Modulation of Phospholipase A2 Membrane Activity by Anti-inflammatory Drugs.

The phospholipase A2 (PLA2) superfamily consists of lipolytic enzymes that hydrolyze specific cell membrane phospholipids and have long been considered a central hub of biosynthetic pathways, where their lipid metabolites exert a variety of physiological roles. A misregulated PLA2 activity is associated with mainly inflammatory-derived pathologies and thus has shown relevant therapeutic potential. Many natural and synthetic anti-inflammatory drugs (AIDs) have been proposed as direct modulators of PLA2 activity. However, despite the specific chemical properties that these drugs share in common, little is known about the indirect modulation able to finely tune membrane structural changes at the precise lipid-binding site. Here, we use a novel experimental strategy based on differential scanning calorimetry to systematically study the structural properties of lipid membrane systems during PLA2 cleavage and under the influence of several AIDs. For a better understanding of the AIDs-membrane interaction, we present a comprehensive and comparative set of molecular dynamics (MD) simulations. Our thermodynamic results clearly demonstrate that PLA2 cleavage is hindered by those AIDs that significantly reduce the lipid membrane cooperativity, while the rest of the AIDs oppositely tend to catalyze PLA2 activity to different extents. On the other hand, our MD simulations support experimental results by providing atomistic details on the binding, insertion, and dynamics of each AID on a pure lipid system; the drug efficacy to impact membrane cooperativity is related to the lipid order perturbation. This work suggests a membrane-based mechanism of action for diverse AIDs against PLA2 activity and provides relevant clues that must be considered in its modulation.

Fluidized or not fluidized? Biophysical characterization of biohybrid lipid/protein/polymer liposomes and their interaction with tetracaine.

BACKGROUND: Nanomedicine and the pharmaceutical industry demand the investigation of new biomaterials to improve drug therapies. Combinations of lipids, proteins, and polymers represent innovative platforms for drug delivery. However, little is known about the interactions between such compounds and this knowledge is key to prepare successful drug delivery systems. METHODS: Biophysical properties of biohybrid vesicles (BhVs) composed of phospholipids, proteins, and amphiphilic block copolymers, assembled without using organic solvents, were investigated by differential scanning calorimetry and dynamic light scattering. We studied four biohybrid systems; two of them included the effect of incorporating tetracaine. Thermal changes of phospholipids and proteins when interacting with the amphiphilic block copolymers and tetracaine were analyzed. RESULTS: Lysozyme and the copolymers adsorb onto the lipid bilayer modifying the phase transition temperature, enthalpy change, and cooperativity. Dynamic light scattering investigations revealed relevant changes in the size and zeta potential of the BhVs. Interestingly, tetracaine, a membrane-active drug, can fluidize or rigidize BhVs. CONCLUSIONS: We conclude that positively charged regions of lysozyme are necessary to incorporate the block copolymer chains into the lipid membrane, turning the bilayer into a more rigid system. Electrostatic properties and the hydrophilic-lipophilic balance are determinant for the stability of biohybrid membranes. GENERAL SIGNIFICANCE: This investigation provides fundamental information associated with the performance of biohybrid drug delivery systems and can be of practical significance for designing more efficient drug nanocarriers.

Xenon and Krypton Dissolved in Water Form Nanoblobs: No Evidence for Nanobubbles.

We demonstrate, experimentally and by molecular dynamics simulations, that krypton and xenon form nanostructured water-gas domains. High pressure was applied to force the inert gases to dissolve in water following Henry's law, then the liquid was depressurized, centrifuged, and inspected by dynamic light scattering. The observed objects have similar sizes and electrical properties to nanobubbles, but we found that they have fairly neutral buoyancy even at high gravitational fields. We posit that the formed nano objects are not bubbles but blobs, unique structures conceived as clathrate-hydrate precursors, thus resolving the so-called Laplace pressure bubble catastrophe.

Características clínicas de la valoración fisioterapéutica en pacientes con síndrome subacromial / Clinical characteristics of physiotherapeutic assessment in patients with shoulder impingement syndrome

Antecedentes y objetivo: Existe una gran variabilidad en la situación clínica de los pacientes diagnosticados de síndrome subacromial que acuden a tratamiento de fisioterapia, cuyos resultados terapéuticos son cuestionados. El objetivo de este estudio es obtener datos epidemiológicos que permitan elaborar un protocolo de evaluación y un tratamiento más eficaz. Material y métodos: Estudio observacional descriptivo transversal realizado en 19 pacientes con síndrome subacromial, en el Hospital Universitario Central de Asturias y en Atención Primaria del área IV. Se evalúan aspectos posturales, movilidad escapular y del aparato motor del hombro (Constant-Murley Score) junto con la exploración de puntos gatillo miofasciales activos. Resultados: Los sujetos de la muestra presentaban limitación funcional del hombro afecto con repercusión en las actividades de la vida diaria y dolor asociado (media de 51,2/ 75 puntos del Constant-Murley Score). El 40% tenía limitados todos los movimientos de la escápula y presentaban una actitud postural en flexión con posición antiálgica. Los puntos gatillos explorados estaban presentes en más del 70% de los individuos, con el punto gatillo miofascial del músculo infraespinoso como el más prevalente (94,7%). Conclusiones: Los resultados señalan que la evaluación de los puntos gatillo miofasciales en la musculatura del aparato motor del hombro, la postura y el movimiento escapular deben ser considerados antes de incluir a los pacientes con síndrome subacromial en los protocolos de tratamiento fisioterapéutico.

Background and objective: Often the therapeutic results of patients diagnosed with subacromial syndrome who attend physiotherapy treatment are questioned. The aim of this study is to obtain epidemiological data that will allow a more effective evaluation and thus an improved treatment protocol. Material -and methods: A cross-sectional descriptive observational study was carried out with 19 subacromial syndrome patients at a primary care medical centre of the Central University Hospital of Asturias (area IV). Postural aspects, scapular mobility, and shoulder apparatus (Constant-Murley Score) as well as their myofascial trigger points were explored and evaluated. Results: The subjects of the sample presented functional limitations of the shoulder affecting their activities of daily living and associated pain (51.7/75 Constant-Murley Score). Some 40% of the patients had limits to all their scapular movements and antalgic flexion positions and more than 70% presented trigger points, the infraspinatus muscle being the most prevalent (94.7%). Conclusions: Results suggest that myofascial trigger points in the shoulder apparatus, posture and scapular movement should be evaluated before physiotherapy treatment protocols for patients with subacromial syndrome are designed.

Phase transitions of liposomes: when light meets heat.

Phase transitions of liposomes are normally studied by differential scanning calorimetry. A suspension of liposomes is subjected to an increase (decrease) of temperature and when heat is absorbed (released), the liposomes transit from a gel (liquid) to a liquid (gel) phase. This endothermic (exothermic) process takes place at a temperature called the melting temperatureTm, which is distinctive of the type of lipids forming the vesicles. The vesicles, though, also modify their size in the transition. Indeed, the thickness of the membranes decreases (increases) because carbon tails misalign (align). Concomitant with the modifications in the membrane thickness, the diameter (D) of the liposomes changes too. Therefore, when they are inspected by light, the scattered signal carries information from such dilatation (contraction) process. We performed careful experiments using dynamic light scattering as a function of temperature to detect the size changes of different liposomes. Gaussian fits of the derivatives of theDvsTcurves coincide within 1% with thermograms, which hints to the possibility of performing thermodynamic studies of lipid systems employing light.

On the evolution of the exclusion zone produced by hydrophilic surfaces: A contracted description.

After exciting scientific debates about its nature, the development of the exclusion zone, a region near hydrophilic surfaces from which charged colloidal particles are strongly expelled, has been finally traced back to the diffusiophoresis produced by unbalanced ion gradients. This was done by numerically solving the coupled Poisson equation for electrostatics, the two stationary Stokes equations for low Reynolds numbers in incompressible fluids, and the Nernst-Planck equation for mass transport. Recently, it has also been claimed that the leading mechanism behind the diffusiophoretic phenomenon is electrophoresis [Esplandiu et al., Soft Matter 16, 3717 (2020)]. In this paper, we analyze the evolution of the exclusion zone based on a one-component interaction model at the Langevin equation level, which leads to simple analytical expressions instead of the complex numerical scheme of previous works, yet being consistent with it. We manage to reproduce the evolution of the exclusion zone width and the mean-square displacements of colloidal particles we measure near Nafion, a perfluorinated polymer membrane material, along with all characteristic time regimes, in a unified way. Our findings are also strongly supported by complementary experiments using two parallel planar conductors kept at a fixed voltage, mimicking the hydrophilic surfaces, and some computer simulations.

Nanovesicles drive a tunable dynamical arrest of microparticles.

Vitrification in a dilute colloidal system needs an asymmetric particle composition (a mixture of nano and micro colloids) to materialize. The volume fraction of the large particles increases (up to ≈0.58) driven by depletion forces produced by the smaller colloids. Such entropic forces are short-ranged and attractive. We found a different type of dynamical arrest in an extremely dilute asymmetric mixture of nanovesicles and polystyrene microparticles, where energy, instead of entropy, is the main protagonist to drive the arrest. Furthermore, when the vesicles go through the gel-fluid phase transition, the mean square displacements of the microparticles suffer a sudden splitting indicating a viscous jump. If the vesicles are doped with negatively charged lipids, particles and vesicles repel each other and the rheology of the mixture becomes athermal and Newtonian. Our findings are important to understand caging phenomena in biological systems, where diverse electrostatic distributions are present.

[Bearing surfaces in primary hip arthroplasty. Is there any difference?] / Pares articulares en prótesis primaria de cadera. ¿Existe alguna diferencia?

INTRODUCTION: In primary hip replacement, different materials are used for bearing surfaces. In our medium metal or ceramic heads with highly crossed-linked polyethylene (PA) are the most used. These combinations have good results, but it is not clear which is clinically superior. The objective of this study is to determine whether there is any clinically significant difference based on a systematic review of the literature and national registries of arthroplasty. MATERIAL AND METHODS: We conduct a systematic review of the literature and national registries of arthroplasty and we were looking for studies comparing bearing surfaces: ceramic-highly cross-linked polyethylene (CP) and metal-highly cross-linked polyethylene (MP); describing the revision rate according to the surface type with a minimum 10-year follow-up. The outcome evaluated was: review rate for any cause depending on surface type. RESULTS: Two out of fifteen national registries were included. The Australian registry shows a difference in the 15-year revision rate: CP: 6.3 (IC 5.8, 6.7) vs MP: 5.1 (IC 4.6, 5.7). The New Zealand registry shows no differences in revision rate/100 components/year: CP 0.54 (0.48-0.61) vs MP 0.61 (0.57-0.66). We do not find clinical studies with inclusion criteria that answer the research question. CONCLUSION: The results of this review show a high survival rate with the use of highly cross-linked polyethylene, the results are similar when using ceramic or metal heads.

INTRODUCCIÓN: En un reemplazo total de cadera primario (RTC), diferentes materiales son los que se usan para las superficies de apoyo. En nuestro medio, las cabezas de metal o cerámica con polietileno altamente entrecruzado (PA) son los más utilizados. Estas combinaciones tienen buenos resultados, pero no es claro cuál es superior clínicamente. Basados en una revisión sistemática de la literatura y de los registros nacionales de artroplastía, el objetivo de este estudio es determinar si existe alguna diferencia significativa desde el punto de vista clínico. MATERIAL Y MÉTODOS: Realizamos una revisión sistemática de la literatura y de los registros nacionales de artroplastía. Buscamos estudios en los que se compararan los tipos de superficie de contacto: cerámica-polietileno altamente entrecruzado (CP) y metal-polietileno altamente entrecruzado (MP), además de los registros nacionales de artroplastía que describieran, con un seguimiento mínimo de 10 años, la tasa de revisión según el tipo de superficie. El desenlace evaluado fue: tasa de revisión por cualquier causa según el tipo de superficie. RESULTADOS: Dos de quince registros nacionales fueron incluidos: el registro australiano muestra una diferencia en la tasa de revisión a 15 años, comparando CP: 6.3 (IC 5.8, 6.7) contra MP: 5.1 (IC 4.6, 5.7). El registro de Nueva Zelanda no muestra diferencias en la tasa de revisión/100 componentes/año: CP de 0.54 (0.48-0.61) en comparación con MP de 0.61 (0.57-0.66). No encontramos estudios clínicos con los criterios de inclusión que respondan la pregunta de investigación. CONCLUSIÓN: Los resultados de esta revisión muestran una alta supervivencia cuando se usa polietileno altamente entrecruzado; asimismo, los resultados son similares cuando se utilizan cabezas de cerámica o metálicas.

Cholesterol sequestration by xenon nano bubbles leads to lipid raft destabilization.

Combined coarse-grained (CG) and atomistic molecular dynamics (MD) simulations were performed to study the interactions of xenon with model lipid rafts consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC) and cholesterol (Chol). At a concentration of 2 Xe/lipid we observed an unexpected result: spontaneous nucleation of Xe nano bubbles which rapidly plunged into the bilayer. In this process Chol, essential for raft stabilization, was pulled out from the raft into the hydrophobic zone. When concentration was further increased (3 Xe/lipid), the bubbles increase in size and disrupted both the membrane and raft. We computed the radial distribution functions, pair-wise potentials, second virial coefficients and Schlitter entropy to scrutinize the nature of the interactions. Our findings, concurring with a recent report on the origin of general anaesthesia (M. A. Pavel, E. N. Petersen, H. Wang, R. A. Lerner and S. B. Hansen, Proc. Natl. Acad. Sci. U. S. A., 2020, 117(24), 13757-13766), suggest that the well-known anaesthetic effect of Xe could be mediated by sequestration of Chol, which, in turn, compromises the stability of rafts where specialized proteins needed to produce the nervous signal are anchored.

The cut-off effect of n-alcohols in lipid rafts: A lipid-dependent phenomenon☆.

n-Aliphatic alcohols act as anesthetics only up to a certain chain length, beyond which its biological activity disappears. This is known as the 'cut-off' phenomenon. Although the most accepted explanation is based on action sites in membrane proteins, it is not well understood why alcohols alter their functions. The structural dependence of these protein receptors to lipid domains known as 'lipid rafts', suggests a new approach to tackle the puzzling phenomenon. In this work, by performing molecular dynamic simulations (MDS) to explore the lipid role, we provide relevant molecular details about the membrane-alcohol interaction at the cut-off point regime. Since the high variability of the cut-off points found on protein receptors in neurons may be a consequence of differences in the lipid composition surrounding such proteins, our results could have a clear-cut importance.

Thermal behavior of a lipid-protein membrane model and the effects produced by anesthetics and neurotransmitters.

Despite decades of intense research to understand the phenomenon of anesthesia and its membrane-related changes in neural transmission, where lipids and proteins have been proposed as primary targets of anesthetics, the involved action mechanisms remain unclear. Based on the overall agreement that anesthetics and neurotransmitters induce particular modifications in the plasma membrane of neurons, triggering specific responses and changes in their energetic states, we present here a thermal study to investigate membrane effects in a lipid-protein model made of 1,2-dimyristoyl-sn-glycero-3-phosphocholine and albumin from chicken egg white under the influence of neurotransmitters and anesthetics. First, we observe how ovalbumin, ovotransferrin, and lysozyme (main albumin constituents from chicken egg white) interact with the lipid membrane enhancing their lipophilic character while exposing their hydrophobic domains. This produces a lipid separation and a more ordered hybrid lipid-protein assembly. Second, we measured the thermotropic changes of this assembly induced by acetylcholine, γ-aminobutiric acid, tetracaine, and pentobarbital. Although the protein in our study is not a receptor, our results are striking, for they give evidence of the great importance of non-specific interactions in the anesthesia mechanism.

The effect of green light on the motility of mouse sperm at two different temperatures.

Photo-modulation with visible light has been used to induce gains in the motility of the sperms of rabbits, boars, buffalo, bulls, fish, and humans. Although different hypotheses have been proposed to explain such an effect, the origin and mechanisms by which visible light affects sperm motility are still far from being completely understood. Several groups have observed changes in the intracellular Ca2+ concentration and significant differences in the production of ROS, which are attributed to specific photosensitizers. Also, it has been reported that blue light induces nitric oxide production in sperm cells, which plays a vital role in acrosome reaction and capacitation leading to an augmentation in the fertilisation probability. In the present work, we study the effects of green light (490-540 nm) on the sperm motility of mice. Firstly, we carried out experiments at 37 °C to confirm what previous researchers have observed before using red and blue light: that the overall sperm motility increases. Secondly, we studied the effects of green light at 10 °C and found that the motility drastically diminishes. In order to understand this opposing outcome, we carried out fluorescence measurements to evaluate reactive oxygen species production induced by green light at both temperatures. Our results suggest that the balance between the use and generation of ROS at 37 °C is favorable to the cells, while at 10 °C it is harmful.

Video-tracking and high-speed bright field microscopy allow the determination of swimming and cardiac effects of D. magna exposed to local anaesthetics.

Local anaesthetics are among the most used drugs in clinical practice, but once they are released to the environment, the effects on the aquatic fauna remain uncertain. This study evaluates, for the first time, the impact of tetracaine, lidocaine and bupivacaine on the survival rate and physiological effects of cladocera Daphnia magna. Video-tracking and image processing allowed us to obtain changes in behaviour parameters like swimming average velocity and mean square displacement. We found that tetracaine shows the most severe effect. A high-speed microscopy system was also used to determine the response of D. magna heart to these drugs. Our results show that tetracaine presents dose-dependent area reduction during all cardiac cycle, while bupivacaine and lidocaine did not present significative effects on heart size. The tested drugs, at environmental high concentrations, altered behaviour, heart function and survival of D. magna.

Pattern formation of stains from dried drops to identify spermatozoa motility.

We study how cell motility affects the stains left by the evaporation of droplets of a biofluid suspension containing mouse spermatozoa. The suspension, which contains also a high concentration of salts usually needed by motile cells, forms, upon drying, a crystallized pattern. We examine the structural characteristics of such patterns by optical microscopy. The analysis reveals that cell motility affects the formation of elongated crystals with lateral tips, as well as the creation of interlocked aggregates. We prove that a lacunarity algorithm based on polar symmetry, distinguishes among deposits generated by motile and non-motile cells with an accuracy greater than 95%.

Induced clustering of Escherichia coli by acoustic fields.

Brownian or self-propelled particles in aqueous suspensions can be trapped by acoustic fields generated by piezoelectric transducers usually at frequencies in the megahertz. The obtained confinement allows the study of rich collective behaviours like clustering or spreading dynamics in microgravity-like conditions. The acoustic field induces the levitation of self-propelled particles and provides secondary lateral forces to capture them at nodal planes. Here, we give a step forward in the field of confined active matter, reporting levitation experiments of bacterial suspensions of Escherichia coli. Clustering of living bacteria is monitored as a function of time, where different behaviours are clearly distinguished. Upon the removal of the acoustic signal, bacteria rapidly spread, impelled by their own swimming. Nevertheless, long periods of confinement result in irreversible bacteria entanglements that could act as seeds for levitating bacterial aggregates.

Plant proteases for bioactive peptides release: A review.

Proteins are a potential source of health-promoting biomolecules with medical, nutraceutical, and food applications. Nowadays, bioactive peptides production, its isolation, characterization, and strategies for its delivery to target sites are a matter of intensive research. In vitro and in vivo studies regarding the bioactivity of peptides has generated strong evidence of their health benefits. Dairy proteins are considered the richest source of bioactive peptides, however proteins from animal and vegetable origin also have been shown to be important sources. Enzymatic hydrolysis has been the process most commonly used for bioactive peptide production. Most commercial enzymatic preparations frequently used are from animal (e.g., trypsin and pepsin) and microbial (e.g., Alcalase® and Neutrase®) sources. Although the use of plant proteases is still relatively limited to papain and bromelain from papaya and pineapple, respectively, the application of new plant proteases is increasing. This review presents the latest knowledge in the use and diversity of plant proteases for bioactive peptides release from food proteins including both available commercial plant proteases as well as new potential plant sources. Furthermore, the properties of peptides released by plant proteases and health benefits associated in the control of disorders such as hypertension, diabetes, obesity, and cancer are reviewed.

Ionic interactions determine the morphology of dried alkali/liposome suspension droplets.

We sought to understand why saline drops produce intriguing patterns when drying in the presence of zwitterionic liposomes. Specifically, we would like to comprehend why the nature of such patterns is hierarchically driven by the Hofmeister series. The liposome suspension is made of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) with alkali metal chlorides. A complexity analysis of the patterns gives a fractal dimension around 1.71, which means that the drying process resembles a DLA mechanism. A physicochemical study, including the determination of zeta potential, molecular dynamics simulations, microrheology, and calorimetry, supports the fact that electrostatic interactions among head groups of phospholipids with alkali cations are the driven forces behind the assembling of the observed structures. Moreover, we found that the morphology of the dried droplets is sensitive to the substrate. Our findings could be used in a biological context, for example, to characterize cells in ionic media.

Propagation of a thermo-mechanical perturbation on a lipid membrane.

The propagation of sound waves on lipid monolayers supported on water has been previously studied during the melting transition. Since changes in volume, area, and compressibility in lipid membranes have biological relevance, the observed sound propagation is of paramount importance. However, it is unknown what would occur on a lipid bilayer, which is a more approximate model of a cell membrane. With the aim to answer this relevant question, we built an experimental setup to assemble long artificial lipid membranes. We found that if these membranes are heated in order to force local melting, a thermo-mechanical perturbation propagates a long distance. Our findings may support the existence of solitary waves, postulated to explain the propagation of isentropic signals together with the action potential in neurons.

A technique based on droplet evaporation to recognize alcoholic drinks.

Chromatography is, at present, the most used technique to determine the purity of alcoholic drinks. This involves a careful separation of the components of the liquid elements. However, since this technique requires sophisticated instrumentation, there are alternative techniques such as conductivity measurements and UV-Vis and infrared spectrometries. We report here a method based on salt-induced crystallization patterns formed during the evaporation of alcoholic drops. We found that droplets of different samples form different structures upon drying, which we characterize by their radial density profiles. We prove that using the dried deposit of a spirit as a control sample, our method allows us to differentiate between pure and adulterated drinks. As a proof of concept, we study tequila.

The calorimetric properties of liposomes determine the morphology of dried droplets.

The evaporation of liquid droplets deposited on a substrate is a very complex phenomenon. Driven by capillary and Marangoni flows, particle-particle and particle-substrate interactions, the deposits they leave are vestiges of such complexity. We study the formation of patterns during the evaporation of liposome suspension droplets deposited on a hydrophobic substrate at different temperatures. We observed that as we change the temperature of the substrate, a morphological phase transition occurs at a given temperature Tm. This temperature corresponds to the gel-fluid lipid melting transition of the liposome suspension. Optical microscopy and atomic force microscopy are used to study the morphology of the patterns. Based on the radial density profiles we found that all structures can be classified into two groups: patterns composed by nearly uniform deposition (below Tm) and prominent structures containing randomly distributed voids (above Tm).