Molecular characterization of Chilean patients with a clinical diagnosis of Noonan syndrome.

BACKGROUND: Noonan syndrome (NS) is an autosomal dominant syndrome characterized by typical dysmorphic features, cardiac anomalies as well as postnatal growth retardation, and is associated with Ras-MAPK pathway gene mutations. The purpose of this study was to improve the diagnosis of Chilean patients with suspected NS through molecular analysis. METHODS: We screened 18 Chilean patients with a clinical diagnosis of NS for mutations in PTPN11 by high resolution melting (HRM) and subsequent sequencing. RESULTS: Three PTPN11 missense mutations were detected in 22% of analyzed patients. Of these, two (c.181G>A and c.1510A>G) were previously reported and one was the novel substitution c.328G>A (p.E110K) affecting the linker stretch between the N-SH2 and C-SH2 domains of SHP-2 protein. CONCLUSION: Molecular studies confirmed the clinical diagnosis of NS in 4 of 18 patients, which provided support for therapeutic decisions and improved genetic counseling for their families.

De Novo COL7A1 mutation in a patient with trisomy 21: coexistence of dystrophic epidermolysis bullosa and Down syndrome.

BACKGROUND: Down syndrome (DS) is the most common autosomal chromosomal disorder. Epidermolysis bullosa (EB) is a rare genodermatosis characterized by skin and mucous membrane fragility, with formation of blisters and erosions after minor trauma. Dystrophic EB (DEB) is inherited as an autosomal dominant (DDEB) or recessive (RDEB) trait. Both forms are caused by mutations in COL7A1, the gene coding for the type VII collagen. We report a patient affected by both conditions: DS and DDEB. METHODS: A patient with DS developed generalized blisters at the age of three months. Cytogenetic study was performed to confirm DS. Skin biopsies were examined with immunohistochemical and electron microscopy techniques to determine EB subtype. Genomic DNA was extracted from peripheral blood samples. COL7A1 mutations were screened by heteroduplex analysis using conformation-sensitive gel electrophoresis and sequencing. RESULTS: Karyotype analysis revealed trisomy 21. Histological study agreed with a DEB diagnosis. Mutational analysis showed a heterozygous c.6127G>T mutation in COL7A1, which is compatible with DDEB. Parental study suggests that c.6127G>T arises as a de novo mutation. CONCLUSIONS: This report demonstrates that EB can be associated with other common conditions and reports the case of a patient who suffered two de novo independent genetic conditions. It also contributes to expanding the knowledge and database of clinical and molecular aspects of DDEB.

Hilbert transform analysis of a time series of speckle interferograms with a temporal carrier.

We present an optical phase measurement method based on the Hilbert transform for the analysis of a time series of speckle interferograms modulated by a temporal carrier. We discuss the influence of nonmodulating pixels, modulation loss, and noise that affect the bias and modulation intensities of the interferometric signal and propose the application of the empirical mode decomposition method for its minimization. We also show the equivalence between the phase recovery approaches that are based on the Hilbert and the Fourier transforms. Finally, we present a numerical comparison between these methods using computer-simulated speckle interferograms modulated with a temporal carrier.

Protein kinase CK2 as an ectokinase: the role of the regulatory CK2beta subunit.

Protein kinase CK2 (also known as casein kinase 2) is present in the cytoplasm, nuclei, and several other organelles. In addition, this enzyme has been found bound to the external side of the cell membrane where it acts as an ectokinase phosphorylating several extracellular proteins. Previous experiments with transfection of HEK-293T cells demonstrated that expression of both subunits, CK2alpha (catalytic) and CK2beta (regulatory), was necessary for the appearance of the ectopic enzyme as an ectokinase. In this work, using deletion and point mutations of CK2beta, it was possible to demonstrate that the region between amino acids 20 and 33 was necessary for the export of the enzyme as an ectokinase. Phenylalanines 21 and 22 and acidic residues in positions 26-28 are involved in the structural aspects that are required for export. However, the region encompassing amino acids 20-33 of CK2beta is not sufficient to make the carboxyl half of this subunit functional in bringing CK2 to the ectokinase locus. In cells transfected with only CK2beta, it was demonstrated that 3-4% of the subunit is exported to the cell medium, but the subunit is not bound to the external membrane.

Brevetoxin-3 (PbTx-3) on mouse liver slices: a histological study

Brevetoxin-3 (PbTx-3) is a marine toxin produced by the dinoflagellate Ptychodiscus brevis. Its effects on excitable tissues have been the main subject of studies, but little is known about how it affects non-excitable tissues. To study possible non-neural effects of PbTx-3 (78nM), its effects on hepatic cell structure in vitro were evaluated. PbTx-3 caused hypertrophy and increased vacuolation of hepatocytes, and an increase in basophilia in the perivenous area of the lobules. Ultrastructurally, it was evident that the vacuolation was related to swelling of the endoplasmic reticulum, changes that probably account for the increased basophilic reaction of the cells. The swelling in smooth endoplasmic reticulum, degranulation of rough endoplasmic reticulum, the deformities and lytic cristae in the mitochondria, and the presence of active lysosomes are evidence of the PbTx-3 effects upon liver cells. These responses are probably caused by the liver's detoxification role on the PbTx-3.