Multivalent glycocalixarenes for lectin inhibition

The growing knowledge of the biological role played by the glycoside cluster effect prompted us to design and synthesize compounds able to inhibit target lectins with high selectivity and efficiency by combining a multivalent presentation of ligands with carbohydrates tailored for the best-possible contact complementarity with the proteins. To this aim calixarenes were chosen as multivalent scaffolds because they offer the unique opportunity to easily change the valency, the molecular shape, the conformation and the symmetry of the glycocluster. The work reported in this Ph.D. thesis concerns the synthesis and properties of a series of calix[n]arenes containing units of galactose, lactose, LacNAc or LacNAc modified at 2-N or 3’ positions and connected to the macrocyclic structure via thioureido or triazole groups. These compounds showed high affinity for different types of galectins. Quite interestingly, the selectivity for the different lectins was confirmed to be also dependent on the calixarene conformation, pointing out the importance of the multivalent presentation in space of the glycosyl units. Enzymatic galactosylation and 2,3- and 2,6-sialylation reactions were also explored using glycosylcalixarenes as substrate. Moreover, NMR and computational studies were performed on selected glycocalixarenes in order to better understand the nature of the carbohydrate-lectin interactions in solution on an atomic level. At last, pentavalent glycocalix[5]arenes functionalized with carbohydrates of increasing complexity, including GM2 and GM1 oligosaccharides, were synthesized to allow a perfect match in valency with the target lectin, the pentavalent B subunit of cholera toxin. Preliminary ELISA tests remarkably showed an IC50 value in low nanomolar range indicating the GM1-derivatized glycocalix[5]arene to be one of the most efficient cholera toxin inhibitors known so far

FACILE FUNCTIONALIZATION OF sp2 CARBON ALLOTROPES WITH A BIOBASED JANUS MOLECULE

A simple, versatile, sustainable, not expensive method for the functionalization of sp2 carbon allotropes, both nanosized and nano-structured, without altering their bulk crystalline organization, is presented. Carbon materials available at the commercial scale were used: furnace carbon black (CB), nano-sized graphite with high surface area, and multiwalled carbon nanotubes. A bio-sourced molecule, 2-(2,5-dimethyl-1H-pyrrol-1-yl)-1,3-propanediol (serinol pyrrole), was used for the functionalization. Serinol pyrrole (SP) was obtained from serinol through a reaction with atomic efficiency of about 82%, performed in the absence of solvents or catalysts. Synthesis of serinol pyrrole was performed as well on carbon allotropes as the solid support. Adducts of serinol pyrrole with a carbon allotrope were prepared with the help of either thermal or mechanical energy. Functionalization yield was in all cases larger than 90%. With such adducts, stable dispersions in water and inNRlatex were prepared.Afew layers of graphene were isolated from the water dispersions, and NR-based composites precipitated from the latex revealed very even distribution of fine graphitic particles. Composites were prepared, based on NR, IR, andBRas the rubbers andCBand silica as the fillers, with different amounts of CB–SPadduct, and were cross-linked with a sulfur-based system without observing appreciable effect of functionalization on vulcanization kinetics. The CB–SP adduct led to appreciable reduction of the Payne effect

Evaluating Cyclist Patterns Using GPS Data from Smartphones

In recent years the availability of GPS data has seen as a significant improvement in data accuracy, continuity and quality, due to the spread of smartphones and mobile applications for self-localization and navigation. GPS datasets provide analysts with geo-referenced information about users’ mobility and habits. The first part of the thesis consisted of an analysis of the context of bicycle facilities for the city of Bologna, Italy, made through experimental measures of cycleway and road usage rates, and cycling speed. The following part of the study focused on the use of GPS traces datasets, which allow to record a wide amount of bike trips. The main advances in the last years’ relevant literature have been described. Subsequently, two case studies have been analyzed: GPS traces datasets recorded through mobile devices, both for the city of Bologna and for the whole Netherlands. First, the original dataset of GPS points have been properly filtered in order to exclude instrumental errors. The GPS points have then been matched to a high-detail network database, in order to obtain the actual routes chosen by cyclists. The considered road networks included both attributes of the roadway and attributes of the bicycle facilities, when existing. The percentage of the trips done on a bicycle facility versus the roadway could thus be compared with the results from the first part of the study, and be used as a measure of the attractiveness and effectiveness of the bicycle facilities available. Furthermore, the chosen routes could be compared with the shortest routes for each origin-destination pair, and route choice models could be calibrated, based on different relevant attributes of the networks, of users, and of trips

Development of an in vitro model on cellular adhesion on granular natural bone mineral under dynamic seeding condition- A pilot study

Adhesion of osteogenic cells on biomaterials can be studied with static in vitro models, whereas models representing dynamic seeding conditions are rare. Herein, we present an in vitro model to study cell adhesion on granular biomaterials under dynamic seeding conditions. Radiolabeled osteogenic MC3T3-E1 cells were allowed to adhere to granules of natural bovine bone mineral (NBM) under constant rotation. Adhesion of MC3T3-E1 cells was determined by liquid scintillation counting, and cell morphology was visualized by scanning electron microscopy. Cell viability was determined by MTT assay under static and dynamic conditions, at room and body temperature, and in the presence or absence of serum. We show here that MC3T3-E1 cells rapidly adhere to NBM, reaching a peak 3 h after seeding. Attached cells display characteristic signs of spreading. Five to ten percent of total radioactivity remained on NBM after the removal of nonadherent cells. Viability is maintained at room temperature and under rotation for upto 3 h. This data suggests that the dynamic in vitro model presented here provides a tool to study cell adhesion on granular biomaterial

Epilepsy Phenotypes of Vitamin B6-Dependent Diseases: An Updated Systematic Review

Background: Vitamin B6-dependent epilepsies include treatable diseases responding to pyridoxine or pyridoxal-5Iphosphate (ALDH7A1 deficiency, PNPO deficiency, PLP binding protein deficiency, hyperprolinemia type II and hypophosphatasia and glycosylphosphatidylinositol anchor synthesis defects). Patients and methods: We conducted a systematic review of published pediatric cases with a confirmed molecular genetic diagnosis of vitamin B6-dependent epilepsy according to PRISMA guidelines. Data on demographic features, seizure semiology, EEG patterns, neuroimaging, treatment, and developmental outcomes were collected. Results: 497 published patients fulfilled the inclusion criteria. Seizure onset manifested at 59.8 ± 291.6 days (67.8% of cases in the first month of life). Clonic, tonic-clonic, and myoclonic seizures accounted for two-thirds of the cases, while epileptic spasms were observed in 7.6%. Burst-suppression/suppression-burst represented the most frequently reported specific EEG pattern (14.4%), mainly in PLPB, ALDH7A1, and PNPO deficiency. Pyridoxine was administered to 312 patients (18.5% intravenously, 76.9% orally, 4.6% not specified), and 180 also received antiseizure medications. Pyridoxine dosage ranged between 1 and 55 mg/kg/die. Complete seizure freedom was achieved in 160 patients, while a significant seizure reduction occurred in 38. PLP, lysine-restricted diet, and arginine supplementation were used in a small proportion of patients with variable efficacy. Global developmental delay was established in 30.5% of a few patients in whom neurocognitive tests were performed. Conclusions: Despite the wide variability, the most frequent hallmarks of the epilepsy phenotype in patients with vitamin B6-dependent seizures include generalized or focal motor seizure semiology and a burst suppression/suppression burst pattern in EEG

Obsessive compulsive disorder comorbidity in DBA

Diamond-Blackfan Anemia (DBA) is a congenital erythroid aplasia characterized as a normochromic macrocytic anemia with a selective deficiency in red blood cell precursors in otherwise normocelullar bone marrow. DBA is known to be associated with mental retardation and learning disabilities. Although comorbidities with other psychiatric conditions have not been reported in the existing literature, we report in this paper a case of a DBA patient with previously undiagnosed comorbidity of obsessive compulsive disorder (OCD), successfully treated with sertaline 200 mg/day and valproic acid 600 mg/day. This case of comorbid presentation has clinical, therapeutic and pathophysiological implications. Given the difficulty of distinguishing among mental retardation, learning disabilities and OCD and the importance of precocious diagnosis in treating OCD especially since there are treatment methods interfering with anemia symptoms, physicians should adapt an adequate screening tool treating a child with DBA and comorbid mental disorder

Extracellular vesicles in the Chronic Myeloid Leukemia scenario: an update about the shuttling of disease markers and therapeutic molecules

Extracellular vesicles (EVs) are various sets of cell-derived membranous structures containing lipids, nucleic acids, and proteins secreted by both eukaryotic and prokaryotic cells. It is now well recognized that EVs are key intercellular communication mediators, allowing the functional transfer of bioactive chemicals from one cell to another in both healthy and pathological pathways. It is evident that the condition of the producer cells heavily influences the composition of EVs. Hence, phenotypic changes in the parent cells are mirrored in the design of the secreted EVs. As a result, EVs have been investigated for a wide range of medicinal and diagnostic uses in different hematological diseases. EVs have only recently been studied in the context of Chronic Myeloid Leukemia (CML), a blood malignancy defined by the chromosomal rearrangement t(9;22) and the fusion gene BCR-ABL1. The findings range from the impact on pathogenesis to the possible use of EVs as medicinal chemical carriers. This review aims to provide for the first time an update on our understanding of EVs as carriers of CML biomarkers for minimal residual disease monitoring, therapy response, and its management, as well as the limited reports on the use of EVs as therapeutic shuttles for innovative treatment approaches

Master curves for the mechanical reinforcement of diene elastomers with sp2 carbon allotropes

sp2 carbon allotropes are efficient reinforcing fillers for polymer melt and elastomers: carbon black (CB) has been used since early 1900’s and nanofillers such as carbon nanotubes (CNT), graphene and graphene related materials (GRM) have increased their importance over the last decades. Nanofillers can definitely establish larger interfacial area with the polymer matrix than CB and great impact on material properties is thus expected. However, it is widely acknowledged that they will not be able to completely replace CB. Hence, increasing research efforts are on hybrid systems based on CB-CNT and CB-GRM [1]. Research objective is to identify common features and behaviour of nano (CNT, GRM) and nanostructured (CB) sp2 carbon allotropes. In this work, initial modulus was determined by means of dynamic-mechanical shear measurements of composites based on either poly(1,4-cis-isoprene) or poly(styrene-co-butadiene) as the rubber and either CB or CNT or GRM or hybrid systems as the reinforcing fillers. Filler-polymer interfacial area (i.a.), calculated as the product of filler surface area, density and volume fraction, was used to establish a common correlation with the composite initial modulus. A sort of master curve was derived, able to fit all the points up to interfacial area of about 27 μm-1, corresponding to remarkable filler content. Much better efficiency was shown by carbon fillers, when composites were prepared through latex blending. To allow easy dispersion in rubber latex, sp2 carbon allotropes were functionalized with a serinol derivative: 2-(2,5-dimethyl- 1H-pyrrol-1-yl)-1,3-propanediol (serinol pyrrole, SP) [2, 3], shown in Figure 1

C6orf10 low-frequency and rare variants in italian multiple sclerosis patients

In light of the complex nature of multiple sclerosis (MS) and the recently estimated contribution of low-frequency variants into disease, decoding its genetic risk components requires novel variant prioritization strategies. We selected, by reviewing MS Genome Wide Association Studies (GWAS), 107 candidate loci marked by intragenic single nucleotide polymorphisms (SNPs) with a remarkable association (p-value <= 5 x 10(-6)). A whole exome sequencing (WES)-based pilot study of SNPs with minor allele frequency (MAF) <= 0.04, conducted in three Italian families, revealed 15 exonic low-frequency SNPs with affected parent-child transmission. These variants were detected in 65/120 Italian unrelated MS patients, also in combination (22 patients). Compared with databases (controls gnomAD, dbSNP150, ExAC, Tuscany-1000 Genome), the allelic frequencies of C6orf10 rs 16870005 and IL2RA rs12722600 were significantly higher (i.e., controls gnomAD, p = 9.89 x 10(-7) and p < 1 x 10(-20)). TET2 rs61744960 and TRAF3 rs138943371 frequencies were also significantly higher, except in Tuscany-1000 Genome. Interestingly, the association of C6orf10 rs16870005 (Ala431Thr) with MS did not depend on its linkage disequilibrium with the HLA-DRB1 locus. Sequencing in the MS cohort of the C6orf10 3' region revealed 14 rare mutations (10 not previously reported). Four variants were null, and significantly more frequent than in the databases. Further, the C6orf10 rare variants were observed in combinations, both intra-locus and with other low-frequency SNPs. The C6orf10 Ser389Xfr was found homozygous in a patient with early onset of the MS. Taking into account the potentially functional impact of the identified exonic variants, their expression in combination at the protein level could provide functional insights in the heterogeneous pathogenetic mechanisms contributing to MS.In light of the complex nature of multiple sclerosis (MS) and the recently estimated contribution of low-frequency variants into disease, decoding its genetic risk components requires novel variant prioritization strategies. We selected, by reviewing MS Genome Wide Association Studies (GWAS), 107 candidate loci marked by intragenic single nucleotide polymorphisms (SNPs) with a remarkable association (p-value ≤ 5 × 10−6). A whole exome sequencing (WES)-based pilot study of SNPs with minor allele frequency (MAF) ≤ 0.04, conducted in three Italian families, revealed 15 exonic low-frequency SNPs with affected parent-child transmission. These variants were detected in 65/120 Italian unrelated MS patients, also in combination (22 patients). Compared with databases (controls gnomAD, dbSNP150, ExAC, Tuscany-1000 Genome), the allelic frequencies of C6orf10 rs16870005 and IL2RA rs12722600 were significantly higher (i.e., controls gnomAD, p = 9.89 × 10−7 and p < 1 × 10−20). TET2 rs61744960 and TRAF3 rs138943371 frequencies were also significantly higher, except in Tuscany-1000 Genome. Interestingly, the association of C6orf10 rs16870005 (Ala431Thr) with MS did not depend on its linkage disequilibrium with the HLA-DRB1 locus. Sequencing in the MS cohort of the C6orf10 3′ region revealed 14 rare mutations (10 not previously reported). Four variants were null, and significantly more frequent than in the databases. Further, the C6orf10 rare variants were observed in combinations, both intra-locus and with other low-frequency SNPs. The C6orf10 Ser389Xfr was found homozygous in a patient with early onset of the MS. Taking into account the potentially functional impact of the identified exonic variants, their expression in combination at the protein level could provide functional insights in the heterogeneous pathogenetic mechanisms contributing to MS