Are p.I148T, p.R74W and p.D1270N cystic fibrosis causing mutations ?

BACKGROUND: To contribute further to the classification of three CFTR amino acid changes (p.I148T, p.R74W and p.D1270N) either as CF or CBAVD-causing mutations or as neutral variations. METHODS: The CFTR genes from individuals who carried at least one of these changes were extensively scanned by a well established DGGE assay followed by direct sequencing and familial segregation analysis of mutations and polymorphisms. RESULTS: Four CF patients (out of 1238) originally identified as carrying the p.I148T mutation in trans with a CF mutation had a second mutation (c.3199del6 or a novel mutation c.3395insA) on the p.I148T allele. We demonstrate here that the deletion c.3199del6 can also be associated with CF without p.I148T. Three CBAVD patients originally identified with the complex allele p.R74W-p.D1270N were also carrying p.V201M on this allele, by contrast with non CF or asymptomatic individuals including the mother of a CF child, who were carrying p.R74W-p.D1270N alone. CONCLUSION: These findings question p.I148T or p.R74W-p.D1270N as causing by themselves CF or CBAVD and emphazises the necessity to perform a complete scanning of CFTR genes and to assign the parental alleles when novel missense mutations are identified

Large genomic rearrangements in the CFTR gene contribute to CBAVD

<p>Abstract</p> <p>Background</p> <p>By performing extensive scanning of whole coding and flanking sequences of the <it>CFTR (Cystic Fibrosis Transmembrane Conductance Regulator</it>) gene, we had previously identified point mutations in 167 out of 182 (91.7%) males with isolated congenital bilateral absence of the vas deferens (CBAVD). Conventional PCR-based methods of mutation analysis do not detect gross DNA lesions. In this study, we looked for large rearrangements within the whole <it>CFTR </it>locus in the 32 CBAVD patients with only one or no mutation.</p> <p>Methods</p> <p>We developed a semi-quantitative fluorescent PCR assay (SQF-PCR), which relies on the comparison of the fluorescent profiles of multiplex PCR fragments obtained from different DNA samples. We confirmed the gross alterations by junction fragment amplification and identified their breakpoints by direct sequencing.</p> <p>Results</p> <p>We detected two large genomic heterozygous deletions, one encompassing exon 2 (c.54-5811_c.164+2186del8108ins182) [or <it>CFTRdele2</it>], the other removing exons 22 to 24 (c.3964-3890_c.4443+3143del9454ins5) [or <it>CFTRdele 22_24</it>], in two males carrying a typical CBAVD mutation on the other parental <it>CFTR </it>allele. We present the first bioinformatic tool for exon phasing of the <it>CFTR </it>gene, which can help to rename the exons and the nomenclature of small mutations according to international recommendations and to predict the consequence of large rearrangements on the open reading frame.</p> <p>Conclusion</p> <p>Identification of large rearrangements further expands the <it>CFTR </it>mutational spectrum in CBAVD and should now be systematically investigated. We have designed a simple test to specifically detect the presence or absence of the two rearrangements identified in this study.</p

Utilizing heterogeneous network formation to tune surface roughness: A method to control coating wettability

International audienc

Varietates fortunae : religion et mythologie à Rome : hommage à Jacqueline Champeaux

Publié dans la collection Roma antiqua , ISSN 1953-9215 ; 201

Varietates fortunae : religion et mythologie à Rome : hommage à Jacqueline Champeaux

Publié dans la collection Roma antiqua , ISSN 1953-9215 ; 201

Spontaneous, phase-separation induced surface roughness: A new method to design parahydrophobic polymer coatings with rose petal-like morphology

International audienceWhile the development of polymer coatings with controlled surface topography is a growing research topic, a fabrication method that does not rely on lengthy processing times, bulk solvent solution, or secondary functionalization has yet to be identified. This study presents a facile, rapid, in situ method to develop parahydrophobic coatings based on phase separation during photopolymerization. A comonomer resin of ethylene glycol diacrylate (EGDA) and 1H,1H,2H,2H-perfluorodecyl acrylate (PFDA) is modified with a thermoplastic additive (PVDF) to induce phase separation during polymerization. If applied to a glass substrate and photopolymerized, the EGDA:PFDA copolymer forms a homogeneous network with a single glass transition temperature (Tg) and slight hydrophobicity (θw ~ 114°). When the resin is modified with PVDF, phase separation occurs during photopolymerization producing a heterogeneous network with two Tg’s. The phase separation causes differences in composition and cross-link density within the network, which leads to local variations in polymerization shrinkage across the non-constrained material interface. Domains with higher cross-link densities shrink and contract towards the bulk material more dramatically, permitting the formation of rough surfaces with sub-micron sized spheres enriched in PVDF dispersed in a continuous matrix of EGDA:PFDA copolymer. Both the surface roughness and hydrophobic components in the resin render these surfaces parahydrophobic with θw ~ 150°, high water adhesion, and a similar morphology to rose petals observed in nature

Next-generation sequencing and recombinant expression characterized aberrant splicing mechanisms and provided correction strategies in factor VII deficiency

Despite the exhaustive screening of F7 gene exons and exon-intron boundaries and promoter region, a significant proportion of mutated alleles remains unidentified in patients with coagulation factor VII deficiency. Here, we applied next-generation sequencing to thirteen FVII-deficient patients displaying genotype-phenotype discrepancies upon conventional sequencing, and identified six rare intronic variants. Computational analysis predicted splicing effects for three of them, which would strengthen (c.571+78G&gt;A; c.806-329G&gt;A) or create (c.572-392C&gt;G) intronic 5' splice sites (5'ss). In F7 minigenes assays the c.806-329G&gt;A was ineffective while the c.571+78G&gt;A change led to usage of the +79 cryptic 5'ss with only trace levels of correct transcripts (3% of wild-type), in accordance with factor VII activity levels in homozygotes (1-3% of normal). The c.572-392C&gt;G change led to pseudo-exonization and frame-shift, but also substantial levels of correct transcripts (~70%). However, this variant was associated with the common F7 polymorphic haplotype predicted to further decrease factor VII levels, thus roughly explaining the factor VII levels of 10% in the homozygous patient. Intriguingly, the effect of the c.571+78G&gt;A and c.572-392C&gt;G changes, and particularly of the former, the severest and well-represented in our cohort, was counteracted by antisense U7snRNA variants targeting the intronic 5'ss, thus demonstrating their pathogenic role. In conclusion, the combination of next-generation sequencing of the entire F7 gene with the minigene expression studies elucidated the molecular bases of factor VII deficiency in ten out of thirteen patients, thus improving diagnosis and genetic counselling, and provided a potential therapeutic approach based on antisense molecules, successfully exploited in other disorders

Leukemic non-nodal mantle cell lymphomas have a distinct phenotype and are associated with deletion of PARP1 and 13q14

International audienceLeukemic non-nodal mantle cell lymphoma (lMCL) is a particular subtype of mantle cell lymphoma (MCL), characterized by leukemic non-nodal disease and slow progression. Recognition of this entity is relevant to avoid overtreatment. Despite indolent clinical behaviour, lMCL might transform to a more aggressive disease. The purpose of this study was to compare lMCL with classical MCL (cMCL) and aggressive MCL (aMCL) using immunohistochemistry, interphase fluorescence in situ hybridization (FISH), and array-based comparative genomic hybridization, in order to identify biomarkers for lMCL diagnosis and prognosis. Seven lMCL patients were included. All had bone marrow involvement without lymphadenopathy. An lMCL phenotype was distinct from that of cMCL and aMCL: SOX11-, ATM+, PARP1+/-, and low KI67 (average 2 %). Beyond the t(11;14) translocation, fewer secondary cytogenetic alterations were found in lMCL compared to cMCL and aMCL, including deletion of PARP1 and 13q14. At last follow-up, one patient with lMCL had died of disease and another had progressive disease. These patients were respectively 13q14 deletion- and PARP1-positive. One other case of lMCL harbored a 13q14 deletion associated with PARP1 deletion. This patient had indolent disease. lMCL has a particular phenotype and fewer secondary cytogenetic alterations than cMCL and aMCL. PARP1 protein expression and 13q14 deletion are associated with a progressive clinical course of lMCL and should be included in initial diagnostic studies as predictors of unfavorable outcome. PARP1 deletion is involved in lMCL pathogenesis and might confer advantage

Variation of Goliathus orientalis (Moser, 1909) Elytra Nanostructurations and Their Impact on Wettability

Among the different species of flower beetles, there is one of particular notoriety: the Goliath beetle. This large insect can grow up to 11 cm long and is well-known for its distinctive black and white shield. In this paper, we focus on a particular Goliathus species: G. orientalis (Moser, 1909). We investigated the variations in properties of both the black and white parts of the upper face of G. orientalis; more precisely, the variation in surface properties with respect to the wettability of these two parts. This work reveals that the white parts of the shield have a higher hydrophobic character when compared to the black regions. While the black parts are slightly hydrophobic (&theta; = 91 &plusmn; 5&deg;) and relatively smooth, the white parts are highly hydrophobic (&theta; = 130 &plusmn; 3&deg;) with strong water adhesion (parahydrophobic); similar to the behavior observed for rose petals. Roughness and morphology analyses revealed significant differences between the two parts, and, hence, may explain the change in wettability. The white surfaces are covered with horizontally aligned nanohairs. Interestingly, vertically aligned microhairs are also present on the white surface. Furthermore, the surfaces of the microhairs are not smooth, they contain nanogrooves that are qualitatively similar to those observed in cactus spines. The nanogrooves may have an extremely important function regarding water harvesting, as they preferentially direct the migration of water droplets; this process could be mimicked in the future to capture and guide a large volume of water