino acid contents of different nitrogen sources were analysed, which indicated that hydrophobic amino acids may inhibit B. bifidum growth. Among different hydrophobic amino acids, methionine was identified as the strongest growth inhibitor. Furthermore, the intracellular pH and NH4+ contents of B. bifidum CCFM16 were higher in methionine-free medium. Among the NH3-producing enzymes related to amino acid metabolism, the metC gene was significantly transcribed in methionine-free medium. In addition, tcyP, metB, metC and metE genes encoding cysteine transporter proteins and the conversion pathway of cysteine to methionine were transcribed 2.03-fold, 7.03-fold, 3.05-fold and 2.92-fold, respectively, in methionine-free medium compared to methionine-containing medium. Compared with methionine-containing medium, the activity of tricarboxylic acid cycle rate-limited enzyme isocitrate dehydrogenase in methionine-free medium also increased from 198.89 to 410.56 nmol/min/mg protein, which probably enhanced the carbon flux and promoted growth. Meanwhile, the activity of pyrroline-5-carboxylate reductase, an enzyme that synthesises proline, increased from 14.76 to 35.28 nmol/min/mg protein, which promoted proline synthesis in response to osmotic stress. Cyclopropane fatty acids were also significantly higher in methionine-free medium, which may promote growth by decreasing cell membrane permeability to protons and enhancing cell membrane fluidity. This study provides a theoretical basis for high-density culture of B. bifidum.

Protective effect of wheat gluten peptides against ethanol-stress damage in yeast cell and identification of anti-ethanol peptides

In the study, the protective effects of wheat gluten hydrolysates (WGH) on the ethanol-induced injury in yeast cells were investigated, and the peptides that enhanced the resistance to 10% (v/v) ethanol-stress were identified. Results showed that the growth and viability of yeast under ethanol-stress were significantly enhanced by WGH, and it indicated that there were high activity fractions in WGH. Interestingly, eight wheat gluten peptides (WGP) were obtained and identified as EP, LL, LW, LPL, LML, LLL, LLW, and EFPL, respectively, and five (LL, LLL, LW, LML and EP) of them worked best. Furthermore, WGP could enhance the yeast resistance to ethanol-stress, and the WGP apparently maintained the morphology and structure (especially the cell walls and membranes) of yeast under ethanol-stress. Further studied found that WGP (especially LL and LW) contributed to increasing cell wall integrity and cell membrane fluidity while also reducing cell membrane permeability. TEM analysis found that due to the existence of LL and LW, both the thickness of cell wall and the integrity of cell membrane were enhanced, on the contrary, the cell membrane degradation and cytoplasmic efflux were reduced. These results revealed that WGP could be considered as an effective protector of yeast against ethanol-stress damage.

Miltefosine impacts small molecule transport in Gram-positive bacteria.

Miltefosine (MLT) is an alkylphosphocholine with clinical success as an anticancer and antiparasitic drug. Although the mechanism of action of MLT is highly debated, the interaction of MLT with the membrane, specifically lipid rafts of eukaryotes, is well-documented. Recent reports suggest MLT impacts the functional membrane microdomains in bacteria - regions of the membrane structurally and functionally similar to lipid rafts. There have been conflicting reports, however, as to whether MLT impacts the overall fluidity of cellular plasma membranes. Here, we apply steady-state fluorescence techniques, generalized polarization of laurdan and anisotropy of diphenylhexatriene, to discern how MLT impacts the global ordering and lipid packing of Staphylococcus aureus membranes. Additionally, we investigate how the transport of a range of small molecules is impacted by MLT for S. aureus and Bacillus subtilis by employing time-resolved second harmonic scattering. Overall, we observe MLT does not have an influence on the overall ordering and packing of S. aureus membranes. Additionally, we show that the transport of small molecules across the membrane can be significantly altered by MLT - although this is not the case for all molecules studied. The results presented here illustrate the potential use of MLT as an adjuvant to assist in the delivery of drug molecules in bacteria.

Highly sensitive and accurate detection of cholesterol based on a single red upconversion biosensor.

Cholesterol (CHOL) is an important clinical biochemical indicator that plays an important role in the regulation of the fluidity, permeability, and microstructure of cell membranes. Therefore, it is necessary to accurately monitor CHOL levels in biological samples for the early prevention and diagnosis of various diseases. The single-band red upconversion nanoparticle (UCNP) emits light within the optical transmission window of biological tissues, and can penetrate deeper biological tissues and cause less energy loss due to scattering and thus have higher sensitivity and accuracy. Here, using the nontoxic, sensitive, and photochemically stable 3,3',5,5'-tetramethylbenzidine (TMB) as the quenching agent and single red UCNP as the fluorescent donor, a dual-readout colorimetric and fluorescent sensor was developed to detect CHOL. The detection mechanism and feasibility were discussed in detail, and experimental conditions such as Fe2+ concentration, TMB concentration and reaction time were explored. Under optimal conditions, the limits of CHOL detection by colorimetry and fluorescence were 0.85 μM and 0.63 μM. The sensing system was used to measure CHOL in serum samples and the values obtained by these two modes were close, and the spiked recoveries were 97.2-102.2% and 97.1-103.7%, respectively, which holds great potential in clinical diagnosis and health management.

Analysis of single-molecule diffusion movement in cell membrance based on unsupervised learning methods: Different effects of cholesterol on flowability of model membrane and living cell membrane

<sec>Single molecular tracking is a valuable approach to investigate the dynamic processes and molecular interactions in soft matter systems, particularly in biological systems. However, understanding the complexity of single molecule motion behaviors in biological systems remains a significant challenge. To address this issue, we propose a two-step classification method based on unsupervised learning to efficiently identify and classify single molecule trajectories. Firstly, we employ an entropy-constrained least square method to distinguish between confined (e.g., immobile) and unconfined diffusion trajectories. Subsequently, statistical tests are utilized to categorize the unconfined trajectories into different diffusion modes such as sub-diffusion, normal diffusion, and super-diffusion.</sec><sec>By applying this method, we analyze the diffusion motion of single molecules in both DOPC model cell membranes and living cell membranes while uncovering their distinct responses to cholesterol composition. Our findings demonstrate that both model membranes and living cell membranes exhibit diverse molecular diffusion modes. Specifically, in the DOPC model membrane system, the presence of cholesterol components impedes lipid diffusion within the membrane. The degree of inhibition is positively correlated with the amount of cholesterol present. For instance, as the cholesterol content in the membrane increases from 0 to 20% (DOPC:Chol = 4∶1) and 50% (DOPC:Chol = 1∶1), there is an increase in the proportion of molecules, exhibiting confined diffusion and sub-diffusion (from 55% to 45%), while there is a decrease in the proportion of molecules, displaying normal diffusion and super-diffusion (from 45% to 35%). The ensemble diffusion coefficient of molecules in the membrane significantly decreases, which can be attributed to both a decrease in velocity among fast-moving molecules. Interestingly, after using MeβCD to remove cholesterol, the single-molecule mobility within the DOPC/Chol composite membrane system is restored to a level similar to that of the pure DOPC membrane.</sec><sec>Conversely, in the living cell membrane system, the diffusion coefficient values of molecules are significantly lower than those observed in the model membrane system; furthermore, the removal of cholesterol further slows down the molecular diffusion rate. This study contributes to understanding the intricacies of biomolecular motility and its dependence on environmental factors from a perspective of single molecular motion.</sec>

Cell-membrane engineering strategies for clinic-guided design of nanomedicine.

Cells are the fundamental units of life. The cell membrane primarily composed of two layers of phospholipids (a bilayer) structurally defines the boundary of a cell, which can protect its interior from external disturbances and also selectively exchange substances and conduct signals from the extracellular environment. The complexity and particularity of transmembrane proteins provide the foundation for versatile cellular functions. Nanomedicine as an emerging therapeutic strategy holds tremendous potential in the healthcare field. However, it is susceptible to recognition and clearance by the immune system. To overcome this bottleneck, the technology of cell membrane coating has been extensively used in nanomedicines for their enhanced therapeutic efficacy, attributed to the favorable fluidity and biocompatibility of cell membranes with various membrane-anchored proteins. Meanwhile, some engineering strategies of cell membranes through various chemical, physical and biological ways have been progressively developed to enable their versatile therapeutic functions against complex diseases. In this review, we summarized the potential clinical applications of four typical cell membranes, elucidated their underlying therapeutic mechanisms, and outlined their current engineering approaches. In addition, we further discussed the limitation of this technology of cell membrane coating in clinical applications, and possible solutions to address these challenges.

Extreme makeover: the incredible cell membrane adaptations of extremophiles to harsh environments.

The existence of life beyond Earth has long captivated humanity, and the study of extremophiles-organisms surviving and thriving in extreme environments-provides crucial insights into this possibility. Extremophiles overcome severe challenges such as enzyme inactivity, protein denaturation, and damage of the cell membrane by adopting several strategies. This feature article focuses on the molecular strategies extremophiles use to maintain the cell membrane's structure and fluidity under external stress. Key strategies include homeoviscous adaptation (HVA), involving the regulation of lipid composition, and osmolyte-mediated adaptation (OMA), where small organic molecules protect the lipid membrane under stress. Proteins also have direct and indirect roles in protecting the lipid membrane. Examining the survival strategies of extremophiles provides scientists with crucial insights into how life can adapt and persist in harsh conditions, shedding light on the origins of life. This article examines HVA and OMA and their mechanisms in maintaining membrane stability, emphasizing our contributions to this field. It also provides a brief overview of the roles of proteins and concludes with recommendations for future research directions.

Are nature's strategies the solutions to the rational design of low-melting, lipophilic ionic liquids?

Ionic liquids (ILs) have emerged as a new class of materials, displaying a unique capability to self-assemble into micelles, liposomes, liquid crystals, and microemulsions. Despite evident interest, advancements in the controlled formation of amphiphilic ILs remain in the early stages. Taking inspiration from nature, we introduced the concept of lipid-like (or lipid-inspired) ILs more than a decade ago, aiming to create very low-melting, highly lipophilic ILs that are potentially bio-innocuous - a combination of attributes that is frequently antithetical but highly desirable from several application-specific standpoints. Lipid-like ILs are a subclass of functional organic liquid salts that include a range of lipidic side chains such as saturated, unsaturated, linear, branched, and thioether while retaining melting points below room temperature. It was observed in several homologous series of [Cnmim] ILs that elongation of N-appended alkyl chains to greater than seven carbons leads to a substantial increase in melting point (Tm) - which is the most characteristic feature of ILs. Accordingly, it is challenging to develop ILs with low Tm values while preserving their hydrophobicity and self-organizing properties. We found that two alternative Tm depressive approaches are useful. One of these is the replacement of the double bonds with thioether moieties in the alkyl chains, as detailed in several published papers detailing the chemistry of these ILs. Employing thiol-ene and thiol-yne click reactions is a facile, robust, and orthogonal method to overcome the challenges associated with the synthesis of alkyl thioether-functionalized ILs. The second approach involves replacing the double bonds with the cisoid cyclopropyl motif, mimicking the strategy used by certain organisms to modulate cell membrane fluidity. This discovery has the potential to greatly impact the utilization of lipid-like ILs in various applications, including gene delivery, lubricants, heat transfer fluids, and haloalkane separations, among others. This feature article presents a concise, historical overview, highlighting key findings from our work while offering speculation about the future trajectory of this de novo class of soft organic-ion materials.

Molecularly imprinted sensor based on cascade enzyme system supported by metal-organic framework (UiO-66-NH2) for sensitive colorimetric detection of cholesterol

Cholesterol is a crucial cellular molecule in the human body, and a neutral lipid necessary for maintaining the integrity and fluidity of cell membranes. Additionally, it acts as a precursor for the synthesis of essential substances for living organisms, including steroid hormones, bile acids and vitamins. Nonetheless, cholesterol is not always advantageous, and both excess and deficiency of cholesterol can result in serious diseases. Therefore, maintaining cholesterol at a normal level is of paramount importance for the maintenance of normal cellular activity. In this study, an amino-functionalized metal-organic framework (UiO-66-NH2, abbreviated as U6NH2) was used as a carrier, on the surface of which gold nanoparticles (AuNPs) was embedded and cholesterol oxidase (ChOx) was immobilized, and then cholesterol was employed as a template molecule, followed by the synthesis of molecularly imprinted polymers (MIPs). Remarkably, the obtained U6NH2@AuNPs-ChOx@MIPs can be used to facilitate efficacious and selective detection of blood cholesterol through a facile colorimetric method over a range of 2.9 mM - 6.7 mM with a limit of detection (LOD) = 2.4 mM, while the normal level of blood cholesterol should be less than 5.18 mM. Moreover, the sensing system demonstrates excellent stability and satisfactory selectivity. The constructed cholesterol selective detection system reported in this paper is anticipated to have a crucial impact on bioanalysis.

Action mechanism of the potential biocontrol agent Brevibacillus laterosporus SN19-1 against Xanthomonas oryzae pv. oryzae causing rice bacterial leaf blight.

The causal agent of rice bacterial leaf blight (BLB) is Xanthomonas oryzae pv. oryzae (Xoo), which causes serious damage to rice, leading to yield reduction or even crop failure. Brevibacillus laterosporus SN19-1 is a biocontrol strain obtained by long-term screening in our laboratory, which has a good antagonistic effect on a variety of plant pathogenic bacteria. In this study, we investigated the efficacy and bacterial inhibition of B. laterosporus SN19-1 against BLB to lay the theoretical foundation and research technology for the development of SN19-1 as a biopesticide of BLB. It was found that SN19-1 has the ability to fix nitrogen, detoxify organic phosphorus, and produce cellulase, protease, and siderophores, as well as IAA. In a greenhouse pot experiment, the control efficiency of SN19-1 against BLB was as high as 90.92%. Further investigation of the inhibitory mechanism of SN19-1 on Xoo found that the biofilm formation ability of Xoo was inhibited and the pathogenicity was weakened after the action of SN19-1 sterile supernatant on Xoo. The activities of enzymes related to respiration and the energy metabolism of Xoo were significantly inhibited, while the level of intracellular reactive oxygen species was greatly increased. Scanning electron microscopy observations showed folds on the surface of Xoo. A significant increase in cell membrane permeability and outer membrane permeability and a decrease in cell membrane fluidity resulted in the extravasation of intracellular substances and cell death. The results of this study highlight the role of B. laterosporus SN19-1 against the pathogen of BLB and help elucidate the underlying molecular mechanisms.

A Physiogenomic Study of the Tolerance of Saccharomyces cerevisiae to Isoamyl Alcohol

Isoamyl alcohol is a clear, unpleasantly odorous, colorless liquid of higher alcohol that emits a fruity aroma when heavily diluted. It has received much attention in recent years as a new fuel with a high energy density. Isoamyl alcohol can be produced industrially by microbial fermentation. Still, its toxicity to host cells has limited its potential for industrial production, and the molecular mechanism of its toxic effects has not yet been elucidated. In this study, RNA-Seq technology was used to analyze the transcripts of Saccharomyces cerevisiae under normal conditions and in the presence of isoamyl alcohol (0.5 g/L and 2.5 g/L). The results showed that the expression of the cell wall (CCW12, BGL2, NCW2 and SUN4), cell membrane (ELO1, ERG2, FAA1, and OPI3), translation and other structural genes were significantly down-regulated. The expression of genes related to ATP biosynthesis, NADPH biosynthesis (ZWF1), and metal ion transport (PMC1) proteins were up-regulated. Strains with key genes knocked out were cultured without isoamyl alcohol. Combined results suggested that isoamyl alcohol may affect cell wall stability and cell membrane fluidity, and the expression of genes related to ion homeostasis and energy production may play a protective role against isoamyl alcohol stress. By maintaining cell wall stability/membrane fluidity under isoamyl alcohol pressure, improving certain ion homeostasis, and generating energy/NADPH, it is possible to overcome the toxicity of isoamyl alcohol in industrial fermentation processes to a certain extent.

P140Cap modulates the mevalonate pathway decreasing cell migration and enhancing drug sensitivity in breast cancer cells

p140Cap is an adaptor protein involved in assembling multi-protein complexes regulating several cellular processes. p140Cap acts as a tumor suppressor in breast cancer (BC) and neuroblastoma patients, where its expression correlates with a better prognosis. The role of p140Cap in tumor metabolism remains largely unknown. Here we study the role of p140Cap in the modulation of the mevalonate (MVA) pathway in BC cells. The MVA pathway is responsible for the biosynthesis of cholesterol and non-sterol isoprenoids and is often deregulated in cancer. We found that both in vitro and in vivo, p140Cap cells and tumors show an increased flux through the MVA pathway by positively regulating the pace-maker enzyme of the MVA pathway, the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR), via transcriptional and post-translational mechanisms. The higher cholesterol synthesis is paralleled with enhanced cholesterol efflux. Moreover, p140Cap promotes increased cholesterol localization in the plasma membrane and reduces lipid rafts-associated Rac1 signalling, impairing cell membrane fluidity and cell migration in a cholesterol-dependent manner. Finally, p140Cap BC cells exhibit decreased cell viability upon treatments with statins, alone or in combination with chemotherapeutic at low concentrations in a synergistic manner. Overall, our data highlight a new perspective point on tumor suppression in BC by establishing a previously uncharacterized role of the MVA pathway in p140Cap expressing tumors, thus paving the way to the use of p140Cap as a potent biomarker to stratify patients for better tuning therapeutic options.

Inactivation of Carotenogenic-Biosynthesizing Genes Altered Lipids Composition and Intensity in Cronobacter sakazakii.

Cronobacter sakazakii, an opportunistic milk-borne pathogen responsible for severe neonatal meningitis and bacteremia, can synthesize yellow pigment (various carotenoids) benefiting for bacterial survival, while little literature was available about the influence of various carotenoids on bacterial resistance to a series of stresses and the characteristics of cell membrane, obstructing the development of novel bactericidal strategies overcoming the strong tolerance of C. sakazakii. Thus in this study, for the first time, five carotenogenic genes of C. sakazakii BAA-894 were inactivated, respectively, to construct a series of mutants producing various carotenoids and their effects on the cell membrane properties, and resistances to food- and host-related stresses, were investigated systematically. Furthermore, to explore its possible mode of action, comparative lipidomics analysis was performed to reveal the change of lipids that were mainly located at cell membranes. The results showed that five mutants (ΔcrtB, ΔcrtI, ΔcrtY, ΔcrtZ, and ΔcrtX) displayed negligible change in growth rate but higher permeability of the outer membrane and lower fluidity of cell membrane compared to the wild type. Besides, these mutants exhibited poorer ability of biofilm formation and lower resistances to acid, oxidative, osmotic, and desiccation stresses, indicating that different carotenoid composition significantly affected environmental tolerance of C. sakazakii. To discover the possible causes, lipidomics analysis of C. sakazakii was conducted and more than 500 lipid species belonging to 27 classes had been identified at first. Compared to that of BAA-894, the composition and relative intensity of lipid species in five mutants varied significantly, especially the monounsaturated and biunsaturated phosphatidylethanolamine. The evidence presented in this study demonstrated that the varied composition of carotenoids in C. sakazakii significantly altered the lipid profile and intensity, which maybe a crucial means to influencing the characteristics of cell membranes and resistance to environmental stresses.

Impact of Conventional and Potential New Metal-Based Drugs on Lipid Metabolism in Osteosarcoma MG-63 Cells.

This work investigated the mechanisms of action of conventional drugs, cisplatin and oxaliplatin, and the potentially less deleterious drug Pd2Spermine (Spm) and its Pt(II) analog, against osteosarcoma MG-63 cells, using nuclear-magnetic-resonance metabolomics of the cellular lipidome. The Pt(II) chelates induced different responses, namely regarding polyunsaturated-fatty-acids (increased upon cisplatin), suggesting that cisplatin-treated cells have higher membrane fluidity/permeability, thus facilitating cell entry and justifying higher cytotoxicity. Both conventional drugs significantly increased triglyceride levels, while Pt2Spm maintained control levels; this may reflect enhanced apoptotic behavior for conventional drugs, but not for Pt2Spm. Compared to Pt2Spm, the more cytotoxic Pd2Spm (IC50 comparable to cisplatin) induced a distinct phospholipids profile, possibly reflecting enhanced de novo biosynthesis to modulate membrane fluidity and drug-accessibility to cells, similarly to cisplatin. However, Pd2Spm differed from cisplatin in that cells had equivalent (low) levels of triglycerides as Pt2Spm, suggesting the absence/low extent of apoptosis. Our results suggest that Pd2Spm acts on MG-63 cells mainly through adaptation of cell membrane fluidity, whereas cisplatin seems to couple a similar effect with typical signs of apoptosis. These results were discussed in articulation with reported polar metabolome adaptations, building on the insight of these drugs' mechanisms, and particularly of Pd2Spm as a possible cisplatin substitute.

Monoterpenes as potential antifungal molecules against Candida cell membranes: in-vitro and in-silico studies

Azoles are the frequently used antifungal drugs that target the enzyme lanosterol 14 α-demethylase (erg11p). This enzyme plays a vital role in ergosterol biosynthesis and hence maintainenance of cell membrane fluidity and integrity. The emergence of resistance to azoles and their fungistatic nature against several fungal pathogens is the major challenge to combat invasive candidiasis. Therefore, there is an urgent need to discover new antifungals with better efficacy. This study targets erg11 protein using in silico approach and identifies the monoterpene compounds (α-terpineol, carveol, and terpinene-4-ol) based on docking score and ligand interaction analysis. Further dynamic behavior of best-docked compounds with erg11p was analyzed by various parameters of MD simulation. The binding free energy of selected compounds towards the definitive pocket was also calculated. To further investigate the antifungal activity of selected compounds, in vitro studies were conducted on C. albicans. Studies thus suggest that the proposed the mechanism of antifungal action of test compounds involves targeting the ergosterol biosynthetic pathway. The compounds were explored for their effect on the disruption of membrane integrity by studying ERG11gene expression analysis, scanning electron microscopy, PI uptake (fluorescence microscopy,) and H+-extrusion. The results suggest that the selected monoterpenes are safer natural antifungals that disrupt membrane integrity by inhibiting ergosterol biosynthesis and other membrane associated structures. Communicated by Ramaswamy H. Sarma

The role of the intrinsic pathway of apoptosis in human ejaculated sperm damage under a state of scrotal heat stress

PurposeThe study aimed to determine the associations among standard sperm characteristics and oxidative/apoptotic markers in ejaculated sperm of men exposed to prolonged scrotal hyperthermia of either environmental or clinical origin.MethodsThe original study design included four research groups: professional drivers (n = 54), infertile men with varicocele (n = 78), infertile men not exposed to prolonged genital heat stress (n = 37), and fertile individuals serving as the control group (n = 29). Standard semen analysis was performed according to the 5th WHO laboratory manual. The following oxidative and apoptotic parameters of sperm were investigated: mitochondrial superoxide anion generation (MitoSOX Red dye), phosphatidylserine externalization (Annexin V binding assay), mitochondrial membrane potential (JC-1 dye), DNA fragmentation (TUNEL/PI assay), and membrane fluidity (merocyanine 540 dye).ResultsAll the studied groups presented a strong deterioration in routine sperm parameters and a strongly apoptotic phenotype in sperm, characterized by both decreased mitochondrial membrane potential and enhanced DNA fragmentation, regardless of the thermal insult. Significant induction of mitochondrial superoxide anion generation was noted only in the groups exposed to genital heat stress. A positive correlation between the production of superoxide anion in the mitochondrial chain and the level of DNA fragmentation in drivers was also noted.ConclusionLong-term exposure to scrotal hyperthermia in real-life situations is sufficient to reduce sperm quality in humans. The thermal stress directly induces the oxidative stress cascade in ejaculated sperm, affecting the plasma membrane fluidity, mitochondrial homeostasis, and sperm DNA integrity.

10-Hydroxydec-2-enoic acid enhances the erythrocyte membrane fluidity via interacting with phosphatidylcholine and phosphatidylethanolamine

10-Hydroxydec-2-enoic acid (10-HDA), the unique substance in the natural food royal jelly, is an unsaturated hydroxyl fatty acid with the bio-activity to guard against cell or tissue damages. However, the relevant mechanisms still remain largely unknown. Here, using a mouse erythrocyte model, whether the fluidity of plasma membrane is influenced by 10-HDA was investigated. 10-HDA was shown to enhance the erythrocyte membrane fluidity and to rescue the fluidity from •OH toxicity. In such occasions, 10-HDA promoted the dissolved O2 level in erythrocyte suspension thereof. The levels of β-actin and band 4.1 protein were not affected by 10-HDA in erythrocytes. In the framework of density functional theory, 10-HDA may interact with phosphatidylcholine and phosphatidylethanolamine, the major components of membrane lipid bilayer. This was evidenced by the fact that 10-HDA was detectable in erythrocytes after its addition to the erythrocyte suspension. Moreover, the fluorescent intensity of erythrocytes stained by the membrane probe DiIC18(5) increased with 10-HDA treatment. Hence, 10-HDA could regulate the erythrocyte membrane fluidity and the release of O2 from erythrocytes via targeting lipids rather than proteins. Our data cast light on the elucidation of the mechanism of 10-HDA modulating cell functionality and the utilization of 10-HDA in remedying the erythrocyte membrane fluidity-associated symptoms.

Cholesterol Behind the Whole Life of Humans

Cholesterol is a sterol, synthesized by animal cells and vital lipid molecule for human cells (animal) and is also a component of the diet, being present in food of animal origin. Its main function is to maintain the integrity and fluidity of cell membranes and to serve as a precursor for the synthesis of substance that are vital for the organism including steroid hormones, bile acids and vitamin D. animal cells acquire cholesterol from extracellular lipoprotein and form new synthesis of cells. Dietary cholesterol comes exclusively from animal sources, it is naturally present in our diet and tissues. Cholesterol is synthesized in the human body in order to maintain a stable peel when dietary intake is low, given the necessity for cholesterol, very effective intestinal uptake mechanisms bile acid and cholesterol reabsorption cycles exist1. Conversely carbohydrates are poorly absorbed and indeed rapidly excreted. Dietary cholesterol content does not significantly influence cholesterol value, which are regulated by various nutritional. Factors that influence cholesterol absorption (or) synthesis. Recent study data do not support a link between dietary cholesterol and cardio vascular diseases2.

Assessment of oxidative stress biomarkers in the threatened annual killifish Austrolebias charrua exposed to Roundup

This study aimed to analyze the toxic effects of Roundup Transorb® on the endangered Neotropical annual killifish Austrolebias charrua through the assessment of molecular and biochemical biomarkers. The fish were collected in temporary ponds and exposed to environmentally realistic concentrations of the herbicide (5 mg.L‐1 for 96 h). The production of ROS, lipid peroxidation, DNA damage, and membrane fluidity were evaluated in the blood cells by flow cytometry. The mRNA expression of the antioxidant-related genes sod2, cat, gstα, atp1a1, gclc, and ucp1 across the brain, liver, and gills was quantified. The acute exposure of annual killifish to Roundup significantly increased ROS production, lipid peroxidation, and DNA damage in their erythrocytes. Likewise, Roundup Transorb® decreased membrane fluidity in the blood cells of the exposed fish. Gene expression analysis revealed that Roundup exposure alters the relative expression of genes associated with oxidative stress and antioxidant defense. Our results give rise to new insights into adaptive mechanisms of A. charrua in response to Roundup. Since Brazilian annual killifishes strongly risk extinction, this study paves the way for developing novel biotechnologies applied to environmental monitoring and aquatic toxicology assessment.

Thermoprotection by a cell membrane-localized metacaspase in a green alga.

Caspases are restricted to animals, while other organisms, including plants, possess metacaspases (MCAs), a more ancient and broader class of structurally related yet biochemically distinct proteases. Our current understanding of plant MCAs is derived from studies in streptophytes, and mostly in Arabidopsis (Arabidopsis thaliana) with 9 MCAs with partially redundant activities. In contrast to streptophytes, most chlorophytes contain only 1 or 2 uncharacterized MCAs, providing an excellent platform for MCA research. Here we investigated CrMCA-II, the single type-II MCA from the model chlorophyte Chlamydomonas (Chlamydomonas reinhardtii). Surprisingly, unlike other studied MCAs and similar to caspases, CrMCA-II dimerizes both in vitro and in vivo. Furthermore, activation of CrMCA-II in vivo correlated with its dimerization. Most of CrMCA-II in the cell was present as a proenzyme (zymogen) attached to the plasma membrane (PM). Deletion of CrMCA-II by genome editing compromised thermotolerance, leading to increased cell death under heat stress. Adding back either wild-type or catalytically dead CrMCA-II restored thermoprotection, suggesting that its proteolytic activity is dispensable for this effect. Finally, we connected the non-proteolytic role of CrMCA-II in thermotolerance to the ability to modulate PM fluidity. Our study reveals an ancient, MCA-dependent thermotolerance mechanism retained by Chlamydomonas and probably lost during the evolution of multicellularity.