Functional analysis of cancer-associated EGFR mutants using a cellular assay with YFP-tagged EGFR intracellular domain

Background: The presence of EGFR kinase domain mutations in a subset of NSCLC patients correlates with the response to treatment with the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Although most EGFR mutations detected are short deletions in exon 19 or the L858R point mutation in exon 21, more than 75 different EGFR kinase domain residues have been reported to be altered in NSCLC patients. The phenotypical consequences of different EGFR mutations may vary dramatically, but the majority of uncommon EGFR mutations have never been functionally evaluated. Results: We demonstrate that the relative kinase activity and erlotinib sensitivity of different EGFR mutants can be readily evaluated using transfection of an YFP-tagged fragment of the EGFR intracellular domain (YFP-EGFR-ICD), followed by immunofluorescence microscopy analysis. Using this assay, we show that the exon 20 insertions Ins770SVD and Ins774HV confer increased kinase activity, but no erlotinib sensitivity. We also show that, in contrast to the common L858R mutation, the uncommon exon 21 point mutations P848L and A859T appear to behave like functionally silent polymorphisms. Conclusion: The ability to rapidly obtain functional information on EGFR variants of unknown relevance using the YFP-EGFR-ICD assay might prove important in the future for the management of NSCLC patients bearing uncommon EGFR mutations. In addition, our assay may be used to determine the response of resistant EGFR mutants to novel second-generation TKIs.

Comparativa de metodologías de cálculo de faltas en redes eléctricas de baja tensión residenciales e industriales.

[ES]Este trabajo se basa en la aplicación de distintas metodologías (norma europea, método fácil y norma americana) para la obtención de las corrientes de cortocircuito en un sistema eléctrico de baja tensión, de forma que se sepa de antemano el valor de dichas corrientes en cualquier punto del circuito para el correcto dimensionamiento de los dispositivos de protección a emplear.

Study on the activation mechanism of BCL-2 related ovarian killer (BOK)

BCL-2 family proteins are key regulators of the mitochondrial apoptotic machinery, controlling the mitochondrial outer membrane (MOM) permeabilization (MOMP). BCL-2 related Ovarian Killer (BOK) is a poorly understood pro-apoptotic member of this protein family. It has been reported that BOK localizes predominantly (although not exclusively) at membranes of the endoplasmic reticulum and of the Golgi apparatus. However, it is unclear whether BOK also operates at the MOM to promote apoptosis, as other pro-apoptotic BCL-2 family members do. Basing on the fact that the other two BAX-like pro-apoptotic members have been reported to oligomerize in order to induce MOMP, site-directed mutagenesis was used to generate two point mutations that predictably eliminated BOK’s oligomerization capacity. Then, the effect of such mutations on BOK’s membrane activity was examined using fluorescence spectroscopy.

Coincidence and common fixed point theorems for Suzuki type hybrid contractions and applications

Coincidence and common fixed point theorems for a class of Suzuki hybrid contractions involving two pairs of single-valued and multivalued maps in a metric space are obtained. In addition, the existence of a common solution for a certain class of functional equations arising in a dynamic programming is also discussed.

A New Type of Coincidence and Common Fixed Point Theorem with Applications

Coincidence and common fixed point theorems for a class of 'Ciric-Suzuki hybrid contractions involving a multivalued and two single-valued maps in a metric space are obtained. Some applications including the existence of a common solution for certain class of functional equations arising in a dynamic programming are also discussed..