Phenothiazine normalizes the NADH/NAD+ ratio, maintains mitochondrial integrity and protects the nigrostriatal dopamine system in a chronic rotenone model of Parkinson's disease

Producción CientíficaImpaired mitochondrial function has been associated with the etiopathogenesis of Parkinson's disease (PD). Sustained inhibition of complex I produces mitochondrial dysfunction, which is related to oxidative injury and nigrostriatal dopamine (DA) neurodegeneration. This study aimed to identify disease-modifying treatments for PD. Unsubstituted phenothiazine (PTZ) is a small and uncharged aromatic imine that readily crosses the blood-brain barrier. PTZ lacks significant DA receptor-binding activity and, in the nanomolar range, exhibits protective effects via its potent free radical scavenging and anti-inflammatory activities. Given that DAergic neurons are highly vulnerable to oxidative damage and inflammation, we hypothesized that administration of PTZ might confer neuroprotection in different experimental models of PD. Our findings showed that PTZ rescues rotenone (ROT) toxicity in primary ventral midbrain neuronal cultures by preserving neuronal integrity and reducing protein thiol oxidation. Long-term treatment with PTZ improved animal weight, survival rate, and behavioral deficits in ROT-lesioned rats. PTZ protected DA content and fiber density in the striatum and DA neurons in the SN against the deleterious effects of ROT. Mitochondrial dysfunction, axonal impairment, oxidative insult, and inflammatory response were attenuated with PTZ therapy. Furthermore, we have provided a new insight into the molecular mechanism underlying the neuroprotective effects of PTZ

Isotope-reinforced polyunsaturated fatty acids improve Parkinson’s disease-like phenotype in rats overexpressing α-synuclein

Producción CientíficaLipid peroxidation is a key to a portfolio of neurodegenerative diseases and plays a central role in α-synuclein (α-syn) toxicity, mitochondrial dysfunction and neuronal death, all key processes in the pathogenesis of Parkinson's disease (PD). Polyunsaturated fatty acids (PUFAs) are important constituents of the synaptic and mitochondrial membranes and are often the first molecular targets attacked by reactive oxygen species (ROS). The rate-limiting step of the chain reaction of ROS-initiated PUFAs autoxidation involves hydrogen abstraction at bis-allylic sites, which can be slowed down if hydrogens are replaced with deuteriums. In this study, we show that targeted overexpression of human A53T α-syn using an AAV vector unilaterally in the rat substantia nigra reproduces some of pathological features seen in PD patients. Chronic dietary supplementation with deuterated PUFAs (D-PUFAs), specifically 0.8% D-linoleic and 0.3% H-linolenic, produced significant disease-modifying beneficial effects against α-syn-induced motor deficits, synaptic pathology, oxidative damage, mitochondrial dysfunction, disrupted trafficking along axons, inflammation and DA neuronal loss. These findings support the clinical evaluation of D-PUFAs as a neuroprotective therapy for PD

Determinación de una técnica de referencia para la susceptibilidad antimicrobiana de bacterias anaeróbicas de importancia clínica

The selection of antibiotics for the treatment of anaerobic infections is generally based on purely empirical criteria, with antimicrobial sensitivity tests being carried out only in reference centres. For this reason, it is important to investigate a technique appropnate for use in less specialized clinical bacteriology laboratories. In this study, four antimicrobial sensitivity tests (elution in a broth disk, macrodilution, rnicrodilution and dilution in agar) were carried out using four reference strains with known sensitivity patterns. The reproducibility, concordance, speed and ease of use of each test was compared. The results obtained showed that the most reproducibletest was dilution in agar, followed by microdilution, macrodilution and elution. In terms of concordance the best test was dilution in agar, followed by macrodilution, elution and microdilution. The most complex test was dilution in agar, followed by microdilution, macrodilution and elution. As this study did not find any single test which, aione, proved better than any other in all the criteria analized, we recommend that a combination of at least two tests be used in the study of antimicrobial susceptibility of anaerobes.La selección de antibióticos para el tratamiento de infecciones por microorganismos anaeróbicos se hace en forma empírica en la mayoría de los casos; por tanto, el empleo de técnicas para determinar la susceptibilidad antimicrobiana es controvertido y sólo se realiza en centros de referencia. Con la aparición de cepas resistentes a los antimicrobianos de elección, es necesario determinar entre las técnicas estandarizadas una que pueda ser realizada en un laboratorio con cierta infraestructura para el trabajo en bacteriología clínica. Se realizaron cuatro pruebas estandarizadas por el NCCLS: elución de disco en caldo, macrodilución, microdilución y dilución en agar, empleando cuatro cepas de referencia. Los datos obtenidos fueron analizados para determinar la reproducibilidad, la concordancia y la complejidad de las pruebas. Empleando los criterios analizados, se obtuvo en orden descendente: en reproducibilidad, la dilución en agar, microdilución, macrodilución y elución; en concordancia. la dilución en agar, macrodilución, elución y microdilución; y en complejidad, dilución en agar, microdilución, macrodilución y elución. Debido a la variedad de los resultados obtenidos no se pudo recomendar una sola técnica para la determinación de la susceptibilidad de microorganismos anaeróbicos de importancia clínica; en algunos casos y teniendo en cuenta el microorganismo y el antibiótico a probar, la realización de por lo menos dos de ellas permitiría esa determinación

ABCA1 Deficiency Affects Basal Cognitive Deficits and Dendritic Density in Mice

Producción CientíficaATP-binding cassette transporter A1 (ABCA1) mediates cholesterol efflux to lipid-free apolipoproteins and regulates the generation of high density lipoproteins. Previously, we have shown that lack of Abca1 significantly increases amyloid deposition and cognitive deficits in Alzheimer’s disease model mice expressing human amyloid-β protein precursor (APP). The goal of this study was to determine if ABCA1 plays a role in memory deficits caused by amyloid-β (Aβ) oligomers and examine neurite architecture of pyramidal hippocampal neurons. Our results confirm previous findings that Abca1 deficiency significantly impairs spatial memory acquisition and retention in the Morris water maze and long-term memory in novel object recognition of APP transgenic mice at a stage of early amyloid pathology. Neither test demonstrated a significant difference between Abca1ko and wild-type (WT) mice. We also examined the effect of intra-hippocampal infused Aβ oligomers on cognitive performance of Abca1ko mice, compared to control infusion of scrambled Aβ peptide. Age-matched WT mice undergoing the same infusions were also used as controls. In this model system, we found a statistically significant difference between WT and Abca1ko mice infused with scrambled Aβ, suggesting that Abca1ko mice are vulnerable to the effect of mild stresses. Moreover, examination of neurite architecture in the hippocampi revealed a significant decrease in neurite length, number of neurite segments, and branches in Abca1ko mice when compared to WT mice. We conclude that mice lacking ABCA1 have basal cognitive deficits that prevent them from coping with additional stressors, which is in part due to impairment of neurite morphology in the hippocampus

Performance evaluation of a new gamma imager for small animal SPECT applications

Proceeding of: 2007 IEEE Nuclear Science Symposium Conference Record (NSS '07), Honolulu, Hawaii, USA, Oct. 27 - Nov. 3, 2007Abstract–In this work we characterized a recently developed gamma imager for small animal SPECT applications. The Hamamatsu C9177 is a mini-gamma camera that integrates the detector and all the electronics, including the acquisition system, in a compact and portable housing. The detector is based on a high resolution parallel hole collimator, a CsI(NaI) crystal array and a PS-PMT (flat panel type). The active field of view is 41.9 x 41.9 mm2 and the assembly is optimized for 60 to 200 keV. The electronics in the housing includes the high voltage divider, an ASIC which converts the 64 anodes into Anger-like signals, ADCs which are fed with these signals and position and energy lookup tables which allow digital information for each detected photon to be obtained directly from the imager. In order to be able to obtain tomographic data and to improve the measurement protocols, we mounted the detector in a custom-built motorized gantry. We evaluated detector uniformity and energy resolution using a flood field image. Planar intrinsic spatial resolution and spatial linearity were assessed by stepping a capillary source across the detector surface and plotting the count profile for each individual crystal of the array. We additionally performed phantom studies to preliminarily characterize the tomographic performance. Energy resolution is 11.6% (mean), sensitivity is 2.54 cps/μCi and planar spatial resolution is 2.4 mm (CFOV 20% energy window) when the source is placed on the detector surface.This work was supported in part by the Spanish Ministerio de Educación y Ciencia under Grant No. TEC2004-07052-C02, Comunidad de Madrid Grant No. GR/SAL/024104, and the CDTEAM project of the CENIT program (Spanish Ministerio de Industria)

A super- resolution feasibility study in small-animal SPECT imaging

Proceeding of: 2008 IEEE Nuclear Science Symposium Conference Record (NSS '08), Dresden, Germany, 19-25 Oct. 2008Lack of spatial resolution is a major drawback in small-animal SPECT imaging, particularly when parallel hole collimators are employed. This work evaluates the feasibility of enhancing the spatial resolution of a small-animal SPECT scanner by applying an approach based on a super-resolution technique combined with dedicated hardware. The detectors are based on PS-PMTs (Hamamatsu, H8500), NaI(TI) crystal arrays and parallel hole collimators. The system enables the mechanical shifting of the gamma-cameras and the object under study during the acquisition in steps smaller than the detector intrinsic sampling pitch. This fact allows using a super-resolution technique without the necessity of blind movement estimators between projections. In the absence of noise the algorithm converged to the original image, and with real noisy data it achieved improvements in resolution between 28%(tomographic) and 42% (planar) in phantom studies.This work is partially funded by the CD-TEAM Project, CENIT Program, Spanish Ministerio de Industria, and with grants from the Ministerio de Educaci6n y Ciencia, projects TEC2007-64731 and TEC2008-06715-C02-01

Performance evaluation of SiPM detectors for PET imaging in the presence of magnetic fields

Proceeding of: 2008 IEEE Nuclear Science Symposium Conference Record (NSS '08), Dresden, Germany, 19-25 Oct. 2008The multi-pixel photon counter (MPPC) or silicon photo-multiplier (SiPM), recently introduced as a solid-state photodetector, consists of an array of Geiger-mode photodiodes(microcells). is a promising device for PET thanks to its potential for high photon detection efficiency (PDE) and immunity to high magnetic fields. is also very easy to use, with simple electronic read-out, high gain and small size. In this work we evaluate the performance of three 1 x 1 mm2 and one 6 x 6 mm2 (2 x 2 array) SiPMs offered by Hamamatsu for their use in PET. We examine the dependence of the energy resolution and the gain of these devices on the thermal and reverse bias when coupled to LYSO scintillator crystals. We find that the 400 and 1600 microcells models and the 2 x 2 array are suitable for small size crystals, like those employed in high resolution small animal scanners. The good performance of these devices up to 7 Tesla has also been confirmed.This work was supported in part by the MEC (FPA2007-07393), CDTEAM (CENIT-Ingenio 2010) Ministerio de Industria, Spain, UCM (Grupos UCM: 910059), CPAN (ConsoliderIngenio 2010) CSPD-2007-00042 projects, and the RECAVA-RETIC network

Geoarqueología y paleontología de los depósitos de Pleistoceno Superior del antiguo Arroyo Abroñigal (Cuenca de Manzanares, Madrid): el yacimiento de Puente de los Tres Ojos

La excavación arqueológica del yacimiento del Puente de los Tres Ojos, próximo a la calle Cerro Negro (Madrid), ha aportado nuevos datos para profundizar en el conocimiento de la ocupación humana y en la reconstrucción paleoambiental del valle del antiguo arroyo Abroñigal, cuyo curso fluvial funcionó como afluente del río Manzanares posiblemente desde antes del Pleistoceno Superior hasta la segunda mitad del siglo XX. En el presente artículo se incluyen los aspectos geomorfológicos y cronoestratigráficos de los niveles excavados, documentándose varias secuencias fluviales y aluviales de relleno de la margen derecha del fondo de valle del Abroñigal en su tramo inferior. Durante la excavación se registró un amplio conjunto lítico, formado en su mayoría por piezas recuperadas en niveles de arenas y gravas correspondientes a episodios fluviales de media energía, a las que hay que añadir un porcentaje menor localizado en niveles de limos arcillosos y arenas finas. La mayor parte del conjunto lítico responde a sistemas técnicos propios del Paleolítico Medio, aunque destaca la presencia testimonial de piezas del Paleolítico Superior, además de restos de fauna de mamíferos correspondientes al Pleistoceno Superior, todo ello en niveles datados por OSL entre 14.400 y 11.170 años BP aproximadament

Variabilidad intraespecífica de la respuesta electroantenográfica en Cerambyx welensii Küster (Coleoptera, Cerambycidae). Influencia de factores anatómicos, fisiológicos y experimentales

Intraspecific variability of EAG responses in Cerambyx welensii has been studied.We have not found significant differences of EAG responses neither between sexes nor types of solvent, paraffin oil and hexane, but there was a highly significant variability of responses between individuals (0,92 mV +/- 0,6. Mean +/- DT) to (E)-2-hexenal (50% v/v). More than 68% of individual variability can be explained by the combination of the previous aptitude of insects, its physiological state (vitality at the moment of study) and the capillary's diameter in the reference electrode. Anatomical features of the individuals had not influence in the response. In order to avoid or minimize the effect caused for this variability, it is advisable to carry out EAG tests with insects obtained under controlled conditions. In other case, the EAG responses must be standardized using a standard stimulus, as well as to homogenize the tip diameter of the reference electrode.En este trabajo se estudia la variabilidad intraespecífica de la respuesta electroantenográfica de Cerambyx welensii. No se han encontrado diferencias de respuestas entre sexos, ni entre el tipo de solvente empleado, hexano o aceite de parafina; pero sí se encontró variabilidad entre individuos (0,92 mV +/- 0,6. Media +/- DT) frente al (E)-2-hexenal (50% v/v). Más del 68% de esta variabilidad pudo ser explicada considerando tres factores: la aptitud previa de los insectos, su estado fisiológico (vigor en el momento del estudio) y el diámetro en punta fina del capilar del electrodo de referencia. Las características anatómicas de los individuos no influyeron sobre su respuesta electroantenográfica. La variabilidad de respuesta intraespecífica encontrada en C. welensii puede distorsionar la caracterización del comportamiento olfativo de esta especie, por lo que es deseable realizar las pruebas electroantenográficas a partir de insectos obtenidos en condiciones controladas o bien estandarizar las respuestas a partir de un estímulo estándar, además de normalizar el diámetro del capilar del electrodo de referencia

Assessment of a New High-Performance Small-Animal X-Ray Tomograph

We have developed a new X-ray cone-beam tomograph for in vivo small-animal imaging using a flat panel detector (CMOS technology with a microcolumnar CsI scintillator plate) and a microfocus X-ray source. The geometrical configuration was designed to achieve a spatial resolution of about 12 lpmm with a field of view appropriate for laboratory rodents. In order to achieve high performance with regard to per-animal screening time and cost, the acquisition software takes advantage of the highest frame rate of the detector and performs on-the-fly corrections on the detector raw data. These corrections include geometrical misalignments, sensor non-uniformities, and defective elements. The resulting image is then converted to attenuation values. We measured detector modulation transfer function (MTF), detector stability, system resolution, quality of the reconstructed tomographic images and radiated dose. The system resolution was measured following the standard test method ASTM E 1695 -95. For image quality evaluation, we assessed signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) as a function of the radiated dose. Dose studies for different imaging protocols were performed by introducing TLD dosimeters in representative organs of euthanized laboratory rats. Noise figure, measured as standard deviation, was 50 HU for a dose of 10 cGy. Effective dose with standard research protocols is below 200 mGy, confirming that the system is appropriate for in vivo imaging. Maximum spatial resolution achieved was better than 50 micron. Our experimental results obtained with image quality phantoms as well as with in-vivo studies show that the proposed configuration based on a CMOS flat panel detector and a small micro-focus X-ray tube leads to a compact design that provides good image quality and low radiated dose, and it could be used as an add-on for existing PET or SPECT scannersIEEE Nuclear and Plasma Sciences SocietyPublicad