298 research outputs found
Identificazione del corecettore utilizzato da HIV per l'ingresso nella cellula: confronto tra saggio fenotipico in vitro e fenotipo virtuale
No full textOBIETTIVI: L’introduzione nella pratica clinica degli antagonisti del CCR5 come nuova classe di inibitori dell’entry ha determinato la necessità di conoscere il coreceptor usage del virus prima di iniziare la terapia. Molti clinici ottengono questa informazione utilizzando il test Trofile nell’ ultima versione ultrasensibile (ESTA).Il largo utilizzo di questo saggio è limitato da alti costi e da lunghi tempi di attesa per la risposta. Sono perciò stati fatti numerosi sforzi per ottenere risultati in minor tempo possibile e con bassi costi migliorando i test genotipici che prevedono l’utilizzo di algoritmi interpretativi.
A causa dell’elevata variabilità del gene env i saggi genotipici sono tuttavia inaffidabili mentre i saggi biologici sono quasi sempre in grado di definire il tropismo attuale. A tal fine abbiamo messo a punto un vettore di espressione fluorescente che si caratterizza per consentire il clonaggio delle sequenze V3 amplificate dal paziente di interesse. Questo plasmide è dotato di un gene per la proteina fluorescente GFP che viene espressa quando il virus replica nelle cellule, è pertanto possibile conoscere sia il fenotipo conferito da quella specifica sequenza V3 che la capacità replicativa della variante V3 attraverso l’analisi fluorimetrica delle cellule dopo un saggio di transfezionne-infezione.
Obiettivo di questo lavoro è confrontare il saggio fenotipico home-brew con il saggio commerciale Trofile nella sua forma più sensibile l’Enhanced Trofile (ESTA) e conseguire una valutazione comparativa tra i diversi saggi fenotipici e tra i saggi fenotipici e saggi virtuali nella definizione del coreceptor usage.
MATERIALI E METODI: E’ stata utilizzata una strategia molecolare ricombinante che ha permesso di clonare in pNLmodΔV3GFP la sequenza V3 amplificata dal virus di 38 pazienti analizzati generando chimere virali replicative.
La regione V3 delle varianti virali è stata sequenziata e analizzata al geno2pheno, al fine di ottenere la predizione del fenotipo.
E’ importante determinare il livello di significatività, cioè di selezionare quanto conservativa è la determinazione del virus X4, questo livello di significatività è determinato dal tasso di falsa positività per la classificazione di un virus X4. Questo tasso detto FPR (False Positive Rate) è un cut off, una soglia al di sopra della quale i virus sono definiti R5 e al di sotto X4. Attualmente l’FPR utilizzato dalla comunità scientifica è del 10%; questo valore sembra raggiungere un buon compromesso tra sensibilità e specificità.
RISULTATI: sono stati ottenuti 117 cloni dal plasma di 38 pazienti HIV-1 positivi, il saggio ricombinante home brew ha identificato 90 cloni con fenotipo R5 corrispondenti a 28 pazienti, 20 cloni con fenotipo D/M corrispondenti a 9 pazienti e tutte le chimere virali ottenute da 1 paziente sono risultate essere non replicative (risultato DM con ESTA).
Il saggio ESTA ha identificato 25 pazienti con fenotipo R5, 9 pazienti con fenotipo D/M e 4 NR (due di questi sono risultai D/M e uno R5 con il nostro saggio).
CONCLUSIONI: il saggio fenotipico home brew descritto si mostra più sensibile del saggio ESTA ed inoltre offre la possibilità di analizzare la quasi specie virale del paziente consentendo di studiare l’influenza che ogni residuo aminoacidico della regione V3 ha sul fenotipo.
I nostri risultati mostrano che il 33% delle sequenze V3 clonate identificate come X4 dall’algoritmo geno2pheno quando inserite nel backbone di NL 4-3 conferiscono alla chimera virale un fenotipo R5. Il livello di FPR dovrebbe pertanto essere scelto con molta attenzione
Economic factors affecting obesity: an application in Italy
No full textThe World Health Organization has stated that obesity is spreading around the world like a “global epidemic”. In 2004 the percentage of obese people in the Italian population was 9%, but the trend s increasing in recent years. Focusing on this country, the purpose of the paper is to analyze the socio-economic variables affecting obesity by means of a survey conducted in a consumer sample. Our analysis is based on a survey conducted in Italy, and the sample was composed of 999 consumers. We used a binary logit model and the dependent variable is body mass index (BMI), expressed in a dichotomic way (seriously overweight and obese, value 1, and normal weight, value 0). The results show that the condition of the seriously overweight and obese increases with age, especially in people over 65 of age. Also gender is correlated with the pathology: being seriously overweight and obese is far more likely for men than for women. An inverse relation was shown between obesity and education, and between obesity and the level of food knowledge. The results highlight that disadvantaged social categories are more susceptible to the problem of overweight and obesity. A policy implication of the analysis, to limit the spread of obesity, could lie in programs aimed at improving health and food awareness and focused on these minority groups.economics of obesity, BMI and consumer, logit model, Food Consumption/Nutrition/Food Safety, Health Economics and Policy,
Comparison of HIV-1 recombinant phenotypic assays and virtual phenotype in the definition of coreceptor usage
No full textAuthor response
No full textDetecting pathogens and mounting immune responses upon infection is crucial for animal health. However, these responses come at a high metabolic price (McKean and Lazzaro, 2011, Kominsky et al., 2010), and avoiding pathogens before infection may be advantageous. The bacterial endotoxins lipopolysaccharides (LPS) are important immune system infection cues (Abbas et al., 2014), but it remains unknown whether animals possess sensory mechanisms to detect them prior to infection. Here we show that Drosophila melanogaster display strong aversive responses to LPS and that gustatory neurons expressing Gr66a bitter receptors mediate avoidance of LPS in feeding and egg laying assays. We found the expression of the chemosensory cation channel dTRPA1 in these cells to be necessary and sufficient for LPS avoidance. Furthermore, LPS stimulates Drosophila neurons in a TRPA1-dependent manner and activates exogenous dTRPA1 channels in human cells. Our findings demonstrate that flies detect bacterial endotoxins via a gustatory pathway through TRPA1 activation as conserved molecular mechanism.sponsorship: Vlaams Instituut voor Biotechnologie Alessia Soldano Luis Franco Guangda Liu Natalia Mora Emre Yaksi Bassem A Hassanr Fonds Wetenschappelijk Onderzoek G.0702.12 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0077.15 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0680.10 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0681.10 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0503.12 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0654.15 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0761.10N Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0596.12 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar Fonds Wetenschappelijk Onderzoek G.0565.07 Alessia Soldano Yeranddy A Alpizar Brett Boonen Alejandro Lopez-Requena Natalia Mora Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar KU Leuven GOA/14/011 Alessia Soldano Yeranddy A Alpizar Brett Boonen Luis Franco Alejandro Lopez-Requena Guangda Liu Natalia Mora Emre Yaksi Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar European Commission IUAP P7/13 Alessia Soldano Yeranddy A Alpizar Brett Boonen Luis Franco Alejandro Lopez-Requena Guangda Liu Natalia Mora Emre Yaksi Thomas Voets Rudi Vennekensr KU Leuven OT/12/091 Alessia Soldano Yeranddy A Alpizar Brett Boonen Luis Franco Alejandro Lopez-Requena Guangda Liu Natalia Mora Emre Yaksi Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talaverar KU Leuven PF-TRPLe Alessia Soldano Yeranddy A Alpizar Brett Boonen Luis Franco Alejandro Lopez-Requena Guangda Liu Natalia Mora Emre Yaksi Thomas Voets Rudi Vennekens Bassem A Hassan Karel Talavera (Vlaams Instituut voor Biotechnologie, Fonds Wetenschappelijk Onderzoek|G.0702.12, Fonds Wetenschappelijk Onderzoek|G.0077.15, Fonds Wetenschappelijk Onderzoek|G.0680.10, Fonds Wetenschappelijk Onderzoek|G.0681.10, Fonds Wetenschappelijk Onderzoek|G.0503.12, Fonds Wetenschappelijk Onderzoek|G.0654.15, Fonds Wetenschappelijk Onderzoek|G.0761.10N, Fonds Wetenschappelijk Onderzoek|G.0596.12, KU Leuven|GOA/14/011, KU Leuven|OT/12/091, European Commission|IUAP P7/13, KU Leuven PF-TRPLe)status: Publishe
Analysis of the functional relationship between V3 loop and gp120 context with regard to human immunodeficiency virus coreceptor usage using naturally selected sequences and different viral backbones
No full textAbstractThe human immunodeficiency virus type 1 (HIV-1) gp120 V3 loop plays a predominant role in chemokine receptor usage; however, other linear and nonlinear gp120 domains are involved in this step of the HIV-1 replication cycle. At present, the functional relationship between V3 and these domains with regard to coreceptor usage is unclear. To gain insights into the nature of this relationship in naturally selected viral variants, we developed a recombinant strategy based on two different gp120 backbones derived from CXCR4 (X4)- and CCR5 (R5)-tropic viral strains, respectively. Using this recombinant model system, we evaluated the phenotype patterns conferred to chimeric viruses by exogenous V3 loops from reference molecular clones and samples from infected subjects. In 13 of 17 recombinants (76%), a comparable phenotype was observed independently of the gp120 backbone, whereas in a minority of the recombinant viruses (4/17, 24%) viral infectivity depended on the gp120 context. No case of differential tropism using identical V3 sequence in the two gp120 contexts was observed. Site-directed mutagenesis experiments were performed to evaluate the phenotypic impact of specific V3 motifs. The data indicate that while the interaction of HIV-1 with chemokine receptors is driven by V3 loop and influenced by its evolutionary potential, the gp120 context plays a role in influencing the replication competence of the variants, suggesting that compensatory mutations occurring at sites other than V3 are necessary in some cases
HCV core antigen and HCV-RNA in HIV/HCV co-infected patients with different HCV genotypes.
No full textAbstract. Background: A good correlation between HCV core antigen (HCVAg) and different HCV-RNA assays has been
described, but little data are available in HCV/HIV co-infection. We aimed to evaluate HCVAg in comparison with
HCV-RNA and to determine their kinetics during antiviral treatment in selected HCV/HIV co-infected patients.
Methods: 355 samples from 286 HCV/HIV co-infected subjects for whom HCV-RNA (Abbott RealTime) was
requested were analysed also for HCVAg (Abbott ARCHITECT) in order to evaluate the correlation between the two
parameters both in patients treated or untreated for chronic hepatitis C and according to different HCV genotypes.
The differences between percentages were evaluated by chi square or Fisher’s exact test, while mean and median
values were compared by Student’s t test or the Mann–Whitney test, respectively. All differences were considered
significant for a p value <0.05.
Results: HCVAg was detectable on 288/315 sera (91.4%) positive for HCV-RNA and in 5 out of40 (12.5%) sera with
undetectable HCV-RNA for a total concordance of 90.1%. The correlation was fair both in untreated (r = 0.742) and
in treated (r = 0.881) patients and stronger for genotypes 1 and 4 than for genotype 3. Both HCV-RNA and HCVAg
levels were significantly higher (p = 0.028 and p = 0.0098, respectively) in patients infected by genotype 1 than by
genotype 3. The mean ratio of Log values between HCV-RNA (IU/mL) and HCVAg (fmol/liter) was 2.27 ± 1.09 in
untreated and 2.20 ± 0.82 in treated patients (p = n.s.),consistent with a sensitivity of HCVAg corresponding to about
1,000 IU/mL of HCV-RNA, and ranged from 2.21 to 2.32 among HCV genotypes with no significant differences; five
samples (1.4%; 2 genotype 1a or 1c, 3 genotype 3a) showed highly divergent values. The analysis of 18 monitoring
profiles from patients treated with PEG-IFN and Ribavirin showed similar trends, except in one case in which relapse
could be predicted by HCVAg and not by HCV-RNA.
Conclusion: These results suggest that HCVAg represents an adequate tool for determining an ongoing HCV
infection also in HIV co-infected patients, with lower costs and faster turnaround time than HCV-RNA
Social Network to analyse the relationship between ‘victim-author’ and ‘motivation’ of violence against women in Italy.
No full textThe paper aims to analyse the phenomenon of Violence against women in the Italian context during 2020. It proposes to study the relationship between ‘victim-author’ and ‘motivation’ in femicides committed in domestic environment. By means of the properties of the Social Network Analysis on bimodal data, the study detected main actors and motivations that generated the homicides with female victims. At the same time, the structural relationships allowed to investigate the existence of motivations that better characterized the action of the various actors. The bipartite graph visualization and centrality scores calculated have demonstrated the effectiveness of the methodology for the pursued objectives
Human case of autochthonous West Nile virus lineage 2 infection in Italy, September 2011.
No full textLouis-Philippe Dalembert, «vagabond jusqu’au bout de la fatigue»
No full textThe Haitian novelist and poet Louis-Philippe Dalembert (Port-au-Prince, 1962) has developed in his works of fiction the concept of vagabondage as a literary projection of his biographical wandering through multiple spaces. The aim of this essay is to study the presence of vagabondage and its distinctive features in those novels written by Dalembert that reflect the writer’s perpetual motion: Le Crayon du bon Dieu n’a pas de gomme (1996), L’Autre face de la mer (1998), L’Île du bout des rêves (2003), Les dieux voyagent la nuit (2006). The main characters are constantly moving, they are cosmopolitan wanderers who belong to many places at the same time, just like Dalembert himself. By analyzing the representation of movement in these fictions, we will show that the notion of vagabondage is depicted by the author as a positive and meaningful opportunity for the vagabond who travels across countries, languages and cultures
Una ridiscussione dei concetti di home e identity nell’Asia globalizzata: il caso di These Foolish Things (2004) e How to Get Filthy Rich in Rising Asia (2013)
No full textThe evolution of postcolonial studies in the last thirty years and the development of a transnational approach in literary studies (Jay 2010) have led to a renewed interest towards the subaltern voices, especially in relation to the phenomena of migration and diaspora and their global effects. In this light, the idea of 'home' is characterized by a sort of porosity and by a new geographical and emotional conceptualization which inevitably influences the personal and collective identities of migrant communities.
The aim of the paper is to analyze these topics from a cultural and literary standpoint through the examination of the two different kinds of migrant flows and postcolonial scenarios depicted in These Foolish Things (2004) by the English author Deborah Moggach, and How to Get Filthy Rich in Rising Asia (2013) by the Pakistan novelist Mohsin Hamid. In these novels, the chaotic Indian framework – the former margin of the British empire – is torn between its colonial past and the current effects of the permeability of its borders. It is, therefore, a perfect global context, wherein the experiences and the feelings of the modern Indian identities are reinterpreted by the two authors
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