Cell death induced by the application of alternating magnetic fields to nanoparticle-loaded dendritic cells

In this work, the capability of primary, monocyte-derived dendritic cells (DCs) to uptake iron oxide magnetic nanoparticles (MNPs) is assessed and a strategy to induce selective cell death in these MNP-loaded DCs using external alternating magnetic fields (AMFs) is reported. No significant decrease in the cell viability of MNP-loaded DCs, compared to the control samples, was observed after five days of culture. The amount of MNPs incorporated into the cytoplasm was measured by magnetometry, which confirmed that 1 to 5 pg of the particles were uploaded per cell. The intracellular distribution of these MNPs, assessed by transmission electron microscopy, was found to be primarily inside the endosomic structures. These cells were then subjected to an AMF for 30 min, and the viability of the blank DCs (i.e., without MNPs), which were used as control samples, remained essentially unaffected. However, a remarkable decrease of viability from approximately 90% to 2-5% of DCs previously loaded with MNPs was observed after the same 30 min exposure to an AMF. The same results were obtained using MNPs having either positive (NH2+) or negative (COOH-) surface functional groups. In spite of the massive cell death induced by application of AMF to MNP-loaded DCs, the amount of incorporated magnetic particles did not raise the temperature of the cell culture. Clear morphological changes at the cell structure after magnetic field application were observed using scanning electron microscopy. Therefore, local damage produced by the MNPs could be the main mechanism for the selective cell death of MNP-loaded DCs under an AMF. Based on the ability of these cells to evade the reticuloendothelial system, these complexes combined with an AMF should be considered as a potentially powerful tool for tumour therapy.Comment: In Press. 33 pages, 11 figure

Trajectories of objectively measured physical activity in free-living older men.

BACKGROUND: The steep decline in physical activity (PA) among the oldest old is not well understood; there is little information about the patterns of change in PA and sedentary behaviour (SB) in older people. Longitudinal data on objectively measured PA data can give insights about how PA and SB change with age. METHODS: Men age 70-90 yr, from a United Kingdom population-based cohort wore a GT3X accelerometer over the hip annually on up to three occasions (56%, 50%, and 51% response rates) spanning 2 yr. Multilevel models were used to estimate change in activity. Men were grouped according to achieving ≥150 min·wk of MVPA in bouts of ≥10 min (current guidelines) at two or three time points. RESULTS: A total of 1419 ambulatory men had ≥600 min wear time on ≥3 d at ≥2 time points. At baseline, men took 4806 steps per day and spent 72.5% of their day in SB, 23.1% in light PA, and 4.1% in moderate-to-vigorous PA (MVPA). Mean change per year was -341 steps, +1.1% SB, -0.7% light PA, and -0.4% MVPA each day (all P 30 min increased from 5.1 by 0.1 per year (P = 0.02). CONCLUSIONS: Among older adults, the steep decline in total PA occurred because of reductions in MVPA, while light PA is relatively spared and sedentary time and long sedentary bouts increase

Transferência gênica assistida por campos magnéticos: Estudos em células gliais

Las nanopartículas magnéticas (MNP) complejadas con vectores génicos pueden, en presencia de un campo magnético externo, amplificar sustancialmente la eficiencia de la transferencia génica. Esta técnica, denominada magnetofección, es de gran interés en el campo de la terapia génica. En este estudio se caracterizó la mejora de transferencia génica en células gliales B92 utilizando complejos constituidos por diferentes proporciones de MNP asociadas a dos vectores adenovirales, a saber: los complejos entre las MNP denominadas PEI-Mag2 asociadas al adenovector RAd-GFP que expresa la proteína fluorescente verde GFP o al adenovector RAd-DsRed que expresa la proteína fluorescente roja DsRed2. Se demostró que para ambos vectores, a medida que la relación MNP/partícula viral física (PVF) va aumentando, la amplificación de la transfección también aumenta hasta que se llega a una relación MNP/PVF a partir de la cual el factor de amplificación alcanza un plateau. Se determinó que para el complejo PEI-Mag2/RAd-GFP la relación a partir de la cual se alcanza el plateau es de aproximadamente 0,5 fg Fe/PVF mientras que para el complejo PEI-Mag2/RAd-DsRed, esta relación corresponde a aproximadamente 71 fg Fe/PVF. Se concluye que los dos complejos magnéticos estudiados representan promisorias herramientas para mejorar la eficiencia en la terapia génica en células cerebrales.Fil: Pardo, Joaquin. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - la Plata. Instituto de Invest.bioquimicas de la Plata;Fil: Sosa, Yolanda Elena. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - la Plata. Instituto de Invest.bioquimicas de la Plata;Fil: Reggiani, Paula Cecilia. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - la Plata. Instituto de Invest.bioquimicas de la Plata;Fil: Arciniegas, M. I.. Universidad Nacional de la Plata. Facultad de Ciencias Exactas; Argentina;Fil: Sanchez, Francisco Homero. Universidad Nacional de la Plata. Facultad de Ciencias Exactas; Argentina;Fil: Goya, Rodolfo Gustavo. Consejo Nacional de Invest.cientif.y Tecnicas. Centro Cientifico Tecnol.conicet - la Plata. Instituto de Invest.bioquimicas de la Plata

Fuzzy-based forest fire prevention and detection by wireless sensor networks

Forest fires may cause considerable damages both in ecosystems and lives. This proposal describes the application of Internet of Things and wireless sensor networks jointly with multi-hop routing through a real time and dynamic monitoring system for forest fire prevention. It is based on gathering and analyzing information related to meteorological conditions, concentrations of polluting gases and oxygen level around particular interesting forest areas. Unusual measurements of these environmental variables may help to prevent wildfire incidents and make their detection more efficient. A forest fire risk controller based on fuzzy logic has been implemented in order to activate environmental risk alerts through a Web service and a mobile application. For this purpose, security mechanisms have been proposed for ensuring integrity and confidentiality in the transmission of measured environmental information. Lamport's signature and a block cipher algorithm are used to achieve this objective

Insulin-like growth factor-I gene therapy reverses morphologic changes and reduces hyperprolactinemia in experimental rat prolactinomas

<p>Abstract</p> <p>Background</p> <p>The implementation of gene therapy for the treatment of pituitary tumors emerges as a promising complement to surgery and may have distinct advantages over radiotherapy for this type of tumors. Up to now, suicide gene therapy has been the main experimental approach explored to treat experimental pituitary tumors. In the present study we assessed the effectiveness of insulin-like growth factor I (IGF-I) gene therapy for the treatment of estrogen-induced prolactinomas in rats.</p> <p>Results</p> <p>Female Sprague Dawley rats were subcutaneously implanted with silastic capsules filled with 17-β estradiol (E₂) in order to induce pituitary prolactinomas. Blood samples were taken at regular intervals in order to measure serum prolactin (PRL). As expected, serum PRL increased progressively and 23 days after implanting the E₂capsules (Experimental day 0), circulating PRL had undergone a 3–4 fold increase. On Experimental day 0 part of the E₂-implanted animals received a bilateral intrapituitary injection of either an adenoviral vector expressing the gene for rat IGF-I (RAd-IGFI), or a vector (RAd-GFP) expressing the gene for green fluorescent protein (GFP). Seven days post vector injection all animals were sacrificed and their pituitaries morphometrically analyzed to evaluate changes in the lactotroph population. RAd-IGFI but not RAd-GFP, induced a significant fall in serum PRL. Furthermore, RAd-IGFI but not RAd-GFP significantly reversed the increase in lactotroph size (CS) and volume density (VD) induced by E₂treatment.</p> <p>Conclusion</p> <p>We conclude that IGF-I gene therapy constitutes a potentially useful intervention for the treatment of prolactinomas and that bioactive peptide gene delivery may open novel therapeutic avenues for the treatment of pituitary tumors.</p

Space-time defects and teleparallelism

We consider the class of space-time defects investigated by Puntigam and Soleng. These defects describe space-time dislocations and disclinations (cosmic strings), and are in close correspondence to the actual defects that arise in crystals and metals. It is known that in such materials dislocations and disclinations require a small and large amount of energy, respectively, to be created. The present analysis is carried out in the context of the teleparallel equivalent of general relativity (TEGR). We evaluate the gravitational energy of these space-time defects in the framework of the TEGR and find that there is an analogy between defects in space-time and in continuum material systems: the total gravitational energy of space-time dislocations and disclinations (considered as idealized defects) is zero and infinit, respectively.Comment: 22 pages, no figures, to appear in the Class. Quantum Gravit

Transdiagnostic commonalities and differences in resting state functional connectivity of the default mode network in schizophrenia and major depression

Schizophrenia and depression are prevalent psychiatric disorders, but their underlying neural bases remains poorly understood. Neuroimaging evidence has pointed towards the relevance of functional connectivity aberrations in defaultmode network (DMN) hubs, dorso-medial prefrontal cortex and precuneus, in both disorders, but commonalities and differences in resting state functional connectivity of those two regions across disorders has not been formally assessed. Here, we took a transdiagnostic approach to investigate resting state functional connectivity of those two regions in 75 patients with schizophrenia and 82 controls from 4 scanning sites and 102 patients with depression and 106 controls from 3 sites. Our results demonstrate common dysconnectivity patterns as indexed by a significant reduction of functional connectivity between precuneus and bilateral superior parietal lobe in schizophrenia and depression. Furthermore, our findings highlight diagnosis-specific connectivity reductions of the parietal operculum in schizophrenia relative to depression. In light of evidence that points towards the importance of the DMN for social cognitive abilities and well documented impairments of social interaction in both patient groups, it is conceivable that the observed transdiagnostic connectivity alterations may contribute to interpersonal difficulties, but this could not be assessed directly in our study as measures of social behavior were not available. Given the operculum's role in somatosensory integration, diagnosis-specific connectivity reductions may indicate a pathophysiological mechanism for basic self-disturbances that is characteristic of schizophrenia, but not depression. (C) 2015 The Authors. Published by Elsevier Inc

PTTG expression in different experimental and human prolactinomas in relation to dopaminergic control of lactotropes

Background: Pituitary tumor transforming gene (pttg) is a novel oncogene that is expressed at higher level in most of the tumors analyzed to date compared to normal tissues. Nevertheless, its expression in prolactinomas and its relation with the pituitary dopamine receptor 2 (D2R) are not well defined. We sought to determine the pituitary level of pttg in three different experimental models of prolactinomas with altered dopaminergic control of the pituitary: the dopaminergic D2R knockout female mouse, the estrogen-treated rat, and the senescent female rat. These three models shared the characteristics of increased pituitary weight, hyperprolactinemia, lactotrope hyperplasia and reduced or absent dopaminergic action at the pituitary level. We also studied samples from human macroprolactinomas, which were characterized as responsive or resistant to dopamine agonist therapy. Results: When compared to female wild-type mice, pituitaries from female D2R knockout mice had decreased PTTG concentration, while no difference in pttg mRNA level was found. In senescent rats no difference in pituitary PTTG protein expression was found when compared to young rats. But, in young female rats treated with a synthetic estrogen (Diethylstylbestrol, 20 mg) PTTG protein expression was enhanced (P = 0.029). Therefore, in the three experimental models of prolactinomas, pituitary size was increased and there was hyperprolactinemia, but PTTG levels followed different patterns. Patients with macroprolactinomas were divided in those in which dopaminergic therapy normalized or failed to normalize prolactin levels (responsive and resistant, respectively). When pituitary pttg mRNA level was analyzed in these macroprolactinomas, no differences were found. We next analyzed estrogen action at the pituitary by measuring pituitary estrogen receptor α levels. The D2R knockout female mice have low estrogen levels and in accordance, pituitary estrogen receptors were increased (P = 0.047). On the other hand, in senescent rats estrogen levels were slightly though not significantly higher, and estrogen receptors were similar between groups. The estrogen-treated rats had high pharmacological levels of the synthetic estrogen, and estrogen receptors were markedly lower than in controls (P < 0.0001). Finally, in patients with dopamine resistant or responsive prolactinomas no significant differences in estrogen receptor α levels were found. Therefore, pituitary PTTG was increased only if estrogen action was increased, which correlated with a decrease in pituitary estrogen receptor level. Conclusion: We conclude that PTTG does not correlate with prolactin levels or tumor size in animal models of prolactinoma, and its pituitary content is not related to a decrease in dopaminergic control of the lactotrope, but may be influenced by estrogen action at the pituitary level. Therefore it is increased only in prolactinomas generated by estrogen treatment, and not in prolactinomas arising from deficient dopamine control, or in dopamine resistant compared with dopamine responsive human prolactinomas. These results are important in the search for reliable prognostic indicators for patients with pituitary adenomas which will make tumor-specific therapy possible, and help to elucidate the poorly understood phenomenon of pituitary tumorigenesis.Facultad de Ciencias Médica

Objectively measured physical activity, sedentary behaviour and all-cause mortality in older men: does volume of activity matter more than pattern of accumulation?

OBJECTIVES: To understand how device-measured sedentary behaviour and physical activity are related to all-cause mortality in older men, an age group with high levels of inactivity and sedentary behaviour. METHODS: Prospective population-based cohort study of men recruited from 24 UK General Practices in 1978-1980. In 2010-2012, 3137 surviving men were invited to a follow-up, 1655 (aged 71-92 years) agreed. Nurses measured height and weight, men completed health and demographic questionnaires and wore an ActiGraph GT3x accelerometer. All-cause mortality was collected through National Health Service central registers up to 1 June 2016. RESULTS: After median 5.0 years' follow-up, 194 deaths occurred in 1181 men without pre-existing cardiovascular disease. For each additional 30 min in sedentary behaviour, or light physical activity (LIPA), or 10 min in moderate to vigorous physical activity (MVPA), HRs for mortality were 1.17 (95% CI 1.10 to 1.25), 0.83 (95% CI 0.77 to 0.90) and 0.90 (95% CI 0.84 to 0.96), respectively. Adjustments for confounders did not meaningfully change estimates. Only LIPA remained significant on mutual adjustment for all intensities. The HR for accumulating 150 min MVPA/week in sporadic minutes (achieved by 66% of men) was 0.59 (95% CI 0.43 to 0.81) and 0.58 (95% CI 0.33 to 1.00) for accumulating 150 min MVPA/week in bouts lasting ≥10 min (achieved by 16% of men). Sedentary breaks were not associated with mortality. CONCLUSIONS: In older men, all activities (of light intensity upwards) were beneficial and accumulation of activity in bouts ≥10 min did not appear important beyond total volume of activity. Findings can inform physical activity guidelines for older adults

Application of six detection methods for analysis of paralytic shellfish toxins in shellfish from four regions within Latin America

With the move away from use of mouse bioassay (MBA) to test bivalve mollusc shellfish for paralytic shellfish poisoning (PSP) toxins, countries around the world are having to adopt non-animal-based alternatives that fulfil ethical and legal requirements. Various assays have been developed which have been subjected to single-laboratory and multi-laboratory validation studies, gaining acceptance as official methods of analysis and approval for use in some countries as official control testing methods. The majority of validation studies conducted to date do not, however, incorporate shellfish species sourced from Latin America. Consequently, this study sought to investigate the performance of five alternative PSP testing methods together with the MBA, comparing the PSP toxin data generated both qualitatively and quantitatively. The methods included a receptor binding assay (RBA), two liquid chromatography with fluorescence detection (LC-FLD) methods including both pre-column and post-column oxidation, liquid chromatography with tandem mass spectrometry (LC-MS/MS) and a commercial lateral flow assay (LFA) from Scotia. A total of three hundred and forty-nine shellfish samples from Argentina, Mexico, Chile and Uruguay were assessed. For the majority of samples, qualitative results compared well between methods. Good statistical correlations were demonstrated between the majority of quantitative results, with a notably excellent correlation between the current EU reference method using pre-column oxidation LC-FLD and LC-MS/MS. The LFA showed great potential for qualitative determination of PSP toxins, although the findings of high numbers of false-positive results and two false negatives highlighted that some caution is still needed when interpreting results. This study demonstrated that effective replacement methods are available for countries that no longer wish to use the MBA, but highlighted the importance of comparing toxin data from the replacement method using local shellfish species of concern before implementing new methods in official control testing programs