21 research outputs found

    Structure alteration of a sandy-clay soil by biochar amendments

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    The aim of the present study was to investigate structure alterations of a sandy-clay soil upon addition of different amounts of biochar (f (bc) ). All the f (bc) samples were analyzed by high energy moisture characteristic (HEMC) technique and H-1 nuclear magnetic resonance (NMR) relaxometry. HEMC was applied in order to evaluate aggregate stability of biochar-amended soil samples. H-1 NMR relaxometry experiments were conducted for the evaluation of the pore distributions through the investigation of water dynamics of the same samples. The HEMC technique revealed improvement in aggregate stability through measurements of the amount of drainable pores and the stability ratio. The latter increased as the amount of biochar was raised up. The H-1 NMR relaxometry revealed a unimodal T (1) distribution for both the sole sandy-clay soil and the biochar. Conversely, a bimodal T (1) distribution was acquired for all the different f (bc) samples. Improvement in aggregate stability was obtained as biochar was progressively added to the sandy-clay soil. A dual mechanism of water retention has been hypothesized. In particular, intra-aggregate porosity was indicated as the main responsible for molecular water diffusion when f (bc) comprised between 0 and 0.33. Conversely, inter-aggregate porosity resulted predominant, through swelling processes, when f (bc) overcame 0.33

    321. Sea Urchin sns Chromatin Insulator Prevents Silencing and Positional Effect Variegation of Oncoretroviral Vectors Transgene Expression in Murine Erythroid Cell Line

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    Silencing and position effect are considered significant obstacles to obtain a consistent level of transgene expression in viral gene therapy. Furthermore recent studies had shown that retroviruses tend to land on active genes with the potential consequence of insertional mutagenesis. The inclusion of elements, such as chromatin insulators, capable to insulate a gene from the surrounding chromatin effects at the integration site should improve both efficacy and safety of gene therapy vectors. We have previously characterized a 265 bp insulator element, termed sns, localized at the 3' end of the early histone H2A gene of the sea urchin Paracentrotus lividus. This sequence contains three cis-acting elements (Box A, Box B, and Box C+T) all needed for the enhancer blocking activity in both sea urchin and human cells. By colony assays, in human (K562) and mouse (Mel) erythroid cell lines, we have recently demonstrated that the sns insulator displays directional enhancer-blocking activity in that it interferes with the communication between the human beta-globin enhancer (LCR) and the gamma-globin promoter. By electrophoretic mobility shift assays (EMSA) we found bindings of sns insulator with the erythroid specific GATA1 and the ubiquitous Oct1, and Sp1 transcription factors

    Dysembryoplastic Neuroepithelial Tumor of the brainstem

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    Dysembryoplastic neuroepithelial tumor (DNT) is a clinically benign stable lesion, most frequently located in the temporal and frontal lobes, often responsible for epilepsy in young adults. We describe an unusual case of DNT in the brainstem of a 45-year-old woman. Brain MRI showed a multicystic-like lesion localized in the left inferior pons, involving the ipsilateral cerebellar peduncle and partially dislocating the fourth ventricle. The specific pattern of MRI and CT appearance of DNT and its benign course (our patient is clinically stable with unchanged MRI images at two year follow-up) may help differentiate this tumor from other lesions, i.e. gangliogliomas and glioneural malformations

    Development of plasmids for quantitative detection of integrated lentiviral vectors and evaluation of culture time to perform vector titer by real-time quantitative polymerase chain reaction assay

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    The accurate assessment of provirus copy number per cell (VCN/cell) is a fundamental issue in transgenesis as well as in gene therapy studies based on stably integrated vectors. To this end, real-time quantitative polymerase chain reaction (qPCR) is a powerful method but it is sensible to differences in quality or concentration of the two-plasmid preparations used for the construction of the standard curves. In order to minimize technical errors we included genome specific sequences (mouse or human) and vector specific sequences in the same plasmid. We evaluated the specificity and sensitivity of these bivalent plasmids by qPCR analysis on mouse and human genomic DNA containing a known number of a reporter lentiviral vector and we found that the system is reliable to measure up to 0.1 VCN/cell. Here we have applied this assay to measure vector titer of virus stock preparations and to determine the optimal cell passages at which viral titration effectively reflects the number of integrated vectors

    Development of plasmids for quantitative detection of integrated lentiviral vectors and evaluation of culture time to perform vector titer by real-time quantitative polymerase chain reaction assay

    No full text
    The accurate assessment of provirus copy number per cell (VCN/cell) is a fundamental issue in transgenesis as well as in gene therapy studies based on stably integrated vectors. To this end, real-time quantitative polymerase chain reaction (qPCR) is a powerful method but it is sensible to differences in quality or concentration of the two-plasmid preparations used for the construction of the standard curves. In order to minimize technical errors we included genome specific sequences (mouse or human) and vector specific sequences in the same plasmid. We evaluated the specificity and sensitivity of these bivalent plasmids by qPCR analysis on mouse and human genomic DNA containing a known number of a reporter lentiviral vector and we found that the system is reliable to measure up to 0.1 VCN/cell. Here we have applied this assay to measure vector titer of virus stock preparations and to determine the optimal cell passages at which viral titration effectively reflects the number of integrated vectors

    REM sleep behavior disorder in a patient with frontotemporal dementia

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    We describe a patient with frontotemporal dementia (FTD), a tauopathy, who also showed clinical and polysomnographic features of REM sleep behavior disorder (RBD). The patient is a 78-year-old male with a 1 year history of behavioral dysfunction involving emotion, character and social functioning. Brain imaging and the results of neuropsychological testing were consistent with a diagnosis of FTD. Sleep symptom onset occurred some years before the behavioral changes, and consisted of unpleasant dreams, vocalization, and prominent motor behaviors. A polysomnography confirmed the diagnosis of RBD. Our findings support the hypothesis that RBD, although more frequent in synucleinopathies, might be a pathological stage in the development of almost every neurodegenerative disorder in which the pathological process involves the cerebral structures that regulate muscle tone during REM slee

    Cognitive impairment in Behcet’s disease patients without overt neurological involvement

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    We investigated the prevalence of cognitive impairment in patients with Behc¸et’s disease (BD) without overt neurological involvement. The influence of disease duration, disease activity, prednisone dosage, and anxiety and depression levels was evaluated. Twenty-six consecutive BD outpatients and 26 healthy controls matched for age, education and sex completed a comprehensive neuropsychological battery including tests of memory, visuospatial and constructional abilities, language, attention and psychomotor speed, non-verbal reasoning and executive functioning. The Hamilton scales for anxiety and depression were administered. Disease activity was assessed using the Behc¸et’s Disease Current Activity Form (BDCAF). Compared to controls, BD patients were significantly impaired on tasks evaluating long-term verbal and non-verbal memory, and visuospatial skills. In addition, BD patients were significantly more anxious and depressed than controls. Cognitive impairment was evident in 46.1% of BD patients compared with none of control subjects ( p < 0.0001), with memory representing the cognitive domain most affected. Both high disease activity (OR 1.3, 95% CI 1.0–1.5, p < 0.04) and high prednisone dosage (OR 1.3, 95% CI 1.0–1.7, p < 0.03) were independently associated with cognitive impairment in BD after adjustment for demographic variables. Cognitive impairment, involving mainly memory functions, occurs frequently in BD patients. It may occur independently of clinically overt neurological involvement, and is more common in patients with an active disease and in those receiving prednisone

    Nicotine dependence produces hyperalgesia: Role of corticotropin-releasing factor-1 receptors (CRF1Rs) in the central amygdala (CeA)

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    Because tobacco use has a large negative health and financial impact on society, it is critical to identify the factors that drive excessive use. These factors include the aversive withdrawal symptoms that manifest upon cessation of tobacco use, and may include increases in nociceptive processing. Corticotropin-releasing factor (CRF) signalling in the central amygdala (CeA) has been attributed an important role in: (1) central processing of pain, (2) excessive nicotine use that results in nicotine dependence, and (3) in mediating the aversive symptoms that manifest following cessation of tobacco exposure. Here, we describe three experiments in which the main hypothesis was that CRF/CRF1 receptor (CRF1R) signalling in the CeA mediates nicotine withdrawal-induced increases in nociceptive sensitivity in rats that are dependent on nicotine. In Experiment 1, nicotine-dependent rats withdrawn from chronic intermittent (14-h/day) nicotine vapor exhibited decreased hind paw withdrawal latencies in response to a painful thermal stimulus in the Hargreaves test, and this effect was attenuated by systemic administration of the CRF1R antagonist, R121919. In Experiment 2, nicotine-dependent rats withdrawn from nicotine vapor exhibited robust increases in mRNA for CRF and CRF1Rs in CeA. In Experiment 3, intra-CeA administration of R121919 reduced thermal nociception only in nicotine-dependent rats. Collectively, these results suggest that nicotine dependence increases CRF/CRF1R signaling in the CeA that mediates withdrawal-induced increases in sensitivity to a painful stimulus. Future studies will build on these findings by exploring the hypothesis that nicotine withdrawal-induced reduction in pain thresholds drive excessive nicotine use via CRF/CRF1R signalling pathways
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