Engineered Sleeping Beauty Transposon as Efficient System to Optimize Chimp Adenoviral Production

Sleeping Beauty (SB) is the first DNA transposon employed for efficient transposition in vertebrate cells, opening new applications for genetic engineering and gene therapies. A transposon-based gene delivery system holds the favourable features of non-viral vectors and an attractive safety profile. Here, we employed SB to engineer HEK293 cells for optimizing the production of a chimpanzee Adenovector (chAd) belonging to the Human Mastadenovirus C species. To date, chAd vectors are employed in several clinical settings for infectious diseases, last but not least COVID-19. A robust, efficient and quick viral vector production could advance the clinical application of chAd vectors. To this aim, we firstly swapped the hAd5 E1 with chAd-C E1 gene by using the CRISPR/Cas9 system. We demonstrated that in the absence of human Ad5 E1, chimp Ad-C E1 gene did not support HEK293 survival. To improve chAd-C vector production, we engineered HEK293 cells to stably express the chAd-C precursor terminal protein (ch.pTP), which plays a crucial role in chimpanzee Adenoviral DNA replication. The results indicate that exogenous ch.pTP expression significantly ameliorate the packaging and amplification of recombinant chAd-C vectors thus, the engineered HEK293ch.pTP cells could represent a superior packaging cell line for the production of these vectors.     

Chronic Hyperkaliemia in Chronic Kidney Disease: An Old Concern with New Answers

Increasing potassium intake ameliorates blood pressure (BP) and cardiovascular (CV) prognoses in the general population; therefore the World Health Organization recommends a high-potassium diet (90–120 mEq/day). Hyperkalaemia is a rare condition in healthy individuals due to the ability of the kidneys to effectively excrete dietary potassium load in urine, while an increase in serum K⁺ is prevalent in patients with chronic kidney disease (CKD). Hyperkalaemia prevalence increases in more advanced CKD stages, and is associated with a poor prognosis. This scenario generates controversy on the correct nutritional approach to hyperkalaemia in CKD patients, considering the unproven link between potassium intake and serum K⁺ levels. Another concern is that drug-induced hyperkalaemia leads to the down-titration or withdrawal of renin-angiotensin system inhibitors (RASI) and mineralocorticoids receptors antagonists (MRA) in patients with CKD, depriving these patients of central therapeutic interventions aimed at delaying CKD progression and decreasing CV mortality. The new K⁺-binder drugs (Patiromer and Sodium-Zirconium Cyclosilicate) have proven to be adequate and safe therapeutic options to control serum K⁺ in CKD patients, enabling RASI and MRA therapy, and possibly, a more liberal intake of fruit and vegetables.     

Fighting the Huntington’s Disease with a G-Quadruplex-Forming Aptamer Specifically Binding to Mutant Huntingtin Protein: Biophysical Characterization,In Vitro and In Vivo Studies

A set of guanine-rich aptamers able to preferentially recognize full-length huntingtin with an expanded polyglutamine tract has been recently identified, showing high efficacy in modulating the functions of the mutated protein in a variety of cell experiments. We here report a detailed biophysical characterization of the best aptamer in the series, named MS3, proved to adopt a stable, parallel G-quadruplex structure and show high nuclease resistance in serum. Confocal microscopy experiments on HeLa and SH-SY5Y cells, as models of non-neuronal and neuronal cells, respectively, showed a rapid, dose-dependent uptake of fluorescein-labelled MS3, demonstrating its effective internalization, even in the absence of transfecting agents, with no general cytotoxicity. Then, using a well-established Drosophila melanogaster model for Huntington’s disease, which expresses the mutated form of human huntingtin, a significant improvement in the motor neuronal function in flies fed with MS3 was observed, proving the in vivo efficacy of this aptamer.     

Formalizing REST APIs for web-based communication and SIP interworking

This paper proposes an instantaneous recovery route design scheme using multiple coding-aware link protection scenarios to achieve higher link cost reduction in the network. In this scheme, two protection scenarios, namely, (i) traffic splitting (TS), and (ii) two sources and a common destination (2SD) are used to integrate multiple sources and a common destination. The proposed scheme consists of two phases. In the first phase, the proposed scheme determines routes for 2SD and TS scenarios of all possible source-destination pairs to minimize the total link cost. In this phase, the network coding is applied to the common path within each scenario, individually. In the second phase, network coding is applied to the common path between two scenarios (or a scenario pair) in order to enhance the resource saving. This phase develops conditions that select the most appropriate combination of scenario pairs, such as TS–TS, 2SD–2SD, and 2SD–TS for network coding, including their proofs. Using these conditions, a heuristic algorithm is introduced in order to select the most appropriate combination of scenario pairs for further enhancing of resource saving. Simulation results show that the proposed scheme outperforms the conventional 1 + 1 protection scheme, the TS scenario, and the 2SD scenario in terms of link cost reduction in the network.     

Electrosynthesized CuMgAl Layered Double Hydroxides as New Catalysts for the Electrochemical Reduction of CO2

In this study, new nanostructured CuMgAl Layered Double Hydroxide (LDH) based materials are synthesized on a 4 cm² sized carbonaceous gas diffusion membrane. By means of microscopic and spectroscopic techniques, the catalysts are thoroughly investigated, revealing the presence of several species within the same material. By a one-step, reproducible potentiodynamic deposition it is possible to obtain a composite with an intimate contact between a ternary CuMgAl LDH and Cu⁰/Cu₂O species. The catalyst compositions are investigated by varying: the molar ratio between the total amount of bivalent cations and Al³⁺, the amount of loading, and the molar ratios among the three cations in the electrolyte. Each electrocatalyst has been evaluated based on the catalytic performances toward the electrochemical CO₂ reduction to CH₃COOH at −0.4 V versus reversible hydrogen electrode  in liquid phase. The optimized catalyst, that is, CuMgAl 2:1:1 LDH exhibits a productivity of 2.0 mmol_CH3COOH g_cat⁻¹ h⁻¹. This result shows the beneficial effects of combining a material like the LDHs, alkaline in nature, and thus with a great affinity to CO₂, with Cu⁰/Cu⁺ species, which couples the increase of carbon sources availability at the electrode with a redox mediator capable to convert CO₂ into a C₂ product.     

Transition‐Metal Dichalcogenide NiTe2: An Ambient‐Stable Material for Catalysis and Nanoelectronics

By means of theory and experiments, the application capability of nickel ditelluride (NiTe₂) transition‐metal dichalcogenide in catalysis and nanoelectronics is assessed. The Te surface termination forms a TeO₂ skin in an oxygen environment. In ambient atmosphere, passivation is achieved in less than 30 min with the TeO₂ skin having a thickness of about 7 Å. NiTe₂ shows outstanding tolerance to CO exposure and stability in water environment, with subsequent good performance in both hydrogen and oxygen evolution reactions. NiTe₂‐based devices consistently demonstrate superb ambient stability over a timescale as long as one month. Specifically, NiTe₂ has been implemented in a device that exhibits both superior performance and environmental stability at frequencies above 40 GHz, with possible applications as a receiver beyond the cutoff frequency of a nanotransistor.     

Radical Scavenging,Total Antioxidant Capacity,and Antiproliferative Activity of Phenolic Extracts from Extra Virgin Olive Oil by Cultivar ‘Frantoio’

The beneficial health effects of extra virgin olive oil are due to both its high content of monounsaturated fatty acids and its high content of phenolic compounds, which have recently attracted research interest. In this context, the aim of this work was to examine the in vitro antioxidant and antiproliferative activities of the phenolic extract obtained from extra virgin olive oil from O. europea cultivar ‘Frantoio’ (samples 1–4), one of the main varieties cultivated in Italy. The total phenolic content was determined by Folin–Ciocalteu method and instead the phenolic profile was obtained by HPLC coupled to a diode array detector and mass spectrometry. Extra virgin olive oil extracts exhibited high antioxidant activity through different mechanisms of action and these activities are related to the phenolic content. Sample 3 demonstrated the strongest free radical scavenging activity with IC₅₀ value of 56.5 μg/mL. The reducing ability measured with FRAP assay revealed that samples ranged from 91.3 to 156 μM Fe(II)/g. The same interesting trend was observed with Trolox equivalent antioxidant capacity value. Moreover, the virgin olive oils showed a good oxidative stability ranging between 19 to 32 h. Antiproliferative activity evaluated by SRB assay revealed that phenolic extracts from the cultivar ‘Frantoio’ showed a strong antiproliferative activity against CORL-23 cell line with an IC₅₀ value of 14.5 and 55.9 μg/mL for samples 3 and 1, respectively, and these results are comparable to the positive control vinblastine. Overall, these results showed that extra virgin olive oils from the cultivar ‘Frantoio’, may represent an interesting source of phenolic compounds as functional components that could be consumed in diets and/or used for the elaboration of functional food and pharmaceutical industries.     

δ18O of Ethanol in Wine and Spirits for Authentication Purposes

Since 1986 the European Union has established official isotopic analysis methods for detecting the illegal addition of sugar and water to wine and to enable geographical traceability. In this paper we investigate the possibility of using analysis of the ¹⁸O/¹⁶O stable isotope ratio (expressed as δ¹⁸O) of ethanol to improve detection of the watering of wine and to determine the origin of ethanol. Sixty‐nine authentic wine samples from all over Italy, 59 spirits from fruit and cereals, 5 chemically synthesized ethanols, one concentrated and rectified must, one beet and one cane sugar, one fresh must, and 6 waters with increasing δ¹⁸O values were considered. Ethanol was recovered by distillation, using a Cadiot spinning band column, following the official OIV methods. The residual water was trapped by storing the distillate for at least 24 h on a molecular sieve. The ¹⁸O/¹⁶O ratio was measured using a pyrolyser interfaced with an isotope ratio mass spectrometer. The δ^‐18O of ethanol is significantly related to the δ¹⁸O of the fermentation water and can be considered as a reliable internal reference. The values ranged from +24‰ to +36‰ in wine (years 2008 to 2012), +10‰ to +26‰ in fruit and cereal distillates, and from –2‰ to +12‰ in synthetic ethanol. The method was shown to be effective in improving detection of the watering of wine and determining the origin of ethanol (from grapes, other fruit, or synthesis), but not in detecting the addition of cane or beet sugar to wine.     

Fat globule membranes in ewes' milk: The main enzyme activities during lactation

A trial was carried out on seven Massese ewes, from which individual milk samples were taken during lactation. All the enzyme activities investigated (oxidase activity of xanthine oxidoreductase, gamma-glutamyltranspeptidase, alkaline phosphatase and 5′-nucleotidase) were affected by the lactation phase, with the exception of the dehydrogenase activity of xanthine oxidoreductase (XOR). A higher oxidase activity (XO) of XOR was found when the diameter of milk fat globules was smaller. In addition, the absence of XO in colostrum led to the hypothesis that its antibacterial role is independent of its activity. The increase in alkaline phosphatase activity during lactation requires further research to better define the criteria for pasteurised sheep milk.