Gut microbiota related to Giardia duodenalis, Entamoeba spp. and Blastocystis hominis infections in humans from Côte d'Ivoire.

INTRODUCTION: Literature data provide little information about protozoa infections and gut microbiota compositional shifts in humans. This preliminary study aimed to describe the fecal bacterial community composition of people from Côte d'Ivoire harboring Giardia duodenalis, Entamoeba spp., and Blastocystis hominis, in trying to discover possible alterations in their fecal microbiota structure related to the presence of such parasites. METHODOLOGY: Twenty fecal samples were collected from people inhabiting three different localities of Côte d'Ivoire for copromicroscopic analysis and molecular identification of G. duodenalis, Entamoeba spp., and B. hominis. Temporal temperature gradient gel electrophoresis (TTGE) was used to obtain a fingerprint of the overall bacterial community; quantitative polymerase chain reaction (qPCR) was used to define the relative abundances of selected bacterial species/group, and multivariate statistical analyses were employed to correlate all data. RESULTS: Cluster analysis revealed a significant separation of TTGE profiles into four clusters (p < 0.0001), with a marked difference for G. duodenalis-positive samples in relation to the others (p = 5.4×10-6). Interestingly, qPCR data showed how G. duodenalis-positive samples were related to a dysbiotic condition that favors potentially harmful species (such as Escherichia coli), while Entamoeba spp./B. hominis-positive subjects were linked to a eubiotic condition, as shown by a significantly higher Faecalibacterium prausnitzii-Escherichia coli ratio. CONCLUSIONS: This preliminary investigation demonstrates a differential fecal microbiota structure in subjects infected with G. duodenalis or Entamoeba spp./B. hominis, paving the way for using further next-generation DNA technologies to better understand host-parasite-bacteria interactions, aimed at identifying potential indicators of microbiota changes

Two hydroxy pyridinecarboxylic acid derivatives as a possible chelating agents in neurodegenerative disease; equilibrium complexation studies with Cu(II), Zn(II).

The metal ion chelators 4-hydroxy-5-methyl-3-pyridinecarboxylic acid (DQ5) and 1,5-dimethyl-4-hydroxy-3-pyridinecarboxylic acid (DQ715) and Cu(II) and Zn(II) were investigated with the aim to restore the homeostasis of the brain Cu(II) and Zn(II) in neurodegenerative diseases. The proton dissociation constants of the ligands, the stability constants, and the coordination modes of the metal complexes formed were determined by pH-potentiometric, and spectral (UV–Vis and EPR or 1H NMR) methods. The results show that in slightly acidic and neutral pH range mono and bis complexes are formed through bidentate coordination of the ligands. The biological MTT-test reveals that the DQ715 ligand is able to lower the cytotoxic effect of Cu(II) in human embryonic kidney HEK-293 cells. Our studies revealed, however, that none of the chelators were efficient enough to withdraw these metal ions from the amyloid aggregates

“In vitro” osteogenic and angiogenic potential evaluation of a coculture of dental pulp stem and endothelial cells grown on the BisGMA/TEGDMA Chitlac coated thermosets

Securing an adequate blood supply for survival of cell transplants is critical for a successful outcome in tissue engineering. Moreover during regeneration of weaken teeth, which is susceptible to reinfection, fracture and loss, the teeth apical canal is open and a limited blood supply is allowed. Thus the interactions between endothelial and dental pulp progenitor stem cells are important for vascularization of regenerating tissue cells. In particular, the interplay of dental pulp stem cells and endothelial cells can enhance “in vitro” osteo/odontogenic and angiogenic potential (Dissanayaka J Endod 2012, 38,454-463) and “in vivo” ensure angiogenesis and pulp regeneration (Dissanayaka Tissue Engin Part A 2015, 3-4, 550-563). Since dental pulp microenvironment supports HUVEC survival and capillary network formation in the absence of scaffolding material and external angiogenic stimulation , “in vitro” osteogenic and angiogenic potential of dental pulp stemcells cocultured with endothelial cells grown on BisGMA/TEGDMA Chitlac coated thermosets was evaluated. Results: DPSCs were grown on BisGMA/TEGDMA Chitlac coated thermosets, a composite material used in dental restoration, in the presence of two different concentrations of endothelial cells (1:1 e 1:5) for 28 days and their metabolic activity and cytotoxic response were evaluated. MTT analysis discloses that cell metabolic activity significantly increases in the presence of endothelial cells, mainly at 21 days of culture, along with cytotoxic response, while at 28 days of culture a light cytotoxic response occurs. An increasing ALP activity is evidenced in the coculture systems up to 28 days, both in the presence and in absence of Chitlac thermosets and this evidence is further supported by Alizarin red staining, which does not detect mineralization in the early stages of differentiation, but is significantly increased at 28 days of culture in both the conditions (1:1 e 1:5). Even though the positive effect on DPSC differentiation, Chitlac thermosets could induce an inflammatory response in the system and thus an ELISA IL6 assay reveals an increased inflammatory response in 1:1 coculture system after 28 days of culture, furtherly increased in 1:5 coculture system. In parallel an increased PGE2 release is evidenced in 1:1 coculture system in the presence of thermosets, reduced in 1:5 coculture system, suggesting the potential occurrence of neoangiogenesis, furtherly supported by a tubular network formation when DPSC are grown on matrigel. These results evidencing that endothelial cells enhance “in vitro” osteo/odontogenic differentiation of DPSCs and angiogenesis, and that this response is revealed in the presence of Chitlac, usually used in dental restorative practice, indicate a coculture of DPSC and endothelial cells as a promising source for regenerative endodontics

Birth size, growth trajectory and later cardio-metabolic risk

There is increasing evidence of a strong association between intrauterine growth and subsequent development of chronic disease in adult life. Birth size and growth trajectory have been demonstrated to have an impact on cardio-metabolic health, both in childhood and adult life. Hence, careful observation of the children’s growth pattern, starting from the intrauterine period and the first years of life, should be emphasized to detect the possible onset of cardio-metabolic sequelae. This allows to intervene on them as soon as they are detected, first of all through lifestyle interventions, whose efficacy seems to be higher when they are started early. Recent papers suggest that prematurity may constitute an independent risk factor for the development of cardiovascular disease and metabolic syndrome, regardless of birth weight. The purpose of the present review is to examine and summarize the available knowledge about the dynamic association between intrauterine and postnatal growth and cardio-metabolic risk, from childhood to adulthood

Effects of methacrilyc thermosets coated with Silver-polysaccharide nanocomposite on HGFs adhesion in a S. mitis co-culture system

Silver based medical products have been proven to be effective in retarding and preventing bacterial growth, being silver reported to control infections since ancient times (1). In the field of dentistry, the use of silver ions/nanoparticles has been explored to counteract bacteria in resins and implants, as silver can destroy bacterial cell walls by reacting with the thiol groups (–SH) of proteins exposed to the extracellular portion of the bacterial membrane. Conversely, eukaryotic cells lack these exterior binding sites, so nanoparticles are supposed to interact with them only upon metal internalization (2). To reduce both bacterial adhesion to dental devices and cytotoxicity against eukaryotic cells, we coated BisGMA/TEGDMA methacrylic thermosets with a new material, Chitlac-nAg, formed by stabilized silver nanoparticles with a polyelectrolyte solution containing Chitlac. Here we analyzed the proliferative and adhesive ability of human gingival fibroblasts (HGFs) on BisGMA/TEGDMA thermosets uncoated and coated with AgNPs in a co-culture model system with Streptococcus mitis. After 48 h, HGFs well adhered onto both surfaces, while S. mitis cytotoxic response was higher in the presence of AgNPs coated thermosets. After 24 h thermosets coated with Chitlac as well as those coated with Chitlac-nAg exerted a minimal cytotoxic effect on HGFs, while after 48 h LDH release rised up to 20%. Moreover, the presence of S. mitis reduced this release mainly when HGFs adhered to Chitlac-nAg coated thermosets. The reduced secretion of collagen type I was significant in the presence of both surfaces even more when saliva is added. Integrin β1 localized closely to cell membranes onto Chitlac-nAg thermosets and PKC α translocated into nuclei. These data confirm that Chitlac-nAg thermosets have a promising utilization in the field of restorative dentistry exerting their antimicrobial activity due to AgNPs without cytotoxicity for eukaryotic cells.This work was supported by grants from MIUR FIRB 2010 and MIUR PRIN-2009

E-cigarettes fluids trigger molecular and morphological response in oral fibroblasts

Electronic-cigarettes (e-cigarettes) have been recently advertised as a safe alternative to the traditional ones and a possible smoking cessation tool. This electronic device was designed to transform a solution of variable compounds (some of them approved as food additives), in an inhalable aerosol. However, their safety is still not fully know (Lerner et al. 2016). The cytotoxicity of the fluids on human gingival fibroblasts (HGFs) was demonstrated on a previous study by Sancilio et al. (2016) where the occurrence of oxidative stress and apoptosis was found following the exposure to nicotine containing fluids. The aim of this study was to investigate the HGF biological response to e-cigarettes liquids (with and without nicotine) and to clarify the molecular mechanisms driving the cytotoxicity exerted by fluids themselves. To this purpose, cells were treated with e-cigarette fluids containing nicotine (final concentration 1mg/mL) and the equivalent volume of a fluid without nicotine, for times up to 48 h. Lactate Dehydrogenase Assay (LDH), electronic microscopy analysis, collagen I production, flow cytometry lysosome compartment evaluation and western blotting LC3 (microtubule-associated protein 1A/1B-light chain 3) expression were performed. Fluids containing nicotine exerted cytotoxicity as demonstrated by the increased levels of LDH, in parallel to the formation of numerous vacuoles in the cytoplasm, as well as a decrease in collagen I production and an augmented LC3 II expression which characterized autophagy occurrence In conclusion E-cigarette fluids (with and without nicotine) trigger modification ultrastructure, collagen production and lysosomal compartment in HGFs, suggesting an involvement in the pathogenesis of oral diseases

Convenzione CNR-MiSE fondo per la crescita sostenibile

Nel presente documento vengono illustrate in dettaglio le finalità del FCS e le attività rea- lizzate dal RTI nell’ambito del servizio realizzato per il primo intervento che il MiSE ha bandito sul Fondo con Decreto del 20 giugno 2013 e attuato con procedura valutativa “a sportello”, definita nel Decreto del 25 luglio 2014. In particolare, oltre alle caratteristiche principali dei progetti di ricerca che il MiSE intende finanziare attraverso tale intervento, nel presente documento viene altresì descritta l’attività che l’Ufficio Supporto alla Programmazione Operativa della Direzione Centrale alla Rete Scientifica e Infrastrutture del CNR ha condotto per coordinare e gestire il processo di valutazione delle proposte progettuali. Viene infatti descritta la compagine di valutazione del CNR, che è stata attivata grazie alla partecipazione dei ricercatori e tecnologi della rete scientifica dell’Ente, e le attività che questi, in veste di esperti valutatori, hanno condotto per determinare la qualità tecnico-scientifica delle proposte progettuali presentate

A distinctive 'microbial signature' in celiac pediatric patients

<p>Abstract</p> <p>Background</p> <p>Celiac Disease (CD) is an autoimmune disorder of the small intestine in which dietary gluten ingestion leads to a chronic enteropathy. Recently, scientific evidence suggested a potential role of gut microbiota in CD. To have a snapshot of dominant duodenal microbiota we analyzed the mucosa-associated microbiota of 20 children with CD, before and after a gluten-free diet (GFD) regimen, and of 10 controls. Total DNA was extracted from duodenal biopsies and amplification products of 16S ribosomal DNA were compared by temporal temperature gradient gel electrophoresis (TTGE). TTGE profiles were analyzed by statistical multivariate analysis.</p> <p>Results</p> <p>The average number of bands in TTGE profiles was significantly higher (<it>P </it>< 0.0001) in active (n.b. 16.7 ± 0.7) and inactive states (n.b. 13.2 ± 0.8) than in controls (n.b. 3.7 ± 1.3). Mean interindividual similarity index was 54.9% ± 14.9% for active disease, 55.6% ± 15.7% for remission state and 21.8% ± 30.16% for controls. Similarity index between celiac children before and after GFD treatment was 63.9% ± 15.8%. Differences in microbiota biodiversity were among active and remission state (<it>P </it>= 0.000224) and amid active CD and controls (<it>P </it>< 0.001). <it>Bacteroides vulgatus </it>and <it>Escherichia coli </it>were detected more often in CD patients than in controls (<it>P </it>< 0.0001).</p> <p>Conclusions</p> <p>Overall, the results highlighted a peculiar microbial TTGE profile and a significant higher biodiversity in CD pediatric patients' duodenal mucosa. The possible pathophysiological role of these microbial differences needs further characterization.</p