Life-long course and molecular characterization of the original Dutch family with epidermolysis bullosa simplex with muscular dystrophy due to a homozygous novel plectin point mutation

Plectin is one of the largest and most versatile cytolinker proteins known. Cloned and sequenced in 1991, it was later shown to have nonsense mutations in recessive epidermolysis bullosa with muscular dystrophy. A dominant mutation in the gene was found to cause epidermolysis bullosa simplex Ogna without muscular dystrophy. Here we report the DNA sequencing of the plectin gene (PLEC1) in a Dutch family originally described in 1972 as having epidermolysis bullosa with muscular dystrophy. The results revealed homozygosity for a new plectin nonsense mutation at position 13187 and its specific 8q24 marker haplotype profile. Western blotting of cultured fibroblasts and immunofluorescence microscopy of skin biopsy confirm that the plectin protein expression is grossly reduced or absent. A summary of the life-long clinical course of the two affected brothers homozygous for the new E1914X mutation is given

Mutation in exon 1a of PLEC, leading to disruption of plectin isoform 1a, causes autosomal-recessive skin-only epidermolysis bullosa simplex

PLEC, the gene encoding the cytolinker protein plectin, has eight tissue-specific isoforms in humans, arising by alternate splicing of the first exon. To date, all PLEC mutations that cause epidermolysis bullosa simplex (EBS) were found in exons common to all isoforms. Due to the ubiquitous presence of plectin in mammalian tissues, EBS from recessive plectin mutations is always associated with extracutaneous involvement including muscular dystrophy, pyloric atresia and cardiomyopathy. We studied a consanguineous family with sisters having isolated blistering suggesting EBS. Skin disease started with foot blisters at walking age and became generalized at puberty while sparing mucous membranes. DNA sequencing revealed a homozygous nonsense mutation (c.46C>T; p.Arg16X) in the first exon of the plectin variant encoding plectin isoform 1a (P1a). Immunofluorescence antigen mapping, transmission electron microscopy, western blot analysis and qRT-PCR were performed on patient skin and cultured keratinocytes, control myocardium and striated muscle samples. We found hypoplastic hemidesmosomes and intra-epidermal ‘pseudo-junctional' cleavage fitting EBS. Screening for cardiomyopathy and muscle dystrophy showed no abnormalities. We report the first cases of autosomal-recessive EBS from P1a deficiency affecting skin, while mucous membranes, heart and muscle are spared. The dominant expression of the P1a isoform in epidermal basal cell layer and cultured keratinocytes suggests that mutations in the first exon of isoform 1a cause skin-only EBS without extracutaneous involvement. Our study characterizes yet another of the eight isoforms of plectin and adds a tissue-specific phenotype to the spectrum of ‘plectinopathies' produced by mutations of unique first exons of this gen

Biodiversity and dynamics of plant groups of Chebket El Melhassa region (Algeria)

This article examines phytoecological aspects of plant groups in the Chebket El Melhassa region (Tiaret-Ouest Algérien) by several types of analysis: biological, biogeographic and statistical. From the plant analysis, a list of 103 taxa distributed in 36 families was compiled, biologically characterized by a dominance of therophytes (45.6%) with species of the Mediterranean biogeographic type 20 species (19.8%) assuming particular importance. The ordination of the plant community was performed in the search for the optimum solution based on correlation with environmental factors, estimated using the phytoindication approach. The results reveal that the plants represented in the community, mostly prefer strongly lit places. Phytoindication shows a warm sub Mediterranean climate. Continentality corresponds to the sub-oceanic climate. The moisture regime ranges from strongly dry to dry soil. The plant community is formed on moderately acid soils. Plants are able to grow on both strongly acidic and neutral soils. The soil is very poor or poor in nitrogen. A four dimension variant of multidimensional scaling procedure was selected as the most appropriate decision. Dimensions selected after nonmetric multidimensional scaling were interpreted by computing weighted average scores of ecological factors for ordination configuration. Four of the plant species clusters were found to be the optimal solution on the basis of the Calinsky-Harabasz criteria. The clusters can be viewed as a functional group. Functional group A is quite diverse and represented by 42 species. This functional group is closest to the community optimum because in all four space dimensions the corresponding cluster is near the origin, which corresponds to the most typical ecological conditions. Species that constitute the group B prefer minimum values of dimension 2. This indicates a preference for illuminated sites with high temperature regime and low soil humidity. This group includes 32 species. A feature of group С is that it is located in the area of maximum values for dimension 2. Thus, this functional group is opposite to functional group B. This indicates a preference of species included in the functional group C for wetter soils. Functional group C comprises 21 species. Functional group D differs considerably from all others in its ecological characteristics. The difference is in the preference for minimum values for measurement 1. This suggests that more acid soils are optimal for a given functional group. Functional group E comprises 8 species.This article examines phytoecological aspects of plant groups in the Chebket El Melhassa region (Tiaret-Ouest Algérien) by several types of analysis: biological, biogeographic and statistical. From the plant analysis, a list of 103 taxa distributed in 36 families was compiled, biologically characterized by a dominance of therophytes (45.6%) with species of the Mediterranean biogeographic type 20 species (19.8%) assuming particular importance. The ordination of the plant community was performed in the search for the optimum solution based on correlation with environmental factors, estimated using the phytoindication approach. The results reveal that the plants represented in the community, mostly prefer strongly lit places. Phytoindication shows a warm sub Mediterranean climate. Continentality corresponds to the sub-oceanic climate. The moisture regime ranges from strongly dry to dry soil. The plant community is formed on moderately acid soils. Plants are able to grow on both strongly acidic and neutral soils. The soil is very poor or poor in nitrogen. A four dimension variant of multidimensional scaling procedure was selected as the most appropriate decision. Dimensions selected after nonmetric multidimensional scaling were interpreted by computing weighted average scores of ecological factors for ordination configuration. Four of the plant species clusters were found to be the optimal solution on the basis of the Calinsky-Harabasz criteria. The clusters can be viewed as a functional group. Functional group A is quite diverse and represented by 42 species. This functional group is closest to the community optimum because in all four space dimensions the corresponding cluster is near the origin, which corresponds to the most typical ecological conditions. Species that constitute the group B prefer minimum values of dimension 2. This indicates a preference for illuminated sites with high temperature regime and low soil humidity. This group includes 32 species. A feature of group С is that it is located in the area of maximum values for dimension 2. Thus, this functional group is opposite to functional group B. This indicates a preference of species included in the functional group C for wetter soils. Functional group C comprises 21 species. Functional group D differs considerably from all others in its ecological characteristics. The difference is in the preference for minimum values for measurement 1. This suggests that more acid soils are optimal for a given functional group. Functional group E comprises 8 species.This article examines phytoecological aspects of plant groups in the Chebket El Melhassa region (Tiaret-Ouest Algérien) by several types of analysis: biological, biogeographic and statistical. From the plant analysis, a list of 103 taxa distributed in 36 families was compiled, biologically characterized by a dominance of therophytes (45.6%) with species of the Mediterranean biogeographic type 20 species (19.8%) assuming particular importance. The ordination of the plant community was performed in the search for the optimum solution based on correlation with environmental factors, estimated using the phytoindication approach. The results reveal that the plants represented in the community, mostly prefer strongly lit places. Phytoindication shows a warm sub Mediterranean climate. Continentality corresponds to the sub-oceanic climate. The moisture regime ranges from strongly dry to dry soil. The plant community is formed on moderately acid soils. Plants are able to grow on both strongly acidic and neutral soils. The soil is very poor or poor in nitrogen. A four dimension variant of multidimensional scaling procedure was selected as the most appropriate decision. Dimensions selected after nonmetric multidimensional scaling were interpreted by computing weighted average scores of ecological factors for ordination configuration. Four of the plant species clusters were found to be the optimal solution on the basis of the Calinsky-Harabasz criteria. The clusters can be viewed as a functional group. Functional group A is quite diverse and represented by 42 species. This functional group is closest to the community optimum because in all four space dimensions the corresponding cluster is near the origin, which corresponds to the most typical ecological conditions. Species that constitute the group B prefer minimum values of dimension 2. This indicates a preference for illuminated sites with high temperature regime and low soil humidity. This group includes 32 species. A feature of group С is that it is located in the area of maximum values for dimension 2. Thus, this functional group is opposite to functional group B. This indicates a preference of species included in the functional group C for wetter soils. Functional group C comprises 21 species. Functional group D differs considerably from all others in its ecological characteristics. The difference is in the preference for minimum values for measurement 1. This suggests that more acid soils are optimal for a given functional group. Functional group E comprises 8 species

Plectin as a prognostic marker in non-metastatic oral squamous cell carcinoma

Background: Oral squamous cell carcinoma (OSCC) is associated with a poor 5-year survival rate. In general, patients diagnosed with small tumors have a fairly good prognosis, but some small tumors have an aggressive behavior leading to early death. There are at present no reliable prognostic biomarkers for oral cancers. Thus, to optimize treatment for the individual patient, there is a need for biomarkers that can predict tumor behavior. Method: In the present study the potential prognostic value of plectin was evaluated by a tissue microarray (TMA) based immunohistochemical analysis of primary tumor tissue obtained from a North Norwegian cohort of 115 patients diagnosed with OSCC. The expression of plectin was compared with clinicopathological variables and 5 year survival. Results: The statistical analysis revealed that low expression of plectin in the tumor cells predicted a favorable outcome for patients with non-metastatic disease (p = 0.008). Furthermore, the expression of plectin was found to correlate (p = 0.01) with the expression of uPAR, which we have previously found to be a potential prognostic marker for T1N0 tumors. Conclusions: Our results indicate that low expression of plectin predicts a favorable outcome for patients with non-metastatic OSCC and the expression level of plectin may therefore be used in the treatment stratification for patients with early stage disease

Keeping the Vimentin Network under Control: Cell–Matrix Adhesion–associated Plectin 1f Affects Cell Shape and Polarity of Fibroblasts

Mature focal adhesions and fibrillar adhesions act as anchorage sites for vimentin filaments, with plectin isoform 1f being the crucial linker protein. Plectin serves as a nucleation and assembly center for the de novo formation of vimentin networks. Anchored vimentin creates a resilient cage-like core structure that affects cell shape

A combined functional and structural genomics approach identified an EST-SSR marker with complete linkage to the Ligon lintless-2 genetic locus in cotton (Gossypium hirsutum L.)

<p>Abstract</p> <p>Background</p> <p>Cotton fiber length is an important quality attribute to the textile industry and longer fibers can be more efficiently spun into yarns to produce superior fabrics. There is typically a negative correlation between yield and fiber quality traits such as length. An understanding of the regulatory mechanisms controlling fiber length can potentially provide a valuable tool for cotton breeders to improve fiber length while maintaining high yields. The cotton (<it>Gossypium hirsutum </it>L.) fiber mutation Ligon lintless-2 is controlled by a single dominant gene (<it>Li₂</it>) that results in significantly shorter fibers than a wild-type. In a near-isogenic state with a wild-type cotton line, <it>Li₂</it>is a model system with which to study fiber elongation.</p> <p>Results</p> <p>Two near-isogenic lines of Ligon lintless-2 (<it>Li₂</it>) cotton, one mutant and one wild-type, were developed through five generations of backcrosses (BC₅). An F₂population was developed from a cross between the two <it>Li₂</it>near-isogenic lines and used to develop a linkage map of the <it>Li₂</it>locus on chromosome 18. Five simple sequence repeat (SSR) markers were closely mapped around the <it>Li₂</it>locus region with two of the markers flanking the <it>Li₂</it>locus at 0.87 and 0.52 centimorgan. No apparent differences in fiber initiation and early fiber elongation were observed between the mutant ovules and the wild-type ones. Gene expression profiling using microarrays suggested roles of reactive oxygen species (ROS) homeostasis and cytokinin regulation in the <it>Li₂</it>mutant phenotype. Microarray gene expression data led to successful identification of an EST-SSR marker (NAU3991) that displayed complete linkage to the <it>Li₂</it>locus.</p> <p>Conclusions</p> <p>In the field of cotton genomics, we report the first successful conversion of gene expression data into an SSR marker that is associated with a genomic region harboring a gene responsible for a fiber trait. The EST-derived SSR marker NAU3991 displayed complete linkage to the <it>Li₂</it>locus on chromosome 18 and resided in a gene with similarity to a putative plectin-related protein. The complete linkage suggests that this expressed sequence may be the <it>Li₂</it>gene.</p

Evaluation of the Allergenicity Potential of TcPR-10 Protein from Theobroma cacao

Background: The pathogenesis related protein PR10 (TcPR-10), obtained from the Theobroma cacao-Moniliophthora perniciosa interaction library, presents antifungal activity against M. perniciosa and acts in vitro as a ribonuclease. However, despite its biotechnological potential, the TcPR-10 has the P-loop motif similar to those of some allergenic proteins such as Bet v 1 (Betula verrucosa) and Pru av 1 (Prunus avium). The insertion of mutations in this motif can produce proteins with reduced allergenic power. The objective of the present work was to evaluate the allergenic potential of the wild type and mutant recombinant TcPR-10 using bioinformatics tools and immunological assays. Methodology/Principal Findings: Mutant substitutions (T10P, I30V, H45S) were inserted in the TcPR-10 gene by sitedirected mutagenesis, cloned into pET28a and expressed in Escherichia coli BL21(DE3) cells. Changes in molecular surface caused by the mutant substitutions was evaluated by comparative protein modeling using the three-dimensional structure of the major cherry allergen, Pru av 1 as a template. The immunological assays were carried out in 8-12 week old female BALB/c mice. The mice were sensitized with the proteins (wild type and mutants) via subcutaneous and challenged intranasal for induction of allergic airway inflammation. Conclusions/Significance: We showed that the wild TcPR-10 protein has allergenic potential, whereas the insertion of mutations produced proteins with reduced capacity of IgE production and cellular infiltration in the lungs. On the other hand, in vitro assays show that the TcPR-10 mutants still present antifungal and ribonuclease activity against M. perniciosa RNA. In conclusion, the mutant proteins present less allergenic potential than the wild TcPR-10, without the loss of interesting biotechnological properties. (Résumé d'auteur