Phosphodiesterases S-sulfhydration contributes to human skeletal muscle function

The increase in intracellular calcium is influenced by cyclic nucleotides (cAMP and cGMP) content, which rating is governed by phosphodiesterases (PDEs) activity.Despite it has been demonstrated a beneficial effect of PDEs inhibitors in different pathological conditions involving SKM, not much is known on the role exerted by cAMP-cGMP/PDEs axis in human SKM contractility. Here, we show that Ssulfhydration of PDEs modulates human SKM contractility in physiological and pathological conditions. Having previously demonstrated that, in the rare human syndrome Malignant Hyperthermia (MH), there is an overproduction of hydrogen sulfide (H2S) within SKM contributing to hyper-contractility, here we have used MH negative diagnosed biopsies (MHN) as healthy SKM, and MH susceptible diagnosed biopsies (MHS) as a pathological model of SKM hypercontractility. The study has been performed on MHS and MHN human biopsies after diagnosis has been made and on primary SKM cells derived from both MHN and MHS biopsies. Our data demonstrate that in normal conditions PDEs are S-sulfhydrated in both quadriceps' biopsies and primary SKM cells. This post translational modification (PTM) negatively regulates PDEs activity with consequent increase of both cAMP and cGMP levels. In hypercontractile biopsies, due to an excessive H2S content, there is an enhanced Ssulfhydration of PDEs that further increases cyclic nucleotides levels contributing to SKM hyper-contractility. Thus, the identification of a new endogenous PTM modulating PDEs activity represents an advancement in SKM physiopathology understanding

Direttrice sud-ovest: Corsico. Lungo il corso del naviglio. Il recinto della Richard-Ginori

Nel progetto per l'area dismessa della Richard Ginori il naviglio fluisce all'interno del recinto ridisegnato e scandito in fasce che allineano nuovi vuoti e architetture "misurate" per una nuova video città

Sei quello che sei.

Il progetto per la ridefinizione architettonica della piazza C. M. Carafa lavora su corrispondenze formali, metriche e percettive tra gli elementi architettonici che disegnano la figura dell'esagono della piazza. Con lo stesso criterio sono definiti il disegno della pavimentazione e il posizionamento degli elementi di arredo dello spazio vuoto

Sei quello che sei

none6Pochi luoghi, più della piazza centrale di Grammichele testimoniano dello scarto di forma e di senso che può verificarsi tra un “impianto” planimetrico geometricamente compiuto e la sua concretizzazione tipo-morfologica. Del resto, la impressionante “tenuta” del piano di fondazione non è affatto in contraddizione con la “debolezza” della sua costruzione tridimensionale ma ne è probabilmente la logica conseguenza: i contadini e gli artigiani delle piane iblee, chiamati ad abitare “l’elegantissima piazza” esagonale e gli isolati che ne solidificano le tracce concentriche, si limitano a riempire dei contenuti minimi della loro vista quotidiana la singolare opera d’arte voluta dal loro colto principe. Puro spazio geometrico, privo di orientamento, nel suo originario e anomalo rifiuto di ospitare gli edifici del potere che tradizionalmente connotano i luoghi centrali delle città fondate, la piazza di Grammichele ha perso presto anche l’unico elemento di polarizzazione, la meridiana, capriccio del Princi-pe, ad un tempo ridondante e riduttivo tentativo di costringere in un punto materiale l’idea di una centralità vuota e spaziosa. Non accettare la contraddizione tra la discontinuità talvolta stridente dei fronti costruiti e la fluida compiutezza dello spazio della piazza sarebbe non accettare la “qualità” più profonda e complessa, e per questo forse davvero moderna, della sua struttura. Il nostro progetto lavora dunque sulla qualità del vuoto puntando a consolidarne ad un tempo, attraverso analogie discrete, la solidità geometrica e la connotazione di luogo centrale. Se Grammichele esibisce il suo impianto come vero museo della città, la sua piazza centrale, antiquarium all’aperto, può raccogliere i frammenti d’arte sopravvissuti alla furia del terremoto del 1693, spaventoso evento distruttivo che è però al tempo stesso causa prima della costruzione della nuova città. E così come le statue sorrette dai piedistalli che popolano la piazza sono solo copie di oggetti d’arte, violati dal tempo e dalla natura, gli elementi di “arredo urbano”, riassunti in forme geometriche scarne e disposti in ordine sparso nello spazio esagonale, rappresentano ad un tempo la concretizzazione ed il simulacro straniato delle strutture tradizionali di una “piazza centrale”: il sagrato della chiesa, la fontana, il piccolo giardino, le panchine. Gli edifici di progetto si inseriscono compostamente nel perimetro costruito, per colmarne i vuoti planimetrici, ed esibiscono con naturalezza la loro differenza strutturale e formale, appena mitigata dal sobrio riferimento ad una originaria scansione modulare del perimetro costruito, oggi rintracciabile a fatica. Il piccolo edificio che ospita il museo civico sostiene un’alta torre che, nella sua asciutta definizione volumetrica, fa da contrappunto alla composita mole della chiesa: nelle sale del museo i documenti racconteranno la storia di una città “ideale” mentre la città reale potrà raccontare se stessa a chi salirà sulla torre per guardarla dall’alto.Pubblicazione del progetto per la Ridefinizione architettonica di piazza Carlo Maria Carafa in Grammichele (motto: "Sei quello che sei"), Concorso Nazionale di Idee "Grammichele. Una città plurale", bandito dal Comune di Grammichele.mixedR. Amirante; F. Bruni; A. Casillo; A. D'Agostino; S. Pone; L. StendardoR., Amirante; F., Bruni; A., Casillo; A., D'Agostino; S., Pone; Stendardo, Luig

GlcNAc De-N-Acetylase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

Sulfolobus solfataricus is an aerobic crenarchaeal hyperthermophile with optimum growth at temperatures greater than 80°C and pH 2 to 4. Within the crenarchaeal group of Sulfolobales, N-acetylglucosamine (GlcNAc) has been shown to be a component of exopolysaccharides, forming their biofilms, and of the N-glycan decorating some proteins. The metabolism of GlcNAc is still poorly understood in Archaea, and one approach to gaining additional information is through the identification and functional characterization of carbohydrate active enzymes (CAZymes) involved in the modification of GlcNAc. The screening of S. solfataricus extracts allowed the detection of a novel α-N-acetylglucosaminidase (α-GlcNAcase) activity, which has never been identified in Archaea Mass spectrometry analysis of the purified activity showed a protein encoded by the sso2901 gene. Interestingly, the purified recombinant enzyme, which was characterized in detail, revealed a novel de-N-acetylase activity specific for GlcNAc and derivatives. Thus, assays to identify an α-GlcNAcase found a GlcNAc de-N-acetylase instead. The α-GlcNAcase activity observed in S. solfataricus extracts did occur when SSO2901 was used in combination with an α-glucosidase. Furthermore, the inspection of the genomic context and the preliminary characterization of a putative glycosyltransferase immediately upstream of sso2901 (sso2900) suggest the involvement of these enzymes in the GlcNAc metabolism in S. solfataricusIMPORTANCE In this study, a preliminary screening of cellular extracts of S. solfataricus allowed the identification of an α-N-acetylglucosaminidase activity. However, the characterization of the corresponding recombinant enzyme revealed a novel GlcNAc de-N-acetylase, which, in cooperation with the α-glucosidase, catalyzed the hydrolysis of O-α-GlcNAc glycosides. In addition, we show that the product of a gene flanking the one encoding the de-N-acetylase is a putative glycosyltransferase, suggesting the involvement of the two enzymes in the metabolism of GlcNAc. The discovery and functional analysis of novel enzymatic activities involved in the modification of this essential sugar represent a powerful strategy to shed light on the physiology and metabolism of Archaea

Higher Number of Night Shifts Associates with Good Perception of Work Capacity and Optimal Lung Function but Correlates with Increased Oxidative Damage and Telomere Attrition

Sleep deprivation and the consequent circadian clock disruption has become an emergent health question being associated with premature aging and earlier chronic diseases onset. Night-shift work leads to circadian clock misalignment, which is linked to several age-related diseases. However, mechanisms of this association are not well understood. Aim of this study is to explore in night-shift workers early indicators of oxidative stress response and biological aging [oxidized/methylated DNA bases and leukocytes telomere length (LTL)] and late indicators of functional aging [lung function measurements (FEV1 and FVC)] in relation to personal evaluation of work capacity, measured by work ability index (WAI). One hundred fifty-five hospital workers were studied within the framework of a cross-sectional study. We collected physiological, pathological, and occupational history including pack-years, alcohol consumption, physical activity, and night shifts, together with blood and urine samples. Relationships were appraised by univariate and multivariate ordered-logistic regression models. We found that workers with good and excellent WAI present higher FEV1 (p< 0.01) and number of night-work shifts (p<0.05), but they reveal higher urinary levels of 8-oxoGua (p<0.01) and shorter LTL (p<0.05). We confirmed that higher work ability was prevalent among chronological younger workers (p<0.05), who have also a significant reduced number of diseases, particularly chronic (p<0.01) and musculoskeletal diseases (p<0.01). The new findings which stem from our work are that subjects with the highest work ability perception may have more demanding and burdensome tasks; they in fact present the highest number of night-shift work and produce unbalanced oxidative stress response that might induce premature aging

Hydrogen sulfide donor AP123 restores endothelial nitric oxide-dependent vascular function in hyperglycemia via a CREB-dependent pathway

Diabetes is associated with severe vascular complications involving the impairment of endothelial nitric oxide synthase (eNOS) as well as cystathionine γ-lyase (CSE) activity. eNOS function is suppressed in hyperglycaemic conditions, resulting in reduced NO bioavailability, which is paralleled by reduced levels of hydrogen sulfide (H2S). Here we have addressed the molecular basis of the interplay between the eNOS and CSE pathways. We tested the impact of H2S replacement by using the mitochondrial-targeted H2S donor AP123 in isolated vessels and cultured endothelial cells in high glucose (HG) environment, at concentrations not causing any vasoactive effect per se. Aorta exposed to HG displayed a marked reduction of acetylcholine (Ach)-induced vasorelaxation that was restored by the addition of AP123 (10 nM). In HG condition, bovine aortic endothelial cells (BAEC) showed reduced NO levels, downregulation of eNOS expression, and suppression of CREB activation (p-CREB). Similar results were obtained by treating BAEC with propargylglycine (PAG), an inhibitor of CSE. AP123 treatment rescued eNOS expression, as well as NO levels, and restored p-CREB expression in both the HG environment and the presence of PAG. This effect was mediated by a PI3K-dependent activity since wortmannin (PI3K inhibitor) blunted the rescuing effects operated by the H2S donor. Experiments performed in the aorta of CSE−/− mice confirmed that reduced levels of H2S not only negatively affect the CREB pathway but also impair Ach-induced vasodilation, significantly ameliorated by AP123. We have demonstrated that the endothelial dysfunction due to HG involves H2S/PI3K/CREB/eNOS route, thus highlighting a novel aspect of the H2S/NO interplay in the vasoactive response

Interleukin-17 (IL-17) triggers systemic inflammation, peripheral vascular dysfunction, and related prothrombotic state in a mouse model of Alzheimer's disease

Alzheimer's disease (AD) is one of the most prevalent forms of neurodegenerative disorders. Previously, we have shown that in vivo administration of an IL-17 neutralizing antibody (IL-17Ab) rescues amyloid-β-induced neuro-inflammation and memory impairment, demonstrating the pivotal role of IL-17 in AD-derived cognitive deficit. Recently, AD has been recognized as a more intriguing pathology affecting vascular network and platelet function. However, not much is known about peripheral vascular inflammation and how pro-inflammatory circulating cells/mediators could affect peripheral vessels' function. This study aimed to evaluate whether IL-17Ab treatment could also impact peripheral AD features, such as systemic inflammation, peripheral vascular dysfunction, and related pro-thrombotic state in a non-genetic mouse model of AD. Mice were injected intracerebroventricularly with Aβ1-42 peptide (3μg/3μl). To evaluate the systemic/peripheral protective profile of IL-17Ab, we used an intranasal administration of IL-17Ab (1μg/10μl) at 5, 12, and 19 days after Aβ1-42 injection. Circulating Th17/Treg cells and related cyto-chemokines, haematological parameters, vascular/endothelial reactivity, platelets and coagulation function in mice were evaluated. IL-17Ab treatment ameliorates the systemic/peripheral inflammation, immunological perturbance, vascular/endothelial impairment and pro-thrombotic state, suggesting a key role for this cytokine in fostering inflammatory processes that characterize the multifaced aspects of AD