Lentivirus-mediated transgene delivery to the hippocampus reveals sub-field specific differences in expression

<p>Abstract</p> <p>Background</p> <p>In the adult hippocampus, the granule cell layer of the dentate gyrus is a heterogeneous structure formed by neurons of different ages, morphologies and electrophysiological properties. Retroviral vectors have been extensively used to transduce cells of the granule cell layer and study their inherent properties in an intact brain environment. In addition, lentivirus-based vectors have been used to deliver transgenes to replicative and non-replicative cells as well, such as post mitotic neurons of the CNS. However, only few studies have been dedicated to address the applicability of these widespread used vectors to hippocampal cells in vivo. Therefore, the aim of this study was to extensively characterize the cell types that are effectively transduced in vivo by VSVg-pseudotyped lentivirus-based vectors in the hippocampus dentate gyrus.</p> <p>Results</p> <p>In the present study we used Vesicular Stomatitis Virus G glycoprotein-pseudotyped lentivirual vectors to express EGFP from three different promoters in the mouse hippocampus. In contrast to lentiviral transduction of pyramidal cells in CA1, we identified sub-region specific differences in transgene expression in the granule cell layer of the dentate gyrus. Furthermore, we characterized the cell types transduced by these lentiviral vectors, showing that they target primarily neuronal progenitor cells and immature neurons present in the sub-granular zone and more immature layers of the granule cell layer.</p> <p>Conclusion</p> <p>Our observations suggest the existence of intrinsic differences in the permissiveness to lentiviral transduction among various hippocampal cell types. In particular, we show for the first time that mature neurons of the granule cell layer do not express lentivirus-delivered transgenes, despite successful expression in other hippocampal cell types. Therefore, amongst hippocampal granule cells, only adult-generated neurons are target for lentivirus-mediated transgene delivery. These properties make lentiviral vectors excellent systems for overexpression or knockdown of genes in neuronal progenitor cells, immature neurons and adult-generated neurons of the mouse hippocampus in vivo.</p

Ultrasound enhanced prehospital thrombolysis using microbubbles infusion in patients with acute ST elevation myocardial infarction: Rationale and design of the Sonolysis study

Contains fulltext : 70525.pdf (publisher's version ) (Open Access)ABSTRACT: BACKGROUND -: Experimental studies have shown that ultrasound contrast agents enhance the effectiveness of thrombolytic agents in the presence of ultrasound in vitro and in vivo. Recently, we have launched a clinical pilot study, called "Sonolysis", to study this effect in patients with ST-elevation myocardial infarction based on proximal lesions of the infarct-related artery. METHODS/DESIGN -: In our multicenter, randomized, placebo controlled clinical trial we will include patients between 18 and 80 years of age with their first ST-elevation myocardial infarction based on a proximal lesion of the infarct-related artery. After receiving a single bolus alteplase 50 mg IV (Actilyse(R) Boehringer Ingelheim GmbH), a loading dose of aspirin 500 mg, and heparin 5000 IU in the ambulance according to the prehospital thrombolysis protocol, patients, following oral informed consent, are randomized to undergo 15 minutes of pulsatile ultrasound with intravenous administration of ultrasound contrast agent or placebo without ultrasound. Afterwards coronary angiography and, if indicated, percutaneous coronary intervention will take place. A total of 60 patients will be enrolled in approximately 1 year.The primary endpoints are based on the coronary angiogram and consist of TIMI flow, corrected TIMI frame count, and myocardial blush grade. Follow-up includes 12-lead ECG, 2D-echocardiography, cardiac MRI, and enzyme markers to obtain our secondary endpoints, including the infarct size, wall motion abnormalities, and the global left ventricular function. DISCUSSION -: The Sonolysis study is the first multicenter, randomized, placebo controlled clinical trial investigating the therapeutic application of ultrasound and microbubbles in acute ST-elevation myocardial infarction patients. A positive finding may stimulate further research and technical innovations to implement the treatment in the ambulance and maybe obtain even more patency at an earlier stage. TRIAL REGISTRATION -: Trialregister NTR161

ERK-regulated double cortin-like kinase (DCLK)-short phosphorylation and nuclear translocation stimulate POMC gene expression in endocrine melanotrope cells.

Contains fulltext : 96838.pdf (publisher's version ) (Open Access)10 p

The microtubule associated protein doublecortin-like regulates the transport of the glucocorticoid receptor in neuronal progenitor cells

Contains fulltext : 72489.pdf (publisher's version ) (Open Access

Combined comprehensive two-dimensional gas chromatographic analysis of PAH/PASH compounds in complex matrices

A new gas chromatographic method has been developed that is able to quantify polycyclic aromatic hydrocarbons (PAH) and polycyclic aromatic sulfur-containing hydrocarbons (PASH) up to four rings. The method combines the power of both flame ionization detection (FID) and sulfur chemiluminescence detection (SCD) in series on a single comprehensive two-dimensional gas chromatography (GC x GC) system and provides mass fractions of compounds separated by carbon number n (CnHxSy) and class. In addition to PAH and PASH separation, the method is extended toward nonaromatic and monoaromatic (sulfur-containing) compounds (paraffins, naphthenes, monoaromatics, thiols, sulfides, disulfides, and thiophenes). The 95% confidence interval is doubled when a single injection technique is used instead of a more-accurate double injection technique. A flexible correction procedure that combines the advantages of the two-dimensional separation of GC x GC and its ability to easily define overlapping groups between the FID and the SCD chromatograms is applied. The method is validated using theoretical reference mixtures and is applied on three commercial gas oils with sulfur content from 0.16 wt% up to 1.34 wt%. The repeatability is good, with an average of 3.4%, which is in the same range as the much more expensive Fourier transform ion cyclotron resonancemass spectroscopy (FTICR-MS) technique