Transcriptional and Translational Effects of Intronic CAPN3 Gene Mutations

Variants of unknown significance in the CAPN3 gene constitute a significant challenge for genetic counselling. Despite the frequency of intronic nucleotide changes in this gene (15–25% of all mutations), so far their pathogenicity has only been inferred by in-silico analysis, and occasionally, proven by RNA analysis. In this study, 5 different intronic variants (one novel) that bioinformatic tools predicted would affect RNA splicing, underwent comprehensive studies which were designed to prove they are disease-causing. Muscle mRNA from 15 calpainopathy patients was analyzed by RT-PCR and splicing-specific-PCR tests. We established the previously unrecognized pathogenicity of these mutations, which caused aberrant splicing, most frequently by the activation of cryptic splicing sites or, occasionally, by exon skipping. The absence or severe reduction of protein demonstrated their deleterious effect at translational level. We concluded that bioinformatic tools are valuable to suggest the potential effects of intronic variants; however, the experimental demonstration of the pathogenicity is not always easy to do even when using RNA analysis (low abundance, degradation mechanisms), and it might not be successful unless splicing-specific-PCR tests are used. A comprehensive approach is therefore recommended to identify and describe unclassified variants in order to offer essential data for basic and clinical geneticists. ©2010 Wiley-Liss, Inc

Measuring the Spins of Stellar Black Holes: A Progress Report

We use the Novikov-Thorne thin disk model to fit the thermal continuum X-ray spectra of black hole X-ray binaries, and thereby extract the dimensionless spin parameter a* = a/M of the black hole as a parameter of the fit. We summarize the results obtained to date for six systems and describe work in progress on additional systems. We also describe recent methodological advances, our current efforts to make our analysis software fully available to others, and our theoretical efforts to validate the Novikov-Thorne model.Comment: 6 pages, conference proceedings, X-ray Astronomy 2009: Present Status, Multi-Wavelength Approach and Future Perspectives, AIP, eds. A. Comastri et al.; list of authors revise

Identification of an intragenic deletion in the SGCB gene through a re-evaluation of negative next generation sequencing results

A large mutation screening of 504 patients with muscular dystrophy or myopathy has been performed by next generation sequencing (NGS). Among this cohort of patients, we report a case with a severe form of muscular dystrophy with a proximal weakness in the limb-girdle muscles. Her biopsy revealed typical dystrophic features and immunohistochemistry for α- and γ-sarcoglycans showed an absent reaction, addressing the clinical diagnosis toward a sarcoglycanopathy. Considering that no causative point mutation was detected in any of the four sarcoglycan genes, we re-evaluated the NGS data by careful quantitative analysis of the specific reads mapping on the four sarcoglycan genes. A complete absence of reads from the sixth exon of the β-sarcoglycan gene was found. Subsequent array comparative genomic hybridization (CGH) analysis confirmed the result with the identification of a novel 3.3 kb intragenic deletion in the SGCB gene. This case illustrates the importance of a multidisciplinary approach involving clinicians and molecular geneticists and the need for a careful re-evaluation of NGS data

The high official Harkhuf and the inscriptions of his tomb in Aswan (Egypt). An integrated methodological approach.

TECH project (Technology for the Egyptian Cultural Heritage) aimed to document an Egyptian monument for Egyptological studies and researches but, at the same time, to check a new methodological approach for conservation, valorisation and enhancement. In particular, the CNR mission focused the attention on the tomb of Harkhuf, a high official of the VI dynasty (XXIII century BC), who led trading and military expeditions into Nubia. The hieroglyphic texts inscribed on the façade of his tomb are very important and famous documents. The team checked an innovative and integrated methodology. The methodology has been focused mainly on the use of digital photogrammetric systems in order to generate an accurate numerical model (3D) and to facilitate the epigraphic study. Different procedures have been established in the processing and representation steps in order to accomplish the final communication of the results. Moreover climatic measurements have been carried out in order to understand the role of environmental factors on the deterioration of the monument. Finally the data have been crossed in order to check the environmental impact and the decay

UAV-based field phenotyping based on low-cost, open-source hardware and software

Here we present a low-cost platform for field phenotyping based on an unmanned aerial vehicle (UAV), which consists mainly of free-to-use or open-source hardware and software. This platform consists of a multirotor helicopter -with 8 rotorsproviding great stability and maneuvrability, which is able to carry a variety of sensors for trait measurement. The platform is controlled by an electronic autopilot board, which can be programmed for GPS-guided, automatic flight through an open-source ground station software. This platform is currently equipped with a low-cost consumer camera modified for the acquisition of near infrared - green - blue images, thus allowing the capture of high-resolution vegetation index images. A pipeline for the processing, geo-referencing and analysis of the images has been developed, which is based on open-source and/or free-to-use software.Sociedad Argentina de Informática e Investigación Operativa (SADIO

Visually evoked phase synchronization changes of alpha rhythm in migraine: Correlations with clinical features

Objective: This study aimed to compute phase synchronization of the alpha band from a multichannel electroencephalogram (EEG) recorded under repetitive flash stimulation from migraine patients without aura. This allowed examination of ongoing EEG activity during visual stimulation in the pain-free phase of migraine. Methods: Flash stimuli at frequencies of 3, 6, 9, 12, 15, 18, 21, 24, and 27 Hz were delivered to 15 migraine patients without aura and 15 controls, with the EEG recorded from 18 scalp electrodes, referred to the linked earlobes. The EEG signals were filtered in the alpha (7.5� 13 Hz) band. For all stimulus frequencies that we evaluated, the phase synchronization index was based on the Hilbert transformation. Results: Phase synchronization separated the patients and controls for the 9, 24 and 27 Hz stimulus frequencies; hyper phase synchronization was observed in patients, whereas healthy subjects were characterized by a reduced phase synchronization. These differences were found in all regions of the scalp. Conclusions: During migraine, the brain synchronizes to the idling rhythm of the visual areas under certain photic stimulations; in normal subjects however, brain regions involved in the processing of sensory information demonstrate desynchronized activity. Hypersynchronization of the alpha rhythm may suggest a state of cortical hypoexcitability during the interictal phase of migraine. Significance: The employment of non-linear EEG analysis may identify subtle functional changes in the migraine brain. D 2005 Elsevier B.V. All rights reserved

Spain

This report is part of the project The Factbook on the Illicit Trade in Tobacco Products (hereafter ITTP). It focuses on Spain, where the illicit trade in tobacco products is an emerging issue, particularly in certain regions. After tax increases in 2009-2010, Spain became a destination market of illicit tobacco and the ITTP penetration peaked at 8.8% in 2013. Nevertheless, the ITTP level is lower than the European average. In the past, ITTP penetration had decreased from 6% to almost 1% between 1998 and 2010, due to the successful enforcement actions against contraband. Spain used to be a transit country

The distinct role of the four voltage sensors of the skeletal CaV1.1 channel in voltage-dependent activation.

Initiation of skeletal muscle contraction is triggered by rapid activation of RYR1 channels in response to sarcolemmal depolarization. RYR1 is intracellular and has no voltage-sensing structures, but it is coupled with the voltage-sensing apparatus of CaV1.1 channels to inherit voltage sensitivity. Using an opto-electrophysiological approach, we resolved the excitation-driven molecular events controlling both CaV1.1 and RYR1 activations, reported as fluorescence changes. We discovered that each of the four human CaV1.1 voltage-sensing domains (VSDs) exhibits unique biophysical properties: VSD-I time-dependent properties were similar to ionic current activation kinetics, suggesting a critical role of this voltage sensor in CaV1.1 activation; VSD-II, VSD-III, and VSD-IV displayed faster activation, compatible with kinetics of sarcoplasmic reticulum Ca2+ release. The prominent role of VSD-I in governing CaV1.1 activation was also confirmed using a naturally occurring, charge-neutralizing mutation in VSD-I (R174W). This mutation abolished CaV1.1 current at physiological membrane potentials by impairing VSD-I activation without affecting the other VSDs. Using a structurally relevant allosteric model of CaV activation, which accounted for both time- and voltage-dependent properties of CaV1.1, to predict VSD-pore coupling energies, we found that VSD-I contributed the most energy (~75 meV or ∼3 kT) toward the stabilization of the open states of the channel, with smaller (VSD-IV) or negligible (VSDs II and III) energetic contribution from the other voltage sensors (<25 meV or ∼1 kT). This study settles the longstanding question of how CaV1.1, a slowly activating channel, can trigger RYR1 rapid activation, and reveals a new mechanism for voltage-dependent activation in ion channels, whereby pore opening of human CaV1.1 channels is primarily driven by the activation of one voltage sensor, a mechanism distinct from that of all other voltage-gated channels