Trans2Care, 2012. The Partnership and the Objectives of Trans2Care, an Italy-Slovenia cross-border network of science

The main areas of the research at the Blood Transfusion Center of Slovenia are cell therapies, stem cells and new diagnostic reagents, based on monoclonal antibodies (mAbs). Our laboratories are equipped for cell-culture work (certified GLP; GMP in reconstruction), molecular biology, immunological techniques and biochemistry. In the course of our research we conducted studies and applied projects (the last mainly industrial) beside production of reagents, with common topic - the antibodies. They were prepared and used to optimise and validate ELISA tests for quality control assessment of the end-product (polyclonal antibodies, in production of drugs), to study protein structure, for immunodiagnostics and as potential immunotherapeutics. Among different mAbs, which we produced in the last decade, a panel of mAbs against prion protein (PrP) and mAbs against bilitranslocase (prepared in collaboration with the University of Trieste) will be studied with different project partners in the scope of Trans2Care. We are fully dedicated to translate the results of the basic research onto applied, possibly clinical level and to contribute to improved healthcare in a sense of advanced modes of immunotherapeutics development and cell based therapies.

The Pediatric Department of the University of Ferrara has developed a special expertise in the field of hemoglobinopathies and has also an interest in gastrointestinal diseases. It has a long standing collaboration with the Department of Genetics of the University of Verona. The main fields of interest are thalassemia, gluten intolerance, and osteoporosis in its various aspects. Using our previous research experience as a platform, we plan to study Vitamin D metabolism, bone mineral density, the FGF23 and Klotho axis in patients with thalassemia, in patients with adult type lactose intolerance, in patients treated with antiepileptic drugs and in those who suffer from gluten intolerance. Finally, we intend to cooperate with another group (PP9) of the Trans2Care project in order to clarify the role of tissue antitransglutaminases in seronegative patients with symptoms of gluten intolerance.

The researchers involved in the Trans2Care project at the Department of Medical and Biological Sciences of the University of Udine investigate the biological properties of the antimicrobial peptides (AMPs) of the immune system and their therapeutic potential for human and veterinary application. In addition to potent and broad-spectrum antimicrobial activities, some AMPs display anti-inflammatory and immunomodulatory effects and hold promise as novel antiinfective agents combining antibiotic and immunostimulating properties. A detailed knowledge of their physicochemical, biological and pharmacological properties and of their impact on clinical settings is an important prerequisite to this end. The Trans2Care project offers an invaluable opportunity to share knowledge, technical expertise and laboratory facilities to achieve a better understanding of the biological features and therapeutic potential of AMPs.

Prion diseases are incurable and fatal neurodegenerative disorders that affect both humans and animals. The causative agent is an infectious protein called prion (PrPSc), which is the pathological form of a normal protein (PrPC) present on the cell membrane. The molecular mechanisms underlying prion replication and subsequent degeneration of the Central Nervous System (CNS) are still poorly understood and therefore innovative approaches are needed to build diagnostic, therapeutic, taxonomic, and disease surveillance tools. We are going to adopt an unbiased genomic approach and conduct whole transcriptome analyses using microarray gene expression methods in brain and/or blood of infected animals versus healthy controls. We hope to identify a set of genes that can be used for early diagnosis and/or as targets for therapeutic strategies. Within the Trans2Care project we intend to promote collaboration and exchange of knowledge to facilitate all partners’ research objectives, and possibly find a common way to accelerate the process aimed at improving our healthcare system.

Overthe past half-century, there have been many advances in the design, construction, and implantation of joint prostheses, resulting in a high percentage of successful long-term outcomes. One of the most common concerns of both patients and physicians is the problem of joint replacements becoming loose over time. Causes of failure include infections, aseptic loosening, dislocations, and fracture of the prosthesis or bone. Multidisciplinary research team studies are needed for an improvement in understanding in pathophysiological mechanisms of joint implant loosening and failure, which is the key point to improve implant survival and to minimize revisions.

At the Research Department in hospital of Nova Gorica we are faced with new cathlab practice and problems connected with it. Together with other project partners we intend to investigate contrast media induced nephropathy by measuring the release of NGAL, which is considered to be a good biomarker for detecting reduced kidney function. Our first priority is extensive study of NGAL and develop special detecting lab-on-a chip that should be small and simple to use at every day hospital practice. Our second priority is to develop new and intensify pre-existing collaborations and exchange of knowledge between project partners.

Polymerase chain reaction (PCR) has enabled enormous progress in the field of molecular biology in the last twenty-five years. It became popular due to its high sensitivity and specificity and was introduced to numerous scientific fields. Today many different variants of PCR exist, with real-time PCR being the most common. This method is being successfully used at our institution for a variety of different applications, among which gene expression analyzes and development of protocols for clinical diagnostics currently dominate. But the extreme flexibility and customizability of real-time PCR, coupled with our team’s expertise, ensures efficient application of this technique to various new research projects.

Flavonoids are a large class of plant secondary metabolites, belonging to polyphenol family, which possess pharmacological and nutritional properties. Their synthesis takes place only in plants, while mammals can acquire them only with diet. It has been demonstrated that flavonoid uptake occurs in rat also by the activity of bilitranslocase, a carrier that is involved in anion transport in liver cell, vascular endothelium and gastric mucosa. A sequence of bilitranslocase interacting with flavonoid moieties is already known and characterized. Antibody raised against such protein epitope were shown to exhibit cross-reactivity against plant membrane proteins in tissues involved in flavonoid transport and accumulation, such as teguments of carnation petals and skin of grape berries. Further immunolocalization studies allowed to demonstrate the presence of cross-reacting protein not only at the level of tegumental tissues, but also associated to sieve elements and seed teguments in grape berries.

Trans2Care project is an operation of the Cross-Border Cooperation Programme Italy-Slovenia 2007-2013, funded by the European Regional Development Fund and some national funds. This project, started on 1st April 2011, will be implemented through a period of three and half years by a consortium of 13 Partners, lead by the University of Trieste. It aims at translating results of their biomedical research into innovative products and services to improve health. The kick-off meeting of Trans2Care took place in Trieste on 21st-22nd November 2011. The venue was the Narodni Dom, i.e. the Trieste National Hall, a multimodal building of the Slovene minority in the city, which formerly included the Slovene theatre and a hotel (Hotel Balkan). Narodni Dom was built on 1904 on the plan of Max Fabiani, one of the greatest Mittel-European architects. Burnt by the Italian fascists in 1920, it hosted later the Hotel Regina. Ultimately restored in 1990, it now hosts the Slovenski informativni center (Slovene information center) and the Advanced School of Modern Languages for Translators and Interpreters of the University of Trieste. On 21st November, the project was presented to the Partners, the academic and public authorities, and the general public, as reported in Section I. On 22nd November, the Partners had an internal meeting, aimed at laying down the scientific contents and objectives of their collaboration, described in the 21 chapters of Section II. The project’s proposal has had a unique trajectory during the 2-step evaluation process, recapitulated in Section III. The latter also hosts the points of view on the project by external observers.

Biosensors are self-contained analytical devices in which a bioreceptor is integrated with a transducer. The interaction between the bioreceptor and a target analyte generates a signal suitable for analytical purposes. In electrochemical biosensors, a change in the redox state of the biorecognition/analyte system generates a change in an electrochemical quantity which can be monitored by electroanalytical techniques. Electrochemical sensors can be miniaturized using ultramicroelectrodes and nanoelectrodes and their arrays as transducers. These devices are characterized by high specificity and sensitivity and improved detection limits. Biosensors can be used by non-specialist operators at the point of care. For the above reasons, within the frame of the Trans2care project, the Laboratory of Electrochemical Sensors of the University Ca’ Foscari of Venice will collaborate with the project partners to develop electrochemical sensors suitable for specific clinical needs.

Perhaps the most appealing opportunity is represented by gaining inspiration from nature for the precise tailoring of biomaterials with finely tuned unique functional properties. A very promising model is represented by Human Elastin-Like Polypeptides (HELP), repetitive artificial polypeptides based on penta- or hexa- peptidic motifs that characterize elastin. These protein polymers retain several peculiar biophysical properties as, the reversible inverse phase transition, changing the solubility and aggregation state in response to temperature variation. The smart nature of this class of compounds makes them attractive for many applications in the biomedical and biotechnological fields, in particular for biomaterial development. The Trans2Care project aiming to translating scientific and technological expertise into products for the biomedical field represents an exciting and challenging environment to best exploit the potential these biomimetic macromolecules.

At the Immunopathology Laboratory at the IRCCS Burlo Garofolo hospital the research activity is based on autoimmune diseases, above all on celiac disease in order to diagnose it in an early stage. For this reason, we are collecting many serum samples and intestinal biopsies to analyse them with molecular (phage-display) and immunofluorescent (double staining and activated beads) assays. Within the Trans2Care project we intend to apply these methods in several areas related to the problems explored by the Project partners with the aim of promote collaboration, mobility of researchers and exchange of knowledge between partners.

The Research department of the Valdoltra Orthopaedic Hospital is determined to conduct more extensive research studies on modification of materials for orthopaedic implants. The starting research in the newly founded Research laboratory shall include basic and applied studies which results shall be potentially considered and eventually implemented in daily clinical practice of Valdoltra Orthopaedic Hospital. With an accurate inspection of the emerging medical needs in the field of orthopaedics we envisaged the urgent need to provide a long-term protection for orthopaedic prostheses. By means of using nano-engineering approaches for the functionalization of orthopaedic implant surfaces with suitable antimicrobial agents, it is possible to protect orthopaedic implants against harmful bacteria, which trigger the initiation of implantassociated bacterial infection. As the implant-associated bacterial infection can affect the longevity of the prosthesis, thus, the scientific and financial efforts, with the help of the project Trans2care, will be focused substantially on the design and fabrication of protective antibacterial coatings for orthopaedic implants.

Although mathematics finds large application in physical sciences, its use in the medical and biological field is less usual. Nevertheless, mathematical models can help in understanding many medical and biological phenomena and they can be a powerful tool in the designing process of many technological devices connected to the above mentioned phenomena. Of course, fundamental pre-requisite for building up a reliable mathematical model is the deep cooperation of different competences such as those of medical doctors, biologists, pharmacists and engineers. This paper deals with four different examples of mathematical models developed by our group and dealing with restenosis, enzyme action on two dimensional DNA brushes, oral delivery of drug in amorphous or nano-crystalline form and tumour cell behaviour.

At the University of Primorska College of Health Care Izola the researchers are joined in an interdisciplinary team which will perform a research entitled “A multidisciplinary approach in the treatment of obesity”. Obesity and overweight pose a major risk for diet-related chronic diseases. So, understanding the role of obesity is of major importance in the prevention and treatment of these related diseases. It is known that genetic factors related to food intake, basal metabolic rate and adipocyte differentiation contribute, but also life style, psychological factors, and tradition, may provide to the etiology of obesity, that is why obesity will be studied from different aspects. The aim of the research is in addition to nutritional treatment also attempt to understand the regulation of the secretion of adipokines by different food components. Research is versatile and interdisciplinary, and is a challenge for researchers.

Electroporation refers to exposure of cells to external electric field thatresults in transiently or permanently increased permeability of cell membranes. Cancer treatment, where local application of electroporation to tumor nodules is combined with chemotherapeutic drugs bleomycin or cisplatin is called electrochemotherapy. The antitumor effectiveness of electrochemotherapy is primarily based on direct killing of tumor cells due to the increased chemotherapeutic drug uptake, but other mechanisms, such as vascular disrupting action was also demonstrated. Our group is involved in elucidating the underlying mechanism of vascular disrupting action by studying the changes in cytoskeletal proteins after electrochemotherapy in endothelial cells by immunocytochemistry. In addition, metastatic potential and global changes in gene expression of melanoma cells that survived electrochemotherapy were also studied, demonstrating that metastatic potential of cells is not changed and that a very low percentage of genes are down or up regulated after electrochemotherapy, which further supports its safe use in the clinical setting. Another application of electroporation is delivery of plasmid DNA into the cells for gene therapy. Our group is involved in preparation of plasmid DNA with tissue specific promoters and without genes for antibiotic resistance, thus enabling safe use of gene therapy in line with USA Food and Drug Administration (FDA) and European medicines Agency (EMA) recommendations.