Application of high-throughput sequencing for studying genomic variations in congenital heart disease

Congenital heart diseases (CHD) represent the most common birth defect in human. The majority of cases are caused by a combination of complex genetic alterations and environmental influences. In the past, many disease-causing mutations have been identified; however, there is still a large proportion of cardiac malformations with unknown precise origin. High-throughput sequencing technologies established during the last years offer novel opportunities to further study the genetic background underlying the disease. In this review, we provide a roadmap for designing and analyzing high-throughput sequencing studies focused on CHD, but also with general applicability to other complex diseases. The three main next-generation sequencing (NGS) platforms including their particular advantages and disadvantages are presented. To identify potentially disease-related genomic variations and genes, different filtering steps and gene prioritization strategies are discussed. In addition, available control datasets based on NGS are summarized. Finally, we provide an overview of current studies already using NGS technologies and showing that these techniques will help to further unravel the complex genetics underlying CHD

Magnetic trapping of metastable 3P2^3P_2 atomic strontium

We report the magnetic trapping of metastable $^3P_2$ atomic strontium. Atoms are cooled in a magneto-optical trap (MOT) operating on the dipole allowed $^1S_0-^1P_1$ transition at 461 nm. Decay via $^1P_1\to {^1D_2}\to {^3P_2}$ continuously loads a magnetic trap formed by the quadrupole magnetic field of the MOT. Over $10^8$ atoms at a density of $8 \times 10^9$ cm$^{-3}$ and temperature of 1 mK are trapped. The atom temperature is significantly lower than what would be expected from the kinetic and potential energy of atoms as they are transferred from the MOT. This suggests that thermalization and evaporative cooling are occurring in the magnetic trap.Comment: This paper has been accepted by PR

Green competence framework: Evidence from China

© 2015 Taylor & Francis Recently human resources management functions such as recruitment, selection, training and performance evaluation are expected in considering environmental management issues. Environmental protective acts with adequate ecological knowledge and socio-economic behavior and skills are referred to in this paper as green competencies (GCs). However, a systematic approach for developing and understanding key factors that enhance individuals' GCs is lacking. This study contributes to green human resource literature by integrating environmental consumer behavior literature with traditional skills and competencies literature to help firms to select the right individuals to achieve their environmental goals. Using Robert's competencies framework and structural equation modeling, this paper empirically examines the influence of individual GCs on organizations' green practices and performance objectives. Our model is tested using a sample of 1230 employees working in key industries in the Chinese coastal city of Ningbo. The results indicate that acquired GCs are more positively associated with individuals' GCs and green behavior. The study empirically demonstrates that verifying acquired GC attributes such as environmental knowledge, green purchase attitude and intention during employee selection would certainly be helpful for firms to identify individual green performance potential

Long-Time Asymptotics of Perturbed Finite-Gap Korteweg-de Vries Solutions

We apply the method of nonlinear steepest descent to compute the long-time asymptotics of solutions of the Korteweg--de Vries equation which are decaying perturbations of a quasi-periodic finite-gap background solution. We compute a nonlinear dispersion relation and show that the $x/t$ plane splits into $g+1$ soliton regions which are interlaced by $g+1$ oscillatory regions, where $g+1$ is the number of spectral gaps. In the soliton regions the solution is asymptotically given by a number of solitons travelling on top of finite-gap solutions which are in the same isospectral class as the background solution. In the oscillatory region the solution can be described by a modulated finite-gap solution plus a decaying dispersive tail. The modulation is given by phase transition on the isospectral torus and is, together with the dispersive tail, explicitly characterized in terms of Abelian integrals on the underlying hyperelliptic curve.Comment: 45 pages. arXiv admin note: substantial text overlap with arXiv:0705.034

Fast analysis of antibody-derived therapeutics by automated multidimensional liquid chromatography - mass spectrometry

Characterization of post-translational modifications (PTMs) of therapeutic antibodies is commonly performed by bottom-up approaches, involving sample preparation and peptide analysis by liquid chromatography-mass spectrometry (LC-MS). Conventional sample preparation requires extensive hands-on time and can increase the risk of inducing artificial modifications as many off-line steps - denaturation, disulfide-reduction, alkylation and tryptic digestion - are performed. In this study, we developed an on-line multidimensional (mD)-LC-MS bottom-up approach for fast sample preparation and analysis of (formulated) monoclonal antibodies and antibody-derived therapeutics. This approach allows on-column reduction, tryptic digestion and subsequent peptide analysis by RP-MS. Optimization of the 1D -and 2D flow and temperature improved the trapping of small polar peptides during on-line peptide mapping analysis. These adaptations increased the sequence coverage (95-98% versus 86-94% for off-line approaches) and allowed identification of various PTMs (i.e. deamidation of asparagine, methionine oxidation and lysine glycation) within a single analysis. This workflow enables a fast (<2 h) characterization of antibody heterogeneities within a single run and a low amount of protein (10 mu g). Importantly, the new mD-LC-MS bottom-up method was able to detect the polar, fast-eluting peptides: Fc oxidation at Hc-Met-252 and the Fc N-glycosylation at Hc-Asn-297, which can be challenging using mD-LC-MS. Moreover, the method showed good comparability across the different measurements (RSD of retention time in the range of 0.2-1.8% for polar peptides). The LC system was controlled by only a standard commercial software package which makes implementation for fast characterization of quality attributes relatively easy. (C) 2021 The Author(s). Published by Elsevier B.V.Proteomic

Ehlers-Danlos syndromes: state of the art on clinical practice guidelines.

Objective To report the effort of the European Reference Network for Rare and Complex CONnective tissue and musculoskeletal diseases NETwork working group on Ehlers-Danlos syndromes (EDS) and related disorders to assess current available clinical practice guidelines (CPGs) specifically addressed to EDS, in order to identify potential clinician and patient unmet needs. Methods Systematic literature search in PUBMED and EMBASE based on controlled terms (MeSH and Emtree) and keywords of the disease and publication type (CPGs). All the published articles were revised in order to identify existing CPGs on diagnosis, monitoring and treatment of EDS. Results Literature revision detected the absence of papers reporting good quality CPGs to optimise EDS patient care. The current evidence-based literature regarding clinical guidelines for the EDS was limited in size and quality, and there is insufficient research exploring the clinical features and interventions, and clinical decision-making are currently based on theoretical and limited research evidences. Conclusions Many clinician and patient unmet needs have been identifie

Loss of the mammal-specific tectorial membrane component CEA cell adhesion molecule 16 (CEACAM16) leads to hearing impairment at low and high frequencies

The vertebrate-restricted carcinoembryonic antigen gene family evolves extremely rapidly. Among their widely expressed members, the mammal-specific, secreted CEACAM16 is exceptionally well conserved and specifically expressed in the inner ear. To elucidate a potential auditory function we inactivated murine Ceacam16 by homologous recombination. In young Ceacam16-/- mice the hearing threshold for frequencies below 10 kHz and above 22 kHz was raised. This hearing impairment progressed with age. A similar phenotype is observed in hearing-impaired members of Family 1070 with non-syndromic autosomal dominant hearing loss (DFNA4) who carry a missense mutation in CEACAM16. CEACAM16 was found in interdental and Deiters cells and was deposited in the tectorial membrane of the cochlea between postnatal day 12 and 15, when hearing starts in mice. In cochlear sections of Ceacam16-/- mice tectorial membranes were significantly more often stretched out as compared to wild-type mice where they were mostly contracted and detached from the outer hair cells. Homotypic cell sorting observed after ectopic cell surface expression of the carboxy-terminal immunoglobulin variable-like N2 domain of CEACAM16 indicated that CEACAM16 can interact in trans. Furthermore, Western blot analyses of membrane-bound CEACAM16 under reducing and non-reducing conditions demonstrated oligomerization via unpaired cysteines. Taken together, CEACAM16 probably can form higher order structures with other tectorial membrane proteins such as α-tectorin and β-tectorin and influences the physical properties of the tectorial membrane. Evolution of CEACAM16 might have been an important step for the specialization of the mammalian cochlea allowing hearing over an extended frequency range

Application of stochastic programming to reduce uncertainties in quality-based supply planning of slaughterhouses

To match products of different quality with end market preferences under supply uncertainty, it is crucial to integrate product quality information in logistics decision making. We present a case of this integration in a meat processing company that faces uncertainty in delivered livestock quality. We develop a stochastic programming model that exploits historical product quality delivery data to produce slaughterhouse allocation plans with reduced levels of uncertainty in received livestock quality. The allocation plans generated by this model fulfil demand for multiple quality features at separate slaughterhouses under prescribed service levels while minimizing transportation costs. We test the model on real world problem instances generated from a data set provided by an industrial partner. Results show that historical farmer delivery data can be used to reduce uncertainty in quality of animals to be delivered to slaughterhouses

Long-term in vitro expansion ensures increased yield of central memory T cells as perspective for manufacturing challenges

Adoptive T cell therapy (ATT) has revolutionized the treatment of cancer patients. A sufficient number of functional T cells is indispensable for ATT efficacy; however, several ATT dropouts have been reported due to T cell expansion failure or lack of T cell persistence in vivo. With the aim of providing ATT also to those patients experiencing insufficient T cell manufacturing via standard protocol, we evaluated if minimally manipulative prolongation of in vitro expansion (long-term (LT) >3 weeks with IL-7 and IL-15 cytokines) could result in enhanced T cell yield with preserved T cell functionality. The extended expansion resulted in a 39-fold increase of murine CD8(+) T central memory cells (Tcm). LT expanded CD8(+) and CD4(+) Tcm cells retained a gene expression profile related to Tcm and T memory stem cells (Tscm). In vivo transfer of LT expanded Tcm revealed persistence and anti-tumor capacity. We confirmed our in vitro findings on human T cells, on healthy donors and diffuse large B cell lymphoma patients, undergoing salvage therapy. Our study demonstrates the feasibility of an extended T cell expansion as a practicable alternative for patients with insufficient numbers of T cells after the standard manufacturing process thereby increasing ATT accessibility

An exploratory study into the role and interplay of intrinsic and extrinsic cues in Australian consumers’ evaluations of fish.

This study explores the role and interplay of intrinsic and extrinsic cues when evaluating fish quality and in shaping consumers' attitudes toward fish consumption. A sensory analysis of nine different fish including five variants of barramundi was conducted to determine how consumers evaluated the fish on intrinsic cues. Focus groups were then conducted to explore the impact of extrinsic cues on attitudes and purchase intentions. While the sensory analysis revealed distinct differences between barramundi variants on intrinsic cues (notably taste), the focus groups revealed that, as a brand, barramundi is perceived much more favourably and consistently. Consumers used extrinsic cues, particularly country of origin, as surrogate indicators of quality. Aquaculture producers need to ensure intrinsic product quality and consistency, as while consumers use the extrinsic cue of "Australian grown" as a surrogate indicator of quality, as their familiarity and confidence with seafood grows, this overreliance on extrinsic cues may diminish