Comparison of Physical Properties of Untreated and Heat Treated Beech and Hornbeam

Istraživanjem fizikalnih svojstava toplinski obrađene bukovine i grabovine utvrđeno je da je njihova srednja vrijednost manja i signifikantno se razlikuje od srednjih vrijednosti fizikalnih svojstava neobrađene bukovine i grabovine. Srednja vrijednost gustoće u apsolutno suhom stanju toplinski obrađene bukovine manja je za 8,5 % od neobrađene, a za grabovinu je ona manja za 7,5 %. Smanjenje srednjih vrijednosti maksimalnih utezanja toplinski obrađene bukovine i grabovine u odnosu prema neobrađenoj još je veće. Maksimalno radijalno utezanje toplinski obrađene bukovine manje je za 7 %, maksimalno tangencijalno utezanje za 23,5 %, a maksimalno volumno utezanje za 19,3 % od istih fizikalnih svojstava neobrađene bukovine. Toplinski obrađena grabovina ima srednju vrijednost maksimalnoga radijalnog utezanja za 123 %, maksimalnoga tangencijalnog utezanja za 86 % i maksimalnoga volumnog utezanja za 99,5 % manju od istih fizikalnih svojstava neobrađene grabovine. Takvim smanjenjem maksimalnih utezanja u radijalnome i tangencijalnom smjeru toplinskom obradom grabovina postaje znatno prihvatljivija za izradu proizvoda za koje je važna dimenzionalna stabilnost.The investigation of physical properties of heat treated beech wood and hornbeam wood found that their average value is lower and significantly different from average values of physical properties of untreated beech wood and hornbeam wood. The average value of density in absolutely dry condition of heat treated beech wood is smaller by 8.5% from the untreated, and the hornbeam wood is smaller by 7.5%. Reduction of average values of maximum shrinkage of heat treated beech wood and hornbeam wood is even bigger in relation to the untreated wood. Maximum radial shrinkage of heat treated beech wood is smaller by 7%, maximum tangential shrinkage by 23.5% and maximum volumetric shrinkage by 19.3% compared to the same physical properties of untreated beech wood. Heat treated hornbeam wood has an average value of maximum radial shrinkage smaller by 123%, maximum tangential shrinkage by 86% and maximum volume shrinkage by 99.5% compared to the same physical properties of untreated hornbeam wood. With such reduction in the maximum shrinkage in radial and tangential direction using heat treatment, hornbeam becomes particulary suitable for making products where dimensional stability is important

Hidden Mutations in CdLS - Limitations of Sanger Sequencing in Molecular Diagnostics

International audienceCornelia de Lange syndrome (CdLS) is a well characterized developmental disorder. The genetic cause of CdLS is a mutation in one of five associated genes (NIPBL, SMC1A, SMC3, RAD21 and HDAC8) accounting for about 70 % of cases. To improve our current molecular diagnostic and to analyze some of CdLS candidate genes we developed and established a gene panel approach. Because recent data indicate a high frequency of mosaic NIPBL mutations that were not detected by conventional sequencing approaches of blood DNA, we started to collected buccal mucosa samples of our patients that were negative for mutations in the known CdLS genes. Here we report the identification of three mosaic NIPBL mutations by our high-coverage gene panel sequencing approach that were undetected by classical Sanger sequencing analysis of buccal mucosa DNA. All mutations were confirmed by the use of highly sensitive SNaPshot fragment analysis using DNA from buccal mucosa, urine and fibroblast samples. In blood samples we could not detect the respective mutation. Finally, in fibroblast samples from all three patients, Sanger sequencing could identify all the mutations. Thus, our study highlights the need for highly sensitive technologies in molecular diagnostic of CdLS to improve genetic diagnosis and counseling of patients and their families. This article is protected by copyright. All rights reserved

New <i>NR5A1</i> mutations and phenotypic variations of gonadal dysgenesis

<div><p>Mutations in <i>NR5A1</i> have been reported as a frequent cause of 46,XY disorders of sex development (DSD) associated to a broad phenotypic spectrum ranging from infertility, ambiguous genitalia, anorchia to gonadal dygenesis and female genitalia. Here we present the clinical follow up of four 46,XY DSD patients with three novel heterozygous mutations in the <i>NR5A1</i> gene leading to a p.T40P missense mutation and a p.¹⁸DKVSG²²del nonframeshift deletion in the DNA-binding domain and a familiar p.Y211Tfs*83 frameshift mutation. Functional analysis of the missense and nonframeshift mutation revealed a deleterious character with loss of DNA-binding and transactivation capacity. Both, the mutations in the DNA-binding domain, as well as the familiar frameshift mutation are associated with highly variable endocrine values and phenotypic appearance. Phenotypes vary from males with spontaneous puberty, substantial testosterone production and possible fertility to females with and without Müllerian structures and primary amenorrhea. Exome sequencing of the sibling’s family revealed <i>TBX2</i> as a possible modifier of gonadal development in patients with <i>NR5A1</i> mutations.</p></div

Electrophoretic mobility shift assay of NR5A1 mutants.

<p>A) DNA-binding capability of mutant NR5A1-T40P and NR5A1-¹⁸DKVSG²²del were compared to wt-NR5A1 using a SF-1 responsive element of the mouse <i>Amh</i> promoter. Nuclear extracts containing WT-SF1 shifted the labelled probe (arrow). A 200 fold excess of unlabelled competitor abolished the shifted signal, while a 200 fold excess of unlabelled mutated competitor rescues the shifted signal. Both mutant proteins have lost their DNA-binding capability. B) Aliquots of the nuclear extracts used in EMSA were separated by SDS gel electrophoresis, transferred to a nitrocellulose membrane and stained with Ponceau S and anti-myc antibodies to verify equal SF-1 protein concentrations.</p

Hormonal and clinical data of patients.

<p>Hormonal and clinical data of patients.</p

Transactivation of the human <i>AMH</i>- and <i>STAR</i>-promoter.

<p>The transactivation capacity of mutant NR5A1-T40P and NR5A1-¹⁸DKVSG²²del were compared to wt-NR5A1 using human <i>AMH-</i> (A) and <i>STAR</i>-promoter (B) containing reporter genes. The empty pCMV-Myc vector represents background activity. Wild type (WT) activity was set 100%. RLU = relative luciferase activity. Error bars represent standard deviations, ***P = <0.001, t-test comparison of WT and mutant. C: Immunoblot detection of myc-tagged NR5A1 proteins. Equal loadings were verified by detection of total proteins using the TGX Stain Free system (BioRad). D: statistical analysis of 3 western blots of three different experiments showing an approximately equal protein accumulation of NR5A1-WT and mutant ¹⁸DKVSG²²del, while mutant T40P showed an 2–3 fold enhanced accumulation. Images were quantified and normalized to total protein using Image Lab 5.2.1 (BioRad).</p

Electropherogram of <i>NR5A1</i> mutations.

<p>Electropherogram of the heterozygous c.118A>C mutation in patient 1 (A), the nonframeshift deletion c.51_65delGGACAAGGTGTCCGG in patient 2 (B), the frame-shift deletion c.630_636delGTACGGC in patient 3 (C) and the pedigree of the family of patient 3 and 4 (D). Circles denote phenotypic females, squares denote phenotypic males. Filled squares and circles correspond to a DSD condition, dots to a carrier status.</p

Human variation in population-wide gene expression data predicts gene perturbation phenotype.

Funder: German Research FoundationFunder: Dutch Research Council (NWO)Funder: BMBF BonnFunder: Europäische KommissionPopulation-scale datasets of healthy individuals capture genetic and environmental factors influencing gene expression. The expression variance of a gene of interest (GOI) can be exploited to set up a quasi loss- or gain-of-function "in population" experiment. We describe here an approach, huva (human variation), taking advantage of population-scale multi-layered data to infer gene function and relationships between phenotypes and expression. Within a reference dataset, huva derives two experimental groups with LOW or HIGH expression of the GOI, enabling the subsequent comparison of their transcriptional profile and functional parameters. We demonstrate that this approach robustly identifies the phenotypic relevance of a GOI allowing the stratification of genes according to biological functions, and we generalize this concept to almost 16,000 genes in the human transcriptome. Additionally, we describe how huva predicts monocytes to be the major cell type in the pathophysiology of STAT1 mutations, evidence validated in a clinical cohort