Morphometric and Nanomechanical Screening of Peripheral Blood Cells with Atomic Force Microscopy for Label-Free Assessment of Alzheimer's Disease, Parkinson's Disease, and Amyotrophic Lateral Sclerosis.

Neurodegenerative disorders (NDDs) are complex, multifactorial disorders with significant social and economic impact in today's society. NDDs are predicted to become the second-most common cause of death in the next few decades due to an increase in life expectancy but also to a lack of early diagnosis and mainly symptomatic treatment. Despite recent advances in diagnostic and therapeutic methods, there are yet no reliable biomarkers identifying the complex pathways contributing to these pathologies.

Thermodynamic Signatures of Blood Plasma Proteome in Neurodegenerative Pathologies.

Discovery of diagnostic biomarkers for age-related neurodegenerative pathologies (NDDs) is essential for accurate diagnosis, following disease progression and drug development. Blood plasma and blood cells are important peripheral sources for NDDs' biomarkers that, although present in lower concentrations than in cerebrospinal fluid, would allow noninvasive diagnostics. To identify new biomarkers for Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS), in this work we have evaluated the modifications in the thermodynamic behavior of blood plasma proteome exploring differential scanning calorimetry.

Calorimetric Markers for Detection and Monitoring of Multiple Myeloma.

This review summarizes data obtained thus far on the application of differential scanning calorimetry (DSC) for the analysis of blood sera from patients diagnosed with multiple myeloma (MM) with the secretion of the most common isotypes of monoclonal proteins (M-proteins), free light chains (FLC) and non-secretory MM, as well as Waldenström macroglobulinemia and the premalignant state monoclonal gammopathy of undetermined significance. The heterogeneous nature of MM is reflected in the thermal stability profiles of the blood serum proteome of MM patients found to depend on both the level and the isotype of the secreted M-proteins or FLC. Common calorimetric markers feature the vast majority of the different myeloma types, i.

Nonlinear electric response of the diffuse double layer to an abrupt charge displacement inside a biological membrane.

In order to elucidate the old, still unsolved problem of how the diffuse electric double layer responds to an abrupt, intramolecular charge displacement inside a biological membrane, we investigated the fastest components of the light-induced electric signals of bacteriorhodopsin and its mutants, in numerous ionic and buffer solutions. The obtained data for temperature and solute concentration dependence were interpreted as a consequence of changes in the capacity of the diffuse double layer surrounding the purple membrane. The possible physiological consequences of this so far not demonstrated phenomenon are discussed.

Salicylic Acid as Ionic Liquid Formulation May Have Enhanced Potency to Treat Some Chronic Skin Diseases.

In recent years, numerous studies have shown that conversion of conventional drugs in ionic liquid (IL) formulation could be a successful strategy to improve their physicochemical properties or suggest a new route of administration. We report the synthesis and detailed characterization of eight salicylic acid-based ILs (SA-ILs) containing cation non-polar or aromatic amino acid esters. Using in vitro assays, we preliminary evaluated the therapeutic potency of the novel SA-ILs.

Morphometry and Stiffness of Red Blood Cells-Signatures of Neurodegenerative Diseases and Aging.

Human red blood cells (RBCs) are unique cells with the remarkable ability to deform, which is crucial for their oxygen transport function, and which can be significantly altered under pathophysiological conditions. Here we performed ultrastructural analysis of RBCs as a peripheral cell model, looking for specific signatures of the neurodegenerative pathologies (NDDs)-Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD), utilizing atomic force (AFM) and conventional optical (OM) microscopy. We found significant differences in the morphology and stiffness of RBCs isolated from patients with the selected NDDs and those from healthy individuals.

Red Blood Cells' Thermodynamic Behavior in Neurodegenerative Pathologies and Aging.

The main trend of current research in neurodegenerative diseases (NDDs) is directed towards the discovery of novel biomarkers for disease diagnostics and progression. The pathological features of NDDs suggest that diagnostic markers can be found in peripheral fluids and cells. Herein, we investigated the thermodynamic behavior of the peripheral red blood cells (RBCs) derived from patients diagnosed with three common NDDs-Parkinson's disease (PD), Alzheimer's disease (AD), and amyotrophic lateral sclerosis (ALS) and compared it with that of healthy individuals, evaluating both fresh and aged RBCs.

Morphometric and Nanomechanical Features of Platelets from Women with Early Pregnancy Loss Provide New Evidence of the Impact of Inherited Thrombophilia.

Pregnancy is associated with hypercoagulation states and increased thrombotic risk, especially in women with thrombophilia. We combine atomic force microscopy (AFM) and flow cytometry to examine the morphology and nanomechanics of platelets derived from women with early pregnancy loss (EPL) and control pregnant (CP) and non-pregnant (CNP) women. Both control groups exhibit similar morphometric parameters (height and surface roughness) and membrane stiffness of platelets.

Single-Walled Carbon Nanotubes Modify Leaf Micromorphology, Chloroplast Ultrastructure and Photosynthetic Activity of Pea Plants.

Single-walled carbon nanotubes (SWCNTs) emerge as promising novel carbon-based nanoparticles for use in biomedicine, pharmacology and precision agriculture. They were shown to penetrate cell walls and membranes and to physically interact and exchange electrons with photosynthetic complexes in vitro. Here, for the first time, we studied the concentration-dependent effect of foliar application of copolymer-grafted SWCNTs on the structural and functional characteristics of intact pea plants.

Insights into graphene oxide interaction with human serum albumin in isolated state and in blood plasma.

The interactions of graphene oxide (GO), a 2-dimensional nanomaterial with hydrophilic edges, hydrophobic basal plane and large flat surfaces, with biological macromolecules, are of key importance for the development of novel nanomaterials for biomedical applications. To gain more insight into the interaction of GO flakes with human serum albumin (HSA), we examined GO binding to HSA in its isolated state and in blood plasma. Calorimetric data reveal that GO strongly stabilizes free isolated HSA against a thermal challenge at low ionic strength, indicating strong binding interactions, confirmed by the drop in ζ-potential of the HSA/GO assemblies compared to bare GO flakes.

Scop. induces cell cycle arrest, apoptosis, genotoxic effects, thermodynamic and epigenetic events in MCF7 breast cancer cells.

Current plant-derived anticancer therapeutics aim to reach higher effectiveness, to potentiate chemosensitivity and minimize the toxic side effects compared to conventional chemotherapy. Scop. is a herb with high pharmacological potential, widely applied in traditional phytotherapy.

Phycobilisome integrity and functionality in lipid unsaturation and xanthophyll mutants in Synechocystis.

The major light-harvesting system in cyanobacteria, the phycobilisome, is an essential component of the photosynthetic apparatus that regulates the utilization of the natural light source-the Sun. Earlier works revealed that the thylakoid membrane composition and its physical properties might have an important role in antennas docking. Polyunsaturated lipids and xanthophylls are among the most significant modulators of the physical properties of thylakoid membranes.

Spectrokinetic characterization of photoactive yellow protein films for integrated optical applications.

In this paper, the photocycle of the dried photoactive yellow protein film has been investigated in different humidity environments, in order to characterize its nonlinear optical properties for possible integrated optical applications. The light-induced spectral changes of the protein films were monitored by an optical multichannel analyser set-up, while the accompanying refractive index changes were measured with the optical waveguide lightmode spectroscopy method. To determine the number and kinetics of spectral intermediates in the photocycle, the absorption kinetic data were analysed by singular value decomposition and multiexponential fitting methods, whose results were used in a subsequent step of fitting a photocycle model to the data.

Thylakoid membrane unstacking increases LHCII thermal stability and lipid phase fluidity.

Thylakoids are highly protein-enriched membranes that harbor a number of multicomponent photosynthetic complexes. Similarly to other biological membranes the protein constituents are heterogeneously distributed laterally in the plane of the membrane, however the specific segregation into stacked (grana patches) and unstacked (stroma lamellae) membrane layers is a unique feature of the thylakoid. Both the lateral and the vertical arrangements of the integral membrane proteins within the three-dimensional thylakoid ultrastructure are thought to have important physiological function.

Physiological and structural adjustments of two ecotypes of L. from different habitats in response to drought and re-watering.

covers a very fragmented area in Europe and, at the edge of its natural distribution, is considered a relic endangered species near extinction. In our study, it was hypothesized that individuals from the edge of the habitat, with stronger climate constrains (drier and warmer environment, Italy, IT ecotype), developed different mechanisms of adaptation than those growing under optimal conditions at the center of the habitat (more humid and colder environment, Bulgaria, BG ecotype). Indeed, the two ecotypes displayed physiological, structural and functional differences already under control (unstressed) conditions.

Calorimetric markers for monitoring of multiple myeloma and Waldenström's macroglobulinemia patients.

The blood proteome has been studied extensively for identification of novel reliable disease biomarkers. In recent years, differential scanning calorimetry has emerged as a new tool for characterization of the thermodynamic properties of the major serum/plasma proteins and for the establishment of calorimetric markers for a variety of diseases. Here we applied calorimetry to monitor the effect of treatment of patients diagnosed with multiple myeloma and Waldenström's macroglobulinemia on the calorimetric profiles of patients' blood sera.

Structural integrity of Synechocystis sp. PCC 6803 phycobilisomes evaluated by means of differential scanning calorimetry.

Phycobilisomes (PBSs) are supramolecular pigment-protein complexes that serve as light-harvesting antennae in cyanobacteria. They are built up by phycobiliproteins assembled into allophycocyanin core cylinders (ensuring the physical interaction with the photosystems) and phycocyanin rods (protruding from the cores and having light-harvesting function), the whole PBSs structure being maintained by linker proteins. PBSs play major role in light-harvesting optimization in cyanobacteria; therefore, the characterization of their structural integrity in intact cells is of great importance.

Hybrid graphene oxide/polysaccharide nanocomposites with controllable surface properties and biocompatibility.

Herein, a strong interdependence between the composition of hybrid graphene oxide/hyaluronan/chitosan GO/HA/Chi multilayers and their surface properties and biocompatibility was demonstrated that can be used to build up coatings with desirable and precisely tunable properties. Both the position and the abundance of GO-layers into the polymer matrix were systematically varied to draw interconnection with the growth type, thickness, morphology, roughness, hydrophilicity and biocompatibility. It was found that when deposited in-between the HA and Chi layers GO forms diffusion barrier, hindering the mobility of Chi-chains and changing the exponential film growth to linear.

PlA2 Polymorphism in Glycoprotein IIb/IIIa Modulates the Morphology and Nanomechanics of Platelets.

Glycoprotein IIb/IIIa (GPIIb/IIIa) is the most abundant platelet surface receptor for fibrinogen and von Willebrand factor. Polymorphism PlA1/A2 in the gene of GPIIb/IIIa is among the risk factors for the development of arterial and venous thrombosis. The aim of this study is to evaluate the effect of the carriage of PlA1/A2 on the size, topographic features, and membrane stiffness of platelets from healthy controls and patients with deep venous thrombosis (DVT).

Regulation of the growth, morphology, mechanical properties and biocompatibility of natural polysaccharide-based multilayers by Hofmeister anions.

Herein the optimization of the physicochemical properties and surface biocompatibility of polyelectrolyte multilayers of the natural, biocompatible and biodegradable, linear polysaccharides hyaluronan and chitosan by Hofmeister anions was systematically investigated. We demonstrated that there is an interconnection between the bulk and surface properties of HA/Chi multilayers both varying in accordance with the arrangement of the anions in the Hofmeister series. Kosmotropic anions increased the hydration, thickness, micro- and macro-roughness, and hydrophilicity and improved the biocompatibility of the films by reduction (2 orders of magnitude) of the films stiffness and complete anti-thrombogenicity.

Intercriteria analysis of calorimetric data of blood serum proteome.

Background: Biological microcalorimetry has entered into a phase where its potential for disease diagnostics is readily recognized. A wide variety of oncological and immunological disorders have been characterized by differential scanning calorimetry (DSC) and characteristic thermodynamic profiles were reported. Now the challenge before DSC is not the experimental data collection but the development of analysis protocols for reliable data stratification/classification and discrimination of disease specific features (calorimetric markers).

Thermal stability and binding energetics of thymidylate synthase ThyX.

The bacterial thymidylate synthase ThyX is a multisubstrate flavoenzyme that takes part in the de novo synthesis of thymidylate in a variety of microorganisms. Herein we study the effect of FAD and dUMP binding on the thermal stability of wild type (WT) ThyX from the mesophilic Paramecium bursaria chlorella virus-1 (PBCV-1) and from the thermophilic bacterium Thermotoga maritima (TmThyX), and from two variants of TmThyX, Y91F and S88W, using differential scanning calorimetry. The energetics underlying these processes was characterized by isothermal titration calorimetry.