Full probabilistic design of a submerged floating tunnel and format for partial safety factors

A submerged floating tunnel (SFT) can be a promising solution for crossing a deep or wide waterway. This innovative concept however lacks research into its probabilistic design. In this research, the reliability of the tether-stabilized SFT is assessed. A first-order reliability method (FORM) and a Monte Carlo simulation (MCS) are performed for the limit state functions of the most important failure mechanisms. Stochastic variables are chosen so that a target reliability index of 3.8 for a reference period of 50 years is met. The calculated factors from the full probabilistic design are compared with the general recommended partial factors for strength and resistance from Eurocode EN1990. For the strength mechanisms, the calculated factors are smaller than the factors from Eurocode. However, for the equilibrium mechanism, the calculated factor for the unfavorable loading is larger than the factor from Eurocode and should be increased by 10% in order to design a safe enough SFT

FELINE ATRIAL NATRIURETIC PEPTIDE (ANP) HAS A SINGLE COPY GENE

O peptídeo natriurético atrial felino (PNA) é um hormônio sintetizado pelo miocárdio cardíaco atrial que, uma vez liberado na circulação, produz diurese, natriurese e vasodilatação periférica. Nós recentemente isolamos, sequenciamos e estabelemos a expressão cardíaca normal do PNA felino. No presente estudo, nós apresentamos o número de cópias gênicas do PNA em gatos. Foram extraídas amostras de DNA genômico do sangue periférico humano e felino, as quais foram purificadas e digeridas com as enzimas de restrição EcoRI, BamHI e HindIII, com posterior realização da técnica de Southern Blots. Primers para PCR, confeccionados para regiões de alta conservatividade entre espécies, foram utilizados para amplificação, clonagem e sequenciamento de um produto de 900 pares de bases, posteriormente utilizado como sonda de DNA. O gene do PNA felino possui uma única cópia gênica, a exemplo do PNA humano. Devido a sua similaridade, sondas de DNA felino puderam ser utilizadas para a hibridização de DNA genômico tanto do gato como do homem. Abstract Atrial natriuretic peptide (ANP) is a hormone normally synthesized by the cardiac atrial myocardium that once released, produces diuresis, natriuresis and peripheral vasodilation. We have recently isolated, sequenced and assessed the normal cardiac expression of feline ANP. In this study, we report on the number of copies of the feline ANP gene, which could have an effect on future molecular biology studies of ANP gene expression in cat models of cardiovascular disease. Cat and human genomic DNA (gDNA) were extracted for Southern blotting from blood. The purified DNA was digested with the restriction enzymes EcoRI, BamHI and HindIII followed by agarose gel electrophoresis and blotting onto nytran membranes. A feline ANP cDNA probe of approximately 900 base pairs was used for hybridization of the membranes. The results of Southern blotting showed that both the feline and human genomes contain a single copy of the ANP gene

Mining for viral fragments in methylation enriched sequencing data

Most next generation sequencing experiments generate more data than is required for the experimental set up. For example, methyl-CpG binding domain (MBD) affinity purification based sequencing is often used for DNA-methylation profiling, but up to 30% of the sequenced fragments cannot be mapped uniquely to the reference genome. Here we present and evaluate a methodology for the identification of viruses in these otherwise unused paired-end MBD-seq data. Viral detection is accomplished by mapping non-reference alignable reads to a comprehensive set of viral genomes. As viruses play an important role in epigenetics and cancer development, 92 (pre)malignant and benign samples, originating from two different collections of cervical samples and related cell lines, were used in this study. These samples include primary carcinomas (n=22), low- & high-grade cervical intrapeithelial neoplasia (CIN1 & CIN2/3 - n=2/n=30) and normal tissue (n=20), as well as control samples (n=17). Viruses that were detected include phages, adenoviruses, herpesviridae and HPV. HPV, which causes virtually all cervical cancers, was identified in 95% of the carcinomas, 100% of the CIN2/3 samples, both CIN1 samples and in 55% of the normal samples. Comparing the amount of mapped fragments on HPV for each HPV-infected sample yielded a significant difference between normal samples and carcinomas or CIN2/3 samples (adjusted p-values resp. < 10^-5, < 10^-5), reflecting different viral loads and/or methylation degrees in non-normal samples. Fragments originating from different HPV types could be distinguished and were independently validated by PCR-based assays with a specificity of 98% and a sensitivitity of 66%. In conclusion, although limited by the a priori knowledge of viral reference genome sequences, the proposed methodology can provide a first but substantial insight into the presence, concentration and types of methylated viral sequences in MBD-seq data without additional costs

Eye–hand coordination during manual object transport with the affected and less affected hand in adolescents with hemiparetic cerebral palsy

In the present study we investigated eye–hand coordination in adolescents with hemiparetic cerebral palsy (CP) and neurologically healthy controls. Using an object prehension and transport task, we addressed two hypotheses, motivated by the question whether early brain damage and the ensuing limitations of motor activity lead to general and/or effector-specific effects in visuomotor control of manual actions. We hypothesized that individuals with hemiparetic CP would more closely visually monitor actions with their affected hand, compared to both their less affected hand and to control participants without a sensorimotor impairment. A second, more speculative hypothesis was that, in relation to previously established deficits in prospective action control in individuals with hemiparetic CP, gaze patterns might be less anticipatory in general, also during actions performed with the less affected hand. Analysis of the gaze and hand movement data revealed the increased visual monitoring of participants with CP when using their affected hand at the beginning as well as during object transport. In contrast, no general deficit in anticipatory gaze control in the participants with hemiparetic CP could be observed. Collectively, these findings are the first to directly show that individuals with hemiparetic CP adapt eye–hand coordination to the specific constraints of the moving limb, presumably to compensate for sensorimotor deficits

Gene conversion in human rearranged immunoglobulin genes

Over the past 20 years, many DNA sequences have been published suggesting that all or part of the V&lt;sub&gt;H&lt;/sub&gt; segment of a rearranged immunoglobulin gene may be replaced in vivo. Two different mechanisms appear to be operating. One of these is very similar to primary V(D)J recombination, involving the RAG proteins acting upon recombination signal sequences, and this has recently been proven to occur. Other sequences, many of which show partial V&lt;sub&gt;H&lt;/sub&gt; replacements with no addition of untemplated nucleotides at the V&lt;sub&gt;H&lt;/sub&gt;–V&lt;sub&gt;H&lt;/sub&gt; joint, have been proposed to occur by an unusual RAG-mediated recombination with the formation of hybrid (coding-to-signal) joints. These appear to occur in cells already undergoing somatic hypermutation in which, some authors are convinced, RAG genes are silenced. We recently proposed that the latter type of V&lt;sub&gt;H&lt;/sub&gt; replacement might occur by homologous recombination initiated by the activity of AID (activation-induced cytidine deaminase), which is essential for somatic hypermutation and gene conversion. The latter has been observed in other species, but not in human Ig genes, so far. In this paper, we present a new analysis of sequences published as examples of the second type of rearrangement. This not only shows that AID recognition motifs occur in recombination regions but also that some sequences show replacement of central sections by a sequence from another gene, similar to gene conversion in the immunoglobulin genes of other species. These observations support the proposal that this type of rearrangement is likely to be AID-mediated rather than RAG-mediated and is consistent with gene conversion