Liver cirrhosis in HIV/HCV ‐coinfected individuals is related to NK cell dysfunction and exhaustion, but not to an impaired NK cell modulation by CD 4 + T‐cells

Introduction HIV worsens HCV‐related liver disease by accelerating fibrosis progression; however, progression rates are extremely variable among HIV/HCV‐coinfected individuals. NK cells are associated with modulation of liver fibrosis and are profoundly altered during HCV and HIV infections. CD4+ T‐cells modulate NK cell function, and are also affected by HIV infection. Here, we aim to characterize the association of hepatic fibrosis with both the phenotype and function of peripheral NK cells and their regulation by CD4+ T‐cells, in HIV/HCV‐coinfected individuals. Methods Thirty‐four HIV/HCV‐coinfected individuals with minimal (n = 16) and advanced (n = 18) fibrosis (METAVIR F0/F1 and F4 scores respectively) and 20 healthy volunteers were enrolled. PBMC were obtained from peripheral blood samples and NK and CD4+ T‐cells were isolated and analysed. NK cell phenotype (CD25, CD69, Nkp46, NKG2D, PD‐1), degranulation (CD107a) and IFN‐γ and TNF‐α production, as well as CD4+ T‐cell activation (CD69, CD25 and CD38) were measured by flow cytometry. CD4+ T‐cell conditioned medium (CM) derived from F0/F1 or F4 individuals was assessed for IL‐2 levels by ELISA. Modulation of NK cell functionality by these CMs was also analysed. Results When comparing to NK cells from individuals with minimal fibrosis, degranulation and cytokine secretion by NK cells from subjects with F4 scores was significantly impaired, while PD‐1 expression was augmented. On the one hand, neither the expression of activation markers nor IL‐2 secretion was distinctly induced in CD4+ T‐cells from subjects with F0/F1 or F4 METAVIR scores. Finally, NK cell degranulation and cytokine secretion were not differentially modulated by CD4+ T‐cell CM, whether CD4+ T‐cells derived from subjects with minimal or advanced fibrosis. Conclusions Low levels of NK and CD4+ T‐cells in HIV/HCV‐coinfected individuals with advanced liver fibrosis have been previously described. Here, we show that advanced liver fibrosis in coinfected individuals is associated to a defective function of NK cells and an increased expression of the exhaustion/senescence marker PD‐1. This NK signature could not be attributed to changes in the ability of CD4+ T‐cells to modulate NK cell function.Fil: Polo, María L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina. Universidad de Buenos Aires; ArgentinaFil: Ghiglione, Yanina Alexandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Salido, Jimena Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Urioste, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Poblete, Gabriela. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Sisto, Alicia E. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Martinez, Ana. Gobierno de la Ciudad de Buenos Aires. Instituto Multidisciplinario de Investigaciones en Patologías Pediátricas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto Multidisciplinario de Investigaciones en Patologías Pediátricas; ArgentinaFil: Rolón, María Eugenia. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Ojeda, Diego Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Cahn, Pedro. Fundación Huésped; ArgentinaFil: Turk, Gabriela Julia Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina. Universidad de Buenos Aires; ArgentinaFil: Laufer, Natalia Lorna. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina. Universidad de Buenos Aires; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; Argentin

Light transmission measurement of solute dispersion in non-Brownian suspension flow

We characterize and employ a light transmission technique to measure the dispersivity of a solute in the flow of a neutrally-buoyant non-Brownian spherical particle suspension in a Hele-Shaw cell (parallelplate axial flow). Particle radii (a) were 20 and 40 μm, the particle bulk volume fraction ϕbulk was 0.2, and the cell aperture was 420 ± 10 μm. In each displacement experiment a suspension with a colouring solute displaces a transparent one at constant flow rates ranging from 0.721 to 0.928 mL/min (corresponding to solute Péclet numbers (Pes) between 350 and 450). A reference measurement, identical to the displacement ones but without particles in the flow (ϕbulk = 0), were performed in the same experimental assembly for comparison purposes. A light calibration related the transmitted intensity I to the solute concentration c for each combination of ϕbulk and a. The time variation of the solute concentration was found to be well-fitted by the solution of the advection-dispersion equation (ADE) in the range of Pes studied, and consequently a dispersion coefficient D and a dispersivity ld of the solute were measured

Interpopulation variation in nest architecture in a secondary cavity-nesting bird suggests site-specific strategies to cope with heat loss and humidity

© 2017 American Ornithological Society. Nest morphology can affect the breeding success of birds. Thus, birds inhabiting different environments may experience divergent selection for nest structure and composition that results in intraspecific geographic variation in nest architecture. We describe interpopulation differences in nest architecture among Thorn-tailed Rayaditos (Aphrastura spinicauda) in 2 contrasting environments near the species' distribution limits: a temperate and very humid environment in north-central Chile (the forest relicts of Fray Jorge National Park; 30°38′S, 71°40′W) and a cold and windy sub-Antarctic environment in the south of Chile (Isla Navarino; 55°4′S, 67°40′W). We collected a total of 62 nests from Fray Jorge and 61 nests from Navarino in 2013 and 2014, measured their dimensions, and quantified their constitutive materials. We tested the nests' thermal properties (simulating heat loss by convection and conduction) and hygroscopic features (water absorpt

Effects of temperature and time constraints on the seasonal variation in nest morphology of the Thorn tailed Rayadito (Aphrastura spinicauda)

Environmental adjustment is the most invoked explanation for intra-seasonal variation in bird nest morphology. However, time constraints may also be important, coming as a trade-off between the costs of nest building and the requirement to coincide a breeding attempt with maximal food supply. We describe the seasonal variation in nest morphology of the Thorn-tailed Rayadito (Aphrastura spinicauda) in a sub-Antarctic population in southern Chile, and investigate its relation with the seasonal fluctuation of ambient temperature (an 'environmental adjustment' hypothesis) and time constraints (an 'optimal time frame' hypothesis). As the breeding season progressed, rayaditos spent fewer days building their nests, built smaller nests, and used less animal-derived insulating material. After statistically removing the effects of daily temperatures on nest building periods and nest morphological measurements, we observed no seasonal trend in the amount of insulating material used, supporting an 'environmental adjustment' explanation. However, the nest building periods, nest depth, and nest dry weight still showed a seasonal trend, favouring an 'optimal time frame' hypothesis. Our study shows that both temperature fluctuations and time constraints can affect different components of the nesting ecology of birds, and that nest morphology is the consequence of distinct non-mutually exclusive forces.FONDECYT 1100359 1140548 ICM-P05-002 CONICYT-Chile PFB-23 63130100 21130127 COLFUTURO PCB-201

Intraspecific variation in exploratory behavior and elevational affinity in a widely distributed songbird

Populations of the same species can vary substantially in their behavioral and morphometric traits when they are subject to different environmental pressures, which may lead to the development of different adaptive strategies. We quantified variation in exploratory behavior and morphometric traits among two rufous-collared sparrow populations that occur at low and high elevations in central Chile. Moreover, we used census and delta H-2 values of feather and blood to evaluate migration. We found that individual sparrows inhabiting high elevations were larger and showed more intense exploratory behavior in comparison with those that were captured at lower elevation. Moreover, we observed a steady decline in sparrow abundance during the winter and similar delta H-2 values for blood collected in the winter and summer at this site, which were significantly lower than blood delta H-2 values observed at low elevation. This pattern suggests that individuals do not move long distances during winter, and likely they remain at similar elevations in refuge habitats. As predicted, our results support the existent of different adaptive strategies among populations of the same species, and suggest that the combination of behavioral, morphometric, and stable isotope data is a novel and robust integrative approach to assess differences in adaptation across environmental gradients.FONDECYT-Chile 1140548 1160115 Institute of Ecology and Biodiversity ICM-P05-002-Chile PFB-23-CONICYT-Chile Fondo Basal FB 0002-2014 21130127 CONICYT-Chil

Deforestation patterns shape population structure of the Magellanic Woodpecker (Campephilus magellanicus) in southern Chile

One important landscape-scale consequence of deforestation is reduced connectivity, which has the potential to isolate populations in ways that affect genetic diversity and population structure. Among the many regions of the world where this scenario has played out is the South American temperate forest (SATF) in southern Chile, and there is now strong concern about the population viability of forest taxa. We studied one such species, the Magellanic Woodpecker (Campephilus magellanicus), a forest specialist that is now listed as vulnerable in parts of its range in Chile. We characterized genetic variability and population structure from 33 samples of Magellanic Woodpeckers from two large but isolated populations in Nahuelbuta National Park in the Coastal mountain range and the Conguillío National Park in the Andes using ddRAD-seq method. We found lower genetic variability in Nahuelbuta than Conguillío, but inbreeding values (FIS) did not show evidence for inbreeding depression. Results suggest the presence of two genetic clusters, with an average FST value of 0.04. Phylogenetic analysis showed that the Nahuelbuta population forms a clade that is nested within the individuals from Conguillío, suggesting limited gene flow between these populations. Our results support the idea that extensive deforestation has played a role in shaping the genetic patterns that we have identified. Because of this, we emphasize the need for regional planning to increase the structural connectivity between fragments of mature native forests, to provide an opportunity for the persistence of Magellanic Woodpeckers in this region

Markers of Natural Killer Cell Exhaustion in HIV/HCV Coinfection and Their Dynamics After HCV Clearance Mediated by Direct-Acting Antivirals

Background. Liver fibrosis is a leading cause of morbimortality in people with HIV/hepatitis C virus (HCV). Natural killer (NK) cells are linked with amelioration of liver fibrosis; however, NK cells from individuals coinfected with HIV/HCV with cirrhosis display impaired functionality and high PD-1 expression. Here, we aimed to study PD-1, TIGIT, and Tim3 as potential exhaustion markers in NK cells from persons coinfected with HIV/HCV with mild and advanced liver fibrosis. We also evaluated the role of PD-1 expression on NK cells after HCV clearance by direct-acting antivirals (DAAs). Methods. Peripheral blood mononuclear cells were isolated from individuals coinfected with HIV/HCV (N = 54; METAVIR F0/F1, n = 27; F4, evaluated by transient elastography, n = 27). In 26 participants, samples were collected before, at the end of, and 12 months after successful DAA treatment. The frequency, immunophenotype (PD-1, TIGIT, and Tim3 expression), and degranulation capacity (CD107a assay) of NK cells were determined by flow cytometry. Results. Unlike PD-1, Tim3 and TIGIT were comparably expressed between persons with mild and advanced fibrosis. Degranulation capacity was diminished in NK/TIGIT+ cells in both fibrosis stages, while NK/PD-1+ cells showed a lower CD107a expression in cirrhotic cases. Twelve months after DAA treatment, those with advanced fibrosis showed an improved NK cell frequency and reduced NK/PD-1+ cell frequency but no changes in CD107a expression. In individuals with mild fibrosis, neither PD-1 nor NK cell frequency was modified, although the percentage of NK/CD107a+ cells was improved at 12 months posttreatment. Conclusions. Although DAA improved exhaustion and frequency of NK cells in cirrhotic cases, functionality was reverted only in mild liver fibrosis, remarking the importance of an early DAA treatment.Fil: Osegueda, Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Polo, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Baquero, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Urioste, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Ghiglione, Yanina Alexandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Paz, Silvia. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; ArgentinaFil: Poblete, Gabriela. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; ArgentinaFil: Gonzalez Polo, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Turk, Gabriela Julia Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Quiroga, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; ArgentinaFil: Laufer, Natalia Lorna. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentin