Biofluid Biomarkers in Huntington's Disease

Huntington's disease (HD) is a chronic progressive neurodegenerative condition where new markers of disease progression are needed. So far no disease-modifying interventions have been found, and few interventions have been proven to alleviate symptoms. This may be partially explained by the lack of reliable indicators of disease severity, progression, and phenotype.Biofluid biomarkers may bring advantages in addition to clinical measures, such as reliability, reproducibility, price, accuracy, and direct quantification of pathobiological processes at the molecular level; and in addition to empowering clinical trials, they have the potential to generate useful hypotheses for new drug development.In this chapter we review biofluid biomarker reports in HD, emphasizing those we feel are likely to be closest to clinical applicability

Preliminary results for application of culture-independent molecular methods to olive brines bacterial communities.

Transformation of green table olives requires a brining stage during which olives ferment. In naturally transformed olives, both flavour improvement and debittering are associated to fermentation. Because of brine features, naturally occurring bacteria and yeasts are progressively selected by the high salt concentration, low pH and phenolic compounds with antimicrobial activity. The establishment of an adequate fermenting microflora is essential to obtain a good and safe product; then, during transformation, control of chemical and microbiological parameters is crucial. Currently, microflora is periodically monitored by plate-counting of main groups. These routine analyses, however, don’t show neither the complexity of population nor possible differences between various samples, because the most used cultural media are often not selective enough, and fail to differentiate single species. We carried out preliminary trials to evaluate the possibility of application of molecular analyses to describe microflora dynamic and complexity in fermenting green olive brines. We used culture-independent methods, based on analysis of total DNA directly extracted from the whole microbial and blastomycetic community in naturally fermented olive brines. Olives (cv. “Nocellara del Belice”) were transformed with both traditional (NaCl only) and modified brines (with low NaCl content, where the total ionic strength was ensure by KCl and/or CaCl2, to obtain “hyposodic” olives ) which allow regular fermentation. In particular, we carried out analyses on PCR amplified fragments of polymorphic rDNA regions with length or sequence variability among different species, fractionating them by denaturing gel electrophoresis (RISA, Ribosomal Intergenic Sequences Analysis) and by automated capillary electrophoresis (ARISA, Automated-RISA), or by DGGE (Denaturing Gradient Gel Electrophoresis) when analyzing variable regions (but with similar length) of 16S rDNA. Resulting specific profiles can be compared between different samples, allowing to show the dynamic of microbial populations. By DNA cloning and sequencing, we were able to identify dominant species, assigning them to specific bands. Although these are preliminary data, we think these techniques might be rapid and useful tools, complementary to traditional microbiological analyses, to study brine microflora and to follow simultaneously, comparing patterns, the proper growth of a number of species in complex populations (with known profiles) used as natural starters, when more specific markers are not available

Gain-of-function SOS1 mutations cause a distinctive form of Noonan syndrome

Noonan syndrome (NS) is a developmental disorder characterized by short stature, facial dysmorphia, congenital heart defects and skeletal anomalies1. Increased RAS-mitogenactivated protein kinase (MAPK) signaling due to PTPN11 and KRAS mutations cause 50 percent of NS2-6. Here, we report that 22 of 129 NS patients without PTPN11 or KRAS mutation (17 percent) have missense mutations in SOS1, which encodes a RAS-specific guanine nucleotide exchange factor (GEF). SOS1 mutations cluster at residues implicated in the maintenance of SOS1 in its autoinhibited form and ectopic expression of two NS-associated mutants induced enhanced RAS activation. The phenotype associated with SOS1 defects is distinctive, although within NS spectrum, with a high prevalence of ectodermal abnormalities but generally normal development and linear growth. Our findings implicate for the first time gain-of-function mutations in a RAS GEF in inherited disease and define a new mechanism by which upregulation of the RAS pathway can profoundly change human development