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

3D-Printed Lipid Mesophases for the Treatment of Chronic Liver Disease

Although lipid-based formulations are an attractive approach for enhancing the oral bioavailability of lipophilic drugs, their addition into solid oral dosage forms has been proven challenging due to their high viscosity and heat sensitivity. Therefore, unlike the traditional tableting process, this study employed semi-solid extrusion 3D-printing to produce-at room temperature-gastro-resistant printlets containing a high percentage of bioactive lipids for the effective delivery of lipophilic drugs through self-emulsification. The bio-compatible lipidic mesophase ink, owing to a tunable 3D nanostructure, is employed as a starting material to produce printlets via additive manufacturing. An active lipid mixture - with antifibrotic properties - is blended with the antioxidant vitamin E and water, and the ink printability is optimized by carefully tailoring its composition, and thus its phase identity. The obtained printlets disintegrated upon contact with intestinal fluids forming colloidal structures that enhanced the solubility of a poorly water-soluble drug. The printlets exhibited antifibrotic activity on human hepatic stellate cells, LX-2, suggesting that the generated self-emulsified colloidal structures made both the fibrosis-resolving bioactive excipients and the drug promptly available, enhancing their cell uptake and, in turn, their therapeutic activity.Lipidic mesophases are employed as starting material to produce gastro-resistant oral dosage forms via 3Dprinting. The formulation contains a high percentage of bioactive lipids and it is suitable for the effective delivery of lipophilic drugs through self-emulsification. Indeed, upon contact with intestinal fluids printlets disintegrate, forming colloidal structures that enhance the bioavailability of the drug and its therapeutic activity

Variations in the Molecular and Physiological Characteristics and the Virulence of Monilinia fructicola, M. fructigena and M. laxa Isolates

Twenty-three Monilinia isolates of various origin were identified and compared by morphological means and by biomolecular protocols based on random amplified polymorphic DNA (RAPD) analysis, and on PCR detection by species-specific primer pairs. The identification of most Monilinia isolates by PCR and RAPD confirmed the identification by morphological criteria, although two isolates initially identified as M. fructigena by morphological means were attributed to M. fructicola by the biomolecular techniques. Monilinia isolates were also compared in terms of their in vitro pectolytic activity and isoenzyme patterns. All the isolates produced polygalacturonase and pectin methylesterase, but pectin lyase activity was found only in M. fructicola and M. fructigena, and not in most M. laxa isolates. The Monilinia isolates differed significantly in their isoenzyme patterns. Based on RAPD-PCR and PCR amplification and pectolytic isoenzymes, the isolates analysed clustered in three major groups, each corresponding to a Monilinia species. Artificial inoculations in peach and pear fruits revealed differences in virulence between the Monilinia species and within isolates. No correlation was found between the amount of pectolytic enzymes produced in vitro and virulence. Variations between and within the Monilinia species may depend at the physiological level on variations in the isoenzyme patterns of the pectolytic enzymes

Sleep duration and the risk of breast cancer: the Ohsaki Cohort Study

In a prospective study of 23 995 Japanese women, short sleep duration was associated with higher risk of breast cancer (143 cases), compared with women who slept 7 h per day, the multivariate hazard ratio of those who slept ⩽6 h per day was 1.62 (95% confidence interval: 1.05–2.50; P for trend=0.03)

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

Associação de SNPs nos genes para κ-caseína e β-lactoglobulina com curvas de lactação em cabras leiteiras

O objetivo deste trabalho foi identificar o modelo com melhor ajuste aos dados de produção de leite, gordura, proteína e sólidos totais para cabras leiteiras, bem como associar polimorfismos de nucleotídeo único (SNPs) nos genes para κ-caseína (κ-CSN3) e β-lactoglobulina (β-LG) aos parâmetros das curvas de produção e qualidade do leite. Foram avaliados 4.160 registros de produção de leite, gordura, proteína e sólidos totais de cabras das raças Alpina, Saanen e mestiça, no Estado de Antioquia, na Colômbia. Os modelos não lineares com melhor ajuste à estrutura dos dados foram os de Nelder, para produção de leite, e de Cappio-Borlino, para qualidade do leite. As análises de associação mostraram efeito significativo do SNP κ-CSN3 sobre o pico de produção, a produção inicial e a persistência das características qualitativas do leite. Já o SNP β-LG apresentou efeito significativo sobre os picos de produção de leite e gordura, e sobre o tempo necessário para atingir o pico de produção de proteína. A identificação e a estimação da influência dos marcadores SNP avaliados sobre as curvas de lactação e a qualidade do leite podem contribuir para a seleção de caprinos leiteiros