Despite three decades of successful, predominantly phenotype-driven discovery of the genetic causes of monogenic disorders, up to half of children with severe developmental disorders of probable ...genetic origin remain without a genetic diagnosis. Particularly challenging are those disorders rare enough to have eluded recognition as a discrete clinical entity, those with highly variable clinical manifestations, and those that are difficult to distinguish from other, very similar, disorders. Here we demonstrate the power of using an unbiased genotype-driven approach to identify subsets of patients with similar disorders. By studying 1,133 children with severe, undiagnosed developmental disorders, and their parents, using a combination of exome sequencing and array-based detection of chromosomal rearrangements, we discovered 12 novel genes associated with developmental disorders. These newly implicated genes increase by 10% (from 28% to 31%) the proportion of children that could be diagnosed. Clustering of missense mutations in six of these newly implicated genes suggests that normal development is being perturbed by an activating or dominant-negative mechanism. Our findings demonstrate the value of adopting a comprehensive strategy, both genome-wide and nationwide, to elucidate the underlying causes of rare genetic disorders.
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DOBA, IJS, IZUM, KILJ, KISLJ, NUK, PILJ, PNG, SAZU, SBMB, SIK, UILJ, UKNU, UL, UM, UPUK
A "green" protocol was used for the rapid generation of nanoscale zerovalent iron (NZVI) particles using tea polyphenols. The NZVI particles were subsequently examined for
in vitro
biocompatibility ...using the human keratinocyte cell (HaCaT) line as a representative skin exposure model. The cells were exposed to NZVI for time periods of 24 and 48 h. Biocompatibility was assessed using the methyl tetrazolium, or MTS, (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2
H
-tetrazolium)) and lactate dehydrogenase (LDH) assays to determine
in vitro
cytotoxicity. The evaluation of mitochondrial function (MTS) and membrane integrity (LDH) in human keratinocytes showed that these "green" synthesized NZVI particles were nontoxic in the human keratinocytes exposed when compared with control samples synthesized using a borohydride protocol. In fact, in most cases, these "green" nanoparticles induced a prolific response in the cellular function even at the highest concentration (100μg ml
−1
).
A "green" protocol was developed for the rapid generation of nanoscale zerovalent iron (NZVI) particles using tea polyphenols and they were found to be nontoxic in the human keratinocyte cell (HaCaT) line.
Recurrent 15q13.3 microdeletions were recently identified with identical proximal (BP4) and distal (BP5) breakpoints and associated with mild to moderate mental retardation and epilepsy.
To assess ...further the clinical implications of this novel 15q13.3 microdeletion syndrome, 18 new probands with a deletion were molecularly and clinically characterised. In addition, we evaluated the characteristics of a family with a more proximal deletion between BP3 and BP4. Finally, four patients with a duplication in the BP3-BP4-BP5 region were included in this study to ascertain the clinical significance of duplications in this region.
The 15q13.3 microdeletion in our series was associated with a highly variable intra- and inter-familial phenotype. At least 11 of the 18 deletions identified were inherited. Moreover, 7 of 10 siblings from four different families also had this deletion: one had a mild developmental delay, four had only learning problems during childhood, but functioned well in daily life as adults, whereas the other two had no learning problems at all. In contrast to previous findings, seizures were not a common feature in our series (only 2 of 17 living probands). Three patients with deletions had cardiac defects and deletion of the KLF13 gene, located in the critical region, may contribute to these abnormalities. The limited data from the single family with the more proximal BP3-BP4 deletion suggest this deletion may have little clinical significance. Patients with duplications of the BP3-BP4-BP5 region did not share a recognisable phenotype, but psychiatric disease was noted in 2 of 4 patients.
Overall, our findings broaden the phenotypic spectrum associated with 15q13.3 deletions and suggest that, in some individuals, deletion of 15q13.3 is not sufficient to cause disease. The existence of microdeletion syndromes, associated with an unpredictable and variable phenotypic outcome, will pose the clinician with diagnostic difficulties and challenge the commonly used paradigm in the diagnostic setting that aberrations inherited from a phenotypically normal parent are usually without clinical consequences.
The SoLid collaboration has developed a new detector technology to detect electron anti-neutrinos at close proximity to the Belgian BR2 reactor at surface level. A 288kg prototype detector was ...deployed in 2015 and collected data during the operational period of the reactor and during reactor shut-down. Dedicated calibration campaigns were also performed with gamma and neutron sources. This paper describes the construction of the prototype detector with a high control on its proton content and the stability of its operation over a period of several months after deployment at the BR2 reactor site. All detector cells provide sufficient light yields to achieve a target energy resolution of better than 20%/ E(MeV). The capability of the detector to track muons is exploited to equalize the light response of a large number of channels to a precision of 3% and to demonstrate the stability of the energy scale over time. Particle identification based on pulse-shape discrimination is demonstrated with calibration sources. Despite a lower neutron detection efficiency due to triggering constraints, the main backgrounds at the reactor site were determined and taken into account in the shielding strategy for the main experiment. The results obtained with this prototype proved essential in the design optimization of the final detector.
This paper presents a comprehensive optimisation study to maximise the light collection efficiency of scintillating cube elements used in the SoLid detector. Very short baseline reactor experiments, ...like SoLid, look for active to sterile neutrino oscillation signatures in the anti-neutrino energy spectrum as a function of the distance to the core and energy. Performing a precise search requires high light yield of the scintillating elements and uniformity of the response in the detector volume. The SoLid experiment uses an innovative hybrid technology with two different scintillators: polyvinyltoluene scintillator cubes and 6LiF:ZnS(Ag) screens. A precision test bench based on a 207Bi calibration source has been developed to study improvements on the energy resolution and uniformity of the prompt scintillation signal of antineutrino interactions. A trigger system selecting the 1 MeV conversion electrons provides a Gaussian energy peak and allows for precise comparisons of the different detector configurations that were considered to improve the SoLid detector light collection. The light collection efficiency is influenced by the choice of wrapping material, the position of the 6LiF:ZnS(Ag) screen, the type of fibre, the number of optical fibres and the type of mirror at the end of the fibre. This study shows that large gains in light collection efficiency are possible compared to the SoLid SM1 prototype. The light yield for the SoLid detector is expected to be at least 52±2 photo-avalanches per MeV per cube, with a relative non-uniformity of 6 %, demonstrating that the required energy resolution of at least 14 % at 1 MeV can be achieved.
The purpose of this study was to evaluate the clinical feasibility of using a 2-dimensional (2D) video analysis (VA) system compared with visual estimation (VE) for measurement of active cervical ...rotation and lateral flexion in infants with congenital muscular torticollis.
Twelve infants participated in this study. Active cervical motion in rotation and lateral flexion was measured by VE and 2D VA.
Significant differences between VE and VA were found for left lateral flexion, right lateral flexion, and right rotation. Average total time for VA was 23.96 minutes.
This study suggests that the use of VA may improve the measurement of active cervical motion to improve clinical assessment of infants with congenital muscular torticollis. However, VA time is excessive and, therefore, not clinically feasible. Further studies are indicated to explore other software for this application.
A fundamental question in biology is whether the presence of non-reacting macromolecules in the cytoplasm affects the rates and extents of reversible association reactions, a phenomenon often ...referred to as 'macromolecular crowding.' Under certain conditions, crowding has been proposed to dramatically alter the kinetics and thermodynamics of chemical reactions, making it difficult to quantitatively relate rates and extents of reactions measured in vitro to those occurring in vivo. In this work, we use Brownian dynamics simulation and Monte Carlo methods to (1) quantify the overall thermodynamic and kinetic effects of crowding by independently investigating each step of reversible bimolecular association (i.e. translational diffusion, steric specific binding, and dissociation), and (2) provide an explicit, quantitative investigation of how the degree of steric specificity of protein dimerization influences crowding-mediated effects on association and dissociation. We find that kon decreases by ∼2-fold for non-steric specific reactions, and increases by ∼3-fold for highly steric specific reactions. In addition, koff decreases by only ∼30%-60% in the presence of crowders, depending on the strength of the bond between the reactant pair, so that the equilibrium constant is increased by ∼4-fold, at most. These results suggest that crowding-mediated effects on globular protein dimerization reactions in the cytoplasm are modulated by the steric specificity of the reactants, and that reversible protein-protein association is relatively insensitive to the physical presence of crowders (i.e. steric repulsion effects in the cytoplasm) for crowders of similar size and shape to reactants over a range of volume fractions (0-0.3).
A Physical Map of 30,000 Human Genes Deloukas, P.; Schuler, G. D.; Gyapay, G. ...
Science (American Association for the Advancement of Science),
10/1998, Volume:
282, Issue:
5389
Journal Article
Peer reviewed
A map of 30,181 human gene-based markers was assembled and integrated with the current genetic map by radiation hybrid mapping. The new gene map contains nearly twice as many genes as the previous ...release, includes most genes that encode proteins of known function, and is twofold to threefold more accurate than the previous version. A redesigned, more informative and functional World Wide Web site (www.ncbi.nlm.nih.gov/genemap) provides the mapping information and associated data and annotations. This resource constitutes an important infrastructure and tool for the study of complex genetic traits, the positional cloning of disease genes, the cross-referencing of mammalian genomes, and validated human transcribed sequences for large-scale studies of gene expression.
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10.
An STS-Based Map of the Human Genome Hudson, Thomas J.; Stein, Lincoln D.; Gerety, Sebastian S. ...
Science (American Association for the Advancement of Science),
12/1995, Volume:
270, Issue:
5244
Journal Article
Peer reviewed
A physical map has been constructed of the human genome containing 15,086 sequence-tagged sites (STSs), with an average spacing of 199 kilobases. The project involved assembly of a radiation hybrid ...map of the human genome containing 6193 loci and incorporated a genetic linkage map of the human genome containing 5264 loci. This information was combined with the results of STS-content screening of 10,850 loci against a yeast artificial chromosome library to produce an integrated map, anchored by the radiation hybrid and genetic maps. The map provides radiation hybrid coverage of 99 percent and physical coverage of 94 percent of the human genome. The map also represents an early step in an international project to generate a transcript map of the human genome, with more than 3235 expressed sequences localized. The STSs in the map provide a scaffold for initiating large-scale sequencing of the human genome.
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