Atención farmacéutica domiciliaria a pacientes frágiles en Gipuzkoa / Pharmaceutical home care for frail patients in Gipuzkoa

Introducción: la asistencia sanitaria a domicilio es una de las estrategias que impulsan las administraciones para pacientes pluripatológicos que viven en sus domicilios y que debido a su grado de discapacidad o vulnerabilidad no pueden desplazarse a los centros sanitarios. Este estudio consiste en pilotar la Atención Farmacéutica Domiciliaria (AFD) ofreciendo respuestas individualizadas a los pacientes en sus domicilios, dentro del equipo multidisciplinar de atención a los pacientes frágiles de la Fundación Hurkoa.Material y métodos: se convocó a todas las farmacias de los municipios de Irún y Azkoitia a una formación. Los pacientes incluidos en el Plan Integral de Atención a la Fragilidad de la Fundación Hurkoa, que requerían ayuda con la gestión de la medicación y aceptaron incluir a un farmacéutico en el equipo multidisciplinar de cuidados, seleccionaron su farmacia comunitaria. Los farmacéuticos de dichas farmacias realizaron una revisión del botiquín junto con la revisión de la medicación. Resultados: en la revisión del botiquín se detectaron medicamentos caducados o no utilizados y se retiraron entre 2 y 3 medicamentos a todos los pacientes. En la revisión de la medicación se detectaron al menos un Problema Relacionado con la Medicación (PRM) en todos los pacientes del estudio. En la evaluación de la satisfacción el 100 % de los pacientes refirieron un grado de satisfacción alto con el programa. Discusión: el presente estudio refuerza la importancia de la inclusión del farmacéutico comunitario en el equipo multidisciplinar de AFD, ofreciendo respuestas individualizadas a cada paciente, mejorando la gestión de su medicación y así consiguiendo mejoras significativas en la calidad de vida. (AU)

Hiccups in Parkinson's disease: an analysis of cases reported in the European pharmacovigilance database and a review of the literature.

BACKGROUND: Some reports have suggested an association between dopamine agonists and hiccups, involuntary contractions that merit full clinical attention because they can be very debilitating. Many drugs frequently used to treat hiccups are formally contraindicated in Parkinson's disease due to their liability to worsen motor symptoms, making the treatment of hiccups problematic in this disease. The objective of the present study was to analyze all spontaneous reports of hiccups from the European Pharmacovigilance Database in patients with Parkinson's disease and/or on dopaminergic drugs. Finally, we sought to identify evidence-based recommendations on the management of hiccups in Parkinson's disease. METHODS: We searched for all reports of hiccups in the European Pharmacovigilance Database (EudraVigilance) and calculated proportional reporting ratios for dopamine agonists and hiccups. We reviewed the literature on Parkinson's disease, dopamine agonists, and hiccups, searching for specific treatment recommendations for hiccups in this disease. RESULTS: Both rotigotine and pramipexole fulfilled the criteria to generate a safety signal. We found 32 and 13 cases of hiccups associated with dopamine agonists in EudraVigilance and the literature, respectively. There were no specific recommendations for the management of hiccups in Parkinson's disease in the clinical guidelines consulted. CONCLUSIONS: We have found evidence that rotigotine and pramipexole are associated with the appearance of hiccups and that this adverse reaction occurs predominantly in males. Given the scarce information available, specific recommendations are needed in clinical guidelines for the adequate management of hiccups in Parkinson's disease.

Structural recovery of the retina in a retinoschisin-deficient mouse after gene replacement therapy by solid lipid nanoparticles.

X-linked juvenile retinoschisis (XLRS) is a retinal degenerative disorder caused by mutations in the RS1 gene encoding a protein termed retinoschisin. The disease is an excellent candidate for gene replacement therapy as the majority of mutations have been shown to lead to a complete deficiency of the secreted protein in the retinal structures. In this work, we have studied the ability of non-viral vectors based on solid lipid nanoparticles (SLN) to induce the expression of retinoschisin in photoreceptors (PR) after intravitreal administration to Rs1h-deficient mice. We designed two vectors prepared with SLN, protamine, and dextran (DX) or hyaluronic acid (HA), bearing a plasmid containing the human RS1 gene under the control of the murin opsin promoter (mOPS). In vitro, the nanocarriers were able to induce the expression of retinoschisin in a PR cell line. After injection into the murine vitreous, the formulation prepared with HA induced a higher transfection level in PR than the formulation prepared with DX. Moreover, the level of retinoschisin in the inner nuclear layer (INL), where bipolar cells are located, was also higher. Two weeks after vitreal administration into Rs1h-deficient mice, both formulations showed significant improvement of the retinal structure by inducing a decrease of cavities and PR loss, and an increase of retinal and outer nuclear layer (ONL) thickness. HA-SLN resulted in a significant higher increase in the thickness of both retina and ONL, which can be explained by the higher transfection level of PR. In conclusion, we have shown the structural improvement of the retina of Rs1h-deficient mice with PR specific expression of the RS1 gene driven by the specific promoter mOPS, after successful delivery via SLN-based non-viral vectors.

Solid lipid nanoparticle-based vectors intended for the treatment of X-linked juvenile retinoschisis by gene therapy: In vivo approaches in Rs1h-deficient mouse model.

X-linked juvenile retinoschisis (XLRS), which results from mutations in the gene RS1 that encodes the protein retinoschisin, is a retinal degenerative disease affecting between 1/5000 and 1/25,000 people worldwide. Currently, there is no cure for this disease and the treatment is based on the application of low-vision aids. The aim of the present work was the in vitro and in vivo evaluation of two different non-viral vectors based on solid lipid nanoparticles (SLNs), protamine and two anionic polysaccharides, hyaluronic acid (HA) or dextran (DX), for the treatment of XLRS. First, the vectors containing a plasmid which encodes both the reporter green fluorescent protein (GFP) and the therapeutic protein retinoschisin, under the control of CMV promoters, were characterized in vitro. Then, the vectors were subretinally or intravitreally administrated to C57BL/6 wild type mice. One week later, GFP was detected in all treated mice and in all retinal layers except in the Outer Nuclear Layer (ONL) and the Inner Nuclear Layer (INL), regardless of the administration route and the vector employed. Finally, two weeks after subretinal or intravitreal injection to Rs1h-deficient mice, GFP and retinoschisin expression was detected in all retinal layers, except in the ONL, which was maintained for at least two months after subretinal administration. The structural analysis of the treated Rs1h-deficient eyes showed a partial recovery of the retina related to the production of retinoschisin. This work shows for the first time a successful RS1 gene transfer to Rs1h-deficient animals using non-viral nanocarriers, with promising results that point to non-viral gene therapy as a feasible future therapeutic tool for retinal disorders.

Solid Lipid Nanoparticles as Non-Viral Vectors for Gene Transfection in a Cell Model of Fabry Disease.

Here, we demonstrate the ability of solid lipid nanoparticle-based non-viral vectors to increase the α-galactosidase A levels of the IMFE1 cell line, an in vitro model for target cells in Fabry disease. For this purpose, vectors containing the pR-M10-αGal A plasmid, which encodes the α-galactosidase A enzyme, were prepared; the in vitro transfection efficacy was studied in IMFE1 cells, and the results were confirmed by RT-PCR. The cellular uptake of the vectors, intracellular disposition of the plasmid, and probable endocytosis pathways of the nanoparticles were also analyzed. The vectors used for the studies carried protamine (P-DNA-SLN), dextran and protamine (D-P-DNA-SLN), or hyaluronic acid of two different molecular weights and protamine (HA150-P-DNA-SLN or HA500-P-DNA-SLN). The new formulations, which presented a particle size in the range of nanometers (from 218 nm to 348 nm) and a positive superficial charge, were able to increase α-galactosidase A activity up to 4-fold in comparison to non treated IMFE1 cells. The most efficient vectors were those that included HA, and no differences due to changes in the molecular weight of HA were detected. The observed lack of colocalization with each of the four different Nile Red-labeled vectors and transferrin or cholera toxin appears to indicate that clathrin- and caveolae-independent pathways may be involved in their cellular uptake. Additionally, colocalization with LysoTracker indicated that the formulations were exposed to lysosomal activity, which may be responsible for the release of the plasmid from the vector. In conclusion, we reveal the potential of SLN-based vectors to efficiently transfect an immortalized Fabry patient cell line.

Biodistribution of Nanostructured Lipid Carriers (NLCs) after intravenous administration to rats: influence of technological factors.

Nanoparticles for medical applications are frequently administered via parenteral administration. In this study, the tissue distribution of three lipid formulations based on Nanostructured Lipid Carriers (NLCs) after intravenous administration to rats was evaluated. NLCs were prepared by a high pressure homogenization method and varied in terms of particle size, surface charge, and surfactant content. The (99m)Tc radiolabeled NLCs were intravenously administered to rats, and radioactivity levels in blood and tissues were measured. Cmax, AUC0-24, and MRT0-24 were obtained from the radioactivity level versus time profiles. The radiolabeled nanocarriers exhibited a long circulation time since radioactivity was detected in blood even 24 h post-injection. No differences on the MRT values in blood among the NLCs were observed, in spite of the different particle size and surface charge. The highest radioactivity levels were measured in the kidney, followed by the bone marrow, the liver, and the spleen. In the kidney, there was a higher accumulation of the positive nanoparticles, and in the liver, uptake of negative nanoparticles was higher than positive ones. NLCs with the largest particle size showed a higher uptake in the lung and lower accumulation in liver and bone marrow, in comparison with the smaller ones.

Short- and long-term stability study of lyophilized solid lipid nanoparticles for gene therapy.

Most studies in gene therapy are focused on developing more efficient non-viral vectors, ignoring their stability, even though physically and chemically stable vectors are necessary to achieve large easily shipped and stored batches. In the present work, the effect of lyophilization on the morphological characteristics and transfection capacity of solid lipid nanoparticles (LyoSLN) and SLN-DNA vectors (Lyo(SLN-DNA)) has been evaluated. The lyophilized preparations were stored under three different sets of temperature and humidity ICH conditions: 25 degrees C/60%RH, 30 degrees C/65%RH and 40 degrees C/75%RH. After lyophilization we found an increase in particle size which did not imply a reduction of "in vitro" transfection capacity. Stability studies of formulations lyophilized with trehalose showed that SLNs were physically stable during 9 months at 25 degrees C/60%RH and 6 months at 30 degrees C/65%RH. This stability was lost when harder conditions were employed (40 degrees C/75%RH). LyoSLNs maintained or increased the transfection efficacy (from 19% to approximately 40% EGFP positive cells) over time only at 25 degrees C/60%RH and 30 degrees C/65%RH. Lyo(SLN-DNA) resulted in almost no transfection under all conditions. LyoSLNs showed less DNA condensation capacity, whereas in Lyo(SLN-DNA) the plasmid became strongly bound, hampering the transfection. Furthermore, the storage of lyophilized lipoplexes stabilized with the disaccharide trehalose did not affect cell viability.

A proline-rich peptide improves cell transfection of solid lipid nanoparticle-based non-viral vectors.

The aim of this work was to improve the transfection efficacy of solid lipid nanoparticle (SLN)-based non-viral vectors into ARPE-19 cells through the addition of Sweet Arrow Peptide (SAP). First, we prepared SAP-DNA complexes at ratios of at least 50:1, and then incorporated them into the SLNs. All formulations were able to protect DNA, and the peptide favoured the most bioactive form (supercoiled) of open circular DNA turns. In vitro transfection studies of the vectors containing the pCMS-EGFP plasmid in HEK293 and ARPE-19 cell lines revealed that incorporation of SAP led to greater transfection in both cell lines, although via different mechanisms. The presence of SAP in the formulations did not affect the viability of HEK293 or ARPE-19 cells. In HEK293 cells, SAP enabled greater uptake of the vectors, and an SAP to DNA ratio of 50:1 was sufficient for enhancing transfection. In contrast, in ARPE-19 cells, SAP induced a change in the dominant entrance mechanism, from clathrin endocytosis to caveolae/raft-dependent endocytosis, thereby decreasing use of the lysosomal pathway and consequently, reducing vector degradation. The extent to which SAP uses one mechanism or the other largely depends on its concentration in the formulation.

Solid lipid nanoparticles for retinal gene therapy: transfection and intracellular trafficking in RPE cells.

Retinal pigment epithelial (RPE) cells are usually employed to study DNA systems for diseases related to problems in the retina. Solid lipid nanoparticles (SLNs) have been shown to be useful non-viral vectors for gene therapy. The objective of this work was to evaluate the transfection capacity of SLNs in the human retinal pigment epithelial established cell line (ARPE-19) in order to elucidate the potential application of this vector in the treatment of retinal diseases. Results showed a lower transfection level of SLNs in ARPE-19 cells than in HEK293 (2.5% vs. 14.9% EGFP positive cells at 72h post-transfection). Trafficking studies revealed a delay in cell uptake of the vectors in ARPE-19 cells. Differences in internalization process into the two cell lines studied explain, in part, the difference in the gene expression. The clathrin-mediated endocytosis in ARPE-19 cells directs the solid lipid nanoparticles to lysosomes; moreover, the low division rate of this cell line hampers the entrance of DNA into the nucleus. The knowledge of intracellular trafficking is very useful in order to design more efficient vectors taking into account the characteristics of the specific cell line to be transfected.

Solid lipid nanoparticles: formulation factors affecting cell transfection capacity.

Since solid lipid nanoparticles (SLNs) were introduced as non-viral transfection systems, very few reports of their use for gene delivery have been published. In this work different formulations based on SLN-DNA complexes were formulated in order to evaluate the influence of the formulation components on the "in vitro" transfection capacity. SLNs composed by the solid lipid Precirol ATO 5, the cationic lipid DOTAP and the surfactant Tween 80, and SLN-DNA complexes prepared at different DOTAP/DNA ratios were characterized by studying their size, surface charge, DNA protection capacity, transfection and cell viability in HEK293 cultured cells. The incorporation of Tween 80 allowed for the reduction of the cationic lipid concentration. The formulations prepared at DOTAP/DNA ratios 7/1, 5/1 and 4/1 provided almost the same transfection levels (around 15% transfected cells), without significant differences between them (p>0.05). Other assayed formulations presented lower transfection. Transfection activity was dependent on the DOTAP/DNA ratio since it influences the DNA condensation into the SLNs. DNA condensation is a crucial factor which conditions the transfection capacity of SLNs, because it influences DNA delivery from nanoparticles, gene protection from external agents and DNA topology.