Cellular Membrane Function Research Articles

Impact of Omega-3 on Hypercholesteremia Induced Male Rabbits

Atherosclerosis is a disorder of vasculature wall resulting mainly from the inflammatory processes. This condition involves vascular endothelial dysfunction, recruitment and activation of phagocytic cells in early stages. Omega-3 polyunsaturated fatty acids comprise part of the cellular membrane function many roles including increased development and functionality of neuronal synapses. This study assesses the significance of omega-3 fatty acids on atherosclerosis events via lowering oxidative and inflammatory insults. In this study, twenty-four domestic rabbits (male) were randomly distributed into three groups. The first one was the negative control group in which, rabbits were received normal diet (Oxide) for twelve week-period. The second group was the hypercholesteremia-induced, untreated rabbits fed with cholesterol (1%) enriched diet. The third group consumed cholesterol (1%) enriched diet supplemented with 5% omega-3 fatty acids. Blood samples were collected after twelve weeks to measure high-density lipoprotein-C (HDL-C), total cholesterol (TC), serum triglycerides (TG), endothelin-1 (ET-1), intracellular adhesion molecule-1 (ICAM-1) and highly sensitive C-reactive protein (hs-CRP) levels. After twelve weeks, aorta of each rabbit was isolated to identify glutathione (GSH),malondialdehyde (MDA) and intimal thickness. Administration of omega-3 was un-capable to modify lipid parameters significantly when compared with high cholesterol-fed rabbits. Administration of omega-3 results in improvement of ICAM-1, hs-CRP, ET-1, aortic MDA and intimal thickness of aorta significantly when compared with untreated high cholesterol-fed rabbits (P

Features of disturbances of functions of cellular membranes in patients with chronic kidney disease, treated with programmed hemodialysis

Features of disturbances of functions of cellular membranes in patients with chronic kidney disease, treated with programmed hemodialysis

Embracing Biological Complexity in Atomistic Simulations of Cellular Membranes

In all biological systems, complexity is a defining feature that is inherently coupled to function. For example, the function of cellular membranes relies, in part, on an intricate interplay between their geometries and their lipid and protein compositions. Molecular dynamics (MD) simulations have the power to reveal the atomistic structural basis of this interplay, but modeling cellular membranes has previously been intractable due to their inherent complexity and immense spatial scale. New software we have developed called xMAS (Experimentally-Derived Membranes of Arbitrary Shape) Builder overcomes these challenges. Starting from experimental data (e.g., structures from electron microscopy and lipid compositions from chromatography), xMAS Builder builds realistic cellular membrane models through the application of a series of automated, robust, and efficient modeling algorithms (e.g., to appropriately place lipids and proteins in the models). Using xMAS Builder, we have built the first atomistic cell-scale (∼1.9μm³) model of a helicoidal membrane structure from the endoplasmic reticulum called a Terasaki Ramp, and we have also built several models of a smaller synthetic system with equivalent complexity. From extensive MD simulations of these models, we have gained fundamental insights into the general behavior of cellular membranes, including, for example, the principles that determine the number of lipid molecules that can be accommodated by curved membranes and the natural response of cellular membranes to perturbations to their structures. As we apply xMAS Builder to other cellular membrane systems, MD simulations of the resulting models will continue to reveal how their constituent lipids and proteins work together in the context of crowded, curved environments. Complemented by other experimental and computational techniques, these simulations will enable detailed understanding of how the functions of cellular membranes are influenced by and derived from their innate biological complexity.

Metabolism of phospholipids in the yeast Schizosaccharomyces pombe.

The fission yeast Schizosaccharomyces pombe is an important model organism for the study of fundamental questions in eukaryotic cell and molecular biology. A plethora of cellular processes are membrane associated and/or dependent on the proper functioning of cellular membranes. Phospholipids are not only the basic building blocks of cellular membranes; they also serve as precursors to numerous signaling molecules. In this review, we describe the biosynthetic pathways leading to major S. pombe phospholipids, how these pathways are regulated, and what is known about degradation and turnover of fission yeast phospholipids. This review also addresses the synthesis, regulation and the role of water-soluble phospholipid precursors. The last chapter of the review is devoted to the use of S. pombe for the biotechnological production of value-added lipid molecules.

Plasmonic Nanoparticle-Interfaced Lipid Bilayer Membranes.

Plasmonic nanoparticles are widely exploited in diverse bioapplications ranging from therapeutics to biosensing and biocomputing because of their strong and tunable light-matter interactions, facile and versatile chemical/biological ligand modifications, and biocompatibility. With the rapid growth of nanobiotechnology, understanding dynamic interactions between nanoparticles and biological systems at the molecular or single-particle level is becoming increasingly important for interrogating biological systems with functional nanostructures and for developing nanoparticle-based biosensors and therapeutic agents. Therefore, significant efforts have been devoted to precisely design and create nano-bio interfaces by manipulating the nanoparticles' size, shape, and surface ligand interactions with complex biological systems to maximize their performance and avoid unwanted responses, such as their agglomeration and cytotoxicity. However, investigating physicochemical interactions at the nano-bio interfaces in a quantitative and controllable manner remains challenging, as the interfaces involve highly complex networks between nanoparticles, biomolecules, and cells across multiple scales, each with a myriad of different chemical and biological interactions. A lipid bilayer is a membrane made of two layers of lipid molecules that forms a barrier around cells and plays structural and functional roles in diverse biological processes because they incorporate and present functional molecules (such as membrane proteins) with lateral fluidity. Plasmonic nanoparticles conjugated on lipid membranes provide reliable analytical labels and functional moieties that allow for studying and manipulating interactions between nanoparticles and molecules with single-particle resolution; they also serve as efficient tools for applying optical, mechanical, and thermal stimuli to biological systems, which stem from plasmonic properties. Recently, new opportunities have emerged by interfacing nanoparticle-modified lipid bilayers (NLBs) with complex systems such as molecular circuits and living systems. In this Account, we briefly review how plasmonic properties can be beneficially harnessed on lipid bilayer membranes to investigate the structures and functions of cellular membranes and to develop new platforms for biomedical applications. In particular, we discuss the versatility of supported lipid bilayers (SLBs), which are planar lipid bilayers on hydrophilic substrates, as dynamic biomaterials that provide lateral fluidity and cell membrane-like environments. We then summarize our efforts to create a quantitative analytical platform utilizing nanoparticles as active building blocks and SLBs as integrative substrates. Through this bottom-up approach, various functionalized nanoparticles have been introduced onto lipid bilayers to render nanoparticle-nanoparticle, nanoparticle-lipid bilayer, and biomolecule-lipid bilayer interfaces programmable. Our system provides a new class of tools for studying thermodynamics and kinetics in complex networks of nanostructures and for realizing unique applications in biosensing and biocomputing.

Terpenes, hormones and life: isoprene rule revisited.

The year 2019 marks the 80th anniversary of the 1939 Nobel Prize in Chemistry awarded to Leopold Ruzicka (1887-1976) for work on higher terpene molecular structures, including the first chemical synthesis of male sex hormones. Arguably his crowning achievement was the 'biogenetic isoprene rule', which helped to unravel the complexities of terpenoid biosynthesis. The rule declares terpenoids to be enzymatically cyclized products of substrate alkene chains containing a characteristic number of linear, head-to-tail condensed, C5 isoprene units. The number of repeat isoprene units dictates the type of terpene produced (i.e., 2, monoterpene; 3, sesquiterpene; 4, diterpene, etc.). In the case of triterpenes, six C5 isoprene units combine into C30 squalene, which is cyclized into one of the signature carbon skeletons from which myriad downstream triterpenoid structures are derived, including sterols and steroids. Ruzicka also had a keen interest in the origin of life, but the pivotal role of terpenoids has generally been overshadowed by nucleobases, amino acids, and sugars. To redress the balance, we provide a historical and evolutionary perspective. We address the potential abiotic generation of isoprene, the crucial role that polyprene terpenoids played in early membranes and cellular life, and emphasize that endocrinology from microbes to plants and vertebrates is firmly grounded on Ruzicka's pivotal insights into the structure and function of terpenes. A harmonizing feature is that all known lifeforms (including bacteria) biosynthesize triterpenoid substances that are essential for cellular membrane formation and function, from which signaling molecules such as steroid hormones and cognate receptors are likely to have evolved.

Quantification of phospholipid fatty acids by chemical isotope labeling coupled with atmospheric pressure gas chromatography quadrupole- time-of-flight mass spectrometry (APGC/Q-TOF MS)

Phospholipid fatty acids play the crucial role in biophysical properties and the function of cellular membranes. In the present study, an accurate and sensitive method was developed to quantify phospholipid fatty acids in biological samples by using chemical isotope labeling coupled with atmospheric pressure gas chromatography quadrupole-time-of-flight mass spectrometry (APGC/Q-TOF MS). APGC, a soft ionization source, was operated under proton-transfer condition by introducing methanol into the ionization source as a modifier, which provided high quantifiable molecular ion peaks to substantially enhance the sensitivity. Fatty acid standards were methylated with methanol-d4 to yield FAMEs-d3 that were used as one-to-one internal standards to ensure accurate quantification. Thirty fatty acids in phospholipids were accurately quantified in wide linear range with limit of quantification ranging from 84.6 to 113.2 pg/mL. The newly developed method was successfully applied to quantify phospholipid fatty acids in brain and liver tissues from both fat-1 and WT mice. This method might be expanded to quantify free fatty acids or other conjugated fatty acids in biological samples or other matrices.

Dissolved Organic Matter Modulates Algal Oxidative Stress and Membrane System Responses to Binary Mixtures of Nano-Metal-Oxides (nCeO2, nMgO and nFe3O4) and Sulfadiazine.

Joint biomarker responses, oxidative stress and membrane systems, were determined for nano-metal-oxides (nMeO, i.e., nCeO2, nMgO, and nFe3O4) and sulfadiazine (SDZ) exposed at relevant low concentrations to two freshwater microalgae Scenedesmus obliquus and Chlorella pyrenoidosa. The impacts of dissolved organic matter (DOM) on the joint biomarker responses were also investigated. Results indicated that the presence of SDZ significantly decreased the level of intercellular reactive oxygen species (ROS) in the algal cells exposed to each nMeO. Reduction of cell membrane permeability (CMP) and mitochondrial membrane potential (MMP) in the algal cells was observed when the algae were exposed to the mixture of SDZ and the nMeO. The degree of reduction of the ROS level, CMP, and MMP significantly went down with the addition of DOM to a certain extent. Changes in cellular oxidative stress and membrane function depended on the types of both nMeO and algal species. This contribution provides an insight into the hazard assessment of a mixture consisting of emerging contaminants and DOM, as they can coexist in the aquatic environment.

Ecotoxicological effects on Scenedesmus obliquus and Danio rerio Co-exposed to polystyrene nano-plastic particles and natural acidic organic polymer

The importance of attention to unravel the interaction of nano-plastic particles (NPs) with natural acidic organic polymer (NAOP) in freshwater environment should not be neglected. However, toxicological data available for the interaction between NPs and NAOP remain limited. Here, we investigate the toxicological effects of three model polystyrene (PS) NPs with different functional groups (unmodified, amino- and carboxyl-modified PS NPs) on two freshwater organisms of different trophic levels (Scenedesmus obliquus and Danio rerio) in the absence and presence of two classes of NAOP, namely fulvic acid and humic acid. The NAOP interaction with the NPs is shown to alter oxidative stress and disturb membrane function in S. obliquus cells to a certain extent. Combined oxidative stress responses to the NPs and NAOP in D. rerio as a function of their mixture levels showed inhibition, alleviation, and reinforce. Changes in cellular oxidative stress and membrane function depended on the concentration and types of both NPs and NAOP. Furthermore, the characterization parameters of the NPs were important for the explanation of the ecotoxicological mechanism of the NPs in the presence of NAOP. Our findings emphasized the critical role of NAOP in the fate and toxicity of plastic particles in freshwater environment.

Preferential Equilibrium Partitioning of Positively Charged Tryptophan into Phosphatidylcholine Bilayer Membranes.

The interactions between small molecules and lipid bilayers play a critical role in the function of cellular membranes. Understanding how a small molecule interacts with the lipid bilayer differently based on its charge reveals primordial mechanisms of transport across membranes and assists in the design of drug molecules that can penetrate cells. We have previously reported that tryptophan permeated through a phosphatidylcholine lipid bilayer membrane at a faster rate when it was positively charged (Trp+) than when negatively charged (Trp−), which corresponded to a lower potential of mean force (PMF) barrier determined through simulations. In this report, we demonstrate that Trp+ partitions into the lipid bilayer membrane to a greater degree than Trp– by interacting with the ester linkage of a phosphatidylcholine lipid, where it is stabilized by the electron withdrawing glycerol functional group. These results are in agreement with tryptophan’s known role as an anchor for transmembrane proteins, though the tendency for binding of a positively charged tryptophan is surprising. We discuss the implications of our results on the mechanisms of unassisted permeation and penetration of small molecules within and across lipid bilayer membranes based on molecular charge, shape, and molecular interactions within the bilayer structure.

Novel Insights into Concepts and Directionality of Maternal⁻Fetal Cholesterol Transfer across the Human Placenta.

Cholesterol is indispensable for cellular membrane composition and function. It is also a precursor for the synthesis of steroid hormones, which promote, among others, the maturation of fetal organs. A role of the ATP-binding-cassette-transporter-A1 (ABCA1) in the transport of maternal cholesterol to the fetus was suggested by transferring cholesterol to apolipoprotein-A-1 (apo-A1), but the directionality of the apoA-1/ABCA1-dependent cholesterol transport remains unclear. We isolated primary trophoblasts from term placentae to test the hypotheses that (1) apoA-1/ABCA1 dispatches cholesterol mainly towards the fetus to support fetal developmental maturation at term, and (2) differentiated syncytiotrophoblasts (STB) exert higher cholesterol transport activity than undifferentiated cytotrophoblasts (CTB). As experimental models, we used (1) trophoblast monolayers grown on Transwell® system consisting of apical (maternal-like) and basal (fetal-like) compartments, and (2) trophoblasts grown on conventional culture plates at CTB and STB stages. Surprisingly, apoA-1-mediated cholesterol efflux operated almost exclusively at the apical-maternal side, where ABCA1 was also localized by immunofluorescence. We found greater cholesterol efflux capacity in STB, which was increased by liver-X-receptor agonist treatment and decreased by ABCA1 inhibition. We conclude that at term the apoA-1/ABCA1 pathway is rather involved in cholesterol transport to the mother than in transfer to the fully developed fetus.

Effects of Maternal Vitamin D Supplementation on the Maternal and Infant Epigenome.

Mothers and infants are at high risk for inadequate vitamin D status. Mechanisms by which vitamin D may affect maternal and infant DNA methylation are poorly understood. This study quantified the effects of vitamin D3 supplementation on DNA methylation in pregnant and lactating women and their breastfed infants. In this randomized controlled pilot study, pregnant women received vitamin D3 400 international units (IU) (n = 6; control) or 3,800 IU (n = 7; intervention) daily from late second trimester through 4-6 weeks postpartum. Epigenome-wide DNA methylation was quantified in leukocytes collected from mothers at birth and mother-infant dyads at 4-6 weeks postpartum. At birth, intervention group mothers showed DNA methylation gain and loss at 76 and 89 cytosine-guanine (CpG) dinucleotides, respectively, compared to controls. Postpartum, methylation gain was noted at 200 and loss at 102 CpGs. Associated gene clusters showed strongest biologic relevance for cell migration/motility and cellular membrane function at birth and cadherin signaling and immune function at postpartum. Breastfed 4-6-week-old infants of intervention mothers showed DNA methylation gain and loss in 217 and 213 CpGs, respectively, compared to controls. Genes showing differential methylation mapped most strongly to collagen metabolic processes and regulation of apoptosis. Maternal vitamin D supplementation during pregnancy and lactation alters DNA methylation in mothers and breastfed infants. Additional work is needed to fully elucidate the short- and long-term biologic effects of vitamin D supplementation at varying doses, which could hold important implications for establishing clinical recommendations for prenatal and offspring health promotion.

An Amphipathic Helix Directs Cellular Membrane Curvature Sensing and Function of the BAR Domain Protein PICK1

BAR domains are dimeric protein modules that sense, induce, and stabilize lipid membrane curvature. Here, we show that membrane curvature sensing (MCS) directs cellular localization and function of the BAR domain protein PICK1. In PICK1, and the homologous proteins ICA69 and arfaptin2, we identify an amphipathic helix N-terminal to the BAR domain that mediates MCS. Mutational disruption of the helix in PICK1 impaired MCS without affecting membrane binding per se. In insulin-producing INS-1E cells, super-resolution microscopy revealed that disruption of the helix selectively compromised PICK1 density on insulin granules of high curvature during their maturation. This was accompanied by reduced hormone storage in the INS-1E cells. In Drosophila, disruption of the helix compromised growth regulation. By demonstrating size-dependent binding on insulin granules, our finding highlights the function of MCS for BAR domain proteins in a biological context distinct from their function, e.g., at the plasma membrane during endocytosis.

Tissue-specific molecular and cellular toxicity of Pb in the oyster (Crassostrea gigas): mRNA expression and physiological studies

Lead (Pb) is one of the ubiquitous and toxic elements in aquatic environment. In oysters, gills and digestive glands are the main target organs for Pb-induced toxicity, but there is limited information on the molecular mechanisms underlying its toxicity. The present study investigated the Pb-induced toxicity mechanisms in the Pacific oyster (Crassostrea gigas) based on transcriptome, phenotypic anchoring, and validation of targeted gene expression. Gene ontology and pathway enrichment analyses revealed the differential Pb toxicity mechanisms in the tissues. In the gills, Pb disturbed the protein metabolism, with the most significant enrichment of the “protein processing in endoplasmic reticulum” pathway. The main mechanism comprised of a Pb-stimulated calcium (Ca2+) increase by the up-regulation of transporter-Ca-ATPase expression. The disturbed Ca2+ homeostasis then further induced high expressions of endoplasmic reticulum (ER) chaperones, leading to ER stress in the oysters. Unfolded proteins induced ER associated degradation (ERAD), thereby preventing the accumulation of folding-incompetent glycoproteins. However, Pb mainly induced oxidative reduction reactions in the digestive gland with high accumulation of lipid peroxidation products and high expression of antioxidant enzymes. Further, Pb induced fatty acid β-oxidation and CYP450 catalyzed ω-oxidation due to increased metabolic expenditure for detoxification. The increased content of arachidonic acid indicated that Pb exposure might alter unsaturated fatty acid composition and disturb cellular membrane functions. Taken together, our results provided a new insight into the molecular mechanisms underlying Pb toxicity in oysters.

Nanopore-Confined Bilayers: A Model of Biomembranes with Defined Curvature and a Tool for Oriented Sample Magnetic Resonance

It is well established that biological function of cellular membranes is determined by both membrane proteins and the membrane biochemical composition. Moreover, the membrane shape/local curvature may also play a role in modulating membrane properties and the proteins’ function. While specific biomembrane compositions are more readily replicated in biophysical bench experiments, formation of bilayers of specific shapes and curvature represents a more challenging task. Macroscopic alignment of such membranes – an important prerequisite of high resolution magnetic resonance and other studies – is even more difficult especially over a broad range of experimental conditions such pH, ionic strength, and temperature. Here we describe methods for forming self-assembled lipid nanotubular bilayers inside cylindrical nanopores composed of anodic aluminum oxide (AAO). Such hybrid nanostructures, named lipid nanotube arrays, represent a new type of substrate-supported and macroscopically-aligned lipid bilayers of defined curvature that have many attractive features for both membrane biophysics and structure-function protein studies by spectroscopic techniques. Optical properties of AAO allow for assessing the integrity of membrane protein complexes by UV-vis while high density of the deposited lipids and proteins enable examination by other biophysical methods, including DSC, QCM, and magnetic resonance. The latter studies have shown that the individual lipids in such nanopore-confined structures maintain fast uniaxial diffusion and a high degree of macroscopic alignment. The macroscopic alignment enables detailed studies of effects of lipid composition on structure of integral membrane proteins by solid state oriented sample NMR, EPR and DEER. Accessibility of either both or mainly inner leaflet of the nanotubular bilayers to water-soluble species provides for studies of protein, peptides, and drug binding. Supported by DE-FG02-02ER15354 to AIS.

СТВОРЕННЯ ФОРМУЛИ СКРИНІНГУ ГІПОВІТАМІНОЗУ РЕТИНОЛУ ПРИ ХРОНІЧНОМУ ПАНКРЕАТИТІ В АМБУЛАТОРНИХ УМОВАХ

У хворих на хронічний панкреатит часто розвивається гіповітаміноз ретинолу, який бере участь в окисно-відновних процесах, регуляції синтезу білків, сприяє нормальному обміну речовин, функції клітинних і субклітинних мембран, відіграє важливу роль у формуванні кісток і зубів, а також жирових відкладень. Метою роботи було створення формулу прогнозування дефіциту ретинолу на основі встановленими нами предикторних факторів: рівень холестерину, об’єм м’язів плеча, УЗ-критерії та критерії копрограми. Для персоніфікованого прогнозування дефіциту ретинолу в хворих на хронічний панкреатит застосували встановлені попередніми дослідженнями фактори формування і глибини дефіциту ретинолу як показового прояву полінутрієнтної недостатності. Ці показники є доступними для визначення у практиці лікарів первинної медичної допомоги. Запропонована математична модель прогнозування дефіциту ретинолу в хворих на хронічний панкреатит створена на основі доступних для визначення характеристик хронічний панкреатит. Формулу можна використовувати для виділення груп ризику за виникненням зниження рівня вітаміну А серед хворих на ХП для своєчасного проведення профілактичних і лікувальних заходів по корекції втрати ретинолу, що особливо актуально для рівня первинної медичної допомоги.

The dynamics of magnetic nanoparticles exposed to non-heating alternating magnetic field in biochemical applications: theoretical study

In the past decade, magneto-nanomechanical approach to biochemical systems stimulation has been studied intensively. This method involves macromolecule structure local deformation via mechanical actuation of functionalized magnetic nanoparticles (f-MNPs) by non-heating low frequency (LF) alternating magnetic field (AMF). Specificity at cellular or molecular level and spatial locality in nanometer scale are its key advantages as compared to magnetic fluid hyperthermia. However, current experimental studies have weak theoretical basis. Several models of magneto-nanomechanical actuation of macromolecules and cells in non-heating uniform LF AMF are presented in the article. Single core-shell spherical, rod-like, and Janus MNPs, as well as dimers consisting of two f-MNPs with macromolecules immobilized on their surfaces are considered. AMF-induced rotational oscillations of MNPs can affect properties and functioning of macromolecules or cellular membranes attached to them via periodic deformations in nanometer scale. This could be widely used in therapy, in particular for targeted drug delivery, controlled drug release, and cancer cell killing. An aggregate composed of MNPs can affect associated macromolecules by force up to several hundreds of piconewton in the case of MNPs of tens of nanometers in diameter and LF AMF below 1 T. AMF parameters and MNP design requirements for effective in vitro and in vivo magneto-nanomechanical treatment are presented.

Morphological Effect of the New Antifungal Agent ME1111 on Hyphal Growth of Trichophyton mentagrophytes, Determined by Scanning and Transmission Electron Microscopy.

The effects of ME1111, a novel antifungal agent, on the hyphal morphology and ultrastructure of Trichophyton mentagrophytes were investigated by using scanning and transmission electron microscopy. Structural changes, such as pit formation and/or depression of the cell surface, and degeneration of intracellular organelles and plasmolysis were observed after treatment with ME1111. Our results suggest that the inhibition of energy production by ME1111 affects the integrity and function of cellular membranes, leading to fungal cell death.

Fungal and Prokaryotic Activities in the Marine Subsurface Biosphere at Peru Margin and Canterbury Basin Inferred from RNA-Based Analyses and Microscopy.

The deep sedimentary biosphere, extending 100s of meters below the seafloor harbors unexpected diversity of Bacteria, Archaea, and microbial eukaryotes. Far less is known about microbial eukaryotes in subsurface habitats, albeit several studies have indicated that fungi dominate microbial eukaryotic communities and fungal molecular signatures (of both yeasts and filamentous forms) have been detected in samples as deep as 1740 mbsf. Here, we compare and contrast fungal ribosomal RNA gene signatures and whole community metatranscriptomes present in sediment core samples from 6 and 95 mbsf from Peru Margin site 1229A and from samples from 12 and 345 mbsf from Canterbury Basin site U1352. The metatranscriptome analyses reveal higher relative expression of amino acid and peptide transporters in the less nutrient rich Canterbury Basin sediments compared to the nutrient rich Peru Margin, and higher expression of motility genes in the Peru Margin samples. Higher expression of genes associated with metals transporters and antibiotic resistance and production was detected in Canterbury Basin sediments. A poly-A focused metatranscriptome produced for the Canterbury Basin sample from 345 mbsf provides further evidence for active fungal communities in the subsurface in the form of fungal-associated transcripts for metabolic and cellular processes, cell and membrane functions, and catalytic activities. Fungal communities at comparable depths at the two geographically separated locations appear dominated by distinct taxa. Differences in taxonomic composition and expression of genes associated with particular metabolic activities may be a function of sediment organic content as well as oceanic province. Microscopic analysis of Canterbury Basin sediment samples from 4 and 403 mbsf produced visualizations of septate fungal filaments, branching fungi, conidiogenesis, and spores. These images provide another important line of evidence supporting the occurrence and activity of fungi in the deep subseafloor biosphere.

Cholesterol Effect on the Elastic Properties of Unsaturated Lipid Bilayers

Cholesterol is one of the major determinants of the structure and function of cellular membranes. Interaction of cholesterol with lipid molecules affects diverse membrane properties, such as elasticity, fluidity, dynamics of membrane proteins and lipids. Despite decades of studies, the effects of cholesterol on lipid bilayer are not fully understood, especially in the context of liquid disordered bilayers omnipresent in cellular membranes. We demonstrate here that cholesterol can effectively interact with unsaturated lipid molecules and, via these interactions, regulate bending stiffness of lipid bilayers made of unsaturated lipids of different molecular geometry. Using highly curved lipid nanotubes to measure the mean curvature bending modulus, we found that cholesterol could produce up to 5-fold increase of the stiffness of membranes containing lyso-lipids. Cholesterol also inhibited curvature-driven redistribution of lyso-lipids, indicating formation of lyso-lipid/cholesterol quasi-dimers. The magnitude of the membrane stiffening depended on lipid composition, with both polar heads and fatty acid tails modulating the cholesterol effect. Addition of chaotropic agents affecting hydrogen bonding dramatically decreased the cholesterol-induced membrane stiffening, indicating that strong hydrogen bonding between cholesterol and neighbor lipids contribute to bending rigidity. We discuss how these molecular interactions could affect the cholesterol orientation in the membrane and augment its bending rigidity.