Direct measurement of DNA-mediated adhesion between lipid bilayers

Multivalent interactions between deformable mesoscopic units are ubiquitous in biology, where membrane macromolecules mediate the interactions between neighbouring living cells and between cells and solid substrates. Lately, analogous artificial materials have been synthesised by functionalising the outer surface of compliant Brownian units, for example emulsion droplets and lipid vesicles, with selective linkers, in particular short DNA sequences. This development extended the range of applicability of DNA as a selective glue, originally applied to solid nano and colloidal particles. On very deformable lipid vesicles, the coupling between statistical effects of multivalent interactions and mechanical deformation of the membranes gives rise to complex emergent behaviours, as we recently contributed to demonstrate [Parolini et al., Nature Communications, 2015, 6, 5948]. Several aspects of the complex phenomenology observed in these systems still lack a quantitative experimental characterisation and fundamental understanding. Here we focus on the DNA-mediated multivalent interactions of a single liposome adhering to a flat supported bilayer. This simplified geometry enables the estimate of the membrane tension induced by the DNA-mediated adhesive forces acting on the liposome. Our experimental investigation is completed by morphological measurements and the characterisation of the DNA-melting transition, probed by in-situ F\"{o}rster Resonant Energy Transfer spectroscopy. Experimental results are compared with the predictions of an analytical theory that couples the deformation of the vesicle to a full description of the statistical mechanics of mobile linkers. With at most one fitting parameter, our theory is capable of semi-quantitatively matching experimental data, confirming the quality of the underlying assumptions.Comment: 16 pages, 7 figure

Effects of an antioxidant protective topical formulation on eye exposed to ultraviolet-irradiation: A study in rabbit animal model

Ultraviolet-radiation exerts a well-known role in the development of various ocular diseases and may contribute to the progress of age-related macular degeneration. Therefore, the use of compounds able to protect the eyes from UV-induced cellular damage is challenging. The aim of this study has been to test the protective effects of an antioxidant topical formulation against UV-induced damage in rabbit eyes. Twelve male rabbits were used. Animals were divided into 4 groups of 3 animals each. Control group (CG) did not receive any irradiation and/or eye drop. The other three experimental groups were treated as follows: the first group received only UVR irradiation for 30 min, without eye drop supplementation (Irradiation group, IG), the second (G30) and the third (G60) groups received UV irradiation for 30' and 60', respectively, and eye drop supplementation (riboflavin, d-α-tocopheryl polyethylene glycol, proline, glycine, lysine and leucine solution) every 15 min for three hours. In the IG group a significant increase of oxidized glutathione (GSSG) and hydrogen peroxide (H2O2) was recorded in the aqueous humor, whereas ascorbic acid levels were significantly lower when compared to control eyes. In the groups exposed to UVR rays for 30 min, and treated with the topical antioxidant formulation, the GSSG, H2O2 and ascorbic acid levels were similar to those recorded in controls, whereas in the G60 group the three markers significantly differ from control group. In the lens, a significant decrease of alpha tocopherol and total antioxidant capacity (TAC) was recorded in IG-animals as compared to control group, whereas malondialdehyde (MDA) levels were significantly higher in UV-induced eye than in control eyes. In the G30 groups the alpha tocopherol, MDA and TAC levels do not significantly differ from those recorded in controls, whereas in the G60 group these three markers significantly differ from control group. Present findings demonstrate that topical treatment with the antioxidant formulation used herein protects ocular structures from oxidative stress induced by UV exposure in in vivo animal model

effects of an antioxidant protective topical formulation on eye exposed to ultraviolet irradiation a study in rabbit animal model

Ultraviolet-radiation exerts a well-known role in the development of various ocular diseases and may contribute to the progress of age-related macular degeneration. Therefore, the use of compounds able to protect the eyes from UV-induced cellular damage is challenging. The aim of this study has been to test the protective effects of an antioxidant topical formulation against UV-induced damage in rabbit eyes. Twelve male rabbits were used. Animals were divided into 4 groups of 3 animals each. Control group (CG) did not receive any irradiation and/or eye drop. The other three experimental groups were treated as follows: the first group received only UVR irradiation for 30 min, without eye drop supplementation (Irradiation group, IG), the second (G30) and the third (G60) groups received UV irradiation for 30' and 60', respectively, and eye drop supplementation (riboflavin, d-α-tocopheryl polyethylene glycol, proline, glycine, lysine and leucine solution) every 15 min for three hours. In the IG group a significant increase of oxidized glutathione (GSSG) and hydrogen peroxide (H2O2) was recorded in the aqueous humor, whereas ascorbic acid levels were significantly lower when compared to control eyes. In the groups exposed to UVR rays for 30 min, and treated with the topical antioxidant formulation, the GSSG, H2O2 and ascorbic acid levels were similar to those recorded in controls, whereas in the G60 group the three markers significantly differ from control group. In the lens, a significant decrease of alpha tocopherol and total antioxidant capacity (TAC) was recorded in IG-animals as compared to control group, whereas malondialdehyde (MDA) levels were significantly higher in UV-induced eye than in control eyes. In the G30 groups the alpha tocopherol, MDA and TAC levels do not significantly differ from those recorded in controls, whereas in the G60 group these three markers significantly differ from control group. Present findings demonstrate that topical treatment with the antioxidant formulation used herein protects ocular structures from oxidative stress induced by UV exposure in in vivo animal model

Polystyrene microplastics ingestion induced behavioral effects to the cladoceran Daphnia magna

Microplastic (\u3bcPs) contamination represents a dramatic environmental problem threatening both aquatic and terrestrial organisms. Although several studies have highlighted the presence of \u3bcPs in aquatic environments, the information regarding their toxicity towards organisms is still scant. Moreover, most of the ecotoxicological studies of \u3bcPs have focused on marine organisms, largely neglecting the effects on freshwater species. The present study aimed at exploring the effects caused by 21-days exposure to three concentrations (0.125, 1.25 and 12.5 \u3bcg/mL) of two differently sized polystyrene microplastics (P\u3bcPs; 1 and 10 \u3bcm) to the Cladoceran Daphnia magna. The ingestion/egestion capability of daphnids (<24 h) and adults, the changes in individual growth and behavior, in terms of changes in swimming activity, phototactic behavior and reproduction, were investigated. Both particles filled the digestive tract of daphnids and adults within 24 h of exposure at all the tested concentrations. Ingested P\u3bcPs remained in the digestive tract even after 96 h in a clean medium. For both particles, an overall increase in body size of adults was noted at the end of the exposure to the highest tested concentrations, accompanied by a significant increase in swimming activity, in terms of distance moved and swimming velocity, and by an alteration of the phototactic behavior. A significant increase in the mean number of offspring after the exposure to the highest P\u3bcPs concentrations of different size was recorded. Polystyrene \u3bcPs can affect behavioral traits of D. magna leading to potentially harmful consequences on population dynamics of this zooplanktonic species

European Vitreoretinal Society Macular Hole Study, Prognostic Factors for Anatomical and Functional Success

Background: To identify prognostic preoperative and intraoperative factors for anatomical and visual success of idiopathic macular hole (MH) surgery. Methods: We conducted a non-randomized, collaborative multicenter study using data of 4207 MH surgery from 140 surgeons. Main study outcomes were anatomical closure and best corrected visual acuity (BCVA) improvement postoperative at 6-12 months. Results: Information on anatomical success was available for 4138 eyes of 4207 operations. Anatomical closure of MH was achieved in 85.7% (3546 eyes). Closure was higher in smaller MH (stages 1-2 versus stage 3: OR=0.35; stage 2 versus stage 4: OR=0.16, and in MH with shorter duration before the operation (OR=0.94). Macular Holes were more likely to close when dyes were used to facilitate internal limiting membrane (ILM) peeling (odds ratio=1.73 to 3.58). The most important predictors of postoperative BCVA were the preoperative BCVA (estimate=0.39, p<0.001) and closure of the macular hole (estimate=0.34, p<0.001). We observed Larger improvement in BCVA in combined vitrectomy and phacoemulsification (estimate = 0.10) and post cataract surgery in phakic eyes (estimate=0.05). Retinal tears occurred in 5.1% of eyes, and were less with use of trocars (OR= -1.246) and in combined vitrectomy/ phacoemulsification surgery (OR= -0.688). Conclusion: This international survey confirmed that staining with dyes improves anatomical results but not visual outcomes. After surgery, visual acuity improved during the first year, and final visual acuity was better in both pseudophakic eyes and eyes that underwent cataract surgery during the first year following MH repair

Genetic Variants of the Renin-Angiotensin-Aldosterone System and Reverse Remodeling After Cardiac Resynchronization Therapy

Background: Reverse remodeling (RR) after cardiac resynchronization therapy (CRT) is associated with favorable clinical outcomes in heart failure (HF). The renin-angiotensin-aldosterone system (RAAS) is involved in the remodeling process. Methods and Results: We assessed the association between RR and 8 common RAAS gene variants, which were determined by TaqMan assays, in 156 outpatients with chronic HF. RR was defined as a O15% decrease in left ventricular end systolic volume (LVESV) at 9 (interquartile range 7e12) months after CRT. We matched 76 patients who did not show RR (RR) to 80 RR? control subjects by age, sex, HF etiology, New York Heart Association (NYHA) functional class and left ventricular ejection fraction (LVEF). The frequency of the minor allele of the NR3C2 gene (rs5522 C/T), encoding the mineralocorticoid receptor, was higher in RR than in RR (24/126 vs 10/150; P value after false discovery rate correction: <.0193). Conversely, LVESV decreased significantly less after CRT in carriers of the NR3C2 minor C allele (P 5 .02). After adjustment for age, sex, NYHA functional class, previous myocardial infarction, atrial fibrillation, and LVEF, RR remained independently associated with NR3C2 C allele carriage (odds ratio 3.093, 95% confidence interval 1.253e7.632). Conclusions: The association of RR after CRT with a common polymorphism in the mineralocorticoid receptor gene involved in aldosterone signaling suggests a possible role for variants in RAAS genes in progressive LV function decline, despite apparently effective CRT

NFKB1 regulates human NK cell maturation and effector functions

12siopenopenLougaris, Vassilios; Patrizi, Ornella; Baronio, Manuela; Tabellini, Giovanna; Tampella, Giacomo; Damiati, Eufemia; Frede, Natalie; van der Meer, Jos W.M.; Fliegauf, Manfred; Grimbacher, Bodo; Parolini, Silvia; Plebani, AlessandroLougaris, Vassilios; Patrizi, Ornella; Baronio, Manuela; Tabellini, Giovanna; Tampella, Giacomo; Damiati, Eufemia; Frede, Natalie; van der Meer, Jos W. M.; Fliegauf, Manfred; Grimbacher, Bodo; Parolini, Silvia; Plebani, Alessandr

Predicting PCB concentrations in cow milk: validation of a fugacity model in high-mountain pasture conditions

A fugacity model reported in the literature was applied to a high - altitude pasture in the Italian Alps. The model takes into account three compartments (digestive tract, blood and fat tissues) in unsteady-state conditions using food as the contamination source. Disregarding biotransformation inside cow tissues, the predicted concentrations of 14 polychlorinated biphenyls (PCBs) in milk were in good agreement with the observed data, especially for congeners known for their resistance to biotransformation (e.g., CB-138 and 153). In contrast, the predicted concentrations were clearly overestimated for congeners with high biotransformation susceptibilities. Therefore data measured in milk and faeces were used to calculate the first-order-biotransformation rate constants in dairy cows. The PCB absorption efficiency observed for pasture conditionswas lower than that observed in the cowshed. The final version of the model included biotransformation and observed PCB absorption andwas able to predict PCB concentrations in cow milkwith mean differences between the predicted and measured data below \ub120% for most congener

A Catalog of Coding Sequence Variations in Salivary Proteins’ Genes Occurring during Recent Human Evolution

Saliva houses over 2000 proteins and peptides with poorly clarified functions, including proline-rich proteins, statherin, P-B peptides, histatins, cystatins, and amylases. Their genes are poorly conserved across related species, reflecting an evolutionary adaptation. We searched the nucleotide substitutions fixed in these salivary proteins’ gene loci in modern humans compared with ancient hominins. We mapped 3472 sequence variants/nucleotide substitutions in coding, noncoding, and 5′-3′ untranslated regions. Despite most of the detected variations being within noncoding regions, the frequency of coding variations was far higher than the general rate found throughout the genome. Among the various missense substitutions, specific substitutions detected in PRB1 and PRB2 genes were responsible for the introduction/abrogation of consensus sequences recognized by convertase enzymes that cleave the protein precursors. Overall, these changes that occurred during the recent human evolution might have generated novel functional features and/or different expression ratios among the various components of the salivary proteome. This may have influenced the homeostasis of the oral cavity environment, possibly conditioning the eating habits of modern humans. However, fixed nucleotide changes in modern humans represented only 7.3% of all the substitutions reported in this study, and no signs of evolutionary pressure or adaptative introgression from archaic hominins were found on the tested genes