Niosomes based on synthetic cationic lipids for gene delivery: the influence of polar head-groups on the transfection efficiency in HEK-293, ARPE-19 and MSC-D1 cells.

We designed niosomes based on three lipids that differed only in the polar-head group to analyze their influence on the transfection efficiency. These lipids were characterized by small-angle X-ray scattering before being incorporated into the niosomes which were characterized in terms of pKa, size, zeta potential, morphology and physical stability. Nioplexes were obtained upon the addition of a plasmid. Different ratios (w/w) were selected to analyze the influence of this parameter on size, charge and the ability to condense, release and protect the DNA. In vitro transfection experiments were performed in HEK-293, ARPE-19 and MSC-D1 cells. Our results show that the chemical composition of the cationic head-group clearly affects the physicochemical parameters of the niosomes and especially the transfection efficiency. Only niosomes based on cationic lipids with a dimethyl amino head group (lipid 3) showed a transfection capacity when compared with their counterparts amino (lipid 1) and tripeptide head-groups (lipid 2). Regarding cell viability, we clearly observed that nioplexes based on the cationic lipid 3 had a more deleterious effect than their counterparts, especially in ARPE-19 cells at 20/1 and 30/1 ratios. Similar studies could be extended to other series of cationic lipids in order to progress in the research on safe and efficient non-viral vectors for gene delivery purposes.

Comparative study of extracellular recording methods for analysis of afferent sensory information: Empirical modeling, data analysis and interpretation.

BACKGROUND: Physiological studies of sensorial systems often require the acquisition and processing of data extracted from their multiple components to evaluate how the neural information changes in relation to the environment changes. In this work, a comparative study about methodological aspects of two electrophysiological approaches is described. NEW METHOD: Extracellular recordings from deep vibrissal nerves were obtained by using a customized microelectrode Utah array during passive mechanical stimulation of rat´s whiskers. These recordings were compared with those obtained with bipolar electrodes. We also propose here a simplified empirical model of the electrophysiological activity obtained from a bundle of myelinated nerve fibers. RESULTS: The peripheral activity of the vibrissal system was characterized through the temporal and spectral features obtained with both recording methods. The empirical model not only allows the correlation between anatomical structures and functional features, but also allows to predict changes in the CAPs morphology when the arrangement and the geometry of the electrodes changes. COMPARISON WITH EXISTING METHOD(S): This study compares two extracellular recording methods based on analysis techniques, empirical modeling and data processing of vibrissal sensory information. CONCLUSIONS: This comparative study reveals a close relationship between the electrophysiological techniques and the processing methods necessary to extract sensory information. This relationship is the result of maximizing the extraction of information from recordings of sensory activity.

Multiscale dynamics of interstimulus interval integration in visual cortex.

Although the visual cortex receives information at multiple temporal patterns, much of the research in the field has focused only on intervals shorter than 1 second. Consequently, there is almost no information on what happens at longer temporal intervals. We have tried to address this question recording neuronal populations of the primary visual cortex during visual stimulation with repetitive grating stimuli and intervals ranging from 1 to 7 seconds. Our results showed that firing rate and response stability were dependent of interval duration. In addition, there were collective oscillations with different properties in response to changes in intervals duration. These results suggest that visual cortex could encode visual information at several time scales using oscillations at multiple frequencies.

Time-frequency analysis of neuronal populations with instantaneous resolution based on noise-assisted multivariate empirical mode decomposition.

BACKGROUND: Linear analysis has classically provided powerful tools for understanding the behavior of neural populations, but the neuron responses to real-world stimulation are nonlinear under some conditions, and many neuronal components demonstrate strong nonlinear behavior. In spite of this, temporal and frequency dynamics of neural populations to sensory stimulation have been usually analyzed with linear approaches. NEW METHOD: In this paper, we propose the use of Noise-Assisted Multivariate Empirical Mode Decomposition (NA-MEMD), a data-driven template-free algorithm, plus the Hilbert transform as a suitable tool for analyzing population oscillatory dynamics in a multi-dimensional space with instantaneous frequency (IF) resolution. RESULTS: The proposed approach was able to extract oscillatory information of neurophysiological data of deep vibrissal nerve and visual cortex multiunit recordings that were not evidenced using linear approaches with fixed bases such as the Fourier analysis. COMPARISON WITH EXISTING METHODS: Texture discrimination analysis performance was increased when Noise-Assisted Multivariate Empirical Mode plus Hilbert transform was implemented, compared to linear techniques. Cortical oscillatory population activity was analyzed with precise time-frequency resolution. Similarly, NA-MEMD provided increased time-frequency resolution of cortical oscillatory population activity. CONCLUSIONS: Noise-Assisted Multivariate Empirical Mode Decomposition plus Hilbert transform is an improved method to analyze neuronal population oscillatory dynamics overcoming linear and stationary assumptions of classical methods.

The influence of the polar head-group of synthetic cationic lipids on the transfection efficiency mediated by niosomes in rat retina and brain.

The development of novel non-viral delivery vehicles is essential in the search of more efficient strategies for retina and brain diseases. Herein, optimized niosome formulations prepared by oil-in water (o/w) and film-hydration techniques were characterized in terms of size, PDI, zeta potential, morphology and stability. Three ionizable glycerol-based cationic lipids containing a primary amine group (lipid 1), a triglycine group (lipid 2) and a dimethylamino ethyl pendent group (lipid 3) as polar head-groups were part of such niosomes. Upon the addition of pCMS-EGFP plasmid, nioplexes were obtained at different cationic lipid/DNA ratios (w/w). The resultant nioplexes were further physicochemically characterized and evaluated to condense, release and protect the DNA against enzymatic digestion. In vitro experiments were performed to evaluate transfection efficiency and cell viability in HEK-293, ARPE-19 and PECC cells. Interestingly, niosome formulations based on lipid 3 showed better transfection efficiencies in ARPE-19 and PECC cells than the rest of cationic lipids showed in this study. In vivo experiments in rat retina after intravitreal and subretinal injections together with in rat brain after cerebral cortex administration showed promising transfection efficiencies when niosome formulations based on lipid 3 were used. These results provide new insights for the development of non-viral vectors based on cationic lipids and their applications for efficient delivery of genetic material to the retina and brain.

[Personal experience in the surgery of nasal sinus polyps]. / Experiencia personal en la cirugía de la poliposis nasosinusal.

Personal opinions on surgery of polyps of the nasal cavity and paranasal sinuses, including from simple polypectomy to ethmoid microsurgery and nasal endoscopic surgery. The diagnostic importance of tomodensitometry and the essential postsurgical care are emphasized. The results of 29 patients with polyps of the nasal cavity and paranasal sinuses treated with intranasal microsurgery and endoscopic surgery are reported.

[New protocol for the early diagnosis of acoustic schwannoma in the outpatient clinic]. / Nuevo protocolo para el diagnóstico precoz del neurinoma del acústico en consulta ambulatoria.

For the last two years we have been testing a protocol for early diagnosis of acoustic schwannoma. We analyzed the results obtained in outpatients of the ENT clinic for ten months. Using the protocol described 10.7% accurate diagnoses, were obtained although in only one diagnosis was made in the intracanalicular stadium that was susceptible to ENT surgery.