Detection of exon skipping events in BRCA1 RNA using MLPA kit P002

A rapid and easy method to screen for aberrant cDNA would be a very useful diagnostic tool in genetics since a fraction of the DNA variants found affect RNA splicing. The currently used RT-PCR methods require new primer combinations to study each variant that might affect splicing. Since MLPA is routinely used to detect large genomic deletions and successfully detected exon skipping events in Duchenne muscular dystrophy in cDNA, we performed a pilot study to evaluate its value for BRCA1 cDNA. The effect of puromycin, DNase I and two different DNA cleaning protocols were tested in the RNA analysis of lymphocyte cultures. We used two samples from unrelated families with two different BRCA1 exon deletion events, two healthy unrelated controls and six samples from hereditary breast/ovarian cancer syndrome (HBOC) patients without BRCA1/2 mutations. Using RNA treated with DNase I and cleaned in a column system from puromycin-treated fractions, we were able to identify the two BRCA1 deletions. Additional HBOC patients did not show additional splice events. However, we were not able to get reproducible results. Therefore, the cDNA-MLPA technique using kit BRCA1 P002 is in our hands currently not reliable enough for routine RNA analysis and needs further optimization

Proliferative Hypothalamic Neurospheres Express NPY, AGRP, POMC, CART and Orexin-A and Differentiate to Functional Neurons

Some pathological conditions with feeding pattern alterations, including obesity and Huntington disease (HD) are associated with hypothalamic dysfunction and neuronal cell death. Additionally, the hypothalamus is a neurogenic region with the constitutive capacity to generate new cells of neuronal lineage, in adult rodents

Narrative benchmarking in scenario assessments for digital design

This article aims to propose a strategy for narrative development, for photo/ image sharing social tools. By generating scenarios for product assessment through comparing consumption narratives as a contribution to the creation of new communication strategies. Developing as hypothesis that the development of narratives from a brand and its consequent comparison with consumers feedback, allows a more reliable way to generate inputs for the development of more effective communication strategies. The article follows an exploratory study methodology intended “to develop pertinent hypotheses and propositions for further inquiry. The article draws on a qualitative proposal of a method to apply in narratives of consumption. The results of the proposed strategy open avenues for narratives developments, where the consumers contribution receive expansion values from the brands strategy, returning value to first consumer feedbacks. The implications of this study are the creation of a paradigm of curatorship as authorship. The research limitations/ implications address the speculative scope of the proposed strategy and the need for more empirical work in the subject.info:eu-repo/semantics/publishedVersio