Search

Supplementary Figures 1-4, Table 1 from Novel Dyskerin-Mediated Mechanism of p53 Inactivation through Defective mRNA Translation

In up to 60% of human cancers, p53 gene mutations are responsible for direct inactivation of the tumor suppressor function of p53. Alternative mechanisms of p53 inactivation described thus far mainly affect its posttranslational regulation. In X-linked dyskeratosis congenita, a multisystemic syndrome characterized by increased cancer susceptibility, mutations of the DKC1 gene encoding dyskerin cause a selective defect in the translation of a subgroup of internal ribosome entry site (IRES)–containing cellular mRNAs. In this study, we show that impairment of dyskerin function can cause p53 inactivation due to a defect in p53 mRNA translation. siRNA-mediated reduction of dyskerin levels caused a decrease of p53 mRNA translation, protein levels, and functional activity, both in human breast cancer cells and in primary mammary epithelial progenitor cells. These effects seemed to be independent of the known role of dyskerin in telomerase function, and they were associated with a specific impairment of translation initiation mediated by IRES elements present in p53 mRNA. In a series of human primary breast cancers retaining wild-type p53, we found that low levels of dyskerin expression were associated with reduced expression of p53-positive target genes. Our findings suggest that a dyskerin-mediated mechanism of p53 inactivation may occur in a subset of human tumors. Cancer Res; 70(11); 4767–77. ©2010 AACR.

According to established cognitive neuroscience knowledge based on studies on disabled and typically developing readers, reading is based on a dual-stream model in which a phonological-dorsal stream (left temporo-parietal and inferior frontal areas) processes unfamiliar words and pseudowords, whereas an orthographic-ventral stream (left occipito-temporal and inferior frontal areas) processes known words. However, correlational neuroimaging, causal longitudinal, training, and pharmacological studies have suggested the critical role of visuo-spatial attention in reading development. In a double blind, crossover within-subjects experiment, we manipulated the neuromodulatory effect of a short-term bilateral stimulation of posterior parietal cortex (PPC) by using active and sham tRNS during reading tasks in a large sample of young adults. In contrast to the dual-stream model predicting either no effect or a selective effect on the stimulated phonological-dorsal stream (as well as to a general multisensory effect on both reading streams), we found that only word-reading performance improved after active bilateral PPC tRNS. These findings demonstrate a direct neural connectivity between the PPC, controlling visuo-spatial attention, and the ventral stream for visual word recognition. These results support a neurobiological model of reading where performance of the orthographic-ventral stream is boosted by an efficient deployment of visuo-spatial attention from bilateral PPC stimulation.

Although developmental reading disorders (developmental dyslexia) have been mainly associated with auditory-phonological deficits, recent longitudinal and training studies have shown a possible causal role of visuo-attentional skills in reading acquisition. Indeed, visuo-attentional mechanisms could be involved in the orthographic processing of the letter string and the graphemic parsing that precede the grapheme-to-phoneme mapping. Here, we used a simple paper-and-pencil task composed of three labyrinths to measure visuo-spatial attention in a large sample of primary school children (n = 398). In comparison to visual search tasks requiring visual working memory, our labyrinth task mainly measures distributed and focused visuo-spatial attention, also controlling for sensorimotor learning. Compared to typical readers (n = 340), children with reading difficulties (n = 58) showed clear visuo-spatial attention impairments that appear not linked to motor coordination and procedural learning skills implicated in this paper and pencil task. Since visual attention is dysfunctional in about 40% of the children with reading difficulties, an efficient reading remediation program should integrate both auditory-phonological and visuo-attentional interventions.

Human perception of a visual scene is hierarchically organized. Such rapid, albeit coarse, global processing allows people to create a useful context in which local details can be successively allocated. Lack of the typical hierarchical global-to-local visual processing is longitudinally predictive of future reading difficulties in pre-readers, which suggests that an atypical local perception can interfere with reading skill acquisition. Global and local Navon tasks were used to induce a transient perceptual priming before a reading-aloud task. We tested the effect of an atypical local perception on lexical and sublexical reading routes in typical adult readers. Local (vs. global) priming resulted in a slower phonological access to irregular, relative to regular, words. By contrast, pseudoword reading was not affected by local (vs. global) perceptual priming. Our findings demonstrate that, in typical adult readers, local priming impairs the fast processing of the letter string useful for lexical reading.

According to established background knowledge, playing is essential in human development and a power remediation tool in clinical populations. In clinical interventions, the beneficial roles of playing have often been sought and investigated in the specific features of the game, rather than in the positive emotions generated by playing. However, regardless of game specifications, cognitive enhancement could be driven by the emotions linked to play. Establishing the causal connections between play and cognitive enhancement should allow us to determine how to involve play in therapy, prevention and educational programmes. Today, video-gaming is one of the most diffused forms of play. In the first crossover randomized controlled trial, we compared the short-term effects induced by shooting and puzzle video-games in visual perception, sensorimotor and reading skills in children with developmental coordination disorder and dyslexia. The funnier and more activating game enhanced breadth of visual perception and reduced sensorimotor and reading disorders. Visual perception, sensorimotor and reading improvements correlated with fun. In the second crossover randomized controlled trial, comparing the effects of the same shooting with a fighting video-game in healthy young adults, we show that regardless of game characteristics, changes in positive emotions correlated with contextual reading enhancement, while play-driven biochemical activation boosted single word and pseudoword reading. The short-term effects induced by play could be a useful clinical tool for the prevention and treatment of multiple cognitive disorders.

For about 10% of children reading acquisition is extremely difficult because they are affected by a heritable neurobiological disorder called developmental dyslexia (DD), mainly associated to an auditory-phonological disorder. Visual crowding is a universal phenomenon that impairs the recognition of stimuli in clutter, such as a letter in a word or a word in a text. Several studies have shown an excessive crowding in individuals with DD, but the causal link between excessive crowding and DD is not yet clearly established. An excessive crowding might be, indeed, a simple effect of DD due to reduced reading experience. The results of five experiments in 181 children reveal that: (i) an excessive crowding only at unattended locations characterizes an unselected group of children with DD (Experiment 1); (ii) an extra-large spaced text increases reading accuracy by reducing crowding in an unselected group of children with DD (Experiment 2); (iii) efficient attentional action video game trainings reduce crowding and accelerate reading speed in two unselected groups of children with DD (Experiment 3 and 4), and; (iv) pre-reading crowding longitudinally predicts future poor readers (Experiment 5). Our results show multiple causal links between visual crowding and learning to read. These findings provide new insights for a more efficient remediation and prevention for DD.

Learning to read is extremely difficult for about 10% of the children because they are affected by a heritable neurobiological disorder called developmental dyslexia (DD). The causal role of cognitive deficits typically associated to DD can be investigated through intervention studies. It has been demonstrated that visual-attention and reading speed could be simultaneously improved by using action video game (AVG) training both in shallow and deep alphabetic orthographies. Here, in a clinical study we showed that after this general-domain behavioral intervention both the phonological decoding speed and phonological short-term memory were increased only in DD children in which their video game score was improved. These findings confirm that an AVG training enhances the efficiency of both visual and auditory processing. The plasticity of the multi-sensory attentional network could explain the reading and reading-related improvements induced by the AVG training in children with DD.

Background: Reading is a unique human skill. Several brain networks involved in this complex skill mainly involve the left hemisphere language areas. Nevertheless, nonlinguistic networks found in the right hemisphere also seem to be involved in sentence and text reading. These areas do not deal with phonological information, but are involved in verbal and nonverbal pattern information processing. The right hemisphere is responsible for global processing of a scene, which is needed for developing reading skills. Aims: Caffeine seems to affect global pattern processing specifically. Consequently, our aim was to discover if it could enhance text reading skill. Methods: In two mechanistic studies ( n=24 and n=53), we tested several reading skills, global and local perception, alerting, spatial attention and executive functions, as well as rapid automatised naming and phonological memory, using a double-blind, within-subjects, repeated-measures design in typical young adult readers. Results: A single dose of 200 mg caffeine improved global processing, without any effect on local information processing, alerting, spatial attention and executive or phonological functions. This improvement in global processing was accompanied by faster text reading speed of meaningful sentences, whereas single word/pseudoword or pseudoword text reading abilities were not affected. These effects of caffeine on reading ability were enhanced by mild sleep deprivation. Conclusions: These findings show that a small quantity of caffeine could improve global processing and text reading skills in adults.

Individuals perceive the wor(l)d hierarchically. Firsty, the global visual scene is processed by the right hemisphere, and later, the local features are perceived by the left hemisphere. Based on this hierarchical analysis, humans evolved unique communication ability: reading. However, for about 10% of people reading acquisition is extremely difficult, they are affected by a heritable neurodevelopmental disorder called dyslexia. Differences in perceiving the wor(l)d might be one of the causes of reading disabilities. Here we show multiple causal links between the global before local perception and learning to read. Five behavioral experiments in 353 children reveal that: (i) a local before global perception characterizes three independent groups of unselected children with dyslexia; (ii) two global before local perception trainings improve reading skills in children with dyslexia; and stringently (iii) pre-reading local before global perception longitudinally predicts future poor readers. Challenging the uni-causal and left-lateralized phonological explanation of dyslexia, our results demonstrate that learning to read depends also on an efficient right neural network for the global analysis of the visual scene. These results provide new insights in learning strategies and pave the way for early identification and possible prevention programs.

Il disturbo da comportamento dirompente e uno dei piu frequenti motivi di consultazione neuropsichiatrica presso i servizi ambulatoriali per l’eta evolutiva in Italia (Frigerio et al., 2006), cosi come in altri paesi (Steiner e Remsing, 2007). Tale disturbo, comprendente il disturbo oppositivo provocatorio e il disturbo della condotta, e contraddistinto da un gruppo eterogeneo di pazienti con una vasta gamma di problemi comportamentali, che variano da un relativamente minore atteggiamento di sfida e scatti d’ira a piu serie violazioni, quali aggressione fisica, distruttivita e furto (American Psychiatric Association, 2000). I problemi maggiori ai quali rischiano di andare incontro i bambini con disturbo oppositivo provocatorio e disturbo della condotta sono principalmente: rifiuto dei pari, fallimento scolastico, abuso di sostanze, e criminalita (Burke, Arkowitz e Dunn, 2002; Odgers et al., 2008), con conseguenti costi sociali elevati (Kolko et al., 2009). La persistenza di tali problematiche comportamentali nei bambini e il loro ruolo nello sviluppo di rilevanti problemi clinici, mette in luce il bisogno di ulteriori studi nel campo dei fattori di rischio del disturbo da comportamento dirompente (Martens, 2000; Simonoff et al., 2004).

Dyslexia is a neurodevelopmental disorder with an atypical activation of posterior left-hemisphere brain reading networks (i.e., temporo-occipital and temporo-parietal regions) and multiple neuropsychological deficits. Transcranial direct current stimulation (tDCS) is a tool for manipulating neural activity and, in turn, neurocognitive processes. While studies have demonstrated the significant effects of tDCS on reading, neurocognitive changes beyond reading modulation have been poorly investigated. The present study aimed at examining whether tDCS on temporo-parietal regions affected not only reading, but also phonological skills, visuo-spatial working memory, visuo-spatial attention, and motion perception in a polarity-dependent way. In a within-subjects design, ten children and adolescents with dyslexia performed reading and neuropsychological tasks after 20 min of exposure to Left Anodal/Right Cathodal (LA/RC) and Right Anodal/Left Cathodal (RA/LC) tDCS. LA/RC tDCS compared to RA/LC tDCS improved text accuracy, word recognition speed, motion perception, and modified attentional focusing in our group of children and adolescents with dyslexia. Changes in text reading accuracy and word recognition speed—after LA/RC tDCS compared to RA/LC—were related to changes in motion perception and in visuo-spatial working memory, respectively. Our findings demonstrated that reading and domain-general neurocognitive functions in a group of children and adolescents with dyslexia change following tDCS and that they are polarity-dependent.

Visual crowding is the difficulty in perceiving a target in the presence of nearby flankers. Most neurophysiological studies of crowding employed functional neuroimaging, but because of its low temporal resolution, no definitive answer can be given to the question: is crowding arising at the earliest or at later stages of visual processing? Here, we used a classic letters crowding paradigm in combination with electroencephalography (EEG). We manipulated the critical space between peripheral target and flankers, while ensuring a proper control of basic stimulus characteristics. Analyses were focused on event-related potentials (ERPs) and oscillatory activity in the alpha (8-12 Hz), beta (15-30 Hz) and gamma (30-80 Hz) bands. At the ERP level, we found that the first sign of a crowding-induced modulation of EEG activity was a suppression of the N1 component. Oscillatory analysis revealed an early stimulus-evoked gamma enhancement and a later alpha reduction that, however, were not influenced by the amount of crowding. Importantly, reduction in the beta band reflected the amount of crowding (i.e., stronger reduction for strong relative to mid crowding condition) and correlated with individual behavioral performance. Collectively, these findings show that crowding for complex objects emerges at later stages of visual processing, possibly as a result of large-scale network interaction.

The hypoxic environment is a crucial component of the cancer stem cell niche and it is capable of eliciting stem cell features in cancer cells. We previously reported that SNAI2 up-regulates the expression of Carbonic Anhydrase iso-enzyme 9 (CA9) in hypoxic MCF7 cells. Here we show that SNAI2 down-regulates miR34a expression in hypoxic MCF7 cell-derived mammospheres. Next, we report on the capability of miR34a to decrease CA9 mRNA stability and CA9 protein expression. We also convey that the over-expression of cloned CA9-mRNA-3'UTR increases the mRNA half-life and protein levels of two miR34a targets JAGGED1 and NOTCH3. The data here reported shows that the SNAI2-dependent down-regulation of miR34a substantially contributes to the post-transcriptional up-regulation of CA9, and that CA9-mRNA-3'UTR acts as an endogenous microRNA sponge. We conclude that CA9/miR34 interplay shares in the hypoxic regulation of mammospheres and therefore, may play a relevant role in the hypoxic breast cancer stem cell niche.

Dyslexia (D) is a neurodevelopmental reading disorder characterized by phonological and orthographic deficits. Before phonological decoding, reading requires a specialized orthographic system for parallel letter processing that assigns letter identities to different spatial locations. The magnocellular-dorsal (MD) stream rapidly process the spatial location of visual stimuli controlling visuo-spatial attention. To investigate the visuo-spatial attention efficiency during orthographic processing, inhibition of return (IOR) was measured in adults with and without D in a lexical decision task. IOR is the delay in responding to stimuli displayed in a cued location after a long cue-target interval. Only adults with D did not showed IOR effect during letter-string recognition, despite the typical left-hemisphere specialization for word identification. A specific deficit in coherent-dot-motion perception confirmed an MD-stream disorder in adults with D. Our results suggest that adults with D might develop an efficient visual word form area, but a dorsal-attentional dysfunction impairs their reading fluency.

This work aims to design and develop a web application for reading and attentional evaluation in schooling children using a standardized tests battery. The application allows the automatic and immediate calculation of child's scores by inserting times and errors committed by the subject. The obtained scores are needed to establish whether the participant's performance is in deficit or not (i.e., Z-score).

Cancer stem cells (CSCs) are affected by the local micro-environment, the niche, in which inflammatory stimuli and hypoxia act as steering factors. Here, two nuclear receptors (NRs) agonists, i.e. pioglitazone (PGZ), a ligand of peroxisome proliferator activated receptor-γ, and 6-OH-11-O-hydroxyphenanthrene (IIF), a ligand of retinoid X receptors, were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment. We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF7 cancer cells, reducing the expression of CSCs regulatory genes (Notch3, Jagged1, SLUG, Interleukin-6, Apolipoprotein E, Hypoxia inducible factor-1α and Carbonic anhydrase IX). Notably, these effects are not observed in normal-MS obtained from human breast tissue. Importantly, NRs agonists abolish the capability of hypoxic MCF7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts. Moreover, NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases (MMP2 and MMP9) expression and activity. In conclusion, NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche.

Background Genetic, neurobiological, environmental and psychosocial mechanisms have received considerable attention in exploring the mechanisms that underlie comorbid PTSD and SUD. PTSD and SUD are not necessarily linked by a causal relationship, as the self-medication hypothesis had supposed. They might, in fact, both be caused by a third factor that predisposes these subjects to develop the two disorders (so allowing a unitary perspective). Methods Using a conceptualization of the PTSD spectrum, we studied the PTSD–SUD unitary perspective by testing the correlation between severity of heroin addiction, dose of opioid medication and severity of PTSD spectrum in 82 methadone-treated, heroin-dependent patients. Results Canonical correlation analysis (Wilks Lambda = 0.125 F = 1.41 p = 0.014), univariate and multivariate comparisons between subgroups, identified on the basis of addiction severity, showed a highly positive correlation between the PTSD spectrum and the severity of heroin addiction. In addition, negative correlations were found between PTSD spectrum severity and methadone dose (r = 0.225; p = 0.042). Conclusions This strength and breadth of the correlations encourage us to move towards a unified vision of the two disorders.

In up to 60% of human cancers, p53 gene mutations are responsible for direct inactivation of the tumor suppressor function of p53. Alternative mechanisms of p53 inactivation described thus far mainly affect its posttranslational regulation. In X-linked dyskeratosis congenita, a multisystemic syndrome characterized by increased cancer susceptibility, mutations of the DKC1 gene encoding dyskerin cause a selective defect in the translation of a subgroup of internal ribosome entry site (IRES)–containing cellular mRNAs. In this study, we show that impairment of dyskerin function can cause p53 inactivation due to a defect in p53 mRNA translation. siRNA-mediated reduction of dyskerin levels caused a decrease of p53 mRNA translation, protein levels, and functional activity, both in human breast cancer cells and in primary mammary epithelial progenitor cells. These effects seemed to be independent of the known role of dyskerin in telomerase function, and they were associated with a specific impairment of translation initiation mediated by IRES elements present in p53 mRNA. In a series of human primary breast cancers retaining wild-type p53, we found that low levels of dyskerin expression were associated with reduced expression of p53-positive target genes. Our findings suggest that a dyskerin-mediated mechanism of p53 inactivation may occur in a subset of human tumors. Cancer Res; 70(11); 4767–77. ©2010 AACR.

Impaired linguistic-phonological processing is the most accepted explanation of developmental dyslexia (DD). However, growing literature shows that DD is the result of the combination of several neurocognitive causes. Visual attention and magnocellular-dorsal (MD) pathway deficits are now considered causes of DD. Interestingly, a large portion of literature showed that action video games (AVG) are able to improve attentional and perceptual skills in typical readers. Consequently, employing AVG trainings in individuals with DD could improve attention and perception, resulting in better reading skills. The aim of our review is to show the benefits of the AVG training on DD through the changes in the neurocognitive functions at the basis of learning to read. Since visual attentional and MD dysfunctions can be diagnosed in infancy, our review paves the way for possible early prevention programs that could use AVG training.

Busulfan liver metabolism depends on glutathione, a crucial mediator of cellular and systemic stress. Here we investigated 40 polymorphisms at 27 loci involved in hepatic glutathione homeostasis, with the aim of testing their impact on the clinical outcome of 185 busulfan-conditioned allogeneic transplants. GSTA2 S112T serine allele homozygosity is an independent prognostic factor for poorer survival (RR=2.388), for increased any time- and 100-day transplant-related mortality (RR=4.912 and RR=5.185, respectively). The genotype also predicts a wider busulfan area under the concentration-time curve (1214.36 ± 570.06 vs. 838.10 ± 282.40 mMol*min) and higher post-transplant bilirubin serum levels (3.280 ± 0.422 vs. 1.874+0.197 mg/dL). In vitro, busulfan elicits pro-inflammatory activation (increased NF-KappaB activity and interleukin-8 expression) in human hepatoma cells. At the same time, the drug down-regulates a variety of genes involved in bilirubin liver clearance: constitutive androstane receptor, multidrug resistance-associated protein, solute carrier organic anion transporters, and even GSTA2. It is worthy of note that GSTA2 also acts as an intra-hepatic bilirubin binding protein. These data underline the prognostic value of GSTA2 genetic variability in busulfan-conditioned allotransplants and suggest a patho-physiological model in which busulfan-induced inflammation leads to the impairment of post-transplant bilirubin metabolism.

Cancer stem cells (CSCs) are affected by the local micro-environment, the niche, in which inflammatory stimuli and hypoxia act as steering factors. Here, two nuclear receptors (NRs) agonists, i.e. pioglitazone (PGZ), a ligand of peroxisome proliferator activated receptor-γ, and 6-OH-11-O-hydroxyphenanthrene (IIF), a ligand of retinoid X receptors, were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment. We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF7 cancer cells, reducing the expression of CSCs regulatory genes (Notch3, Jagged1, SLUG, Interleukin-6, Apolipoprotein E, Hypoxia inducible factor-1α and Carbonic anhydrase IX). Notably, these effects are not observed in normal-MS obtained from human breast tissue. Importantly, NRs agonists abolish the capability of hypoxic MCF7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts. Moreover, NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases (MMP2 and MMP9) expression and activity. In conclusion, NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche.

AIMS: Cancer stem cell biology is tightly connected to the regulation of the pro-inflammatory cytokine network. The concept of cancer stem cells "inflammatory addiction" leads to envisage the potential role of anti-inflammatory molecules as new anti-cancer targets. Here we report on the relationship between nuclear receptors activity and the modulation of the pro-inflammatory phenotype in breast cancer stem cells. METHODS: Breast cancer stem cells were expanded as mammospheres from normal and tumor human breast tissues and from tumorigenic (MCF7) and non tumorigenic (MCF10) human breast cell lines. Mammospheres were exposed to the supernatant of breast tumor and normal mammary gland tissue fibroblasts. RESULTS: In mammospheres exposed to the breast tumor fibroblasts supernatant, autocrine tumor necrosis factor-α signalling engenders the functional interplay between peroxisome proliferator activated receptor-α and hypoxia inducible factor-1α (PPARα/HIF1α). The two proteins promote mammospheres formation and enhance each other expression via miRNA130b/miRNA17-5p-dependent mechanism which is antagonized by PPARγ. Further, the PPARα/HIF1α interplay regulates the expression of the pro-inflammatory cytokine interleukin-6, the hypoxia survival factor carbonic anhydrase IX and the plasma lipid carrier apolipoprotein E. CONCLUSION: Our data demonstrate the importance of exploring the role of nuclear receptors (PPARα/PPARγ) in the regulation of pro-inflammatory pathways, with the aim to thwart breast cancer stem cells functioning.

For children affected by developmental dyslexia learning to read is extremely difficult. Pre-reading visual attention predicts future reading acquisition skills. Action video game (AVG) training increases attentional functioning and induces learning that transfers well beyond the task domain, such as reading. We investigated the effects of AVG training on predictors of future reading acquisition (i.e., visuo-spatial attention, auditory-phonological processing and rapid naming skills) and on the dense-array EEG resting-state in pre-reading children at risk for dyslexia. Three matched groups of pre-readers at risk for dyslexia were tested before and after they played with AVG, non-AVG (for 20 hrs) or no-treatment (Exp. 1). We found that only playing AVG improved children's visuo-spatial attention processing. Phonemes discrimination was also increased only after AVG training. We confirmed this effect of AVG training on phonological processing in a replication study with another independent sample (Exp. 2). Two new samples of pre-schoolers at risk for dyslexia were selected (Exp. 3): half of them were trained with AVG while the other half had no training. We measured visuo-spatial attention and auditory-phonological skills. Eyes-closed resting-state EEG was also recorded in both groups. Results showed that only the AVG training improved visuo-spatial attention as well as auditory-phonological skills. A reduction of the upper alpha band (10-14 Hz) oscillatory activity in posterior areas was found only after the AVG training, showing a possible neural basis of the effect of attentional improvement on auditory-phonological processing. Our results showed, for the first time, that attention improvements can directly translate into better language abilities, providing a new, fast and fun prevention training for dyslexia.