33 research outputs found
A colorimetric strategy based on dynamic chemistry for direct detection of Trypanosomatid species
Leishmaniasis and Chagas disease are endemic in many countries, and re-emerging in the developed
countries. A rapid and accurate diagnosis is important for early treatment for reducing the duration
of infection as well as for preventing further potential health complications. In this work, we have
developed a novel colorimetric molecular assay that integrates nucleic acid analysis by dynamic
chemistry (ChemNAT) with reverse dot-blot hybridization in an array format for a rapid and easy
discrimination of Leishmania major and Trypanosoma cruzi. The assay consists of a singleplex PCR
step that amplifies a highly homologous DNA sequence which encodes for the RNA component of the
large ribosome subunit. The amplicons of the two different parasites differ between them by single
nucleotide variations, known as “Single Nucleotide Fingerprint” (SNF) markers. The SNF markers can
be easily identified by naked eye using a novel micro Spin-Tube device "Spin-Tube", as each of them
creates a specific spot pattern. Moreover, the direct use of ribosomal RNA without requiring the PCR
pre-amplification step is also feasible, further increasing the simplicity of the assay. The molecular
assay delivers sensitivity capable of identifying up to 8.7 copies per ÎĽL with single mismatch specificity.
The Spin-Tube thus represents an innovative solution providing benefits in terms of time, cost, and
simplicity, all of which are crucial for the diagnosis of infectious disease in developing countries.This research work has received funding from Junta de AndalucĂa, ConsejerĂa de EconomĂa e InnovaciĂłn (project
number 2012-BIO1778), the Spanish Ministerio de EconomĂa y Competitividad (Grants CTQ2012-34778,
BIO2016-80519-R, FPI Grant BES-2013- 063020). This research was partially supported by the 7th European
Community Framework Program (FP7-PEOPLE-2012-CIG-Project Number 322276)
Using C. elegans to decipher the cellular and molecular mechanisms underlying neurodevelopmental disorders
Prova tipográfica (uncorrected proof)Neurodevelopmental disorders such as epilepsy, intellectual disability (ID), and autism spectrum disorders (ASDs) occur in over 2 % of the population, as the result of genetic mutations, environmental factors, or combination of both. In the last years, use of large-scale genomic techniques allowed important advances in the identification of genes/loci associated with these disorders. Nevertheless, following association of novel genes with a given disease, interpretation of findings is often difficult due to lack of information on gene function and effect of a given mutation in the corresponding protein. This brings the need to validate genetic associations from a functional perspective in model systems in a relatively fast but effective manner. In this context, the small nematode, Caenorhabditis elegans, presents a good compromise between the simplicity of cell models and the complexity of rodent nervous systems. In this article, we review the features that make C. elegans a good model for the study of neurodevelopmental diseases. We discuss its nervous system architecture and function as well as the molecular basis of behaviors that seem important in the context of different neurodevelopmental disorders. We review methodologies used to assess memory, learning, and social behavior as well as susceptibility to seizures in this organism. We will also discuss technological progresses applied in C. elegans neurobiology research, such as use of microfluidics and optogenetic tools. Finally, we will present some interesting examples of the functional analysis of genes associated with human neurodevelopmental disorders and how we can move from genes to therapies using this simple model organism.The authors would like to acknowledge Fundação para a Ciência e Tecnologia (FCT) (PTDC/SAU-GMG/112577/2009). AJR and CB are recipients of FCT fellowships: SFRH/BPD/33611/2009 and SFRH/BPD/74452/2010, respectively
Lasers in Dermatological Rare Diseases
Laser treatments are widely used in various rare dermatological diseases, such as tuberous sclerosis (TSC), neurofibromatosis type 1, Birt-Hogg-Dubé (BHD) syndrome, idiopathic calcinosis cutis of the scrotum, and Darier disease (DD). Although laser treatment may not help a final resolution of the disease, patient’s life quality is always improved