Antimicrobial Nanoplexes meet Model Bacterial Membranes: the key role of Cardiolipin

Antimicrobial resistance to traditional antibiotics is a crucial challenge of medical research. Oligonucleotide therapeutics, such as antisense or Transcription Factor Decoys (TFDs), have the potential to circumvent current resistance mechanisms by acting on novel targets. However, their full translation into clinical application requires efficient delivery strategies and fundamental comprehension of their interaction with target bacterial cells. To address these points, we employed a novel cationic bolaamphiphile that binds TFDs with high affinity to form self-assembled complexes (nanoplexes). Confocal microscopy revealed that nanoplexes efficiently transfect bacterial cells, consistently with biological efficacy on animal models. To understand the factors affecting the delivery process, liposomes with varying compositions, taken as model synthetic bilayers, were challenged with nanoplexes and investigated with Scattering and Fluorescence techniques. Thanks to the combination of results on bacteria and synthetic membrane models we demonstrate for the first time that the prokaryotic-enriched anionic lipid Cardiolipin (CL) plays a key-role in the TFDs delivery to bacteria. Moreover, we can hypothesize an overall TFD delivery mechanism, where bacterial membrane reorganization with permeability increase and release of the TFD from the nanoplexes are the main factors. These results will be of great benefit to boost the development of oligonucleotides-based antimicrobials of superior efficacy

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

Fungi associated with root rot and collapse of melon in Italy

Melon represents the most widespread cucurbit in Italy. In recent years melon has been subjected to significant losses in yield and quality due to an increasing number of soil-borne fungal diseases. The collapse of melon, caused by a complex of fungal pathogens, including Monosporascus cannonballus , Acremonium cucurbitacearum , Plectosporium tabacinum and Rhizopycnis vagum , represents one of most destructive diseases worldwide. The purpose of this study was to determine the occurrence of collapse throughout melon-producing areas in Italy in recent years, to verify the identification of isolates collected, and to test their pathogenicity on melon and other cucurbits. Several fungi were isolated from symptomatic roots of melons in the Italian production areas. The identification was supported by PCR with a species-specific primer and DNA sequence data. RFLP and sequence analyses showed the existence of a substantial homogeneity among Italian M. cannonballus isolates. Given the self-incompatibility of these isolates it is impossible to ascertain vegetative compatibility groups (VGC) and consequently genetic relatedness cannot be studied. The frequency of isolation of fungal species varied with geographic locations, M. cannonballus being present mainly in Central Italy, while A. cucurbitacearum and P. tabacinum were most common in Apulia. In pathogenicity tests under greenhouse conditions M. cannonballus , A. cucurbitacearum and P. tabacinum caused collapse symptoms and root rots, whereas R. vagum was found to be a weak pathogen

Fungi associated with root rot and collapse of melon in Italy

Melon represents the most widespread cucurbit in Italy. In recent years melon has been subjected to significant losses in yield and quality due to an increasing number of soil-borne fungal diseases. The collapse of melon, caused by a complex of fungal pathogens, including Monosporascus cannonballus , Acremonium cucurbitacearum , Plectosporium tabacinum and Rhizopycnis vagum , represents one of most destructive diseases worldwide. The purpose of this study was to determine the occurrence of collapse throughout melon-producing areas in Italy in recent years, to verify the identification of isolates collected, and to test their pathogenicity on melon and other cucurbits. Several fungi were isolated from symptomatic roots of melons in the Italian production areas. The identification was supported by PCR with a species-specific primer and DNA sequence data. RFLP and sequence analyses showed the existence of a substantial homogeneity among Italian M. cannonballus isolates. Given the self-incompatibility of these isolates it is impossible to ascertain vegetative compatibility groups (VGC) and consequently genetic relatedness cannot be studied. The frequency of isolation of fungal species varied with geographic locations, M. cannonballus being present mainly in Central Italy, while A. cucurbitacearum and P. tabacinum were most common in Apulia. In pathogenicity tests under greenhouse conditions M. cannonballus , A. cucurbitacearum and P. tabacinum caused collapse symptoms and root rots, whereas R. vagum was found to be a weak pathogen