Sustainable Agrochemical Exploitation of Ligurian Aromatic Plant Ecotypes.

The characterization of local improved varieties as well as the reduction of synthetic chemical fertilizers are sustainable approaches in the vision of a new precision Farming. Aim of our study was to improve the geographical characterization of local ecotypes and to identify peculiar features of new crops in terms of bioactive compounds. NMR and LC-MS metabolite profiling approaches followed by multivariate data analysis were applied to characterize local rosemary and garlic ecotypes. With the aim of applying for a protected designation of origin, orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify representative sensory quality indicators for Vessalico garlic and rosemary "Eretto Liguria" local ecotypes, Variable Influence on Projections (VIP) values of OPLS-DA indicated six metabolites as quality indicators for Vessalico garlic and sixteen metabolites as quality indicators for rosemary "Eretto Liguria". Finally, to discover and utilize new ecotypes in a sustainable way, Vessalico garlic extracts antiviral activity, previously evaluated against Tomato brown rugose fruit virus (ToBRFV), a Tobamovirus affecting tomato crops, was extended to Pepino mosaic virus (PepMV) with positive results.

NMR Metabolite Profiling for the Characterization of Vessalico Garlic Ecotype and Bioactivity against Xanthomonas campestris pv. campestris.

The Italian garlic ecotype "Vessalico" possesses distinct characteristics compared to its French parent cultivars Messidor and Messidrôme, used for sowing, as well as other ecotypes in neighboring regions. However, due to the lack of a standardized seed supply method and cultivation protocol among farmers in the Vessalico area, a need to identify garlic products that align with the Vessalico ecotype arises. In this study, an NMR-based approach followed by multivariate analysis to analyze the chemical composition of Vessalico garlic sourced from 17 different farms, along with its two French parent cultivars, was employed. Self-organizing maps allowed to identify a homogeneous subset of representative samples of the Vessalico ecotype. Through the OPLS-DA model, the most discriminant metabolites based on values of VIP (Variable Influence on Projections) were selected. Among them, S-allylcysteine emerged as a potential marker for distinguishing the Vessalico garlic from the French parent cultivars by NMR screening. Additionally, to promote sustainable agricultural practices, the potential of Vessalico garlic extracts and its main components as agrochemicals against Xanthomonas campestris pv. campestris, responsible for black rot disease, was explored. The crude extract exhibited a MIC of 125 µg/mL, and allicin demonstrated the highest activity among the tested compounds (MIC value of 31.25 µg/mL).

Targeted metabolite profiling of Salvia rosmarinus Italian local ecotypes and cultivars and inhibitory activity against Pectobacterium carotovorum subsp. carotovorum.

Introduction: The development of agriculture in terms of sustainability and low environmental impact is, at present, a great challenge, mainly in underdeveloped and marginal geographical areas. The Salvia rosmarinus "Eretto Liguria" ecotype is widespread in Liguria (Northwest Italy), and farmers commonly use it by for cuttings and for marketing. In the present study, this ecotype was characterized in comparison with other cultivars from the same geographical region and Campania (Southern Italy), with a view to application and registration processes for the designation of protected geographical indications. Moreover, the possibility of using the resulting biomass after removing cuttings or fronds as a source of extracts and pure compounds to be used as phytosanitary products in organic farming was evaluated. Specifically, the potential of rosemary extracts and pure compounds to prevent soft rot damage was then tested. Methods: A targeted NMR metabolomic approach was employed, followed by multivariate analysis, to characterize the rosemary accessions. Bacterial soft rot assay and disk diffusion test were carried out to evaluate the activity of extracts and isolated compounds against Pectobacterium carotovorum subsp. carotovorum. Enzymatic assay was performed to measure the in vitro inhibition of the pectinase activity produced by the selected pathogen. Molecular docking simulations were used to explore the possible interaction of the selected compounds with the pectinase enzymes. Results and Discussion: The targeted metabolomic analysis highlighted those different geographical locations can influence the composition and abundance of bioactive metabolites in rosemary extracts. At the same time, genetic factors are important when a single geographical area is considered. Self-organizing maps (SOMs) showed that the accessions of "Eretto Liguria" appeared well characterized when compared to the others and had a good content in specialized metabolites, particularly carnosic acid. Soft rotting Enterobacteriaceae belonging to the Pectobacterium genus represent a serious problem in potato culture. Even though rosemary methanolic extracts showed a low antibacterial activity against a strain of Pectobacterium carotovorum subsp. carotovorum in the disk diffusion test, they showed ability in reducing the soft rot damage induced by the bacterium on potato tissue. 7-O-methylrosmanol, carnosol and isorosmanol appeared to be the most active components. In silico studies indicated that these abietane diterpenoids may interact with P. carotovorum subsp. carotovorum pectate lyase 1 and endo-polygalacturonase, thus highlighting these rosemary components as starting points for the development of agents able to prevent soft rot progression.

From FAIR4Health Project to 1+MG Initiative: A Spain - Italy Collaboration.

Results of two major projects funded by the European Union are taken into consideration: Fair4Health regarding the possibility of sharing clinical data in various environments applying FAIR principles and 1+Million Genome for the in-depth study of the human genome in Europe. Specifically, the Gaslini hospital plans to move on both areas joining the Hospital on FHIR initiative matured within the fair4health project and also collaborate with other Italian healthcare facilities through the implementation of a Proof of Concept (PoC) in the 1+MG. The aim of this short paper is to evaluate the applicability of some of the tools of the fair4health project to the Gaslini infrastructure to facilitate its participation in the PoC. One of the aims is also to prove the possibility of reuse the results of well-performed European funded projects to boost routine research in qualified healthcare facilities.

Healthcare Insights: Evaluating the Access to the Italian Healthcare System.

The Italian health system is organised on a regional basis and services are provided by both public and private operators, affecting the planning of services, access to services by citizens and their health rights. The creation of an observatory monitoring the methods and times of access to healthcare services has been pursued. The preliminary phase of the project is presented, which will lead to the comparison of the data obtained from 2019, with an eye on the Covid-19 pandemic impact.

Interoperability Standards for Data Sharing as a Basis to Fill in a Tailored EHR for Undiagnosed Rare Diseases.

Undiagnosed rare diseases include diseases with a well-characterised phenotype, diseases with unknown molecular causes or due to non-genetic factors, and pathological condition that cannot be named. Several initiatives have been launched for healthcare of patients with undiagnosed rare diseases. A project for development of medical records with special reference to the HL7 standards is being carried out in Genoa (Italy), taking into account regional and national regulations. The project is based on the integration of functionality related to patient diagnostics, taking into account omic sciences for disease prevention and risk assessment. Considering the evolution of standards, the use of FHIR is being considered in order to increase the elasticity of the system also in view of foreseeable adoption of this standard by the Italian healthcare system.

Semantics Management for a Regional Health Information System in Italy by CTS2 and FHIR.

An infrastructure for the management of semantics is being developed to support the regional health information exchange in Veneto - an Italian region which has about 5 million inhabitants. Terminology plays a key role in the management of the information fluxes of the Veneto region, in which the management of electronic health record is given great attention. An architecture for the management of the semantics of laboratory reports has been set up, adopting standards by HL7. The system has been initially developed according to the common terminology service release 2 (CTS2) standard and, in order to overcome complexities of CTS2 is being revised according to the Fast Healthcare Interoperability Resources (FHIR) standard, which has been subsequently introduced. Aspects of CST2 and of FHIR have been considered in order to retain most suitable aspects of both. This integration can be regarded as most worthwhile.

Integrating an Electronic Health Record System into a Regional Health Information System: An HL7 FHIR Architecture.

This paper presents an architecture which has been developed in order to integrate a routinely used cardiologic EHR system into the health information infrastructure system of the region where the EHR is used. A Service Oriented Approach based on HL7 and FHIR was used, achieving interoperability, security and confidentiality at all levels. The system has been used for about a year and has brought about significant improvements in the provision of care.

Healthcare Associated Infections: An Interoperable Infrastructure for Multidrug Resistant Organism Surveillance.

Prevention and surveillance of healthcare associated infections caused by multidrug resistant organisms (MDROs) has been given increasing attention in recent years and is nowadays a major priority for health care systems. The creation of automated regional, national and international surveillance networks plays a key role in this respect. A surveillance system has been designed for the Abruzzo region in Italy, focusing on the monitoring of the MDROs prevalence in patients, on the appropriateness of antibiotic prescription in hospitalized patients and on foreseeable interactions with other networks at national and international level. The system has been designed according to the Service Oriented Architecture (SOA) principles, and Healthcare Service Specification (HSSP) standards and Clinical Document Architecture Release 2 (CDAR2) have been adopted. A description is given with special reference to implementation state, specific design and implementation choices and next foreseeable steps. The first release will be delivered at the Complex Operating Unit of Infectious Diseases of the Local Health Authority of Pescara (Italy).

Medical Equipment Replacement Prioritisation: A Comparison Between Linear and Fuzzy System Models.

In hospital management, health technology assessment techniques are being increasingly developed. This paper presents a comparison of the results obtained using two models for replacement priority value calculation applied to the Galliera hospital in Genoa (Italy). One the models was developed at the Galliera Hospital along the lines of the model by Fennigkoh and addresses four primary replacement issues: equipment service and support, equipment function, cost benefits and clinical efficacy, by a "yes-no" scheme. This model is compared with a model based on fuzzy logic. The comparison between the two models shows a conservative behaviour by the Galliera model, according to which 77.4% of the analysed instrumentation is maintained, whereas the classification by the fuzzy model allows for a better discrimination among the devices.

Information Technology System Including Patient Generated Health Data for Cancer Clinical Care and Research.

This paper presents the extension of a service-oriented architecture framework for precision oncology to the management of patient generated health data from wearables. The solution follows the indication provided by the Health Level 7 (HL7) and Object Management Group (OMG) initiative Healthcare Service Specification Project (HSSP) and is compliant to Retrieve Locate and Update Service (RLUS) Release 1 standard adopting Clinical Document Architecture Release 2 (CDA R2) as semantic signifier. The system which has been developed supports the management of visits, the setting up of a clinical diary and a comprehensive view of the patients from the wearables data for improve clinical care and for research. The system structure is highly modular and the parameters relating to wearables data are only present in one module. Extension of the systems to other aspects, such as genomics and immune therapy, are planned following the same modular design criteria.

Cancer precision medicine today: Towards omic information in healthcare systems.

INTRODUCTION:: This article focuses on the integration of omics data in electronic health records and on interoperability aspects relating to big data analysis for precision medicine. METHODS:: Omics data integration methods for electronic health record and for systems interoperability are considered, with special reference to the high number of specific software tools used to manage different aspects of patient treatment. This is an important barrier against the use of this integrated approach in daily clinical routine. RESULTS:: The correct use of all three levels of interoperability (technical, semantic, and process interoperability) plays a key role in order to achieve an easy access to a significant amount of data, all with correct contextualization, which is the only way to obtain a real value from data for precision medicine. CONCLUSIONS:: The proposed architecture could improve the potentialities of data routinely collected in many health information systems to form a real patient center information environment.

Optimizing Texture Modified Foods for Oro-pharyngeal Dysphagia: A Difficult but Possible Target?

Dysphagia is a swallowing disorder characterized by the difficulty in transferring solid foods and/or liquids from the oral cavity to the stomach, imparing autonomous, and safe oral feeding. The main problems deriving from dysphagia are tracheo-bronchial aspiration, aspiration pneumonia, malnutrition and dehydration. In order to overcome dysphagia-induced problems, over the years water and food thickening has been used, focusing specifically on viscosity increase, but limited results have been obtained. Elastic components and their effects on the cohesiveness on the bolus should be taken into account in the first place. We provide an analysis of dysphagia and suggest possible corrections to the protocols which are being used at present, taking into account rheological properties of food and the effect of saliva on the bolus. We reckon that considering such aspects in the dysphagia management market and healthcare catering would result in significant clinical risk reduction.

Prediction of potential barcoding sites on ITS1 by wavelet transform.

For sustainable development, biodiversity conservation and life-quality improvement must be simultaneously considered. Molecular techniques have greatly impacted biotechnology. These methods have, in particular, improved the capability to investigate the fine differences among organisms and, as a consequence, to better investigate the effects on environmental factors on them. We propose an approach to support the optimal selection of molecular probes for barcoding application in many biotechnological fields. The aim of our work is specificity maximization. To this purpose, we have integrated a filter system based on wavelet transforms with biological knowledge about the sequence proneness to mutation and post-translational modification. Specifically, we have tested the proposed method on ITS1 sequences that are a region of the rRNA locus. Our analysis has shown the presence of other local relative stable conformations in addition to known cleavage site. Their characteristics differ within the group of mammals selected for our analysis. These variations could be used to design new species-specific barcoding probes or other quick molecular screening tools.

Comparative analysis of Rac1 binding efficiency with different classes of ligands: morpholines, flavonoids and imidazoles.

The focal adhesion pathway has a great impact on cellular growth and survival. Its disregulation is correlated with the loss of cellular mechanical properties. Such modifications are, in many cases, associated with pathologies such as cancer and cardiovascular diseases. Actin remodeling is a critical reaction cascade embedded in focal adhesion pathway, and Rac1 is one of the proteins involved in actin remodeling. In order to design highly selective pharmacophores against this target, it is necessary to maximize the binding affinity of chemical entities against Rac1. To this purpose we propose an integrative chemo-bioinformatics tool to screen ligand specificity for a target protein. Our integrative workflow includes chemo-informatics data mining (Chemical System), structural bioinformatics and combined exploratory data analysis. We have applied this integrative chemo-bioinformatics workflow to a comparative analysis of three different classes of ligands (morpholines, flavonoids and imidazoles) against the Rac1 protein. Our analysis emphasizes the presence of several ligands that preferentially dock Rac1 in the domain that seems to be responsible for Rac1-phospholipase C gamma 1 interaction. Recent studies have highlighted the Rac1 and PLC interactions in platelet adhesion. Our study has highlighted the role of Rac1-PLC gamma1 interaction in cytoskeleton remodeling associated with cardiovascular diseases. Rac1 PLC interaction is Calcium dependent. This suggest that some of the analysed ligands, could be used to control the Calcium dependent cytoskeleton remodeling since they dock Rac1 in the switch 2 domain. Our results, in a nanotechnology perspective, also endorse the use Rac1's switch 2 domain suitable for new highly specific biosensors.

Potential MiRNAs recognition site identification in 3' UTR regions by DSP methods.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate fundamental cellular processes in diverse organisms and that have an important function in gene expression regulation. miRNAs seem capable to concurrently modulate hundreds of target genes. Their abnormal expression is emerging as important element in many pathological conditions. The identification of microRNA binding sites on those proteins that can be disease biomarker is fundamental to design synthetic artificial oligomers. In this paper we suggest a method, based on signal processing, to filter out potential miRNA recognition sites in the 3' UTR region of mRNAs.

Drug design for cardiovascular disease: the effect of solvation energy on Rac1-ligand interactions.

'OMICS' techniques have deeply changed the drug discovery process. The availability of many different potential druggable genes, generated by these new techniques, have exploited the complexity of new lead compounds screening. 'Virtual screening', based on the integration of different analytical tools on high performance hardware platforms, has speeded up the search for new chemical entities suitable for experimental validation. Docking is a key step in the screening process. The aim of this paper is the evaluation of binding differences due to solvation. We have compared two commonly used software, one of which takes into account solvation, on a set of small molecules (Morpholines, flavonoids and imidazoles) which are able to target the RAC1 protein--a cardiovascular target. We have evaluated the degree of agreement between the two different programs using a machine learning approach combined with statistical test. Our analysis, on a sample of small molecules, has pointed out that 35% of the molecules seem to be sensitive to solvation. This result, even though quite preliminary, stresses the need to combine different algorithms to obtain a more reliable filtered set of ligands.

SNP analysis of Rac1 For personalized ligand interaction.

This paper addresses mutational events that give rise to differing response to drugs focusing on Rac1, a protein that has been recognized as a target for drug design for cardiovascular disease due its regulatory role of angiogenesis. Rac1 has been considered with reference to Single Nucleotide Polymorphism (SNP), which has become of great value for personalized medicine. We have considered four variation of Rac1 registered in UNIPROTKB. Two of these variations are due to the environmental or population factors and two are mutation that we have selected because they are located near the binding sites of Rac1. Rac1 has been modelled by Rosetta software and by i-Tasser web server. We have chosen i-Tasser based modelling because the Rac1 structure obtained was more closely resembling crystallography data. In silico model have been used as receptors for docking with a set of 20 morpholines. The results that have been obtained on SNPs shows that a single ligand can react very differently with a mutated structure. Our analysis shows that all mutations that have been considered change Rac1 conformation and increase the accessible surface of Rac1. Our analysis highlights the effect of two sources of genetic variability: single base variation and alternative splicing.

In silico screening of Rac1 ligand specificity.

Microtubule (MT) destabilization promotes the formation of actin stress fibers and enhances the contractility of cells. The actin cytoskeleton is bound to each junction and controls the integrity of each through actin remodeling and these junctions can be disassembled or assembled to either increase or decrease cellular permeability. Mediators, such as thrombin, stimulate their respective receptor on endothelial cells to initiate signaling that increases cytosolic Ca2+ and activates myosin light chain kinase (MLCK), as well as monomeric GTPases RhoA, Rac1, and Cdc42. Ca2+ activation of MLCK and RhoA disrupts junctions, whereas Rac1 and Cdc42 promote junctional assembly. In order to develop formal systems biology model of actin remodelling it is necessary to investigate the reciprocal interactions between Rac1 and Cdc42 by using experimental selective inhibition. We have screened, by docking analysis, a new class of compounds for Rac1 and/or Cdc42 inhibition, the Morpholinos, that could be used as alternative tool to switch off a gene.