Role of Chaperone Mediated Autophagy (CMA) in the Degradation of Misfolded N-CoR Protein in Non-Small Cell Lung Cancer (NSCLC) Cells

Nuclear receptor co-repressor (N-CoR) plays important role in transcriptional control mediated by several tumor suppressor proteins. Recently, we reported a role of misfolded-conformation dependent loss (MCDL) of N-CoR in the activation of oncogenic survival pathway in acute promyelocytic leukemia (APL). Since N-CoR plays important role in cellular homeostasis in various tissues, therefore, we hypothesized that an APL like MCDL of N-CoR might also be involved in other malignancy. Indeed, our initial screening of N-CoR status in various leukemia and solid tumor cells revealed an APL like MCDL of N-CoR in primary and secondary tumor cells derived from non-small cell lung cancer (NSCLC). The NSCLC cell specific N-CoR loss could be blocked by Kaletra, a clinical grade protease inhibitor and by genistein, an inhibitor of N-CoR misfolding previously characterized by us. The misfolded N-CoR presented in NSCLC cells was linked to the amplification of ER stress and was subjected to degradation by NSCLC cell specific aberrant protease activity. In NSCLC cells, misfolded N-CoR was found to be associated with Hsc70, a molecular chaperone involved in chaperone mediated autophagy (CMA). Genetic and chemical inhibition of Lamp2A, a rate limiting factor of CMA, significantly blocked the loss of N-CoR in NSCLC cells, suggesting a crucial role of CMA in N-CoR degradation. These findings identify an important role of CMA-induced degradation of misfolded N-CoR in the neutralization of ER stress and suggest a possible role of misfolded N-CoR protein in the activation of oncogenic survival pathway in NSCLC cells

Affective Influences without Approach-Avoidance Actions: On the Congruence Between Valence and Stimulus-Response Mappings

The valence of stimuli can influence performance in the spatial stimulus–response compatibility task, but this observation could arise from the process of selecting responses or selecting stimulus–response mappings. The response-selection account proposes that spatial compatible and incompatible keypress responses serve as approaching and avoiding actions to a valenced target. The mapping-selection account suggests that there is congruence between stimulus valence and stimulus–response mappings; positive-compatible/negative-incompatible is more congruent than negative-compatible/positive-incompatible. Whereas affective valence was part of the target stimuli to which participants responded in previous studies, the present study isolated affective valence from the target by presenting an additional mapping cue separately from the target, so that spatially compatible and incompatible keypress responses could no longer serve as approaching and avoiding actions to valenced target stimuli. The present results revealed that responses were still faster when positive and negative mapping cues were assigned to the spatially compatible and incompatible mappings than when the assignment was reversed. The finding supports the mapping-selection account, indicating that positive and negative cues influence performance without approach–avoidance actions to valenced stimuli. The experiment provides important implications as to how tasks are represented and are dependent on affective processing

MSH3 polymorphisms and protein levels affect CAG repeat instability in huntington's disease mice

Expansions of trinucleotide CAG/CTG repeats in somatic tissues are thought to contribute to ongoing disease progression through an affected individual's life with Huntington's disease or myotonic dystrophy. Broad ranges of repeat instability arise between individuals with expanded repeats, suggesting the existence of modifiers of repeat instability. Mice with expanded CAG/CTG repeats show variable levels of instability depending upon mouse strain. However, to date the genetic modifiers underlying these differences have not been identified. We show that in liver and striatum the R6/1 Huntington's disease (HD) (CAG)~100 transgene, when present in a congenic C57BL/6J (B6) background, incurred expansion-biased repeat mutations, whereas the repeat was stable in a congenic BALB/cByJ (CBy) background. Reciprocal congenic mice revealed the Msh3 gene as the determinant for the differences in repeat instability. Expansion bias was observed in congenic mice homozygous for the B6 Msh3 gene on a CBy background, while the CAG tract was stabilized in congenics homozygous for the CBy Msh3 gene on a B6 background. The CAG stabilization was as dramatic as genetic deficiency of Msh2. The B6 and CBy Msh3 genes had identical promoters but differed in coding regions and showed strikingly different protein levels. B6 MSH3 variant protein is highly expressed and associated with CAG expansions, while the CBy MSH3 variant protein is expressed at barely detectable levels, associating with CAG stability. The DHFR protein, which is divergently transcribed from a promoter shared by the Msh3 gene, did not show varied levels between mouse strains. Thus, naturally occurring MSH3 protein polymorphisms are modifiers of CAG repeat instability, likely through variable MSH3 protein stability. Since evidence supports that somatic CAG instability is a modifier and predictor of disease, our data are consistent with the hypothesis that variable levels of CAG instability associated with polymorphisms of DNA repair genes may have prognostic implications for various repeat-associated diseases

Índice de Massa Corporal, Idade, Maturação Sexual e a Incidência de Hiperlordose Lombar em crianças e adolescentes

Introduction: Hyperlordosis can cause several degenerative spinal pathologies in children and adolescents. Objective: Determine whether body mass index, age and sexual maturation predict the occurrence of hyperlordosis in children and adolescents. Method: The study analyzed 380 students aged between 10 and 18 years. Body mass index was evaluated using the reference values suggested by the Fitnessgram test battery, and sexual maturation through Tanner’s scale of self-assessed pubic hair growth. Postural assessment was conducted using the DIPA photogrammetry method, version 3.1. (Digital Image Based Postural Assessment) The SPSS 24.0 program was used to analyze the data, and the following statistical tests were applied: chi squared, Mann-Whitney, Fisher’s exact and binary logistic regression. Results: There was statistical significance between hyperlordosis, girls’ age and puberty in boys (p 0.05). Conclusion: The girls’ age and boys’ stage of puberty were associated with the occurrence of hyperlordosis.Introdução: A Hiperlordose lombar pode ocasionar diversas patologias degenerativas na coluna vertebral de crianças e adolescentes. Objetivo: Identificar se o Índice de Massa Corporal, a Idade e a Maturação Sexual são previsores da ocorrência da hiperlordose lombar em crianças e adolescentes. Método: O estudo analisou 380 estudantes entre 10 e 18 anos. O Índice de Massa Corporal foi avaliado por meio dos valores de referência sugeridos pela bateria de testes Fitnessgram e a maturação sexual por meio da auto-avaliação da pilosidade pubiana de Tanner. A avaliação postural foi realizada pelo método de fotogrametria DIPA versão 3.1. (Avaliação Postural Baseada em Imagem Digital). Para análise dos dados foi utilizado o programa SPSS 24.0, tendo sido aplicados os testes estatísticos: Qui-Quadrado, Mann Whitney, Exato de Fisher e Regressão Logística Binária. Resultados: Observou-se que houve significância estatística entre a Hiperlordose lombar e a idade das meninas e a puberdade dos meninos (p0,05). Conclusão: A idade das meninas e a puberdade dos meninos foi associada à ocorrência da hiperlordose lombar.This study was funded by CIEC (Center for Investigations in Childhood Studies), Strategic Project UID/CED/00317/2013, via National FCT (Science and Technology Foundation) funds and co-funded by the European Regional Development Fund (FEDER), via COMPETE 2020 – Competitivity and Internalization Operational Program (POCI) under reference number POCI-01-0145-FEDER-007562

Mutation Screening of Multiple Genes in Spanish Patients with Autosomal Recessive Retinitis Pigmentosa by Targeted Resequencing

Retinitis Pigmentosa (RP) is a heterogeneous group of inherited retinal dystrophies characterised ultimately by the loss of photoreceptor cells. RP is the leading cause of visual loss in individuals younger than 60 years, with a prevalence of about 1 in 4000. The molecular genetic diagnosis of autosomal recessive RP (arRP) is challenging due to the large genetic and clinical heterogeneity. Traditional methods for sequencing arRP genes are often laborious and not easily available and a screening technique that enables the rapid detection of the genetic cause would be very helpful in the clinical practice. The goal of this study was to develop and apply microarray-based resequencing technology capable of detecting both known and novel mutations on a single high-throughput platform. Hence, the coding regions and exon/intron boundaries of 16 arRP genes were resequenced using microarrays in 102 Spanish patients with clinical diagnosis of arRP. All the detected variations were confirmed by direct sequencing and potential pathogenicity was assessed by functional predictions and frequency in controls. For validation purposes 4 positive controls for variants consisting of previously identified changes were hybridized on the array. As a result of the screening, we detected 44 variants, of which 15 are very likely pathogenic detected in 14 arRP families (14%). Finally, the design of this array can easily be transformed in an equivalent diagnostic system based on targeted enrichment followed by next generation sequencing

Covert Tracking: A Combined ERP and Fixational Eye Movement Study

Attention can be directed to particular spatial locations, or to objects that appear at anticipated points in time. While most work has focused on spatial or temporal attention in isolation, we investigated covert tracking of smoothly moving objects, which requires continuous coordination of both. We tested two propositions about the neural and cognitive basis of this operation: first that covert tracking is a right hemisphere function, and second that pre-motor components of the oculomotor system are responsible for driving covert spatial attention during tracking. We simultaneously recorded event related potentials (ERPs) and eye position while participants covertly tracked dots that moved leftward or rightward at 12 or 20°/s. ERPs were sensitive to the direction of target motion. Topographic development in the leftward motion was a mirror image of the rightward motion, suggesting that both hemispheres contribute equally to covert tracking. Small shifts in eye position were also lateralized according to the direction of target motion, implying covert activation of the oculomotor system. The data addresses two outstanding questions about the nature of visuospatial tracking. First, covert tracking is reliant upon a symmetrical frontoparietal attentional system, rather than being right lateralized. Second, this same system controls both pursuit eye movements and covert tracking