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Warfarin induces coagulopathy. Guidelines protocolize reversal of supratherapeutic international normalized ratio (INR) in patients dependent on anticoagulation, but practices vary for reversing warfarin-induced coagulopathy after overdose in non-warfarin-dependent patients.This is the report of a 15-year-old female who ingested her father's warfarin (100-200 mg) in a self-harm attempt. At hour 24 post-ingestion, her INR was 2.00 and she was admitted for monitoring. Reversal of coagulopathy was initially deferred pending the INR trend. The INR was 5.10 at hour 60 and 2.5 mg oral vitamin KA restrictive approach to coagulopathy reversal in non-warfarin-dependent patients with intentional warfarin overdose may result in worsening coagulopathy, bleeding, and lengthy hospital stay. Given the risk for significant, prolonged coagulopathy, these patients should be treated early with VK1, with subsequent serial INR monitoring and probable additional VK1 dosing. Delayed peak warfarin concentrations support consideration of gastrointestinal decontamination in late presenters.

A method of ranking the rate of corrosion of metals in molten fluoride or chloride salts is proposed based on a zero-resistance ammeter (ZRA). The metal of interest for the corrosion study is shorted to a relatively noble metal counter electrode. Stainless steel 316 was tested in FLiNaK (LiF-NaF-KF). Haynes 230, stainless steel 316L, and Hastelloy C-22 were tested in MgCl2-NaCl-KCl. The ZRA oxidation current from SS-316L was reduced by 89% in FLiNaK after the addition of Li metal, which is known to reduce the redox potential. Post-test examination of metal surfaces and salt samples were also consistent with the relative ZRA current response in fluoride salt. Consistency was also observed between the relative ZRA response for the different metal alloys in the chloride salt and post-test analysis of metal and salt samples.

CaCl2 has applications for electrochemical processing of nuclear materials. Thermal dehydration leads to formation of oxide ions, which are shown to react and cause precipitation of dissolved CeCl3 that was selected as a surrogate for actinide chlorides. Thus, measurement of residual water in CaCl2 is an essential capability. Thermogravimetric analysis (TGA) was shown to underestimate starting water concentration. Subsequent analysis of solid samples via acid–base titration and cyclic voltammetry (CV) of the molten salt yielded consistent values within 5% for residual water. Hydroxides were shown to be unstable, thus oxygen is retained as oxide ions. Thus, total water in a sample of CaCl2 can be quantified by combining TGA with with either CV of molten salt or titration of salt samples. The importance of quantifying residual water in the salt was demonstrated by showing that cerium chloride (surrogate for actinide chlorides) will react with oxide ions in CaCl2 to form insoluble oxides and oxychlorides.

Patients awaiting lung transplantation face high wait-list mortality, as injury precludes the use of most donor lungs. Although ex vivo lung perfusion (EVLP) is able to recover marginal quality donor lungs, extension of normothermic support beyond 6 h has been challenging. Here we demonstrate that acutely injured human lungs declined for transplantation, including a lung that failed to recover on EVLP, can be recovered by cross-circulation of whole blood between explanted human lungs and a Yorkshire swine. This xenogeneic platform provided explanted human lungs a supportive, physiologic milieu and systemic regulation that resulted in functional and histological recovery after 24 h of normothermic support. Our findings suggest that cross-circulation can serve as a complementary approach to clinical EVLP to recover injured donor lungs that could not otherwise be utilized for transplantation, as well as a translational research platform for immunomodulation and advanced organ bioengineering. In a new strategy for increasing the availability of lungs for transplantation, human lungs declined for transplantation because of their poor quality can be recuperated by connecting them to the circulation of a pig.

Parkinson’s disease (PD) reduces independence and quality of life through deterioration of upper limb motor function. Transcranial direct current stimulation (tDCS) may offer an alternative, adjunctive therapy for PD. However, the efficacy of tDCS for upper limb motor rehabilitation in PD is unknown. In this systematic review, evidence is compiled regarding the effects of tDCS on upper limb motor function in PD. Studies of tDCS applied to PD patients that assessed upper limb motor function, conducted between January 2000 and November 2018, were screened for inclusion via a systematic search of Medline, Cochrane, PsycINFO, EMBASE, CINAHL, and Web of Science. Ten out of 606 studies were included and their findings synthesized into five categories regarding the effects of tDCS on: (1) Unified Parkinson’s Disease Rating Scale motor section (UPDRS III), (2) upper limb motor tasks, (3) manual dexterity, (4) reaction time, and (5) neurophysiology. When applied to the primary motor cortex, tDCS may improve UPDRS III and the speed and force of movement. Considerable variation was found in tDCS parameters and further study is needed to clarify the long-term effects of tDCS on both simple and complex motor tasks and to compile relevant neurophysiological evidence.

4H-SiC alpha detectors were fabricated with a 21-μm thick depletion depth and were packaged into a stainless-steel casing with a mineral insulation cable and a standard BNC connector. The packaged detectors had a resolution of 0.624% FWHM at 5.486 MeV prior to salt immersion. The detectors were then immersed in a LiCl–KCl–UCl3 molten salt at 500 °C, from which a thin layer of depleted uranium was electrodeposited onto the detectors. Alpha particle emission spectra were collected from the electrodeposited source. The energy resolution of the surviving detector was 2.29% FWHM at 4.198 MeV and was sufficient to separate the 234U from 238U alpha emissions (577 keV difference). The 234U/238U activity ratio and the isotopic concentrations of 234U and 238U were determined and are representative of the uranium source used in the electrodeposition.

Recent studies have demonstrated genetic differences between monozygotic (MZ) twins. To test the hypothesis that early post-twinning mutational events associate with phenotypic discordance, we investigated a cohort of 13 twin pairs (n = 26) discordant for various clinical phenotypes using whole-exome sequencing and screened for copy number variation (CNV). We identified a de novo variant in PLCB1, a gene involved in the hydrolysis of lipid phosphorus in milk from dairy cows, associated with lactase non-persistence, and a variant in the mitochondrial complex I gene MT-ND5 associated with amyotrophic lateral sclerosis (ALS). We also found somatic variants in multiple genes (TMEM225B, KBTBD3, TUBGCP4, TFIP11) in another MZ twin pair discordant for ALS. Based on the assumption that discordance between twins could be explained by a common variant with variable penetrance or expressivity, we screened the twin samples for known pathogenic variants that are shared and identified a rare deletion overlapping ARHGAP11B, in the twin pair manifesting with either schizotypal personality disorder or schizophrenia. Parent-offspring trio analysis was implemented for two twin pairs to assess potential association of variants of parental origin with susceptibility to disease. We identified a de novo variant in RASD2 shared by 8-year-old male twins with a suspected diagnosis of autism spectrum disorder (ASD) manifesting as different traits. A de novo CNV duplication was also identified in these twins overlapping CD38, a gene previously implicated in ASD. In twins discordant for Tourette's syndrome, a paternally inherited stop loss variant was detected in AADAC, a known candidate gene for the disorder.

Dechlorination of eutectic LiCl–KCl based electrorefiner (ER) salt is reported via ion-exchange reaction with protonated ultrastable Y-type (USHY) zeolite bound into mechanically fluidized 45–250 μm diameter particles. Evidence of exchange of cations from the salt (Li+, K+, and fission product cations) into the zeolite lattice replacing H+ ions was found based on a change in unit cell size, ICP-MS, XRD and TEM–EDS in addition to detection of HCl off gas. Ion exchange reaction was carried out at 625 and 650 °C, temperatures above the melting point of eutectic LiCl–KCl. Experiments were carried out to optimize zeolite drying temperature, estimate maximum ion-exchange capacity, and determine the thermal stability of USHY zeolite. The results indicate over 90% dechlorination can be achieved without zeolite structure collapse at 625 °C. This provides a promising route to stabilizing waste from radioactive chloride salts into dechlorinated waste forms for permanent geologic disposal.

Idiopathic scoliosis (IS) is a complex 3D deformation of the spine with a strong genetic component, most commonly found in adolescent girls. Adolescent idiopathic scoliosis (AIS) affects around 3% of the general population. In a 5-generation UK family, linkage analysis identified the locus 9q31.2-q34.2 as a candidate region for AIS; however, the causative gene remained unidentified. Here, using exome sequencing we identified a rare insertion c.1569_1570insTT in the tubulin tyrosine ligase like gene, member 11 (TTLL11) within that locus, as the IS causative gene in this British family. Two other TTLL11 mutations were also identified in two additional AIS cases in the same cohort. Analyses of primary cells of individuals carrying the c.1569_1570insTT (NM_194252) mutation reveal a defect at the primary cilia level, which is less present, smaller and less polyglutamylated compared to control. Further, in a zebrafish, the knock down of ttll11, and the mutated ttll11 confirmed its role in spine development and ciliary function in the fish retina. These findings provide evidence that mutations in TTLL11, a ciliary gene, contribute to the pathogenesis of IS.

The alpha emission spectrum of electrodeposited thin film 232Th and depleted uranium sources, including the alpha emitting 232Th decay chain daughters and 238U daughter nuclide 234U, have been measured using a thin film 4H–SiC alpha detector. The energy resolution of this detector was proved suitable for the identification of many of the major characteristic alpha emissions associated with the 232Th daughter isotopes. The identification of the lower energy daughter isotopes made from the 4H–SiC detector measurements was confirmed using a silicon surface barrier detector. A depleted uranium source was fabricated using electrodeposition in a molten salt and the alpha spectrum was acquired with a 4H–SiC detector. The 234U/238U activity ratio and atomic enrichment of 234U in the sample were determined. A method of determining the 235U enrichment in the product stream of the enrichment process used to produce the depleted uranium source was developed. This method makes use of the 234U and 235U waste stream enrichments, as indicated by the depleted uranium source, and a mass balance equation to determine the product 235U enrichment. Since the 235U spectrum was obscured in the electrodeposited source by peak broadening of 238U and 234U, a table of common 235U waste enrichment values was used to compute 235U product enrichments, thereby resulting in a plausible identification of the enrichment process as a Department of Energy HEU enrichment process.

Aqueous two-phase systems and related emulsion-based structures defined within micro- and nanoscale environments enable a bottom-up synthetic biological approach to mimicking the dynamic compartmentation of biomaterial that naturally occurs within cells. Model systems we have developed to aid in understanding these phenomena include on-demand generation and triggering of reversible phase transitions in ATPS confined in microscale droplets, morphological changes in networks of femtoliter-volume aqueous droplet interface bilayers (DIBs) formulated in microfluidic channels, and temperature-driven phase transitions in interfacial lipid bilayer systems supported on micro and nanostructured substrates. For each of these cases, the dynamics were intimately linked to changes in the chemical potential of water, which becomes increasingly susceptible to confinement and crowding. At these length scales, where interfacial and surface areas predominate over compartment volumes, both evaporation and osmotic forces become enhanced relative to ideal dilute solutions. Consequences of confinement and crowding in cell-sized microcompartments for increasingly complex scenarios will be discussed, from single-molecule mobility measurements with fluorescence correlation spectroscopy to spatio-temporal modulation of resource sharing in cell-free gene expression bursting.

Introduction Ductal carcinoma in situ (DCIS) is a non-obligate precursor of invasive ductal breast cancer, and approximately 20% of screen-detected tumours are pure DCIS. Most risk factors for breast cancer have similar associations with DCIS and IDC; however, there is limited data on the prevalence of the known high and moderate penetrance breast cancer predisposition genes in DCIS and which women with DCIS should be referred for genetic screening. The aim of this study was to assess the frequency of germline variants in BRCA2, BRCA1, CHEK2, PALB2 and TP53 in DCIS in women aged less than 50 years of age. Methods After DNA extraction from the peripheral blood, Access Array technology (Fluidigm) was used to amplify all exons of these five known breast cancer predisposition genes using a custom made targeted sequencing panel in 655 cases of pure DCIS presenting in women under the age of 50 years together with 1611 controls. Results Case-control analysis revealed an excess of pathogenic variants in BRCA2 (OR = 27.96, 95%CI 6.56–119.26, P = 2.0 × 10−10) and CHEK2 (OR = 8.04, 95%CI 2.93–22.05, P = 9.0 × 10−6), with weaker associations with PALB2 (P = 0.003), BRCA1 (P = 0.007) and TP53 (P = 0.02). For oestrogen receptor (ER)-positive DCIS the frequency of pathogenic variants was 9% under the age of 50 (14% with a family history of breast cancer) and 29% under the age of 40 (42% with a family history of breast cancer). For ER-negative DCIS, the frequency was 9% (16% with a family history of breast cancer) and 8% (11% with a family history of breast cancer) under the ages of 50 and 40, respectively. Conclusions This study has shown that breast tumourigenesis in women with pathogenic variants in BRCA2, CHEK2, PALB2, BRCA1 and TP53 can involve a DCIS precursor stage and that the focus of genetic testing in DCIS should be on women under the age of 40 with ER-positive DCIS. Electronic supplementary material The online version of this article (10.1186/s13058-019-1143-y) contains supplementary material, which is available to authorized users.

Alpha spectroscopy can be used to quantify actinide (e.g., U, Pu) concentration in a molten salt electrorefining environment. One could electroplate actinide samples directly onto a semiconductor alpha particle detector to obtain representative isotopic concentrations from a measured alpha particle energy spectrum. In this work, we fabricated a SiC Schottky device that can be partially biased to a depletion depth of 8.8 µm that is able to measure the energy spectrum of 4.012 MeV alpha particles emitted from a thorium film with a thickness. We also present a method in calculating the thickness of a medium thick alpha source with a dE/dx detector.

The solubility of lithium metal in molten LiCl–Li2O mixtures has been measured at various concentrations of Li2O ranging from 0 wt.% to 2.7 wt.% at a temperature of approximately 670–680°C. After contacting molten lithium with molten LiCl–Li2O for several hours to achieve equilibrium saturation, samples were taken by freezing the salt onto a room-temperature steel rod and dissolving in water for analysis. Both volume of hydrogen gas generated and volume of titrated HCl were measured to investigate two different approaches to calculating the lithium concentration. There appeared to be no effect of Li2O concentration on the Li solubility in the salt. But the results vary between different methods of deducing the amount of dissolved Li. The H2 collection method is recommended, but care must be taken to ensure all of the H2 has been included.

Although individual differences in intelligence (general cognitive ability) are highly heritable, molecular genetic analyses to date have had limited success in identifying specific loci responsible for its heritability. This study is the first to investigate exome variation in individuals of extremely high intelligence. Under the quantitative genetic model, sampling from the high extreme of the distribution should provide increased power to detect associations. We therefore performed a case–control association analysis with 1409 individuals drawn from the top 0.0003 (IQ >170) of the population distribution of intelligence and 3253 unselected population-based controls. Our analysis focused on putative functional exonic variants assayed on the Illumina HumanExome BeadChip. We did not observe any individual protein-altering variants that are reproducibly associated with extremely high intelligence and within the entire distribution of intelligence. Moreover, no significant associations were found for multiple rare alleles within individual genes. However, analyses using genome-wide similarity between unrelated individuals (genome-wide complex trait analysis) indicate that the genotyped functional protein-altering variation yields a heritability estimate of 17.4% (s.e. 1.7%) based on a liability model. In addition, investigation of nominally significant associations revealed fewer rare alleles associated with extremely high intelligence than would be expected under the null hypothesis. This observation is consistent with the hypothesis that rare functional alleles are more frequently detrimental than beneficial to intelligence.

Background Lobular carcinoma in situ (LCIS) is a non-invasive breast lesion that is typically found incidentally on biopsy and is often associated with invasive lobular carcinoma (ILC). LCIS is considered by some to be a risk factor for future breast cancer rather than a true precursor lesion. The aim of this study was to identify genetic changes that could be used as biomarkers of progression of LCIS to invasive disease using cases of pure LCIS and comparing their genetic profiles to LCIS which presented contemporaneously with associated ILC, on the hypothesis that the latter represents LCIS that has already progressed. Methods Somatic copy number aberrations (SCNAs) were assessed by SNP array in three subgroups: pure LCIS, LCIS associated with ILC and the paired ILC. In addition exome sequencing was performed on seven fresh frozen samples of LCIS associated with ILC, to identify recurrent somatic mutations. Results The copy number profiles of pure LCIS and LCIS associated with ILC were almost identical. However, four SCNAs were more frequent in ILC than LCIS associated with ILC, including gain/amplification of CCND1. CCND1 protein over-expression assessed by immunohistochemical analysis in a second set of samples from 32 patients with pure LCIS and long-term follow up, was associated with invasive recurrence (P = 0.02, Fisher’s exact test). Exome sequencing revealed that PIK3CA mutations were as frequent as CDH1 mutations in LCIS, but were not a useful biomarker of LCIS progression as they were as frequent in pure LCIS as in LCIS associated with ILC. We also observed heterogeneity of PIK3CA mutations and evidence of sub-clonal populations in LCIS irrespective of whether they were associated with ILC. Conclusions Our data shows that pure LCIS and LCIS co-existing with ILC have very similar SCNA profiles, supporting the hypothesis that LCIS is a true precursor lesion. We have provided evidence that over-expression of CCND1 may identify a subgroup of patients with pure LCIS who are more likely to develop invasive disease, in contrast to PIK3CA mutations, which occur too early in lobular tumorigenesis to be informative. Electronic supplementary material The online version of this article (doi:10.1186/s13058-016-0789-y) contains supplementary material, which is available to authorized users.

The relative contribution of immunological dysregulation and impaired epithelial barrier function to allergic diseases is still a matter of debate. Here we describe a new syndrome featuring severe dermatitis, multiple allergies and metabolic wasting (SAM syndrome) caused by homozygous mutations in DSG1. DSG1 encodes desmoglein 1, a major constituent of desmosomes, which connect the cell surface to the keratin cytoskeleton and play a crucial role in maintaining epidermal integrity and barrier function. SAM syndrome-causing mutations resulted in lack of membrane expression of DSG1, leading to loss of cell-cell adhesion. In addition, DSG1 deficiency was associated with increased expression of a number of genes encoding allergy-related cytokines. The deciphering of the pathogenesis of SAM syndrome substantiates the notion that allergy may result from a primary structural epidermal defect.

Elucidating genetic causes of cholestasis has proved to be important in understanding the physiology and pathophysiology of the liver. Here we show that protein-truncating mutations in the tight junction protein 2 gene (TJP2) cause failure of protein localization and disruption of tight-junction structure, leading to severe cholestatic liver disease. These findings contrast with those in the embryonic-lethal knockout mouse, highlighting differences in redundancy in junctional complexes between organs and species.

Childhood general cognitive ability (g) is important for a wide range of outcomes in later life, from school achievement to occupational success and life expectancy. Large-scale association studies will be essential in the quest to identify variants that make up the substantial genetic component implicated by quantitative genetic studies. We conducted a three-stage genome-wide association study for general cognitive ability using over 350,000 single nucleotide polymorphisms (SNPs) in the quantitative extremes of a population sample of 7,900 7-year-old children from the UK Twins Early Development Study. Using two DNA pooling stages to enrich true positives, each of around 1,000 children selected from the extremes of the distribution, and a third individual genotyping stage of over 3,000 children to test for quantitative associations across the normal range, we aimed to home in on genes of small effect. Genome-wide results suggested that our approach was successful in enriching true associations and 28 SNPs were taken forward to individual genotyping in an unselected population sample. However, although we found an enrichment of low P values and identified nine SNPs nominally associated with g (P

Steady-state behavior and bistability have been proposed as mechanisms for decision making in gene circuits. However, transient gene expression has also been proposed to control cell fate, with the decision arbitrated by the duration of a transient gene expression pulse. Here, using an HIV-1 model system, we directly quantify transcriptional feedback strength and its effects on both the duration of HIV-1 Tat transcriptional pulses and the fate of HIV-infected cells. By measuring shifts in the autocorrelation of noise inherent to gene expression, we found that transcriptional positive feedback extends the single-cell Tat expression lifetime two- to sixfold for both minimal Tat circuits and full length, actively replicating HIV-1. Notably, artificial weakening of Tat positive feedback shortened the duration of Tat expression transients and biased the probability in favor of latency. Thus, transcriptional positive feedback can modulate transient expression lifetime to a greater extent than protein half-life modulation, and it has a critical role in the cell-fate decision in HIV.

Fe−Co composition gradient and Fe−Pt multilayer alloy films were tested as catalysts for groving vertically aligned carbon nanofibers (VACNFs) by plasma-enhanced chemical vapor deposition (PECVD). The Fe−Co film yielded nanofibers with alloy tips in a wide compositional range varying from 8.15 pct Fe at the Co-rich end to 46.29 pct Fe in the middle of the wafer as determined by energy-dispersive X-ray analysis. Two Fe−Co cubic phases (SG Pm3m, \({\text{P}}m\overline 3 m\) were identified by preliminary X-ray diffraction (XRD) measurements. Magnetic measurements showed a substantially greater hysteresis loop area and coercivity in Fe−Co catalyst nanoparticles as compared to the asdeposited Fe−Co film. The Fe−Pt film did not break into FePt alloy nanoparticles under the applied processing parameters and thus the utility of FePt as a VACNF catalyst has been inconclusive.

Fusions of genetic regulatory elements with reporter genes have long been used as tools for monitoring gene expression and have become a major component in synthetic gene circuit implementation. A major limitation of many of these systems is the relatively long half-life of the reporter protein(s), which prevents monitoring both the initiation and the termination of transcription in real-time. Furthermore, when used as components in synthetic gene circuits, the long time constants associated with reporter protein decay may significantly degrade circuit performance. In this study, short half-life variants of LuxA and LuxB from Photorhabdus luminescens were constructed in Escherichia coli by inclusion of an 11-amino acid carboxy-terminal tag that is recognized by endogenous tail-specific proteases. Results indicated that the addition of the C-terminal tag affected the functional half-life of the holoenzyme when the tag was added to luxA or to both luxA and luxB, but modification of luxB alone did not have a significant effect. In addition, it was also found that alteration of the terminal three amino acid residues of the carboxy-terminal tag fused to LuxA generated variants with half-lives of intermediate length in a manner similar to that reported for GFP. This report is the first instance of the C-terminal tagging approach for the regulation of protein half-life to be applied to an enzyme or monomer of a multi-subunit enzyme complex and will extend the utility of the bacterial luciferase reporter genes for the monitoring of dynamic changes in gene expression.

Recent work demonstrates that stochastic fluctuations in molecular populations have consequences for gene regulation. Previous experiments focused on noise sources or noise propagation through gene networks by measuring noise magnitudes. However, in theoretical analysis, we showed that noise frequency content is determined by the underlying gene circuits, leading to a mapping between gene circuit structure and the noise frequency range. An intriguing prediction from our previous studies was that negative autoregulation shifts noise to higher frequencies where it is more easily filtered out by gene networks--a property that may contribute to the prevalence of autoregulation motifs (for example, found in the regulation of approximately 40% of Escherichia coli genes). Here we measure noise frequency content in growing cultures of E. coli, and verify the link between gene circuit structure and noise spectra by demonstrating the negative autoregulation-mediated spectral shift. We further demonstrate that noise spectral measurements provide mechanistic insights into gene regulation, as perturbations of gene circuit parameters are discernible in the measured noise frequency ranges. These results suggest that noise spectral measurements could facilitate the discovery of novel regulatory relationships.

Cone-shaped nanostructures of mixed composition (nanocones) and largely graphitic nanofibers were synthesized on silicon substrates using iron/platinum alloy nanoparticles as the catalyst in a direct-current plasma enhanced chemical vapor deposition reactor. The catalyst nanoparticles were monodisperse in size with an average diameter of 3 (±1) nm. The nanocones were produced on laterally widely dispersed catalyst particles and were oriented perpendicular to the substrate surface with an amorphous internal structure. The nanocones were produced by gas phase mixing and deposition of plasma-sputtered silicon, nitrogen, carbon, and oxygen species on a central backbone nucleated by the Fe–Pt catalyst particle. Field emission measurements showed that a very high turn-on electric field was required for electron emission from the nanocones. In contrast, the graphitic nanofibers that were produced when silicon sputtering and redeposition were minimized had the “stacked-cup” structure, and well-defined voids could be observed within nanofibers nucleated from larger catalyst particles.

The hereditary spastic paraplegias (HSPs) are a clinically and genetically heterogeneous group of neurodegenerative disorders characterised by lower limb spasticity and weakness. Mutations in NIPA1 (Nonimprinted in Prader-Willi/Angelman syndrome 1) have recently been identified as a cause of autosomal dominant pure HSP, with one mutation described in two unrelated families. NIPA1 has no known function but is predicted to possess nine transmembrane domains and may function as a receptor or transporter. Here we present a large British pedigree in which linkage analysis conclusively demonstrates linkage to the NIPA1 locus (maximum multipoint LOD score 4.6). Subsequent mutation analysis identified a novel missense substitution in a highly conserved NIPA1 residue (G106R) which further confirms a causative link between NIPA1 mutation and autosomal dominant hereditary spastic paraplegia.

We report an allelic series of eight mutations in GATA2 underlying Emberger syndrome, an autosomal dominant primary lymphedema associated with a predisposition to acute myeloid leukemia. GATA2 is a transcription factor that plays an essential role in gene regulation during vascular development and hematopoietic differentiation. Our findings indicate that haploinsufficiency of GATA2 underlies primary lymphedema and predisposes to acute myeloid leukemia in this syndrome.

Serpentine fibula polycystic kidney syndrome (SFPKS; MIM600330) is a rare skeletal dysplasia that has polycystic kidneys and dysmorphic facies as additional defining phenotypic components. The nosological classification of this disease has been debated as the condition shares features common to other skeletal dysplasias such as Melnick Needles syndrome (MNS; MIM309350) and Hajdu–Cheney Syndrome (HCS; MIM102500). Here, two previously reported cases of SFPKS are presented with emphasis on their phenotypic evolution. With the recent discovery that HCS is caused by mutations in NOTCH2, DNA from the both cases was examined and both were found to have truncating mutations in exon 34 of NOTCH2. The phenotypic evolution of SFPKS and this molecular analysis strongly suggest that SFPKS is part of the phenotypic spectrum of HCS and should no longer be classified as a distinct disease entity.

We used an exome-sequencing strategy and identified an allelic series of NOTCH2 mutations in Hajdu-Cheney syndrome, an autosomal dominant multisystem disorder characterized by severe and progressive bone loss. The Hajdu-Cheney syndrome mutations are predicted to lead to the premature truncation of NOTCH2 with either disruption or loss of the C-terminal proline-glutamate-serine-threonine-rich proteolytic recognition sequence, the absence of which has previously been shown to increase Notch signaling.

Acne vulgaris (acne) is a common inflammatory disorder of the cutaneous pilo-sebaceous unit. Here we perform a genome-wide association analysis in the United Kingdom, comparing severe cases of acne (n=1,893) with controls (n=5,132). In a second stage, we genotype putative-associated loci in a further 2,063 acne cases and 1,970 controls. We identify three genome-wide significant associations: 11q13.1 (rs478304, Pcombined=3.23 × 10(-11), odds ratio (OR) = 1.20), 5q11.2 (rs38055, P(combined) = 4.58 × 10(-9), OR = 1.17) and 1q41 (rs1159268, P(combined) = 4.08 × 10(-8), OR = 1.17). All three loci contain genes linked to the TGFβ cell signalling pathway, namely OVOL1, FST and TGFB2. Transcripts of OVOL1 and TFGB2 have decreased expression in affected compared with normal skin. Collectively, these data support a key role for dysregulation of TGFβ-mediated signalling in susceptibility to acne.

A genetic predisposition to cluster headache (CH) has long been debated. Genetic epidemiological studies have reported an increased risk of CH in relatives of CH sufferers. The familial clustering supports a model of autosomal dominant inheritance with reduced penetrance. Some candidate gene studies have been performed, detecting associations with HCRTR2 and ADH4, but these have been limited by small sample size. We have established a large cohort of CH families in which we have previously reported a genome-wide linkage scan, isolating a number of putative linkage loci[1]. Despite this, a single causative gene is yet to be identified, largely due to substantial genetic heterogeneity.

We have examined the effect of L- and D-2-chloropropionic acid (L-CPA and D-CPA) on the concentrations of pyruvate, lactate, glucose and beta-hydroxybutyrate in the blood at various times after doses which produce cerebellar granule cell necrosis. Blood pyruvate and lactate concentrations were reduced in these animals 4 h after dosing and remained below those of controls for up to 48 h. No changes were seen in concentrations of plasma glucose of beta-hydroxybutyrate at any time point. Similarly, no changes in cerebellar glucose, pyruvate or lactate were seen 24 h after 750 mg/kg L-CPA. Oxidation of [14C] pyruvate or [14C] glucose to [14CO2] by cerebellar slices from control rats was not altered by the presence of L- or D-CPA (1-10 mM for 2 h). Nor were these parameters affected in cerebellar slices from rats killed 24 h after a dose of 750 mg/kg L-CPA. We conclude that the activation of the pyruvate dehydrogenase complex is unlikely to be a critical factor in the selective toxicity of CPA to the cerebellum.

Steady-state behavior and bistability have been proposed as mechanisms for decision-making in gene circuits. However, transient gene expression has also been proposed to control cell fate with the decision arbitrated by the lifetime of the expression transient. Here, we report that transcriptional positive-feedback plays a critical role in determining HIV infected cell-fate by extending the duration of Tat expression transients far beyond what protein half-life modulation can achieve. To directly quantify feedback strength and its effects on the duration of Tat transcriptional pulses, we exploit the noise inherent to gene-expression and measure shifts in the autocorrelation of expression noise. The results indicate that transcriptional positive-feedback extends the single-cell Tat expression lifetime by ~6-fold for both minimal Tat circuits and full-length, actively-replicating HIV-1. Importantly, artificial weakening of Tat positive-feedback shortened the duration of Tat expression transients and biased the probability in favor of latency. Thus, transcriptional positive-feedback appears to modulate transient expression lifetime and thereby control cell-fate in HIV.

In order to achieve multiple intracellular stimulation and recording devices with high electrode density and low manufacturing cost, we have fabricated and characterized an active matrix thin film transistor array with integrated vertically aligned carbon nanofibers. This device has the potential for cell probing and screening that provides great potential to execute direct cell sensing/probing and recording with a high electrode density. Each unit pixel in the array is individually addressed by a thin film transistor (TFT) and the vertically aligned carbon nanofibers (VACNF) used to impale the cell are fabricated on the drain electrode of the TFT. The VACNF is grown by direct current plasma enhanced chemical vapor deposition (DCPECVD) using a nickel catalyst. Electroanalysis or impedance differences between the cell probing site and a reference electrode are recorded with a potentiostat or a semiconductor analyzer connected to the output terminals. Additionally, the impedance change with frequency of the intracellular probe can be measured by applying a frame signal input and the signal can be recorded in storage capacitors after the frame scan of the TFTs. Consequently, actively addressed nanofiber arrays enable bidirectional interfacing with tissue matrices in a format that provides intercellular positioning of electrode elements as well as the potential for intracellular residence of probes within individual cells. In our research, we exploit these non-planar electrode systems for efficient coupling with excitable cell matrices as well as for intracellular biochemical manipulation and sensing of and delivery to single cells. In this paper, we will discuss the fabrication sequence of the inverted metal-oxide-semiconductor (MOS) TFT, and will elaborate the materials issues related to integrating the carbon nanofibers with the TFT.

Single-wall carbon nanotubes are formed by Nd:YAG laser vaporization of a graphite/(1 at. % Ni, 1 at. % Co) target into flowing argon (500 Torr) within a quartz tube furnace (1000 °C). Here, this process is investigated for the first time with time-resolved laser-induced luminescence imaging and spectroscopy of Co atoms, C2 and C3 molecules, and clusters. These measurements under actual synthesis conditions show that the plume of vaporized material is segregated and confined within a vortex ring which maintains a ˜1 cm3 volume for several seconds. Using time-resolved spectroscopy and spectroscopic imaging, the time for conversion of atomic and molecular species to clusters was measured for both carbon (200 μs) and cobalt (2 ms). This rapid conversion of carbon to nanoparticles, combined with transmission electron microscopy analysis of the collected deposits, indicate that nanotube growth occurs during several seconds of time from a feedstock of mixed nanoparticles in the gas-suspended plume. Using these in situ diagnostics to adjust the time spent by the plume within the high-temperature zone, single-walled nanotubes of controlled length were grown at an estimated rate of 0.2 μm/s.

A ceramic waste form is being developed by Argonne National Laboratory* (ANL) as part of the demonstration of the electrometallurgical treatment of spent nuclear fuel [1]. The halide, alkaline earth, alkali, transuranic, and rare earth fission products are stabilized in zeolite which is combined with glass and processed in a hot isostatic press (HIP) to form a ceramic composite. The mineral sodalite is formed in the HIP from the zeolite precursor. The process, from starting materials to final product, is relatively simple. An overview of the processing operations is given. The metrics that have been developed to measure the success or completion of processing operations are developed and discussed. The impact of variability in processing metrics on the durability of the final product is presented. The process is demonstrated to be robust for the type and range of operation metrics considered and the performance metric (PCT durability test) against which the operation metrics are evaluated.

Infantile cholestatic diseases can be caused by mutations in a number of genes involved in different hepatocyte molecular pathways. Whilst some of the essential pathways have a well understood function, such as bile biosynthesis and transport, the role of the others is not known. Here we report the findings of a clinical, biochemical and molecular study of a family with three patients affected with a severe infantile cholestatic disease. A novel homozygous frameshift germline mutation (c.587delG) in the AKR1D1 gene; which encodes the enzyme Δ 4-3-oxosteroid 5β–reductase that is required for synthesis of primary bile acids and is crucial for establishment of normal bile flow, was found in all 3 patients. Although the initial bile acid analysis was inconclusive, subsequent testing confirmed the diagnosis of a bile acid biogenesis disorder. An additional novel homozygous frameshift mutation (c.3391delC) was detected in SKIV2L in one of the patients. SKIV2L encodes a homologue of a yeast ski2 protein proposed to be involved in RNA processing and mutations in SKIV2L were recently described in patients with Tricohepatoenteric syndrome (THES). A combination of autozygosity mapping and whole-exome-sequencing allowed the identification of causal mutations in this family with a complex liver phenotype. Although the initial 2 affected cousins died in the first year of life, accurate diagnosis and management of the youngest patient led to successful treatment of the liver disease and disease-free survival.