Expanding the enzybiotic toolbox by switching catalytic and cell wall binding domains

Motivation: Bacterial resistance to common antimicrobial therapies (i.e., antibiotics) is reaching alarming levels worldwide [1]. This pushes for the development of alternative antimicrobials, such as bacteriophage-encoded endolysins, also called enzybiotics. This antibacterial approach, rooted in phage therapy, could pose a new and valuable adding to the antimicrobial arsenal, with increased specificity and much lower chances of enabling bacterial escape and resistance mechanisms when compared to antibiotics [2]. Endolysins are usually comprised of a cell wall binding domain (CWBD) and one or several catalytic domains that cleave the bacterial cell wall. Both rational design and domain shuffling have proved to be effective approaches towards the improvement of phage endolysins for therapeutic uses [3]. Thus, the aim of this work is to construct new endolysins active against a range of pathogenic bacteria by switching protein domains.Methods: A new lysin, termed Csl2 was designed by fusing the catalytic lysozyme domain of Cpl-7, encoded by pneumococcal phage Cp-7 [3], and the CWBD from LySMP, encoded by Streptococcus suis phage SMP [4]. The gene encoding the chimeric enzyme was cloned in the expression vector pT7-7, overexpressed in the Escherichia coli strain BL21(DE3) and the protein Csl2 purified following a two-step chromatographic procedure. Relevant biochemical properties of Csl2 were studied, and its bacteriolytic activity was tested against several Gram-positive pathogens.Results: Protein Csl2 was efficiently overexpressed in E. coli BL21(DE3) and purified to >90%. Its secondary structure was similar to that of Cpl-7 according to preliminary circular dichroism experiments. Csl2 was found to be active against several streptococci, mainly from mitis group (namely: S. mitis, S. oralis, S. pseudopneumoniae) and Streptococcus suis from serotypes 2 and 9. The bacteriolytic host range of Csl2 differed from that of parental enzymes, with higher activity against S. suis strains.Conclusions: 1) A new bacteriolytic enzyme, Csl2, was constructed by combining Cpl-7 catalytic domain and LySMP putative Cpl-7-like CWBD.2) Csl2 resembles Cpl-7 biochemically and structurally, but has a different bactericidal spectrum.3) Csl2 is specially active against several S. suis strains, so it might be a candidate for therapy in S. suis infections

Budget impact analysis of dabigatran compared with rivaroxaban in the prevention of the thromboembolic risk in patients with non-valvular atrial fibrillation

BACKGROUND: Dabigatran 150 mg BID (D150) and rivaroxaban 20 mg (R20) are indicated for the prevention of thromboembolic events in patients with Non-Valvular Atrial Fibrillation (NVAF). Outcomes from observational study demonstrated that D150 and R20 reduced the rate of thromboembolic events.OBJECTIVE: This analysis estimated the budget impact of the use of D150 and R20 for the treatment of NAFV patients in Italy.METHODS: A budget-impact model (BIM) was developed to estimate the direct costs up to 12 months from an Italian NHS perspective. The resource utilization (drugs and intracranial hemorrhage or major extracranial bleeding event) was derived from an observational study. Only direct medical costs were considered. Ex-factory prices and National Tariffs were considered to estimate the costs of drugs and medical resource used, respectively. The BIM showed the difference of expenditure and clinical events (intracranial hemorrhage or major extracranial bleeding) generated by the base case calculated for current prescription volumes (D150 30%, R20 100%), and for different prescription volume scenarios (D150 at 70% and 100%). Key variables were tested in the sensitivity analysis.RESULTS: D150 was associated with a medical cost offset driven by fewer intracranial hemorrhage and major extracranial bleeding event, these offset the incremental drug cost and results in an annual saving per patient treated (D150: € 1,052.78; R20: € 1,161.23). The present scenario determines an annual cost of € 262,543,583. The impact of total annual costs for the Italian NHS would be lower if D150 prescription volumes would be higher. The total cost is predicted to decrease by 3.8% if the D150 prescription increase to 70% and it is predicted to decrease by 6.7% if the D150 prescription increase to 100%.CONCLUSION: The use of D150, as an alternative to R20 to prevent events in patients with NVAF, could represent a cost-saving option for the Italian NHS

The AFarCloud ECSEL Project

Farming is facing many economic challenges in terms of productivity and cost-effectiveness. Labor shortage partly due to depopulation of rural areas, especially in Europe, is another challenge. Domain specific problems such as accurate identification and proper quantification of pathogens affecting plant and animal health are key factors for minimizing economical risks, and not risking human health. The ECSEL AFarCloud (Aggregate FARming in the CLOUD) project will provide a distributed platform for autonomous farming that will allow the integration and cooperation of agriculture Cyber Physical Systems in real-time in order to increase efficiency, productivity, animal health, food quality and reduce farm labour costs. This platform will be integrated with farm management software and will support monitoring and decision-making solutions based on big data and real-time data mining techniques.The AFarCloud project is funded from the ECSEL Joint Undertaking under grant agreement n° 783221, and from National funding

Aggregate Farming in the Cloud: The AFarCloud ECSEL project

Farming is facing many economic challenges in terms of productivity and cost-effectiveness. Labor shortage partly due to depopulation of rural areas, especially in Europe, is another challenge. Domain specific problems such as accurate monitoring of soil and crop properties and animal health are key factors for minimizing economical risks, and not risking human health. The ECSEL AFarCloud (Aggregate Farming in the Cloud) project will provide a distributed platform for autonomous farming that will allow the integration and cooperation of agriculture Cyber Physical Systems in real-time in order to increase efficiency, productivity, animal health, food quality and reduce farm labor costs. Moreover, such a platform can be integrated with farm management software to support monitoring and decision-making solutions based on big data and real-time data mining techniques.publishedVersio

Geographical information system and environmental epidemiology: a cross-sectional spatial analysis of the effects of traffic-related air pollution on population respiratory health

<p>Abstract</p> <p>Background</p> <p>Traffic-related air pollution is a potential risk factor for human respiratory health. A Geographical Information System (GIS) approach was used to examine whether distance from a main road (the Tosco-Romagnola road) affected respiratory health status.</p> <p>Methods</p> <p>We used data collected during an epidemiological survey performed in the Pisa-Cascina area (central Italy) in the period 1991-93. A total of 2841 subjects participated in the survey and filled out a standardized questionnaire on health status, socio-demographic information, and personal habits. A variable proportion of subjects performed lung function and allergy tests. Highly exposed subjects were defined as those living within 100 m of the main road, moderately exposed as those living between 100 and 250 m from the road, and unexposed as those living between 250 and 800 m from the road. Statistical analyses were conducted to compare the risks for respiratory symptoms and diseases between exposed and unexposed. All analyses were stratified by gender.</p> <p>Results</p> <p>The study comprised 2062 subjects: mean age was 45.9 years for men and 48.9 years for women. Compared to subjects living between 250 m and 800 m from the main road, subjects living within 100 m of the main road had increased adjusted risks for persistent wheeze (OR = 1.76, 95% CI = 1.08-2.87), COPD diagnosis (OR = 1.80, 95% CI = 1.03-3.08), and reduced FEV₁/FVC ratio (OR = 2.07, 95% CI = 1.11-3.87) among males, and for dyspnea (OR = 1.61, 95% CI = 1.13-2.27), positivity to skin prick test (OR = 1.83, 95% CI = 1.11-3.00), asthma diagnosis (OR = 1.68, 95% CI = 0.97-2.88) and attacks of shortness of breath with wheeze (OR = 1.67, 95% CI = 0.98-2.84) among females.</p> <p>Conclusion</p> <p>This study points out the potential effects of traffic-related air pollution on respiratory health status, including lung function impairment. It also highlights the added value of GIS in environmental health research.</p

A Qualitative Exploration of the Use of Contraband Cell Phones in Secured Facilities

Offenders accepting contraband cell phones in secured facilities violate state corrections law, and the possession of these cell phones is a form of risk taking behavior. When offenders continue this risky behavior, it affects their decision making in other domains where they are challenging authorities; and may impact the length of their incarceration. This qualitative phenomenological study examined the lived experience of ex-offenders who had contraband cell phones in secured correctional facilities in order to better understand their reasons for taking risks with contraband cell phones. The theoretical foundation for this study was Trimpop\u27s risk-homeostasis and risk-motivation theories that suggest an individual\u27s behaviors adapt to negotiate between perceived risk and desired risk in order to achieve satisfaction. The research question explored beliefs and perceptions of ex-offenders who chose to accept the risk of using contraband cell phones during their time in secured facilities. Data were collected anonymously through recorded telephone interviews with 8 male adult ex-offenders and analyzed using thematic content analysis. Findings indicated participants felt empowered by possession of cell phones in prison, and it was an acceptable risk to stay connected to family out of concern for loved ones. The study contributes to social change by providing those justice system administrators, and prison managers responsible for prison cell phone policies with more detailed information about the motivations and perspectives of offenders in respect to using contraband cell phones while imprisoned in secured facilities

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 &lt; 60 mL/min/1.73 m2) or eGFR reduction &gt; 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 &lt; 90 ml/min/m2 were independent risk factors for stage 653 CKD and renal function worsening. When compared to patients with eGFR &gt; 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

An architectural approach to semantic Web service for geographical information

This thesis deals with the problem of attempting to integrate the service oriented model with a semantic approach for the geographical information. Semantic Web is a new opportunity for both fields, traditional Web services and the domain of geographical information, for every task with a potential conjunction between them. The main task is the creation of geo-ontologies and harmonisation of existing ones based on standards of geographical communities. For this purpose, Ontology Web Language (OWL) and OWL-S Language are used to provide a set of ontologies to describe services for geographical domain

Enhancing precision agriculture by Internet of Things and cyber physical systems

The recent advances in Internet of Things (IoT) and the proliferation of sensor platforms have allowed the implementation of different applications used to connect physical devices (Things) to the real world, enabling a multi cross domain and multidisciplinary data exchange. The agricultural sector is also greatly benefiting from this progress with several advantages, including the optimal management of resources and the improvement of human labour (i.e., crop growth monitoring and selection, irrigation decision support, fertilizers, pesticide and agrochemicals application, etc.). Moreover, advancements in mechanization and GPS-assisted vehicle guidance in agriculture has established the concept of precision agronomy and precision farming, as well as automation in food production chain. However, current systems still have significant drawbacks in areas such as flexibility, networking, standardization, robustness, skills' requirements and lack of real time data and actuation. A current trend is based on the interaction of machines and autonomous systems, in order to fit into cyber-physical production systems and enabling data collection and networked site-specific monitoring and control. This paper explores all the cutting-edge challenges and solutions required to implement the digital agriculture framework, intended as the evolution from Precision Farming to connected, knowledge-based farm production systems, in a context where digital technologies are first-class elements for the automation of sustainable processes in agriculture

Microbial Biofertilisers in Plant Production and Resistance: A Review

In sustainable agriculture, plant nutrients are the most important elements. Biofertilisers introduce microorganisms that improve the nutrient status of plants and increase their accessibility to crops. To meet the demands of a growing population, it is necessary to produce healthy crops using the right type of fertilisers to provide them with all the key nutrients they need. However, the increasing dependence on chemical fertilisers is destroying the environment and negatively affecting human health. Therefore, it is believed that the use of microbes as bioinoculants, used together with chemical fertilisers, is the best strategy to increase plant growth and soil fertility. In sustainable agriculture, these microbes bring significant benefits to crops. In addition to colonising plant systems (epiphytes, endophytes and rhizospheres), beneficial microbes play a key role in the uptake of nutrients from surrounding ecosystems. Microorganisms, especially fungi, also play a protective function in plants, enhancing the responses of defence systems, and play a key role in situations related to soil iron deficiency or phosphorous solubilisation. Plant-associated microbes can thus promote plant growth regardless of natural and extreme conditions. The most frequently used strategies for growth-promoting microorganisms are nitrogen fixation, the production of growth hormones, siderophores, HCN, various hydrolytic enzymes and the solubilisation of potassium, zinc and phosphorous. Research on biofertilisers has been extensive and available, demonstrating how these microbes can provide crops with sufficient nutrients to increase yields. This review examines in detail the direct and indirect mechanisms of PGPR action and their interactions in plant growth and resistance