Perception of difficulties with vision-related activities of daily living among patients undergoing unilateral posterior capsulotomy

OBJECTIVES: To assess the influence of Nd:YAG (neodymium: yttrium-aluminum- garnet) laser unilateral posterior capsulotomy on visual acuity and patients' perception of difficulties with vision-related activities of daily life. METHODS: We conducted an interventional survey that included 48 patients between 40 and 80 years of age with uni- or bilateral pseudophakia, posterior capsule opacification, and visual acuity <0.30 (logMAR) in one eye who were seen at a Brazilian university hospital. All patients underwent posterior capsulotomy using an Nd:YAG laser. Before and after the intervention, patients were asked to complete a questionnaire that was developed in an exploratory study. RESULTS: Before posterior capsulotomy, the median visual acuity (logMAR) of the included patients was 0.52 (range 0.30-1.60). After posterior capsulotomy, the median visual acuity of the included patients improved to 0.10 (range 0.0-0.52). According to the subjects' perceptions, their ability to perform most of their daily life activities improved after the intervention (p<0.05). CONCLUSIONS: After patients underwent posterior capsulotomy with an Nd:YAG laser, a significant improvement in the visual acuity of the treated eye was observed. Additionally, subjects felt that they experienced less difficulty performing most of their vision-dependent activities of daily living

Design of epidermal growth factor immobilization on 3D biocompatible scaffolds to promote tissue repair and regeneration

Exogenous application of human epidermal growth factor (hEGF) stimulates epidermal wound healing. The aim of this study was to develop bioconjugates based on hEGF mimicking the protein in its native state and thus suitable for tissue engineering applications, in particular for treating skin-related disorders as burns. Ribonuclease A (RNase A) was used to investigate a number of different activated-agarose carriers: cyanogen bromide (CNBr)-activated-agarose and glyoxyl-agarose showed to preserve the appropriate orientation of the protein for receptor binding. EGF was immobilized on these carriers and immobilization yield was evaluated (100% and 12%, respectively). A peptide mapping of unbound protein regions was carried out by LC–MS to take evidence of the residues involved in the immobilization and, consequently, the flexibility and surface accessibility of immobilized EGF. To assess cell proliferative activities, 10, 25, 50, and 100 ng/mL of each immobilized EGF sample were seeded on fibroblast cells and incubated for 24, 48 and 72 h. The immobilized growth factor showed significantly high cell proliferative activity at 50 and 100 ng/mL compared to control and soluble EGF. Although both of the immobilized samples show dose-dependency when seeded with high number of fibroblast cells, CNBr-agarose-EGF showed a significantly high activity at 100 ng/mL and 72 h incubation, compared to glyoxyl-agarose-EGF

[reasons For Cataract Surgery Cancellation].

To study the reasons for cancelling cataract surgeries, and to suggest actions to improve the efficiency of patient care. A cross-sectional study was carried out in a university hospital's ophthalmology clinic of the state of São Paulo, Brazil. Two hundred subjects were randomly selected. The mean age was 68+/- 11.4 years old. The reasons for cancelling surgery were: unpropitious clinical condition (23.1%); tight schedule (35.9%); and patient non-attendance (41%). Most of the reasons related to social issues and the hospital's administrative aspects.35487-

Surface Plasmon Resonance as a Tool for Ligand Binding Investigation of Engineered GPR17 Receptor, a G Protein Coupled Receptor Involved in Myelination

The aim of this study was to investigate the potential of surface plasmon resonance (SPR) spectroscopy for the measurement of real-time ligand-binding affinities and kinetic parameters for GPR17, a G protein-coupled receptor (GPCR) of major interest in medicinal chemistry as potential target in demyelinating diseases. The receptor was directly captured, in a single-step, from solubilized membrane extracts on the sensor chip through a covalently bound anti-6x-His-antibody and retained its ligand binding activity for over 24h. Furthermore, our experimental setup made possible, after a mild regeneration step, to remove the bound receptor without damaging the antibody, and thus to reuse many times the same chip. Two engineered variants of GPR17, designed for crystallographic studies, were expressed in insect cells, extracted from crude membranes and analyzed for their binding with two high affinity ligands: the antagonist Cangrelor and the agonist Asinex 1. The calculated kinetic parameters and binding constants of ligands were in good agreement with those reported from activity assays and highlighted a possible functional role of the N-terminal residues of the receptor in ligand recognition and binding. Validation of SPR results was obtained by docking and molecular dynamics of GPR17-ligands interactions and by functional in vitro studies. The latter allowed us to confirm that Asinex 1 behaves as GPR17 receptor agonist, inhibits forskolin-stimulated adenylyl cyclase pathway and promotes oligodendrocyte precursor cell maturation and myelinating ability

Studying protein–protein affinity and immobilized ligand–protein affinity interactions using MS-based methods

This review discusses the most important current methods employing mass spectrometry (MS) analysis for the study of protein affinity interactions. The methods are discussed in depth with particular reference to MS-based approaches for analyzing protein–protein and protein–immobilized ligand interactions, analyzed either directly or indirectly. First, we introduce MS methods for the study of intact protein complexes in the gas phase. Next, pull-down methods for affinity-based analysis of protein–protein and protein–immobilized ligand interactions are discussed. Presently, this field of research is often called interactomics or interaction proteomics. A slightly different approach that will be discussed, chemical proteomics, allows one to analyze selectivity profiles of ligands for multiple drug targets and off-targets. Additionally, of particular interest is the use of surface plasmon resonance technologies coupled with MS for the study of protein interactions. The review addresses the principle of each of the methods with a focus on recent developments and the applicability to lead compound generation in drug discovery as well as the elucidation of protein interactions involved in cellular processes. The review focuses on the analysis of bioaffinity interactions of proteins with other proteins and with ligands, where the proteins are considered as the bioactives analyzed by MS

Recent developments in protein–ligand affinity mass spectrometry

This review provides an overview of direct and indirect technologies to screen protein–ligand interactions with mass spectrometry. These technologies have as a key feature the selection or affinity purification of ligands in mixtures prior to detection. Specific fields of interest for these technologies are metabolic profiling of bioactive metabolites, natural extract screening, and the screening of libraries for bioactives, such as parallel synthesis libraries and small combichem libraries. The review addresses the principles of each of the methods discussed, with a focus on developments in recent years, and the applicability of the methods to lead generation and development in drug discovery