DETERMINANTS OF HUMAN IMMUNODEFICIENCY VIRUS-1TRANSMISSION TO THE FEMALE GENITAL MUCOSA:ROLE OF CO-INFECTING PATHOGENS AND CYTOKINES IN SEMEN

Background. Nel seme Human immunodeficiency virus (HIV)-1 e co-patogeni sono presenti in una complessa rete di citochine che influenza la replicazione locale e l\u2019infettivita\u2019 dei patogeni sessualmente trasmissibili, e, allo stesso tempo, \ue8 influenzata dalle infezioni del tratto genitale maschile (MGT). La comprensione di queste interazioni richiede un'analisi approfondita dei co-patogeni che infettano il MGT e delle citochine presenti nel liquido seminale, e delle relazioni intercorrenti tra questi e HIV-1. Metodi. La carica virale di 6 herpesvirus e HIV-1 e\u2019 stata valutata nel plasma seminale e sanguigno di 83 soggetti HIV-1 infetti cronici non in terapia, e di 33 soggetti non infetti mediante real-time PCR. Un saggio multiplex basato su beads \ue8 stato utilizzato per misurare i livelli di 21 citochine. I rapporti tra citochine sono stati definiti mediante il calcolo del coefficiente di correlazione di Spearman per tutte le possibili coppie di citochine. Blocchi di tessuto cervico-vaginale e linfoide umano sono stati inoculati con HIV-1 e trattati con interleuchina (IL)-7. La replicazione di varianti HIV-1 R5 o X4 \ue8 stata monitorata mediante misurazione della produzione dell\u2019antigene p24gag. Cellule isolate da blocchi di tessuto al giorno 6 e 9 postinfezione sono state caratterizzate per l'espressione dei marcatori CD3, CD4, CD8,e HIV-1 p24gag, e l\u2019apoptosi \ue8 stata valutata misurando l'espressione delle proteine APO2.7 e Bcl-2 mediante citofluorimetria a flusso. Risultati. La maggioranza dei campioni di seme, ma non di sangue, dei soggetti HIV-1-infetti e\u2019 risultata positiva per EBV e CMV (56% e 70%). Sangue e seme sono compartimenti immunologici separati e con l\u2019infezione da HIV-1 il seme e\u2019 il compartimento interessato dai maggiori cambiamenti nella composizione citochinica (16 vs 9 citochine alterate nel sangue). La riattivazione di CMV nel MGT \ue8 associata ad un aumento dei livelli delle citochine CCL5, CCL11 e CXCL9 nel seme. L\u2019analisi dei rapporti tra citochine ha rivelato un numero maggiore di correlazioni e un aumento dell\u2019intensita\u2019 di correlazione per la maggior parte delle citochine sia nel seme che nel sangue dei soggetti HIV-1-infetti, rispetto ai non infetti. IL-7 e\u2019 risultata essere una tra le citochine pi\uf9 concentrate nel seme e l\u2019infezione da HIV-1 ne aumenta ulteriormente i livelli seminali. Abbiamo dunque utilizzato un sistema di infezione ex vivo di tessuti umani per studiare il ruolo di IL-7 nella trasmissione di HIV-1. IL-7 e\u2019 risultata promuovere la replicazione di varianti R5 e X4 di HIV-1 in modalita\u2019 dose e tempo-dipendente. In blocchi di tessuto trattati con 25 ng/mL di IL-7 e\u2019 stato osservato un numero maggiore di cellule T CD4+ infette e una riduzione della deplezione cellulare, rispetto a blocchi di tessuto non trattati con IL-7. Il trattamento con IL-7 e\u2019 risultato inoltre in una riduzione della frazione di cellule T CD4+ infette esprimenti il marcatore apoptotico APO2.7 e in un aumento dei livelli di espressione del fattore anti-apoptotico Bcl-2. Conclusioni. HIV-1 \ue8 associato ad una alterazione generale dello spettro citochinico nel seme e alla riattivazione locale di CMV e EBV nel MGT, due fenomeni che sembrano essere correlati e potrebbero dunque influenzarsi reciprocamente. La ridotta flessibilita\u2019 dei rapporti tra citochine potrebbe ulteriormente contribuire alla riattivazione di tali infezioni latenti. Tra le citochine nel seme, IL-7 esercita un effetto protettivo sulle cellule T CD4+ infette, favorendo lo stabilirisi di una infezione produttiva nel tratto genitale femminile. La comprensione di come il complesso di citochine, di concerto con i co-patogeni presenti nel seme, influenzano la trasmissione sessuale di HIV-1 sar\ue0 oggetto di ulteriori indagini.Background. In semen Human immunodeficiency virus (HIV)-1 and co-infecting pathogens are immersed in a complex network of cytokines that affects the replication and infectivity of sexually transmitted pathogens, and, at the same time, is affected by local infections in the male genital tract (MGT). Understanding the mechanisms of these interactions requires a comprehensive analysis of coinfecting pathogens and cytokines in semen, and of the relations with HIV-1. Methods. Load of HIV-1 and 6 herpesviruses in semen and blood plasma of 83 HIV-1 chronically infected individuals na\uefve to therapy and 33 HIV-uninfected individuals was evaluated by real-time PCR. A multiplex beads-based assay was used to measure the levels of 21 cytokines. The cytokine network was defined calculating the Spearman`s rank correlation coefficient for all pairwise combinations of cytokines. Human cervico-vaginal and lymphoid tissue blocks were inoculated with HIV-1 and treated with interleukin (IL)-7. Replication of R5 or X4 HIV-1 variants was monitored over a period of 12 days measuring HIV-1 p24gag antigen by a beads-based assay. Cells isolated from tissue blocks at day 6 and 9 post-infection were characterized for the expression of the markers CD3, CD4, CD8, and HIV-1 p24gag antigen, and apoptosis was evaluated measuring the expression of the proteins APO2.7 and Bcl-2 by multicolor flow cytometry. Results. Epstein-Barr virus (EBV) and cytomegalovirus (CMV) DNA was found in semen of the majority of HIV-1-infecetd individuals (56% and 70%), but not in their blood. Blood and semen are separated immunological compartments, and with HIV-1 infection the major changes in cytokine composition occur in semen (16 vs 9 cytokine levels altered in blood). CMV reactivation in the MGT was associated with increased levels of the cytokines CCL5, CCL11 and CXCL9. Analysis of the cytokines network revealed a higher number of correlations and increase in correlation strength for most of the cytokines in semen and blood of HIV-1-infected compared to HIV-uninfected individuals. Of note, interleukin (IL)-7 resulted to be one of the most concentrated cytokines in semen and HIV-1 infection further increased seminal IL-7. Thus we employed our system of human tissues ex vivo to study the effect of IL-7 on HIV-1 replication. IL-7 enhanced the replication of R5 and X4 HIV-1 variants in dose and time-dependent manner. In tissue blocks treated with 25 ng/mL of IL-7 we observed higher number of HIV-1 infected CD4+ T cells and reduced CD4+ T cell depletion, compared with tissue blocks infected and maintained in the absence of IL-7. The fraction of HIV-1 infected CD4+ T cells expressing the apoptotic marker APO2.7 was reduced and the levels of the antiapoptotic factor Bcl-2 in infected cells were increased in the presence of IL-7. Conclusions. HIV-1 infection is associated with a general alteration of the cytokine spectrum in semen, and with the local reactivation of CMV and EBV in the MGT, two phenomena that appear to be related and may affect each other. The reduced flexibility of the cytokine network may favors the reactivation of such latent infections. Among seminal cytokines, IL-7 exerts a protective effect on HIV-1-infected CD4+ T cells in the early stages of infection, preventing their death and thus promoting the establishment of a productive infection in the female genital tract. Understanding how other cytokines in concert with co-infecting pathogens found in semen affects HIV-1 sexual transmission will be object of further investigations

A telescope detection system for direct and high resolution spectrometry of intense neutron fields

A high energy- and spatial-resolution telescope detector was designed and constructed for neutron spectrometry of intense neutron fields. The detector is constituted by a plastic scintillator coupled to a monolithic silicon telescope (MST), in turn consisting of a DE and an E stage. The scintillator behaves as an “active” recoil-proton converter, since it measures the deposited energy of the recoil-protons generated across. The MST measures the residual energy of recoil-protons downstream of the converter and also discriminates recoil-protons from photons associated to the neutron field. The lay-out of the scintillator/MST system was optimized through an analytical model for selecting the angular range of the scattered protons. The use of unfolding techniques for reconstructing the neutron energy distribution was thus avoided with reasonable uncertainty (about 1.6% in neutron energy) and efficiency (of the order of 106 counts per unit neutron fluence). A semi-empirical procedure was also developed for correcting the non-linearity in light emission from the organic scintillator. The spectrometer was characterized with quasi-monoenergetic and continuous fields of neutrons generated at the CN Van De Graaff accelerator of the INFN-Legnaro National Laboratory, Italy, showing satisfactory agreement with literature data

Development of an OpenFOAM multiphysics solver for solid fission products transport in the Molten Salt Fast Reactor

The analysis of innovative reactor concepts such as the Molten Salt Fast Reactor (MSFR) requires the development of new modeling and simulation tools. In the case of the MSFR, the strong intrinsic coupling between thermal-hydraulics, neutronics and fuel chemistry has led to the adoption of the multiphysics approach as a state-of-the-art paradigm. One of the peculiar aspects of liquid-fuel reactors such as the MSFR is the mobility of fission products (FPs) in the reactor circuit. Some FP species appear in form of solid precipitates carried by the fuel flow and can deposit on reactor boundaries (e.g., heat exchangers), potentially representing design issues related to the degradation of heat exchange performance or radioactive hotspots. The integration of transport models for solid particles in multiphysics codes is therefore relevant for the prediction of deposited fractions. To this aim, we develop a multiphysics solver based on the OpenFOAM library to address the issue of solid fission products transport. Single-phase incompressible thermal hydraulics are coupled with neutron diffusion, and advection-diffusion-decay equations are implemented for fission products concentrations. Particle deposition and precipitation are considered as well. The developed solver is tested on two different MSFR application to showcase the capabilities of the solver in steady-state simulation and to investigate the role of precipitation and turbulence modeling in the determination of particle concentration distributions

1D modelling and preliminary analysis of the coupled DYNASTY–eDYNASTY natural circulation loop

In the continuous strive to improve the safety of current-generation and next-generation nuclear power plants, natural circulation can be used to design passive safety systems to remove the decay heat during the shutdown. The Molten Salt Fast Reactor (MSFR) is a peculiar type of Gen-IV nuclear facility, where the fluid fuel is homogeneously mixed with the coolant. This design leads to natural circulation in the presence of an internally distributed heat source during the shutdown. Furthermore, to shield the environment from the highly radioactive fuel, an intermediate loop between the primary and the secondary loops, able to operate in natural circulation, is required. To analyze the natural circulation with a distributed heat source and to study the natural circulation of coupled systems and the influence of the intermediate loop on the behaviour of the primary, Politecnico di Milano designed and built the DYNASTY-eDYNASTY facility. The two facilities are coupled with a double-pipe heat exchanger, which siphons heat from DYNASTY and delivers it to the eDYNASTY loop. This work focuses on modelling the coupled DYNASTY-eDYNASTY natural circulation loops using DYMOLA2023((R)), an integrated development environment based on the Modelica Object-Oriented a-causal simulation language. The 1D Modelica approach allows for building highly reusable and flexible models easing the design effort on a complex system such as the DYNASTY-eDYNASTY case without the need to rewrite the whole model from scratch. The coupled models were developed starting from the already-validated single DYNASTY model and the double-pipe heat exchanger coupling. The models were tested during the whole development process, studying the influence of the numerical integration algorithm on the simulation behaviour. A preliminary analysis of both the adiabatic and the heat loss models analyzed the effect of the secondary natural circulation loop on the behaviour of the DYNASTY loop. The simulation results showed that the eDYNASTY loop dampens the behaviour of the primary DYNASTY loop. Furthermore, a parametric analysis of the DYNASTY and the eDYNASTY coolers highlighted the influence of the cooling configuration on the facility's behaviour. Finally, the simulation results identified the most critical aspects of the models in preparation for an experimental comparison

Characterization of varroa destructor mites in Cuba using mitochondrial and nuclear markers

Varroa destructor has been present in Cuba since 1996, but without the use of acaricidal infestation rates remain at very low levels. The presence of Korean haplotype mites was described in 2007, but there is no information regarding the introgression of the less virulent Japanese haplotype that could account for a low pathogenicity of the mite. In this research, we carried out molecular characterization of Cuban Varroa mites through mitochondrial DNA and hypervariable nuclear loci. We applied an alternative RFLP tech-nique and found that all the analyzed samples corresponded to Korean haplotypes. We analyzed the three STRs loci VD112, VD114 and VD016, previously described as highly variable and found new alleles in all of them, with an absolute allele size very different to those reported worldwide. We also detected genic and genotypic differentiation be-tween samples from two nearby locations (P=0.08). We also tested a new RFLP method for mite haplotype discrimination with an intra-reaction positive control of digestion

Characterization of a Be(p,xn) neutron source for fission yields measurements

We report on measurements performed at The Svedberg Laboratory (TSL) to characterize a proton-neutron converter for independent fission yield studies at the IGISOL-JYFLTRAP facility (Jyv\"askyl\"a, Finland). A 30 MeV proton beam impinged on a 5 mm water-cooled Beryllium target. Two independent experimental techniques have been used to measure the neutron spectrum: a Time of Flight (TOF) system used to estimate the high-energy contribution, and a Bonner Sphere Spectrometer able to provide precise results from thermal energies up to 20 MeV. An overlap between the energy regions covered by the two systems will permit a cross-check of the results from the different techniques. In this paper, the measurement and analysis techniques will be presented together with some preliminary results.Comment: 3 pages, 3 figures, also submitted as proceedings of the International Conference on Nuclear Data for Science and Technology 201

Genetic analyses suggest burrow sharing by Río Negro tuco-tucos (Ctenomys rionegrensis)

Genetic analyses of kinship can generate important insights into social structure, particularly forspecies for which direct observations of social relationships are challenging. We used molecular markersto characterize the kin structure of a population of the Río Negro tuco-tuco (Ctenomys rionegrensis), asubterranean species of rodent that is rarely observed above ground. Previous research has revealed thatadults of this species engage in at least periodic episodes of burrow sharing, indicating thatC. rionegrensismaynot be strictly solitary. To explore the kin structure of this species, we used variability at 10 microsatellite locito determine if (1) adults and juveniles captured at the same burrow entrance were parents and o spring and(2) kinship among adults captured together di ered from that among randomly sampled pairs of individuals inour study population. Our analyses revealed that adults and juveniles captured together were not typicallyparents and o spring, suggesting potential mixing of litters among burrow systems. Relatedness among adultscaptured together did not di er from background levels of genetic similarity, providing no evidence thatspatial proximity was associated with kin structure. Collectively, our ndings support the hypothesis thatC. rionegrensisis not strictly solitary but instead engages in burrow sharing by adults and associated litters ofyoung.Los análisis genéticos de parentesco pueden generar importantes ideas sobre la estructura social, particularmente en especies donde las observaciones directas de las relaciones sociales son aún muy discutidas. Utilizamos marcadores moleculares para caracterizar la estructura de parentesco en una población de los tuco-tucos de Río Negro (Ctenomys rionegrensis), una especie de roedores subterráneos que rara vez se observa sobre la super cie. Investigaciones anteriores han revelado que los adultos de esta especie pueden compartir madrigueras, al menos por periodos, indicando que esta especie puede no ser estrictamente solitaria. Para explorar la estructura de parentesco de esta especie utilizamos la variabilidad presente en 10 loci de microsatélites para determinar si (1) adultos y juveniles capturados en la misma madriguera son padres e hijos y (2) el parentesco entre los adultos capturados en las mismas cuevas di ere del de pares de individuos muestreados al azar en la población estudiada. Nuestros análisis revelaron que los adultos y los juveniles capturados juntos típicamente no están emparentados, lo que sugiere una posible mezcla de camadas dentro del sistema de madrigueras. El parentesco entre los adultos capturados juntos, sin embargo, no di rió de los niveles medios de similitud genética, sin proporcionar evidencia de que la proximidad espacial estuviera asociada con la estructura de parentesco. En conjunto, nuestros hallazgos apoyan la hipótesis de que C. rionegrensis no es una especie estrictamente solitaria, sino que existe intercambio de madrigueras entre los adultos y sus camadas asociadas

Compact thermal neutron sensors for moderator-based neutron spectrometers

In the framework of the NESCOFI@BTF project of the Italian Institute of Nuclear Physics, different types of active thermal neutron sensors were studied by coupling semiconductor devices with a suitable radiator. The objective was to develop a detector of small dimensions with a proper sensitivity to use at different positions in a novel moderating assembly for neutron spectrometry. This work discusses the experimental activity carried out in the framework of the ERINDA program (PAC 3/9 2012) to characterise the performance of a thermal neutron pulse detector based on (6)Li

Definition of indicators of appropriateness in the management of neovascular age-related macular degeneration: An expert opinion

Wet age-related macular degeneration is a chronic condition culminating, in most cases, in blindness. The introduction of anti-angiogenic agents in 2006 has represented a major breakthrough in the treatment of the disease, but timely and effective treatment with regular follow-up and monitoring is mandatory to stabilize and preserve visual acuity. In clinical practice, however, appropriate therapy provision is frequently challenged by economic and organizational issues that result in suboptimal visual outcomes and increased incidence of legal blindness. International Guidelines have defined a diagnostic and therapeutic pathway to ensure the best practice in wet age-related macular degeneration management, but reference parameters to evaluate and compare the performance of Retina Centers are lacking. To address the appropriateness of wet age-related macular degeneration management in Italy, a multidisciplinary panel of ten experts gathered in three meetings. They defined three sets of indicators and relative benchmark values that each Center should comply with to ensure patients optimal care already from the first access: (a) clinical intervention indicators, to determine the possible Center\u2019s deviation from the diagnostic and therapeutic pathway; (b) outcome indicator, to evaluate the socioeconomic impact of the healthcare systems\u2019 performance; (c) management indicators, to test the size of the gap between the Center\u2019s supply and demand. Once the indicators have been analyzed, healthcare systems can plan actions to improve appropriateness and monitor their effects. However, to put this in practice, a concerted effort by all parts involved in healthcare provision is required, together with adequate systems to analyze clinical and administrative documentation