Caratterizzazione di una proteina ABC-transporter di P. x euroamericana clone I-214 sottoposto a stress da zinco: possibile ruolo nel sequestro intracellulare e nella tolleranza

Lo zinco (Zn) è un microelemento essenziale per tutti gli organismi, comprese le piante. Svolge importanti funzioni nel metabolismo cellulare in quanto cofattore di enzimi e componente di proteine come i fattori di trascrizione a dominio Zn-finger. Un eccesso di questo metallo porta a sintomi da tossicità nelle piante, che, quindi, hanno evoluto meccanismi di tolleranza per affrontare lo stress derivante dall’eccesso di zinco nei suoli. Il genere Populus viene utilizzato come modello di piante arboree per la fitoestrazione di metalli pesanti dai suoli inquinati. Il ruolo dei trasportatori è fondamentale per consentire l’omeostasi dei metalli essenziali ed escludere dal citosol i quelli pesanti. Nelle piante sono state identficate diverse famiglie geniche codificanti numerosi trasportatori di membrana e tra questi ci sono anche gli ABC-transporter. Gli ABC-transporter sono una classe di trasportatori molto ampia, in grado di svolgere svariate funzioni in tutti gli organismi. Sono pompe per il trasporto attivo, modulano l’attività di canali ionici, sono proteine canale oppure sono coinvolti in attività diverse dal trasporto, come la trasduzione del segnale. Sono in grado di trasportare un’ampia varietà di substrati come ioni, acidi inorganici, peptidi, zuccheri, polisaccaridi, lipidi e complessi formati da metalli pesanti legati da agenti chelanti come il glutatione. Prendono parte a processi biologici diversi, come la detossificazione cellulare, la crescita e lo sviluppo delle piante, il movimento degli stomi e la tolleranza ai metalli pesanti. Il lavoro sperimentale svolto ha riguardato lo studio funzionale di un gene di P. x euroamericana clone I-214 codificante un putativo ABC-transporter da noi chiamato dr19 che, da studi preliminari, risulta differenzialmente espresso a dosi sub letali di Zn (1 mM). Per indagare il possibile coinvolgimento di questo gene nei meccanismi di tolleranza in risposta a un eccesso di Zn e, in particolare, al sequestro intracellulare di questo metallo, ne è stato eseguito il clonaggio da cDNA P. x euroamericana clone I-214 e P.alba clone Villafranca e la sua caratterizzazione. È stata effettua l’analisi dei livelli di espressione del gene dr19 tramite Real Time qPCR in foglie vecchie e giovani di P. x euroamericana clone I-214 in condizioni di controllo e in presenza di 1 mM di Zn, da cui si notano delle differenze nei livelli di espressione nelle foglie vecchie nelle condizioni di trattamento, rispetto il controllo. È stata studiata la localizzazione subcellulare tramite espressione transiente in protoplasti di A. thaliana in condizioni di controllo e in presenza di 200 µm di Zn, dove la proteina sembra localizzarsi a livello citoplasmatico e, in presenza di Zn, le cellule non manifestano sintomi di sofferenza. Gli studi di caratterizzazione sono proseguiti su P. alba clone Villafranca trattato in vitro con 500 e 1000 µm di Zn e ortovanadato (inibitore degli ABC-transporter): l’analisi dei parametri fisiologici ha evidenziato un aumento generale di biomassa e dello Zn assorbito e una diminuzione della biomassa nei trattamenti con ortovanadato. È stata effettua, infine, l’analisi dei livelli di espressione del gene dr19 tramite Real Time qPCR in foglie di P. alba clone Villafranca da cui non si notano delle differenze tra i trattamenti. L’assenza di stress nei protoplasti trattati con 200 µm di Zn indica un possibile coinvolgimento del gene dr19 nei processi di esclusione e/o omeostasi dello Zn nel pioppo. L’osservazione che nel P. alba fino a 1 mM di Zn (concentrazione in cui non si manifestano effetti di stress) l’espressione di dr19 non cambia, suggerisce il suo coninvolgimento nei processi di omeostasi alle concentrazioni di Zn utilizzate in quanto non compaiono sintomi di tossicità. Nel clone I-214, invece, trattato con 1 mM di Zn (dose subletale) il gene dr19 risulta sovra espresso, e questo sembra indicare un suo coninvolgimento nei processi di detossificazione in presenza di concentrazioni di Zn tali da indurre stress

Effects of syllabic complexity in predicting accuracy of repetition and direction of errors in patients with articulatory and phonological difficulties

The purpose of this paper is to demonstrate the existence of a strong and significant effect of complexity in aphasia independent from other variables including length. Complexity was found to be a strong and significant predictor of accurate repetition in a group of 13 Italian aphasic patients when it was entered in a regression equation either simultaneously or after a large number of other variables. Significant effects were found both when complexity was measured in terms of number of complex onsets (as in a recent paper by Nickels & Howard, 2004) and when it was measured in a more comprehensive way. Significant complexity effects were also found with matched lists contrasting simple and complex words and in analyses of errors. Effects of complexity, however, were restricted to patients with articulatory difficulties. Reasons for this association and for the lack of significant results in Nickels and Howard (2004) are discussed. © 2005 Psychology Press Ltd

Cracking the Toll-like receptor code in fungal infections

Innate control of fungal infection requires the specific recognition of invariant fungal molecular structures by a variety of innate immune receptors, including Toll-like receptors. In addition to the role in inducing protective immune responses, Toll-like receptor engagement may paradoxically favor fungal infections, by inducing inflammatory pathology and impairing antifungal immunity. Although the dissection of complex genetic traits modulating susceptibility to fungal infections is complex, the contribution of host genetics may hold the key to elucidating new risk factors for these severe, often fatal diseases. Understanding host-pathogen interactions at the innate immune interface will eventually lead to the development of new therapeutics and genetic markers in fungal infections.This work was supported by the Specific Targeted Research Projects SYBARIS (FP7-HEALTH-2009), contract number 242220, and by the Italian Project PRIN 2007KLCKP8_004. Cristina Cunha and Agostinho Carvalho were financially supported by fellowships from Fundacao para a Ciencia e Tecnologia, Portugal (contracts SFRH/BD/65962/2009 and SFRH/BPD/46292/2008, respectively). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed

Lexical learning and dysgraphia in a group of adults with developmental dyslexia

We investigated the ability to learn new words in a group of 22 adults with developmental dyslexia/dysgraphia and the relationship between their learning and spelling problems. We identified a deficit that affected the ability to learn both spoken and written new words (lexical learning deficit). There were no comparable problems in learning other kinds of representations (lexical/semantic and visual) and the deficit could not be explained in terms of more traditional phonological deficits associated with dyslexia (phonological awareness, phonological STM). Written new word learning accounted for further variance in the severity of the dysgraphia after phonological abilities had been partialled out. We suggest that lexical learning may be an independent ability needed to create lexical/formal representations from a series of independent units. Theoretical and clinical implications are discussed. © 2005 Psychology Press Ltd

Mucosa-Environment Interactions in the Pathogenesis of Rheumatoid Arthritis

Mucosal surfaces play a central role in the pathogenesis of rheumatoid arthritis (RA). Several risk factors, such as cigarette smoking, environmental pollution, and periodontitis interact with the host at the mucosal level, triggering immune system activation. Moreover, the alteration of microbiota homeostasis is gaining increased attention for its involvement in the disease pathogenesis, modulating the immune cell response at a local and subsequently at a systemic level. Currently, the onset of the clinical manifest arthritis is thought to be the last step of a series of pathogenic events lasting years. The positivity for anti-citrullinated protein antibodies (ACPAs) and rheumatoid factor (RF), in absence of symptoms, characterizes a preclinical phase of RA namely systemic autoimmune phase- which is at high risk for disease progression. Several immune abnormalities, such as local ACPA production, increased T cell polarization towards a pro-inflammatory phenotype, and innate immune cell activation can be documented in at-risk subjects. Many of these abnormalities are direct consequences of the interaction between the environment and the host, which takes place at the mucosal level. The purpose of this review is to describe the humoral and cellular immune abnormalities detected in subjects at risk of RA, highlighting their origin from the mucosa environment interaction

The ability to learn new written words is modulated by language orthographic consistency

Introduction It is well known that a difficulty in forming lexical representations is a strong predictor of reading and spelling difficulties even after controlling for the effects of other cognitive skills. Our study had two main interrelated aims. First, we wanted to examine whether the ability to learn new written words (lexical learning) varies as a function of the orthographic consistency of the language of the learner. Second, we wanted to evaluate the cognitive abilities involved in orthographic lexical learning and whether they differed as a function of language consistency. Method 163 Italian children and 128 English children performed a lexical learning task as well as tasks assessing several cognitive skills potentially related to the ability to establish orthographic representations. Results We found that children learning an orthographic inconsistent orthography (English) were better able to learn novel written words presented in association with pictures than children learning a consistent orthography (Italian). This was true for both younger and older primary school children and also when children were matched for school grade. Lexical learning may be better in English children because the many irregularities of this language promote storing in memory whole-word representations and processing larger orthographic units. In Italian, instead, reading can be accomplished successfully on the basis of grapheme-phoneme conversion rules and on processing smaller orthographic units. This interpretation was supported by the pattern of cognitive skills associated with lexical learning skills in the two languages. Variations in lexical learning were explained by spatial visual memory and phonological awareness tasks in both languages, but phonological STM explained further variance in Italian, while a task tapping visuo-attentional capacity explained further variance in English. Conclusion Learning a language with inconsistent orthography is associated with better lexical learning skills in children at different stages of primary school; the pattern of cognitive skills associated with lexical learning skills is also partially modulated by orthographic consistency

Immunotherapy of aspergillosis

We thank C. M. Benedetti for digital art and editing.Management of invasive aspergillosis in high-risk patients remains challenging. There is an increasing demand for novel therapeutic strategies aimed at enhancing or restoring antifungal immunity in immunocompromised patients. In this regard, modulation of specific innate immune functions and vaccination are promising immunotherapeutic strategies. Recent findings have also provided a compelling rationale for assessment of the contribution of the individual genetic profile to the immunotherapy outcome. Altogether, integration of immunological and genetic data may contribute to the optimization of therapeutic strategies exerting control over immune pathways, ultimately improving the management of fungal infections in high-risk settings.The studies were supported by the Specific Targeted Research Project ALLFUN (FP7-HEALTH-2009 contract number 260338 to L. Romani) and by the Fondazione per la Ricerca sulla Fibrosi Cistica (FFC#21/2010 to L. Romani, with the contribution of Francesca Guadagnin, Coca Cola light Tribute to Fashion and Delegazione FFC di Belluno). C. Cunha and A. Carvalho were financially supported by fellowships from Fundação para a Ciência e Tecnologia, Portugal (contracts SFRH/ BD/65962/2009 and SFRH/BPD/46292/2008, respectively)

Playing a team game improves word production in poststroke aphasia

Background: High intensity, one-to-one rehabilitation therapy is effective in the treatment of poststroke aphasia, but it can put strain on public health providers, as well as lead to high attrition. Working within a group of peers may be efficient for professional speech and language therapists, as well as reduce feelings of isolation and lack of confidence in patients, which can negatively affect progress. Evidence-based, structured group-based approaches, however, are lacking. Aims: We wanted to assess the feasibility a new group-delivered game-based intervention, designed to provide efficacious word-retrieval rehabilitation, in a cost-effective and motivating environment. Method & Procedure: Two cohorts of six participants took part. Each was split into two teams to play language games where pictures were named with the help of team members and facilitation from a speech and language therapist. Facilitation was varied in three different cueing conditions: phonemic, gesture + phonemic, and semantic + phonemic. Overall, 180 words were practiced (90 nouns and 90 verbs). Therapy was delivered 3 days per week, for 6 weeks (for a total of 54 hr). Outcomes & Results: Our intervention was equally effective across the three cueing conditions and for nouns and verbs. Gains were demonstrated in naming the pictures used in training, but also in the description of pictured scenes designed to elicit the same words. With these tasks, there were improvements of 25% and 18% from base-line accuracy, which compares well with gains reported in the literature using individually delivered speech and language therapy based on picture naming. Improvements were mostly maintained at both 4–7 weeks and 6-months post-therapy and were significant in all but the two most severely affected participants. There was some generalization of gains to narrative production, but not to other language tasks, nor to untreated words in picture naming. These positive language outcomes were combined with a high level of engagement and satisfaction (with participants stating a preference for games over standard therapy). Conclusions: Our results support embedding theoretically and empirically based techniques for aphasia rehabilitation within games with a strong social aspect, which may promote linguistic recovery in a way that is both time and cost-efficient and engaging. Future research should explore more formally outcomes in terms of increased well-being and reduced social isolation, as well as language proficiency