Development of a High Oleic Cardoon Cell Culture Platform by SAD Overexpression and RNAi-Mediated FAD2.2 Silencing

The development of effective tools for the sustainable supply of phyto-ingredients and natural substances with reduced environmental footprints can help mitigate the dramatic scenario of climate change. Plant cell cultures-based biorefineries can be a technological advancement to face this challenge and offer a potentially unlimited availability of natural substances, in a standardized composition and devoid of the seasonal variability of cultivated plants. Monounsaturated (MUFA) fatty acids are attracting considerable attention as supplements for biodegradable plastics, bio-additives for the cosmetic industry, and bio-lubricants. Cardoon (Cynara cardunculus L. var. altilis) callus cultures accumulate fatty acids and polyphenols and are therefore suitable for large-scale production of biochemicals and valuable compounds, as well as biofuel precursors. With the aim of boosting their potential uses, we designed a biotechnological approach to increase oleic acid content through Agrobacterium tumefaciens-mediated metabolic engineering. Bioinformatic data mining in the C. cardunculus transcriptome allowed the selection and molecular characterization of SAD (stearic acid desaturase) and FAD2.2 (fatty acid desaturase) genes, coding for key enzymes in oleic and linoleic acid formation, as targets for metabolic engineering. A total of 22 and 27 fast-growing independent CcSAD overexpressing (OE) and CcFAD2.2 RNAi knocked out (KO) transgenic lines were obtained. Further characterization of five independent transgenic lines for each construct demonstrated that, successfully, SAD overexpression increased linoleic acid content, e.g., to 42.5%, of the relative fatty acid content, in the CcSADOE6 line compared with 30.4% in the wild type (WT), whereas FAD2.2 silencing reduced linoleic acid in favor of the accumulation of its precursor, oleic acid, e.g., to almost 57% of the relative fatty acid content in the CcFAD2.2KO2 line with respect to 17.7% in the WT. Moreover, CcSADOE6 and CcFAD2.2KO2 were also characterized by a significant increase in total polyphenolic content up to about 4.7 and 4.1 mg/g DW as compared with 2.7 mg/g DW in the WT, mainly due to the accumulation of dicaffeoyl quinic and feruloyl quinic acids. These results pose the basis for the effective creation of an engineered cardoon cells-based biorefinery accumulating high levels of valuable compounds from primary and specialized metabolism to meet the industrial demand for renewable and sustainable sources of innovative bioproducts

Detection and localization of double compression in MP3 audio tracks

In this work, by exploiting the traces left by double compression in the statistics of quantized modified discrete cosine transform coefficients, a single measure has been derived that allows to decide whether an MP3 file is singly or doubly compressed and, in the last case, to devise also the bit-rate of the first compression. Moreover, the proposed method as well as two state-of-the-art methods have been applied to analyze short temporal windows of the track, allowing the localization of possible tampered portions in the MP3 file under analysis. Experiments confirm the good performance of the proposed scheme and demonstrate that current detection methods are useful for tampering localization, thus offering a new tool for the forensic analysis of MP3 audio tracks

Linee guida per la redazione del progetto di fattibilità tecnica ed economica da porre a base dell’affidamento di contratti pubblici di lavori del PNRR e del PNC (Art. 48, comma 7, del decreto-legge 31 maggio 2021, n. 77, convertito nella legge 29 luglio 2021, n. 108)

Nel quadro dell’innovazione normativa e procedurale introdotta dal PNRR e dal decreto-legge 31 maggio 2021, n. 77, le Linee Guida sono volte a definire il contenuto essenziale dei documenti, degli eventuali modelli informativi digitali e degli elaborati occorrenti alle Stazioni Appaltanti per l’affidamento sulla base del Progetto di Fattibilità Tecnico-Economica - PFTE, secondo quanto stabilito dall’art. 48, comma 7

The application of omics in ruminant production: a review in the tropical and sub-tropical animal production context

The demand for animal products (e.g. dairy and beef) in tropical regions is expected to increase in parallel with the public demand for sustainable practices, due to factors such as population growth and climate change. The necessity to increase animal production output must be achieved with better management and production technologies. For this to happen, novel research methodologies, animal selection and postgenomic tools play a pivotal role. Indeed, improving breeder selection programs, the quality of meat and dairy products as well as animal health will contribute to higher sustainability and productivity. This would surely benefit regions where resource quality and quantity are increasingly unstable, and research is still very incipient, which is the case of many regions in the tropics. The purpose of this review is to demonstrate how omics-based approaches play a major role in animal science, particularly concerning ruminant production systems and research associated to the tropics and developing countriesinfo:eu-repo/semantics/acceptedVersio

Comparative evaluation of analogue front-end designs for the CMS Inner Tracker at the High Luminosity LHC

The CMS Inner Tracker, made of silicon pixel modules, will be entirely replaced prior to the start of the High Luminosity LHC period. One of the crucial components of the new Inner Tracker system is the readout chip, being developed by the RD53 Collaboration, and in particular its analogue front-end, which receives the signal from the sensor and digitizes it. Three different analogue front-ends (Synchronous, Linear, and Differential) were designed and implemented in the RD53A demonstrator chip. A dedicated evaluation program was carried out to select the most suitable design to build a radiation tolerant pixel detector able to sustain high particle rates with high efficiency and a small fraction of spurious pixel hits. The test results showed that all three analogue front-ends presented strong points, but also limitations. The Differential front-end demonstrated very low noise, but the threshold tuning became problematic after irradiation. Moreover, a saturation in the preamplifier feedback loop affected the return of the signal to baseline and thus increased the dead time. The Synchronous front-end showed very good timing performance, but also higher noise. For the Linear front-end all of the parameters were within specification, although this design had the largest time walk. This limitation was addressed and mitigated in an improved design. The analysis of the advantages and disadvantages of the three front-ends in the context of the CMS Inner Tracker operation requirements led to the selection of the improved design Linear front-end for integration in the final CMS readout chip

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2,3,4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Ethical aspects in the management of antibacterial agents utilization

Introduction: Antibacterial prescribing practices between 2004 and 2008 were investigated in the P. Giaccone University Hospital in Palermo, Italy to provide a foundation for critical analysis of the appropriateness of health-care resource usage. Materials and methods: Antibiotic prescribing practices between 2004 and 2008 were analyzed in the hospital as a whole and in different specialty areas. Results were expressed as defined daily doses (DDD) as a function of bed-days, number of admissions, and Diagnosis Related Group (DRG) points. Results: During the study period, increases were observed in the overall DDD per 100 bed-days (68.7 vs. 91.3) and the DDD per admission (4.53 vs. 5.54), but less variation was observed in the DDD/DRG score (3.55 vs. 3.63). Use of metronidazole, carbapenems, and glycopeptides increased, while use of third-generation cephalosporins, quinolones, and oral penicillins remained fairly stable. The drugs most commonly used in 2008 were (% of total DDD): levofloxacin (18.08%), amoxicillin + clavulanic acid (13.32%), ceftriaxone (9.01%), ciprofloxacin (8.21%), clarithromycin (5.74%), metronidazole (5.36%), ceftazidime (5.16%), amoxicillin (4.92%), gentamicin (1.88%), and meropenem (1.80%). An overall trend toward the prescription of newer wide-spectrum antibacterial agents was noted. National guidelines on chemoprophylaxis in surgery were observed in only 3/17 (17.7%) units. The prevalence of nosocomial infections was lower than those reported in the literature. Discussion: Although the cases treated in the hospital have become more complex over the years, the DDD documented in our study are far too high with respect to the actual clinical needs. These findings point to a trend toward the practice of excessively defensive medicine. Greater responsibility among physicians and the promotion of primary and secondary measures of preventive hygiene are fundamental to reduce the prescriptive pressure, a goal that should also have beneficial effects on health-care costs