Design and off-line programming of a robotic tape winding cell

Tape winding is a variation on filament winding, in which a single tape is used instead of a bundle of independently tensioned rovings. At K.U. Leuven, a robotic tape winding cell has been developed. Starting from data of the fiber paths, a collision-free off-line robot program is generated. To avoid collisions a heuristic method is used. The method has been validated by winding of a T-piece.     

Integration of Mutational Signature Analysis with 3D Chromatin Data Unveils Differential AID-Related Mutagenesis in Indolent Lymphomas

Activation-induced deaminase (AID) is required for somatic hypermutation in immunoglobulin genes, but also induces off-target mutations. Follicular lymphoma (FL) and chronic lymphocytic leukemia (CLL), the most frequent types of indolent B-cell tumors, are exposed to AID activity during lymphomagenesis. We designed a workflow integrating de novo mutational signatures extraction and fitting of COSMIC (Catalogue Of Somatic Mutations In Cancer) signatures, with tridimensional chromatin conformation data (Hi-C). We applied the workflow to exome sequencing data from lymphoma samples. In 33 FL and 30 CLL samples, 42% and 34% of the contextual mutations could be traced to a known AID motif. We demonstrate that both CLL and FL share mutational processes dominated by spontaneous deamination, failures in DNA repair, and AID activity. The processes had equiproportional distribution across active and nonactive chromatin compartments in CLL. In contrast, canonical AID activity and failures in DNA repair pathways in FL were significantly higher within the active chromatin compartment. Analysis of DNA repair genes revealed a higher prevalence of base excision repair gene mutations (p = 0.02) in FL than CLL. These data indicate that AID activity drives the genetic landscapes of FL and CLL. However, the final result of AID-induced mutagenesis differs between these lymphomas depending on chromatin compartmentalization and mutations in DNA repair pathways.     

Attaching of flexible chains to novolacs on the basis of bisphenol A

Networks containing both flexible segments and rigid structures were synthesized on the basis of bisphenol A novolacs and diglycidylether of butanediol using imidazole as an accelerator. A stoichiometric ratio between epoxy groups and phenolic groups of the novolacs leads to networks with methylene bridges as network junctions. In contrast to this, the same reaction with bisphenol A leads to completely soluble products. The glass transition temperature of this soluble material is considerably lower than the glass transition temperature of the networks. Increasing content of methylene bridges in the novolacs leads to an increase of the glass transition temperature of the networks and to a decrease of the δc_p value at the glass transition. Furthermore, epoxy excess leads to networks with rubber-structure of the bisphenol A novolac used in the reaction with the diglycidylether. It was found that conformations with intramolecular hydrogen bondings exist between phenolic hydroxyl groups, which considerably influence the reactivity of the novolac with the epoxy group. © 1996 John Wiley & Sons, Inc.     

A proposed multigeneration protocol for Japanese medaka (Oryzias latipes) to evaluate effects of endocrine disruptors.

Definitive data on reproductive impairment of chronically exposed populations may be required to assess the appropriateness of the existing test methods for hazard identification and prioritization of endocrine modulators. Multigeneration toxicity testing protocols for wildlife receptors are lacking. To help address this gap we describe a multigeneration fish assay using the freshwater fish, Japanese medake (Oryzias latipes). This test species has been used for the evaluation of carcinogenic, teratogenic and reproductive effects and is sensitive to estrogen exposure producing ovo-testis, altered biochemical parameters and phenotypic characteristics. Due to the short life cycle, a multigeneration test with medaka can be conducted in 1 year. Endpoints evaluated include: survival, growth, sex ratio, fecundity, embryonic lesion occurrence, embryonic stage development, gonadal and hepatic somatic indices, histopathology and biochemical parameters. As new endpoints are developed they can be incorporated into the protocol. Results of a positive control (17 beta-estradiol) study are presented to give an indication of the baseline associated with various test endpoints and to highlight the importance of nutrition in the experimental design. 17 beta-Estradiol treatment induced vitellogenin production in male and female medaka, feminized males, and disrupted egg production. The proposed protocol provides researchers with an effective multigeneration fish test that can be used to examine potential effects of stressors at the population, individual, cellular and subcellular level.     

Experimental Validation of a Fundamental Model for PCR Efficiency

Recently a theoretical analysis of PCR efficiency has been published by Booth et al. (2010). The PCR yield is the product of three efficiencies: (i) the annealing efficiency is the fraction of templates that form binary complexes with primers during annealing, (ii) the polymerase binding efficiency is the fraction of binary complexes that bind to polymerase to form ternary complexes and (iii) the elongation efficiency is the fraction of ternary complexes that extend fully. Yield is controlled by the smallest of the three efficiencies and control could shift from one type of efficiency to another over the course of a PCR experiment. Experiments have been designed that are specifically controlled by each one of the efficiencies and the results are consistent with the mathematical model. The experimental data has also been used to quantify six key parameters of the theoretical model. An important application of the fully characterized model is to calculate initial template concentration from real-time PCR data. Given the PCR protocol, the midpoint cycle number (where the template concentration is half that of the final concentration) can be theoretically determined and graphed for a variety of initial DNA concentrations. Real-time results can be used to calculate the midpoint cycle number and consequently the initial DNA concentration, using this graph. The application becomes particularly simple if a conservative PCR protocol is followed where only the annealing efficiency is controlling.     

Homogeneous doping of ceramics by infiltration–gelation

Advanced ceramics require small amounts of cation dopants to improve the sintering process or achieve certain properties. Dopant precursors are often dissolved in initial processing liquids, which may adversely affect ceramic colloidal stability, and dopant transport during drying. This would lead to chemical and microstructural inhomogeniety in the eventual ceramics. Here we present a method that circumvents these problems using the example of Al³⁺-doped TiO₂ ceramics. Homogeneous TiO₂ compacts with 34% porosity are first prepared by colloidal casting and calcined at 700 °C. The obtained compacts are infiltrated with an aqueous solution of Al(NO₃)₃, citric acid and ethylene glycol. The solution composition is adjusted such that during drying, a gel forms that covers the internal pore surface. Subsequent decomposition of organics results in a homogeneous dopant distribution in the porous and then dense-sintered TiO₂ compacts. This is verified with SEM-EDX, and laser ablation coupled ICP-MS on dense compacts sintered at 1100 °C.     

Enhanced upconversion study of Er3+-Yb3+ codoped NaYF4 phosphors synthesized by the reverse microemulsion method

Herein, nanocrystals of Er³⁺ and Er³⁺, Yb³⁺ co-doped NaYF₄ upconversion (UC) phosphor were prepared via the reverse-microemulsion method. The impact of different concentrations of Er³⁺ ions on the UC emission intensity after 980?nm diode laser excitation is discussed. The structure, morphology and composition of the nanophosphors were confirmed by X-ray diffraction, transmission electron microscopy, scanning electron microscopy and the results showed the presence of NaYF₄ nanocrystals with hexagonal phases of NaYF₄. The UC spectra revealed two emission bands including a green and a red emission band and the CIE coordinate for the samples were estimated. The present research revealed that the reverse-microemulsion approach will be suitable for the synthesis of efficient upconversion nanophosphors.     

Hemi- and Homozygous Loss-of-Function Mutations in DSG2 (Desmoglein-2) Cause Recessive Arrhythmogenic Cardiomyopathy with an Early Onset

About 50% of patients with arrhythmogenic cardiomyopathy (ACM) carry a pathogenic or likely pathogenic mutation in the desmosomal genes. However, there is a significant number of patients without positive familial anamnesis. Therefore, the molecular reasons for ACM in these patients are frequently unknown and a genetic contribution might be underestimated. Here, we used a next-generation sequencing (NGS) approach and in addition single nucleotide polymor-phism (SNP) arrays for the genetic analysis of two independent index patients without familial medical history. Of note, this genetic strategy revealed a homozygous splice site mutation (DSG2–c.378+1G>T) in the first patient and a nonsense mutation (DSG2–p.L772X) in combination with a large deletion in DSG2 in the second one. In conclusion, a recessive inheritance pattern is likely for both cases, which might contribute to the hidden medical history in both families. This is the first report about these novel loss-of-function mutations in DSG2 that have not been previously identi-fied. Therefore, we suggest performing deep genetic analyses using NGS in combination with SNP arrays also for ACM index patients without obvious familial medical history. In the future, this finding might has relevance for the genetic counseling of similar cases.     

On the optimum mass transfer of flat absorbing falling films

The coupled heat- and mass-transfer of the absorbing falling film is considered in the flat-film approximation. We study the influence of the operating conditions on the absorption efficiency in a fully non-dimensional framework and find an optimal Reynolds number, which can be interpreted as an optimal film thickness. We also test various thermal wall boundary conditions to select the best approximation compared to a realistic test case.     

High Versatility of IPP and DMAPP Methyltransferases Enables Synthesis of C6, C7 and C8 Terpenoid Building Blocks

The natural substance class of terpenoids covers an extremely wide range of different structures, although their building block repertoire is limited to the C₅ compounds DMAPP and IPP. This study aims at the characterization of methyltransferases (MTases) that modify these terpene precursors and the demonstration of their suitability for biotechnological purposes. All seven enzymes tested accepted IPP as substrate and altogether five C₆ compounds and six C₇ compounds were formed within the reactions. A high selectivity for the deprotonation site as well as high stereoselectivity could be observed for most of the biocatalysts. Only the enzyme from Micromonospora humi also accepted DMAPP as substrate, converting it into (2R)-2-methyl-IPP in vitro. In vivo studies demonstrated the production of a C₈ compound and a hydride shift step within the MTase-catalyzed reaction. Our study presents IPP/DMAPP MTases with very different catalytic properties, which provide biosynthetic access to many novel terpene-derived structures.