Repairing folding-defective \u3b1-sarcoglycan mutants by CFTR correctors, a potential therapy for Limb Girdle Muscular Dystrophy 2D

Limb Girdle Muscular Dystrophy type 2D (LGMD2D) is a rare autosomal-recessive disease, affecting striated muscle, due to mutation of SGCA, the gene coding for \u3b1-sarcoglycan. Nowadays more than 50 different SGCA missense mutations have been reported. They are supposed to impact folding and trafficking of \u3b1-sarcoglycan because the defective polypeptide, although potentially functional, is recognized and disposed of by the quality control of the cell. The secondary reduction of \u3b1-sarcoglycan partners, \u3b2-, \u3b3- and \u3b4-sarcoglycan, disrupts a key membrane complex that, associated to dystrophin, contributes to assure sarcolemma stability during muscle contraction. The complex deficiency is responsible for muscle wasting and the development of a severe form of dystrophy.Here, we show that the application of small molecules developed to rescue \u394F508-CFTR trafficking, and known as CFTR correctors, also improved the maturation of several \u3b1-sarcoglycan mutants that were consequently rescued at the plasma membrane. Remarkably, in myotubes from a patient with LGMD2D, treatment with CFTR correctors induced the proper re-localization of the whole sarcoglycan complex, with a consequent reduction of sarcolemma fragility. Although the mechanism of action of CFTR correctors on defective \u3b1-sarcoglycan needs further investigation, this is the first report showing a quantitative and functional recovery of the sarcoglycan-complex in human pathologic samples, upon small molecule treatment. It represents the proof of principle of a pharmacological strategy that acts on the sarcoglycan maturation process and we believe it has a great potential to develop as a cure for most of the patients with LGMD2D

Association of kidney disease measures with risk of renal function worsening in patients with type 1 diabetes

Background: Albuminuria has been classically considered a marker of kidney damage progression in diabetic patients and it is routinely assessed to monitor kidney function. However, the role of a mild GFR reduction on the development of stage 653 CKD has been less explored in type 1 diabetes mellitus (T1DM) patients. Aim of the present study was to evaluate the prognostic role of kidney disease measures, namely albuminuria and reduced GFR, on the development of stage 653 CKD in a large cohort of patients affected by T1DM. Methods: A total of 4284 patients affected by T1DM followed-up at 76 diabetes centers participating to the Italian Association of Clinical Diabetologists (Associazione Medici Diabetologi, AMD) initiative constitutes the study population. Urinary albumin excretion (ACR) and estimated GFR (eGFR) were retrieved and analyzed. The incidence of stage 653 CKD (eGFR < 60 mL/min/1.73 m2) or eGFR reduction > 30% from baseline was evaluated. Results: The mean estimated GFR was 98 \ub1 17 mL/min/1.73m2 and the proportion of patients with albuminuria was 15.3% (n = 654) at baseline. About 8% (n = 337) of patients developed one of the two renal endpoints during the 4-year follow-up period. Age, albuminuria (micro or macro) and baseline eGFR < 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR > 90 ml/min/1.73m2 and normoalbuminuria, those with albuminuria at baseline had a 1.69 greater risk of reaching stage 3 CKD, while patients with mild eGFR reduction (i.e. eGFR between 90 and 60 mL/min/1.73 m2) show a 3.81 greater risk that rose to 8.24 for those patients with albuminuria and mild eGFR reduction at baseline. Conclusions: Albuminuria and eGFR reduction represent independent risk factors for incident stage 653 CKD in T1DM patients. The simultaneous occurrence of reduced eGFR and albuminuria have a synergistic effect on renal function worsening

I fattori socio-demografici ed i processi di cambiamento per smettere di fumare associati agli stadi del cambiamento: confronto tra le donne gravide e non gravide.

Background: Le autorit\ue0 internazionali per il controllo del tabagismo sostengono l\u2019efficacia degli interventi personalizzati. Nonostante numerosi studi eseguiti in tutto il mondo comprovino la pericolosit\ue0 del fumo durante la gravidanza per i danni che provoca alla madre e al feto, poca attenzione \ue8 stata rivolta alla comprensione delle differenze tra donne gravide e non gravide in termini di fattori sociali influenzanti l\u2019uso del tabacco e circa i processi di cambiamento messi in atto per smettere di fumare. Metodi: Lo studio ha coinvolto 177 donne gravide raccolte consecutivamente durante i controlli prenatali del terzo trimestre, che si sono dichiarate fumatrici nei sei mesi prima dell\u2019inizio della gravidanza, attualmente fumatrici o ex-fumatrici. Inoltre sono state intervistate in occasione dei controlli ginecologici di routine( pap test) 177 donne non gravide, che si dichiaravano fumatrici nove mesi prima dell\u2019intervista, attualmente fumatrici o ex-fumatrici. Sono stati somministrati i questionari di Prochaska e DiClemente per valutare gli stadi del cambiamento, i processi del cambiamento, il grado di accettabilit\ue0 e la tentazione di riprendere l\u2019abitudine al fumo in determinate situazioni. Risultati: E\u2019stato dimostrato come per le donne gravide gli stadi del cambiamento sono associati al livello di educazione, allo stato civile; questa associazione non \ue8 stata rilevata nelle non gravide. Inoltre nelle gravide gli stadi del cambiamento sono associati ad avere amici, partner e conviventi fumatori; mentre nelle non gravide solo la convivenza con fumatori \ue8 associata agli stadi del cambiamento . E\u2019 stato inoltre evidenziato come i processi esperienziali siano maggiormente l\u2019utilizzati negli stadi precoci (pre-contemplativo); \ue8 stato osservato che i processi comportamentali sono maggiormente utilizzati dalle donne non gravide rispetto alle gravide. Conclusioni: Nelle donne gravide, diversamente dalle non gravide, gli stadi del cambiamento sono influenzati dai fattori socio-demografici. Inoltre i processi di cambiamento utilizzati per smettere di fumare sono diversi tra future mamme e non: nelle gravide vi \ue8 un minore utilizzo dei processi comportamentali. I programmi d\u2019intervento per aiutare la popolazione femminile a smettere di fumare e prevenirne la ricaduta devono tenere in considerazione queste differenze

Stimulation of P2 receptors causes release of IL-1beta-loaded microvescicles from human dendritic cells

Dendritic cells (DCs) are professional antigen-presenting cells that initiate the immune response by activating T lymphocytes. DCs express plasma membrane receptors for extracellular nucleotides named P2 receptors (P2Rs). Stimulation of P2Rs in these cells is known to cause chemotaxis, cytokine release, and cell death and to modulate LPS-dependent differentiation. Here we show that stimulation of the P2X(7) receptor subtype (P2X(7)R) causes fast microvesicle shedding from DC plasma membrane. Vesicle release occurs from both immature and mature DCs; however, only vesicles from mature DCs, due to their previous exposure to LPS, contain IL-1beta. Microvesicles, whether from immature or mature DCs, also contain caspase-1 and -3 and cathepsin D. They also express the P2X(7)R in addition to other P2Rs and known markers of immune cells such as major histocompatibility complex II (MHC II) and CD39. Activation of the P2X(7)R by extracellular ATP causes IL-1beta release from the vesicle lumen. Previous studies demonstrated that high extracellular K(+) inhibits IL-1beta processing and release; here we show that high ionic strength reduces microvesicle shedding when compared with a low ionic strength medium but strongly increases microvesicle IL-1beta loadin