Preferential electrostatic interactions of phosphatidic acid with arginines.

Phosphatidic acid (PA) is an anionic lipid that preferentially interacts with proteins in a diverse set of cellular processes such as transport, apoptosis, and neurotransmission. One such interaction is that of the PA lipids with the proteins of voltage-sensitive ion channels. In comparison to several other similarly charged anionic lipids, PA lipids exhibit much stronger interactions. Intrigued and motivated by this finding, we sought out to gain deeper understanding into the electrostatic interactions of anionic lipids with charged proteins. Using the voltage sensor domain (VSD) of the KvAP channel as a model system, we performed long-timescale atomistic simulations to analyze the interactions of POPA, POPG, and POPI lipids with arginines (ARGs). Our simulations reveal two mechanisms. First, POPA is able to interact not only with surface ARGs but is able to snorkel and interact with a buried arginine. POPG and POPI lipids on the other hand show weak interactions even with both the surface and buried ARGs. Second, deprotonated POPA with -2 charge is able to break the salt-bridge connection between VSD protein segments and establish its own electrostatic bond with the ARG. Based on these findings, we propose a headgroup size hypothesis for preferential solvation of proteins by charged lipids. These findings may be valuable in understanding how PA lipids could be modulating kinematics of transmembrane proteins in cellular membranes.

A lateral electric field inhibits gel-to-fluid transition in lipid bilayers.

We report evidence of lateral electric field-induced changes in the phase transition temperatures of lipid bilayers. Our atomic scale molecular dynamics simulations show that a lateral electric field increases the melting temperatures of DPPC, POPC and POPE bilayers. Remarkably, these shifts in the melting temperatures are only induced by lateral electric fields, and not normal electric fields. This mechanism could provide new mechanistic insights into lipid-lipid and lipid-protein interactions in the presence of endogenous and exogenous electric fields.

Mechanics of nuclear membranes.

Cellular nuclei are bound by two uniformly separated lipid membranes that are fused with each other at numerous donut-shaped pores. These membranes are structurally supported by an array of distinct proteins with distinct mechanical functions. As a result, the nuclear envelope possesses unique mechanical properties, which enables it to resist cytoskeletal forces. Here, we review studies that are beginning to provide quantitative insights into nuclear membrane mechanics. We discuss how the mechanical properties of the fused nuclear membranes mediate their response to mechanical forces exerted on the nucleus and how structural reinforcement by different nuclear proteins protects the nuclear membranes against rupture. We also highlight some open questions in nuclear envelope mechanics, and discuss their relevance in the context of health and disease.

Border Medicine: The Pediatric Cardiology Perspective.

Pediatric cardiology and cardiovascular surgery have witnessed significant advancements over the last two decades. In spite of this progress, congenital heart disease (CHD) still remains as one of the major causes of death in infants and young children in the United States. Many patient-related and patient-independent factors influence the outcomes in patients with CHD, one of which is the geographical location. In the US-Mexico border, management and outcomes of patients with CHD are further complicated by additional problems stemming from complex interplay between two different health systems, and socioeconomic disparities. In this article, the authors evaluate the various interplaying factors and describe the difficulties facing the practicing pediatric cardiologists in a US-Mexico border city.

Ultradonut topology of the nuclear envelope.

The nuclear envelope is a unique topological structure formed by lipid membranes in eukaryotic cells. Unlike other membrane structures, the nuclear envelope comprises two concentric membrane shells fused at numerous sites with toroid-shaped pores that impart a "geometric" genus on the order of thousands. Despite the intriguing architecture and vital biological functions of the nuclear membranes, how they achieve and maintain such a unique arrangement remains unknown. Here, we used the theory of elasticity and differential geometry to analyze the equilibrium shape and stability of this structure. Our results show that modest in- and out-of-plane stresses present in the membranes not only can define the pore geometry, but also provide a mechanism for destabilizing membranes beyond a critical size and set the stage for the formation of new pores. Our results suggest a mechanism wherein nanoscale buckling instabilities can define the global topology of a nuclear envelope-like structure.

Endocytic proteins drive vesicle growth via instability in high membrane tension environment.

Clathrin-mediated endocytosis (CME) is a key pathway for transporting cargo into cells via membrane vesicles; it plays an integral role in nutrient import, signal transduction, neurotransmission, and cellular entry of pathogens and drug-carrying nanoparticles. Because CME entails substantial local remodeling of the plasma membrane, the presence of membrane tension offers resistance to bending and hence, vesicle formation. Experiments show that in such high-tension conditions, actin dynamics is required to carry out CME successfully. In this study, we build on these pioneering experimental studies to provide fundamental mechanistic insights into the roles of two key endocytic proteins-namely, actin and BAR proteins-in driving vesicle formation in high membrane tension environment. Our study reveals an actin force-induced "snap-through instability" that triggers a rapid shape transition from a shallow invagination to a highly invaginated tubular structure. We show that the association of BAR proteins stabilizes vesicles and induces a milder instability. In addition, we present a rather counterintuitive role of BAR depolymerization in regulating the shape evolution of vesicles. We show that the dissociation of BAR proteins, supported by actin-BAR synergy, leads to considerable elongation and squeezing of vesicles. Going beyond the membrane geometry, we put forth a stress-based perspective for the onset of vesicle scission and predict the shapes and composition of detached vesicles. We present the snap-through transition and the high in-plane stress as possible explanations for the intriguing direct transformation of broad and shallow invaginations into detached vesicles in BAR mutant yeast cells.

Clathrin polymerization exhibits high mechano-geometric sensitivity.

How tension modulates cellular transport has become a topic of interest in the recent past. However, the effect of tension on clathrin assembly and vesicle growth remains less understood. Here, we use the classical Helfrich theory to predict the energetic cost that clathrin is required to pay to remodel the membrane at different stages of vesicle formation. Our study reveals that this energetic cost is highly sensitive to not only the tension in the membrane but also to the instantaneous geometry of the membrane during shape evolution. Our study predicts a sharp reduction in clathrin coat size in the intermediate tension regime (0.01-0.1 mN m-1). Remarkably, the natural propensity of the membrane to undergo bending beyond the Ω shape causes a significant decrease in the energy needed from clathrin to drive vesicle growth. Our studies in mammalian cells confirm a reduction in clathrin coat size in an increased tension environment. In addition, our findings suggest that the two apparently distinct clathrin assembly modes, namely coated pits and coated plaques, observed in experimental investigations might be a consequence of varied tensions in the plasma membrane. Overall, the mechano-geometric sensitivity revealed in this study might also be at play during the polymerization of other membrane remodeling proteins.

Cross-sectional serologic assessment of immunity to poliovirus infection in high-risk areas of northern India.

INTRODUCTION: The objectives of this survey were to assess the seroprevalence of antibodies to poliovirus types 1 and 3 and the impact of bivalent (types 1 and 3) oral poliovirus vaccine (bOPV) use in immunization campaigns in northern India. METHODS: In August 2010, a 2-stage stratified cluster sampling method identified infants aged 6-7 months in high-risk blocks for wild poliovirus infection. Vaccination history, weight and length, and serum were collected to test for neutralizing antibodies to poliovirus types 1, 2, and 3. RESULTS: Seroprevalences of antibodies to poliovirus types 1, 2, and 3 were 98% (95% confidence interval [CI], 97%-99%), 66% (95% CI, 62%-69%), and 77% (95% CI, 75%-79%), respectively, among 664 infants from Bihar and 616 infants from Uttar Pradesh. Infants had received a median of 3 bOPV doses and 2 monovalent type 1 OPV (mOPV1) doses through campaigns and 3 trivalent OPV (tOPV) doses through routine immunization. Among subjects with 0 tOPV doses, the seroprevalences of antibodies to type 3 were 50%, 77%, and 82% after 2, 3, and 4 bOPV doses, respectively. In multivariable analysis, malnutrition was associated with a lower seroprevalence of type 3 antibodies. CONCLUSIONS: This study confirmed that replacing mOPV1 with bOPV in campaigns was successful in maintaining very high population immunity to type 1 poliovirus and substantially decreasing the immunity gap to type 3 poliovirus.

Peripheral giant cell granuloma: a case report.

Peripheral giant cell granuloma (PGCG) is a benign inflammatory hyperplastic lesion of unknown etiology that occurs in the gingiva or alveolar ridge. It normally manifests as a soft tissue nodule, purplish-red in color, consisting of multinucleated giant cells in a background of mononuclear stromal cells and extravasated red blood cells. This article presents a case of a 75-year-old man whose chief complaint was painless swelling in the posterior mandibular region. Intraoral examination revealed a swelling that was red, firm, and sessile, with a smooth surface texture. An orthopantomogram revealed a well-demarcated radiolucency extending from the distal aspect of the mandibular canine to the mesial aspect of the mandibular first molar. Cone beam computed tomography showed the features that suggested a soft tissue lesion causing cupping resorption of the mandible. An excisional biopsy was performed under local anaesthesia, and the tissue was examined histopathologically. The lesion was diagnosed as PGCG after thorough clinical, radiologic, and histopathologic examinations.

Analysis of choroidal structure and vascularity indices with image binarization of swept source optical coherence tomography images: A prospective study of 460 eyes.

PURPOSE: To evaluate choroidal vascularity index (CVI) among normal subjects using image binarization of swept source optical coherence tomography (SS-OCT). MATERIALS AND METHODS: Four hundred and sixty eyes of 230 normal participants were included. Total circumscribed choroidal area, luminal area, stromal area (SA), and CVI were derived from SS-OCT scans using open-source software (ImageJ) with the modified Niblack method. Both CVI and subfoveal choroidal thickness (SFCT) were correlated with age, refractive error, intraocular pressure, and mean ocular perfusion pressure (MOPP) using mixed linear model analysis. Pearson's correlation coefficient was used to determine the relationship between age and each dependent factor. Analyses were performed using the SPSS software version 20.0 (IBM Corp., Armonk USA) and statistical significance was tested at 5%. RESULTS: The mean age was 42.1 (±17.6) years. Mean SFCT was 307 ± 79 µm. Mean CVI was 66.80 (±3.8)%. There was statistically significant positive correlation between CVI and increasing age (r = 0.259, P < 0.0001) and statistically significant negative correlation between SFCT and age (r = -0.361, P < 0.0001). There was positive linear correlation between refractive error and CVI (r = 0.220, P < 0.0001) and negative correlation between SFCT and refractive error. There was no significant effect of MOPP on both CVI (P = 0.07) and SFCT (P = 0.7). CONCLUSION: CVI and SFCT are significantly correlated with age and refractive error in normal Indian eyes.

Lipid bilayer mechanics in a pipette with glass-bilayer adhesion.

Electrophysiology is a central tool for measuring how different driving forces (e.g., ligand concentration, transmembrane voltage, or lateral tension) cause a channel protein to gate. Upon formation of the high resistance seal between a lipid bilayer and a glass pipette, the so-called "giga-seal", channel activity can be recorded electrically. In this article, we explore the implications of giga-seal formation on the mechanical state of a lipid bilayer patch. We use a mechanical model for the free energy of bilayer geometry in the presence of glass-bilayer adhesion to draw three potentially important conclusions. First, we use our adhesion model to derive an explicit relationship between applied pressure and patch shape that is consistent with the Laplace-Young Law, giving an alternative method of calculating patch tension under pressure. With knowledge of the adhesion constant, which we find to be in the range ∼0.4-4 mN/m, and the pipette size, one can precisely calculate the patch tension as a function of pressure, without the difficultly of obtaining an optical measurement of the bilayer radius of curvature. Second, we use data from previous electrophysiological experiments to show that over a wide range of lipids, the resting tension on a electrophysiological patch is highly variable and can be 10-100 times higher than estimates of the tension in a typical cell membrane. This suggests that electrophysiological experiments may be systematically altering channel-gating characteristics and querying the channels under conditions that are not the same as their physiological counterparts. Third, we show that reversible adhesion leads to a predictable change in the population response of gating channels in a bilayer patch.

Intravitreal Ozurdex has no short term influence on choroidal thickness and vascularity index in eyes with diabetic macular edema: A pilot study.

AIM: To analyze choroidal parameters in eyes with diabetic macular edema (DME) treated with intravitreal Ozurdex. PATIENTS AND METHODS: Twenty eyes of 14 patients were included in this prospective study. Optical coherence tomography images were obtained before and 8-10 weeks after intravitreal Ozurdex injection; binarized and subfoveal choroidal thickness (SFCT) and choroidal vascularity index (CVI) were calculated. RESULTS: Mean SFCT (treatment naïve; 242.22 ± 32.87 reduced to 218.10 ± 22.10, P = 0.158 and previously treated; 330.4 ± 56.72 reduced to 328.93 ± 50.55, P = 0.833) and mean CVI (treatment naïve; 0.64 ± 0.03 changed to 0.65 ± 0.04, P = 0.583 and previously treated; 0.65 ± 0.05 reduced to 0.64 ± 0.03, P = 0.208) showed no significant change. CONCLUSION: Intravitreal Ozurdex showed no significant effects on SFCT and CVI in eyes with DME over short term. Larger studies with longer follow-up may allow a better understanding.

Efficient Thin Polymer Coating as a Selective Thermal Emitter for Passive Daytime Radiative Cooling.

Radiative cooling to subambient temperatures can be efficiently achieved through spectrally selective emission, which until now has only been realized by using complex nanoengineered structures. Here, a simple dip-coated planar polymer emitter derived from polysilazane, which exhibits strong selective emissivity in the atmospheric transparency window of 8-13 µm, is demonstrated. The 5 µm thin silicon oxycarbonitride coating has an emissivity of 0.86 in this spectral range because of alignment of the frequencies of bond vibrations arising from the polymer. Furthermore, atmospheric heat absorption is suppressed due to its low emissivity outside the atmospheric transparency window. The reported structure with the highly transparent polymer and underlying silver mirror reflects 97% of the incoming solar irradiation. A temperature reduction of 6.8 °C below ambient temperature was achieved by the structure under direct sunlight, yielding a cooling power of 93.7 W m-2. The structural simplicity, durability, easy applicability, and high selectivity make polysilazane a unique emitter for efficient prospective passive daytime radiative cooling structures.

Geometry of the nuclear envelope determines its flexural stiffness.

During closed mitosis in fission yeast, growing microtubules push onto the nuclear envelope to deform it, which results in fission into two daughter nuclei. The resistance of the envelope to bending, quantified by the flexural stiffness, helps determine the microtubule-dependent nuclear shape transformations. Computational models of envelope mechanics have assumed values of the flexural stiffness of the envelope based on simple scaling arguments. The validity of these estimates is in doubt, however, owing to the complex structure of the nuclear envelope. Here, we performed computational analysis of the bending of the nuclear envelope under applied force using a model that accounts for envelope geometry. Our calculations show that the effective bending modulus of the nuclear envelope is an order of magnitude larger than a single membrane and approximately five times greater than the nuclear lamina. This large bending modulus is in part due to the 45 nm separation between the two membranes, which supports larger bending moments in the structure. Further, the effective bending modulus is highly sensitive to the geometry of the nuclear envelope, ranging from twofold to an order magnitude larger than the corresponding single membrane. These results suggest that spatial variations in geometry and mechanical environment of the envelope may cause a spatial distribution of flexural stiffness in the same nucleus. Overall, our calculations support the possibility that the nuclear envelope may balance significant mechanical stresses in yeast and in cells from higher organisms.

Complimentary action of structured and unstructured domains of epsin supports clathrin-mediated endocytosis at high tension.

Membrane tension plays an inhibitory role in clathrin-mediated endocytosis (CME) by impeding the transition of flat plasma membrane to hemispherical clathrin-coated structures (CCSs). Membrane tension also impedes the transition of hemispherical domes to omega-shaped CCSs. However, CME is not completely halted in cells under high tension conditions. Here we find that epsin, a membrane bending protein which inserts its N-terminus H0 helix into lipid bilayer, supports flat-to-dome transition of a CCS and stabilizes its curvature at high tension. This discovery is supported by molecular dynamic simulation of the epsin N-terminal homology (ENTH) domain that becomes more structured when embedded in a lipid bilayer. In addition, epsin has an intrinsically disordered protein (IDP) C-terminus domain which induces membrane curvature via steric repulsion. Insertion of H0 helix into lipid bilayer is not sufficient for stable epsin recruitment. Epsin's binding to adaptor protein 2 and clathrin is critical for epsin's association with CCSs under high tension conditions, supporting the importance of multivalent interactions in CCSs. Together, our results support a model where the ENTH and unstructured IDP region of epsin have complementary roles to ensure CME initiation and CCS maturation are unimpeded under high tension environments.

Exploring the links between lipid geometry and mitochondrial fission: Emerging concepts.

Mitochondria, the double membrane-walled powerhouses of the eukaryotic cell, are also the seats of synthesis of two critical yet prevalent nonbilayer-prone phospholipids, namely phosphatidylethanolamine (PE) and cardiolipin (CL). Besides their established biochemical roles in the regulation of partner protein function, PE and CL are also key protagonists in the biophysics of mitochondrial membrane remodeling and dynamics. In this review, we address lipid geometry and behavior at the single-molecule level as well as their intimate coupling to whole organelle morphology and remodeling during the concerted events of mitochondrial fission. We present evidence from recent experimental measurements ably supported and validated by computational modeling studies to support our notion that conical lipids play a catalytic as well as a structural role in mitochondrial fission.

Geometric instability catalyzes mitochondrial fission.

The mitochondrial membrane undergoes extreme remodeling during fission. While a few membrane-squeezing proteins are recognized as the key drivers of fission, there is a growing body of evidence that strongly suggests that conical lipids play a critical role in regulating mitochondrial morphology and fission. However, the mechanisms by which proteins and lipids cooperate to execute fission have not been quantitatively investigated. Here, we computationally model the squeezing of the largely tubular mitochondrion and show that proteins and conical lipids can act synergistically to trigger buckling instability and achieve extreme constriction. More remarkably, the study reveals that the conical lipids can act with different fission proteins to induce hierarchical instabilities and create increasingly narrow and stable constrictions. We reason that this geometric plasticity imparts significant robustness to the fission reaction by arresting the elastic tendency of the membrane to rebound during protein polymerization and depolymerization cycles. Our in vitro study validates protein-lipid cooperativity in constricting membrane tubules. Overall, our work presents a general mechanism for achieving drastic topological remodeling in cellular membranes.

Local, transient tensile stress on the nuclear membrane causes membrane rupture.

Cancer cell migration through narrow constrictions generates compressive stresses on the nucleus that deform it and cause rupture of nuclear membranes. Nuclear membrane rupture allows uncontrolled exchange between nuclear and cytoplasmic contents. Local tensile stresses can also cause nuclear deformations, but whether such deformations are accompanied by nuclear membrane rupture is unknown. Here we used a direct force probe to locally deform the nucleus by applying a transient tensile stress to the nuclear membrane. We found that a transient (∼0.2 s) deformation (∼1% projected area strain) in normal mammary epithelial cells (MCF-10A cells) was sufficient to cause rupture of the nuclear membrane. Nuclear membrane rupture scaled with the magnitude of nuclear deformation and the magnitude of applied tensile stress. Comparison of diffusive fluxes of nuclear probes between wild-type and lamin-depleted MCF-10A cells revealed that lamin A/C, but not lamin B2, protects the nuclear membranes against rupture from tensile stress. Our results suggest that transient nuclear deformations typically caused by local tensile stresses are sufficient to cause nuclear membrane rupture.

Vesicle adhesion reveals novel universal relationships for biophysical characterization.

Adhesion plays an integral role in diverse biological functions ranging from cellular transport to tissue development. Estimation of adhesion strength, therefore, becomes important to gain biophysical insight into these phenomena. In this study, we use curvature elasticity to present non-intuitive, yet remarkably simple, universal relationships that capture vesicle-substrate interactions. These relationships not only provide efficient strategies to tease out adhesion energy of biological molecules but can also be used to characterize the physical properties of elastic biomimetic nanoparticles. We validate the modeling predictions with experimental data from two previous studies.

An unresolved LINC in the nuclear envelope.

The nuclear envelope segregates the nucleoplasm from the cytoplasm and is a key feature of eukaryotic cells. Nuclear envelope architecture is comprised of two concentric membrane shells which fuse at multiple sites and yet maintain a uniform separation of 30-50 nm over the rest of the membrane. Studies have revealed the roles for numerous nuclear proteins in forming and maintaining the architecture of the nuclear envelope. However, there is a lack of consensus on the fundamental forces and physical mechanisms that establish the geometry. The objective of this review is to discuss recent findings in the context of membrane mechanics in an effort to define open questions and possible answers.