A multidisciplinary approach to the development of innovative tools for pharmaceutical and technological applications

The research activity carried out during the Ph.D. in Chemical and Pharmaceutical Sciences has regarded the design, synthesis, and characterization of innovative tools relevant to both pharmaceutical and technological fields. Great interest has been dedicated to the investigation of the “affinity polymerization” mechanism of novel polymeric materials founded on repetitive monomeric units based on the 3-phenylbenzofulvene scaffold that spontaneously polymerize by simple removal of the solvent. In this context, the attention has been focused on the synthesis of novel benzofulvene-based derivatives bearing complexed and non-complexed pyridine rings in different positions of the benzofulvene scaffold, to evaluate the effects generated by the insertion of a bulky substituent in the aggregation/polymerization behavior. The experience acquired with this study has been then capitalized on the design, synthesis, and characterization of a novel visible-light-sensitive biomimetic molecular switch inspired by the benzofulvene scaffold and the Green Fluorescent Protein (GFP) chromophore. The chemical-structural manipulation of the benzofulvene structure has made possible the development of a novel set of biomimetic photoswitches inspired by the supramolecular properties of the 3-phenylbenzofulvene scaffold and the molecular features of the GFP chromophore. In the framework of material chemistry, the well-known click-chemistry reaction of hyaluronic acid (HA) derivatives bearing propargylated ferulic groups has been exploited to obtain biomimetic and biocompatible materials useful in different biopharmaceutical fields. In particular, low molecular weight HA has been anchored on the hydrophobic surface of low-generation poly(propylene imine) (PPI) dendrimers by the click-chemistry reaction between their azido-functionalized surfaces and low molecular weight HA derivatives bearing propargylated ferulic groups. The resulting materials have been proposed as biocompatible drug delivery systems (DDSs) of Doxorubicin. Another application of HA has concerned the hyaluronan-based graft copolymers showing low and medium molecular weight values that have been exploited in cross-linking by the click-chemistry reaction. Interestingly, the interaction of resulting HA materials with water led to the formation of hydrogels, and the tunable rheological behavior of these materials led to their applicability in different biomedical fields. Lastly, the knowledge in medicinal chemistry has guided the design and synthesis of innovative bioactive compounds such as novel Cyclooxygenase‑2 (COX-2) inhibitors containing Nitric Oxide (NO) donor moiety (CINODs) endowed with vasorelaxant properties. The outcomes of these studies would provide fertile ground for future projects that will hopefully contribute to accelerate the research in several fields. Indeed, the increased knowledge on the behavior of these innovative tools, by means of a multidisciplinary approach, is the key for boosting the development of novel materials for pharmaceutical and technological applications

Cross-Linked Hyaluronan Derivatives in the Delivery of Phycocyanin

An easy and viable crosslinking technology, based on the “click-chemistry” reaction copper(I)-catalyzed azide-alkyne 1,3-dipolar cycloaddition (click-crosslinking), was applied to graft copolymers of medium molecular weight (i.e., 270 kDa) hyaluronic acid (HA) grafted with ferulic acid (FA) residues bearing clickable propargyl groups, as well as caffeic acid derivatives bearing azidoterminated oligo(ethylene glycol) side chains. The obtained crosslinked materials were characterized from the point of view of their structure and aggregation liability to form hydrogels in a water environment. The most promising materials showed interesting loading capability regarding the antioxidant agent phycocyanin (PC). Two novel materials complexes (namely HA(270)-FA-TEGECCL- 20/PC and HA(270)-FA-HEGEC-CL-20/PC) were obtained with a drug-to-material ratio of 1:2 (w/w). Zeta potential measurements of the new complexes (−1.23 mV for HA(270)-FA-TEGECCL- 20/PC and −1.73 mV for HA(270)-FA-HEGEC-CL-20/PC) showed alterations compared to the zeta potential values of the materials on their own, suggesting the achievement of drug–material interactions. According to the in vitro dissolution studies carried out in different conditions, novel drug delivery systems (DDSs) were obtained with a variety of characteristics depending on the desired route of administration and, consequently, on the pH of the surrounding environment, thanks to the complexation of phycocyanin with these two new crosslinked materials. Both complexes showed excellent potential for providing a controlled/prolonged release of the active pharmaceutical ingredient (API). They also increased the amount of drug that reach the target location, enabling pH-dependent release. Importantly, as demonstrated by the DPPH free radical scavenging assay, the complexation process, involving freezing and freeze-drying, showed no adverse effects on the antioxidant activity of phycocyanin. This activity was preserved in the two novel materials and followed a concentration-dependent pattern similar to pure PC

Click-Chemistry Cross-Linking of Hyaluronan Graft Copolymers

An easy and viable crosslinking procedure by click-chemistry (click-crosslinking) of hyaluronic acid (HA) was developed. In particular, the clickable propargyl groups of hyaluronane-based HA-FA-Pg graft copolymers showing low and medium molecular weight values were exploited in crosslinking by click-chemistry by using a hexa(ethylene glycol) spacer. The resulting HA-FA-HEG-CL materials showed an apparent lack of in vitro cytotoxic effects, tuneable water affinity, and rheological properties according to the crosslinking degree that suggests their applicability in different biomedical fields

Maternal immunity enhances systemic recall immune responses upon oral immunization of piglets with F4 fimbriae

F4 enterotoxigenic Escherichia coli (ETEC) cause diarrhoea and mortality in piglets leading to severe economic losses. Oral immunization of piglets with F4 fimbriae induces a protective intestinal immune response evidenced by an F4-specific serum and intestinal IgA response. However, successful oral immunization of pigs with F4 fimbriae in the presence of maternal immunity has not been demonstrated yet. In the present study we aimed to evaluate the effect of maternal immunity on the induction of a systemic immune response upon oral immunization of piglets. Whereas F4-specific IgG and IgA could be induced by oral immunization of pigs without maternal antibodies and by intramuscular immunization of pigs with maternal antibodies, no such response was seen in the orally immunized animals with maternal antibodies. Since maternal antibodies can mask an antibody response, we also looked by ELIspot assays for circulating F4-specific antibody secreting cells (ASCs). Enumerating the F4-specific ASCs within the circulating peripheral blood mononuclear cells, and the number of F4-specific IgA ASCs within the circulating IgA+ B-cells revealed an F4-specific immune response in the orally immunized animals with maternal antibodies. Interestingly, results suggest a more robust IgA booster response by oral immunization of pigs with than without maternal antibodies. These results demonstrate that oral immunization of piglets with F4-specific maternal antibodies is feasible and that these maternal antibodies seem to enhance the secondary systemic immune response. Furthermore, our ELIspot assay on enriched IgA+ B-cells could be used as a screening procedure to optimize mucosal immunization protocols in pigs with maternal immunity

Guidelines for the use and interpretation of diagnostic methods in adult food allergy

Food allergy has an increasing prevalence in the general population and in Italy concerns 8 % of people with allergies. The spectrum of its clinical manifestations ranges from mild symptoms up to potentially fatal anaphylactic shock. A number of patients can be diagnosed easily by the use of first- and second-level procedures (history, skin tests and allergen specific IgE). Patients with complex presentation, such as multiple sensitizations and pollen-food syndromes, frequently require a third-level approach including molecular diagnostics, which enables the design of a component-resolved sensitization profile for each patient. The use of such techniques involves specialists' and experts' skills on the issue to appropriately meet the diagnostic and therapeutic needs of patients. Particularly, educational programs for allergists on the use and interpretation of molecular diagnostics are needed

Pattern of care and effectiveness of treatment for glioblastoma patients in the real world: Results from a prospective population-based registry. Could survival differ in a high-volume center?

BACKGROUND: As yet, no population-based prospective studies have been conducted to investigate the incidence and clinical outcome of glioblastoma (GBM) or the diffusion and impact of the current standard therapeutic approach in newly diagnosed patients younger than aged 70 years. METHODS: Data on all new cases of primary brain tumors observed from January 1, 2009, to December 31, 2010, in adults residing within the Emilia-Romagna region were recorded in a prospective registry in the Project of Emilia Romagna on Neuro-Oncology (PERNO). Based on the data from this registry, a prospective evaluation was made of the treatment efficacy and outcome in GBM patients. RESULTS: Two hundred sixty-seven GBM patients (median age, 64 y; range, 29-84 y) were enrolled. The median overall survival (OS) was 10.7 months (95% CI, 9.2-12.4). The 139 patients 64aged 70 years who were given standard temozolomide treatment concomitant with and adjuvant to radiotherapy had a median OS of 16.4 months (95% CI, 14.0-18.5). With multivariate analysis, OS correlated significantly with KPS (HR = 0.458; 95% CI, 0.248-0.847; P = .0127), MGMT methylation status (HR = 0.612; 95% CI, 0.388-0.966; P = .0350), and treatment received in a high versus low-volume center (HR = 0.56; 95% CI, 0.328-0.986; P = .0446). CONCLUSIONS: The median OS following standard temozolomide treatment concurrent with and adjuvant to radiotherapy given to (72.8% of) patients aged 6470 years is consistent with findings reported from randomized phase III trials. The volume and expertise of the treatment center should be further investigated as a prognostic factor

Exploring Translocator Protein (TSPO) Medicinal Chemistry: An Approach for Targeting Radionuclides and Boron Atoms to Mitochondria

Translocator protein 18 kDa [TSPO or peripheral-type benzodiazepine receptor (PBR)] was identified in the search of binding sites for benzodiazepine anxiolytic drugs in peripheral regions. In these areas, binding sites for TSPO ligands were recognized in steroid-producing tissues. TSPO plays an important role in many cellular functions, and its coding sequence is highly conserved across species. TSPO is located predominantly on the membrane of mitochondria and is overexpressed in several solid cancers. TSPO basal expression in the CNS is low, but it becomes high in neurodegenerative conditions. Thus, TSPO constitutes not only as an outstanding drug target but also as a valuable marker for the diagnosis of a number of diseases. The aim of the present article is to show the lesson we have learned from our activity in TSPO medicinal chemistry and in approaching the targeted delivery to mitochondria by means of TSPO ligands

Propagación y evolución de epidemias desde la perspectiva de las ciencias sociales

“Propagación y evolución de epidemias desde la perspectiva de las ciencias sociales” es el tercer encuentro del Ciclo de Charlas organizadas por Centro Científico Tecnológico del Consejo Nacional de Investigaciones Científicas y Técnicas de Córdoba, realizado el 11 de junio de 2020. La moderación estuvo a cargo del Dr. Esteban Llamosas (CONICET – Fac Derecho - UNC), los disertantes expusieron e intercambiaron sus abordajes acerca del COVID-19 desde la perspectiva de las Ciencias Sociales. Panelistas: Lorena Saletti, CIECS (CONICET - UNC). “El impacto de las desigualdades del género ante COVID. La importancia de la perspectiva de género en salud” Leandro González, CIECS (CONICET - CEA, FCS, UNC). “Escenarios potenciales de mortalidad por la epidemia de COVID-19 en la Argentina 2020”. Mario Pecheny, Vice Presidente Científico del CONICET, Instituto Gino Germani, FCS, UBA. “Preocupaciones y demandas frente a COVID-19, encuesta al personal de salud”. Natalia Bermudez, IDACOR (CONICET- UNC). “Cómo darle legitimidad a las experiencias de las comunidades en contextos de pandemias, desafíos políticos en la intervención desde las ciencias sociales”. Alfredo Blanco, FCE, UNC. “Coronavirus y economía, lecciones de la pandemia”. Adrián Carbonetti, CIECS (CONICET-CEA, FCS, UNC). “La pandemia de gripe española en la Argentina 1918-1919, lecciones desde el pasado”.Fil: Saletti, Lorena. Universidad Nacional de Córdoba. Facultad de Ciencias Sociales; Argentina.Fil: Saletti, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Estudios sobre Cultura y Sociedad; Argentina.Fil: González, Leandro. Universidad Nacional de Córdoba. Facultad de Ciencias Sociales; Argentina.Fil: González, Leandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Estudios sobre Cultura y Sociedad; Argentina.Fil: Pecheny, Mario. Universidad de Buenos Aires. Facultad de Ciencias Sociales. Instituto de Investigaciones Gino Germani; Argentina.Fil: Pecheny, Mario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Bermudez, Natalia. Universidad Nacional de Córdoba. Facultad de Filosofía y Humanidades. Museo de Antropología; Argentina.Fil: Bermudez, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Antropología de Córdoba; Argentina.Fil: Blanco, Alfredo. Universidad Nacional de Córdoba. Facultad de Ciencias Económicas; Argentina.Fil: Carbonetti, Adrián. Universidad Nacional de Córdoba. Facultad de Ciencias Sociales; Argentina.Fil: Carbonetti, Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Estudios sobre Cultura y Sociedad; Argentina.Fil: LLamosas, Esteban. Universidad Nacional de Córdoba. Facultad de Derecho; Argentina.Fil: LLamosas, Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones Jurídicas y Sociales; Argentina.Fil: Perillo, María Angelica. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales; Argentina.Fil: Perillo, María Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina

Stability Studies of New Caged bis ‐deoxy‐coelenterazine Derivatives and Their Potential Use as Cellular pH Probes

The synthesis of new bis-deoxy-coelenterazine (1) derivatives bearing ester protective groups (acetate, propionate and butyrate esters) was accomplished. Moreover, their hydrolytic stability at room temperature was evaluated in dimethylsulfoxide (DMSO) as solvent, using the nuclear magnetic resonance (NMR) spectra of the key products at different time intervals. The results showed an increasing hydrolysis rate according to longest aliphatic chain, with a half-life of 24 days of the more stable acetate derivative (4a). Furthermore, the analysis of the experimental data revealed the greater stability of the enol tautomer in this aprotic polar solvent. This result was confirmed by theoretical calculations using the density functional theory (DFT) approach, which gave us the opportunity to propose a detailed decomposition mechanism. Additionally, the derivatives obtained were tested by bioluminescence luciferase assays to evaluate their potential use as extracellular pH-sensitive reporter substrates of luciferase. The biological data support the idea that further structural modifications of these molecules may open promising perspectives in this field of research. © 2020 American Society for Photobiolog

Angiostrongylus vasorum in 20 cani della provincia di Chieti, Italia

A seguito di un caso di Angiostrongylus vasorum, diagnosticato all’inizio del 2008 nella provincia di Chieti, è stata organizzata una ricerca parassitologica al fine di indagare la presenza del parassita nei cani nella stessa area. Da gennaio a settembre 2008 sono stati esaminati 178 cani, 56 carcasse e 122 campioni di feci. Nelle carcasse sono stati ricercati i parassiti adulti nel ventricolo destro e nell’arteria polmonare e le forme larvali in tessuti di organi interni e cervello. Nelle feci è stata ricercata la forma larvale L1 con tre metodiche diagnostiche utilizzate correntemente per la ricerca di endoparassiti e larve di strongili broncopolmonari. Sono stati diagnosticati 20 casi (8,9%) con identificazione di parassiti adulti in 5 cani e larve L1 in altri 15 soggetti. L’esame anatomopatologico delle carcasse dei cani con nematodi adulti ha evidenziato polmonite, pleurite, schiuma rossastra in trachea, versamento di liquido sieroemorragico in cavità toracica e ingrossamento di linfonodi medinici e meseraici. L’esame istologico dei tessuti ha evidenziato quadri gravi e sovrapponibili con lesioni da localizzazione dei parassiti in reni, linfonodi e cervello. Il numero cospicuo di casi riscontrati ha reso indispensabile considerare l’angiostrongilosi nelle diagnosi differenziali degli esami clinici e autoptici di cani della provincia di Chieti (Italia) e dei territori confinanti