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These days, the gold standard procedure for otosclerosis treatment is stapes surgery. The endoscopic approach of the procedure is gaining popularity as endoscopic ear surgery develops across the globe. The main objective of this study is to gather and compile well-documented and reliable data regarding surgical outcomes for the endoscopic approach to stapes surgery up to this date. Publications in English were searched in the PUBMED/MEDLINE database and were systematically reviewed. A total of 16 articles were reviewed according to the inclusion criteria, obtaining a total of 573 patients managed surgically for otosclerosis, using an endoscopic approach. Data were systematically extracted and compared across variables. Data were obtained as follows: mean age of 43 years; female proportion of 60%; 3 mm endoscope diameter of 51%, 4 mm of 39%; titanium piston-type prostheses of 52% and Teflon of 48%; length of the prosthesis (mode) was 4.5 mm; 0.6 mm diameter of the piston of 81% and 0.4 mm of 19%; mean surgical time was 55 min. Hearing results, mean preoperative air–bone gap (ABG) 31 dB; mean postoperative ABG 9 dB; ABG improvement of 22 dB; an ABG closure rate to 20 dB or less of 92% and an ABG closure rate to 10 dB or less of 77%. Complication rates: intraoperative tympanic membrane perforation of 5%; postoperative vertigo of 11%; postoperative dysgeusia of 10%; reported a postoperative neurosensorial hearing loss of 0.2%; reported gusher phenomenon of one case (0.2%). Endoscopic stapes surgery is completely achievable using 0o angle and 4-mm-diameter sinus surgery endoscope. Instrument availability should not be an obstacle to the development of this type of surgery in any otolaryngology department. Audiological outcomes are comparable to microscopic approaches.

Objectives: To determine the time to reverse transcription-polymerase chain reaction (RT-PCR) negativity after the first positive RT-PCR test, factors associated with longer time to RT-PCR negativity, proportion of children seroconverting after proven severe acute respiratory syndrome coronavirus 2 infection, and factors associated with the lack of seroconversion. Study design: The Epidemiological Study of Coronavirus in Children of the Spanish Society of Pediatrics is a multicenter study conducted in Spanish children to assess the characteristics of coronavirus disease 2019. In a subset of patients, 3 serial RT-PCR tests on nasopharyngeal swab specimens were performed after the first RT-PCR test, and immunoglobulin G serology for severe acute respiratory syndrome coronavirus 2 antibodies was performed in the acute and follow-up (

Objective: To describe the capacity of a broad spectrum of cytokines and growth factors to predict ICU admission and/or death in patients with severe COVID-19. Design: An observational, analytical, retrospective cohort study with longitudinal follow-up. Setting: Hospital Universitario Príncipe de Asturias (HUPA). Participants: 287 patients diagnosed with COVID-19 admitted to our hospital from 24 March to 8 May 2020, followed until 31 August 2020. Main outcome measures: Profiles of immune response (IR) mediators were determined using the Luminex Multiplex technique in hospitalized patients within six days of admission by examining serum levels of 62 soluble molecules classified into the three groups: adaptive IR-related cytokines (n = 19), innate inflammatory IR-related cytokines (n = 27), and growth factors (n = 16). Results: A statistically robust link with ICU admission and/or death was detected for increased serum levels of interleukin (IL)-6, IL-15, soluble (s) RAGE, IP10, MCP3, sIL1RII, IL-8, GCSF and MCSF and IL-10. The greatest prognostic value was observed for the marker combination IL-10, IL-6 and GCSF. Conclusions: When severe COVID-19 progresses to ICU admission and/or death there is a marked increase in serum levels of several cytokines and chemokines, mainly related to the patient’s inflammatory IR. Serum levels of IL-10, IL-6 and GCSF were most prognostic of the outcome measure., This research was coordinated by ProA Capital and Startlite Foundation, Programa de Actividades de I+D de la Comunidad de Madrid en Biomedicina (B2020/MITICAD-CM).

Territorial behaviour arises as a strategy of ensuring individuals’ access to a variety of potentially limiting resources. While aggressiveness is a well-studied widespread trait across taxa, the mechanisms that allow for a range of aggressive phenotypes to coexist in the wild remains unclear. In this study, we analyse environmental, social and intrinsic variables that can modulate the expression of different strategies of male–male aggressiveness. Furthermore, through network analysis we explore the role of this trait in the establishment of territories during the breeding season as the intensity of different aggressiveness strategies may limit or grant access to resources. Simulating territorial intrusions during the early incubation period, we assessed the aggressiveness of breeding male blue tits (Cyanistes caeruleus). We defined three types of conspecific aggressiveness (nonconfrontational intimidating, nonconfrontational cautious and confrontational) and analysed the effect of habitat structure, territory quality, presence of other breeding species and male condition on the type and intensity of the aggressive display. The results obtained suggest that yearling males rely on intimidating behaviour more than older males, that perform more cautious displays. Furthermore, smaller and heavier males opted for confrontational strategies. The density and nature of neighbours, as well as the territory quality and the habitat structure, also conditioned the intensity and type of display. Surprisingly, the network analysis revealed that the intensity of male–male aggressive displays did not condition the establishment of breeding territories. Our results suggest that aggressiveness is a context-specific trait shaped by a complex array of environmental and intrinsic parameters.

According to current theory, the development of different conspicuous ornaments may provide multiple, redundant or unreliable information. However, multiple visual and acoustic traits have not often been tested at the same time. In this study, we analyse whether black tie size, yellow breast colour indexes (chroma and hue) and song parameters (repertoire size, strophe length, maximum and minimum frequency) are inter-correlated in male great tits (Parus major). We found that males with larger black ties sang greater repertoires and had higher song frequencies, providing redundant information. However, song parameters did not correlate with hue and chroma of the yellow breast coloration, supporting the idea that they convey different units of information. Results therefore stress that since different plumaged patches can transmit very different units of information, bird song will correlate to plumage coloration traits depending on the patch and the colour considered, an aspect rarely taken into account in previous studies relating song to plumage colour traits., This work was supported by funds from the Ministry of Economy and Competitivity, Spanish Research Council to E.S.F. and J.J.S. (CGL-2016-79568-C3-1-P) and to J.C.S. and F.U. (CGL-2016-79568-C3-3-P).

Ambient temperature is assumed to be the major cue used by passerines to synchronize their laying and hatching dates to the expected peak of prey availability. While laying eggs, females are still able to fine-tune their hatching date following increasing or decreasing patterns of ambient temperature, mostly via changes in incubation onse t. The onset of incubation behaviour in relation to the laying sequence could have later conse- quences for the duration of the incubation period and the extent of hatching asynchrony. Clutch size is also known to affect incubation patterns and might therefore condition potential responses to changing temperatures. In this study we assessed the association of ambient temperature and clutch size with the onset of four different incubation behaviours: partial and full nocturnal incubation, and partial and full diurnal incubation. We also evaluated how the onset and duration of each incubation behaviour might predict the duration of diurnal full incubation and the extent of hatching asynchrony. To achieve our aims, we monitored incubation behaviour using temperature data loggers during the egg-laying period in three Mediterranean Great Tit Parus major populations in three consecutive years. Our results showed that increasing temperatures were related to an advance of diurnal partial incubation, but not its duration or the onset of full incubation behaviour. We did not nd any effect of ambient temperatures on nocturnal incubation. However, females lengthened nocturnal partial incubation and delayed the onset of nocturnal full and diurnal partial incubation when laying larger clutches. Longer diurnal incubation before clutch completion was associated with greater hatching asynchrony. Moreover, longer diurnal partial incubation shortened the duration of the full incubation period. In conclusion, increasing ambient temperatures during the egg-laying period advanced diurnal partial incubation, indirectly shortening the full incubation period and increasing hatching asynchrony., This study was supported by projects CGL2013-48001-C2-1-P and CGL2016-79568-C3-1-P (Spanish Ministry of Economy and Competitiveness, MINECO). D.D.-M. benefitted from an FPI fellowship (BES-2014-069191) provided by the MINECO.

Avian embryos need a stable thermal environment to develop optimally, while incubat-ing females need to allocate time to self-maintenance off the nest. In species with female-only incubation, eggs are exposed to ambient temperatures that usually cool them down during female absences. The lower the ambient temperature the sooner females should return to re-warm the eggs. When incubation constraints ease at increasing ambient temperatures, females respond by increasing either incubation effort or self-maintenance time. These responses are population-dependent even within the same species; but it is uncertain whether they are caused by local environmental conditions or they are an artefact from limited datasets, different methodological approaches or the timescale over which incubation behaviour is measured. In this study, we collected incubation data from three Mediterranean great tit Parus major populations during three consecutive years. We measured the duration of each off- and on-bout event, used these variables to compute nest attentiveness at three different timescales (full incubation, daily and hourly periods) and assessed the impact of ambient temperature on bout duration and nest attentiveness. We found that females maximized on-bout duration at different local temperatures, ranging from 10 to 20°C; but lengthened off-bouts linearly across a range of 0–38°C in all three populations. These local differences translated into opposite linear nest attentiveness patterns at the full incubation scale: Females increased either incubation effort, longest on-bouts between 15 and 20°C or self-maintenance time, longest on-bouts at 10°C. It was at daily and hourly periods when we detected non-linear nest attentiveness patterns, as expected from on-bout duration, peaking at different local ambient temperatures. Females first increased incu-bation effort up to a certain temperature value and then increased self-maintenance time at the highest ambient temperatures. Further research is needed to understand which factors are behind the turning points from one behaviour to the other., This study was supported by projects CGL2013-48001-C2-1-P and CGL2016-79568-C3-1-P (Spanish Ministry of Economy and Competitiveness, MINECO). DD-M benefitted from an FPI fellowship (BES-2014-069191) and a short-stay research grant (EEBB-I-16-11668) provided by the MINECO.

This study aimed to create an individualized analysis model of the risk of intensive care unit (ICU) admission or death for coronavirus disease 2019 (COVID-19) patients as a tool for the rapid clinical management of hospitalized patients in order to achieve a resilience of medical resources. This is an observational, analytical, retrospective cohort study with longitudinal follow-up. Data were collected from the medical records of 3489 patients diagnosed with COVID-19 using RT-qPCR in the period of highest community transmission recorded in Europe to date: February&ndash, June 2020. The study was carried out in in two health areas of hospital care in the Madrid region: the central area of the Madrid capital (Hospitales de Madrid del Grupo HM Hospitales (CH-HM), n = 1931) and the metropolitan area of Madrid (Hospital Universitario Pr&iacute, ncipe de Asturias (MH-HUPA) n = 1558). By using a regression model, we observed how the different patient variables had unequal importance. Among all the analyzed variables, basal oxygen saturation was found to have the highest relative importance with a value of 20.3%, followed by age (17.7%), lymphocyte/leukocyte ratio (14.4%), CRP value (12.5%), comorbidities (12.5%), and leukocyte count (8.9%). Three levels of risk of ICU/death were established: low-risk level (&lt, 5%), medium-risk level (5&ndash, 20%), and high-risk level (&gt, 20%). At the high-risk level, 13% needed ICU admission, 29% died, and 37% had an ICU&ndash, death outcome. This predictive model allowed us to individualize the risk for worse outcome for hospitalized patients affected by COVID-19.

Background Angiogenesis participates in re-establishing microcirculation after myocardial infarction (MI). Purpose In this study, we aim to further understand the role of the anti-angiogenic isoform vascular endothelial growth factor (VEGF)-A165b after MI and explore its potential as a co-adjuvant therapy to coronary reperfusion. Methods Two mice MI models were formed: 1) permanent coronary ligation (non-reperfused MI), 2) transient 45-min coronary occlusion followed by reperfusion (reperfused MI); in both models, animals underwent echocardiography before euthanasia at day 21 after MI induction. Serum and myocardial VEGF-A165b levels were determined. In both experimental MI models, functional and structural implication of VEGF-A165b blockade was assessed. In a cohort of 104 ST-segment elevation MI patients, circulating VEGF-A165b levels were correlated with cardiovascular magnetic resonance-derived left ventricular ejection fraction at 6-months and with the occurrence of adverse events (death, heart failure and/or re-infarction). Results In both models, circulating and myocardial VEGF-A165b presence was increased 21 days after MI induction. Serum VEGF-A165b levels inversely correlated with systolic function evaluated by echocardiography. VEGF-A165b blockage increased capillary density, reduced infarct size, and enhanced left ventricular function in reperfused, but not in non-reperfused MI experiments. In patients, higher VEGF-A165b levels correlated with depressed ejection fraction and worse outcomes. Conclusions In experimental and clinical studies, higher serum VEGF-A165b levels associates with a worse systolic function. Its blockage enhances neoangiogenesis, reduces infarct size, and increases ejection fraction in reperfused, but not in non-reperfused MI experiments. Therefore, VEGF-A165b neutralization represents a potential co-adjuvant therapy to coronary reperfusion. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): This study was funded by “Instituto de Salud Carlos III” and “Fondos Europeos de Desarrollo Regional FEDER” (Exp. PIE15/00013, PI17/01836, PI18/00209 and CIBERCV16/11/00486).

The expression of the gene Slc7a11 promotes the antioxidant capacity of cells by providing them with cysteine that can be used for the synthesis of glutathione (GSH), the most important intracellular antioxidant. In melanocytes, intracellular cysteine can also enter melanosomes and get incorporated in the pheomelanin pigment synthesis pathway, thus decreasing cysteine availability for GSH synthesis and potentially creating chronic oxidative stress. We thus hypothesized that a mechanism limiting the use of intramelanocytic cysteine for pheomelanin synthesis in environmental conditions generating oxidative stress may be physiologically advantageous and favored by natural selection. Here we searched for evidence of such a mechanism by comparing the influence of melanocytic Slc7a11 expression on pheomelanin‐based pigmentation in developing Eurasian nuthatch Sitta europaea nestlings from two populations differing in predation risk, a natural source of oxidative stress. Pheomelanin synthesis and pigmentation tended to increase with Slc7a11 expression in the low‐risk population as expected from the activity of this gene, but decreased with Slc7a11 expression in the high‐risk population. The same was not observed in the expression of five other genes influencing pheomelanin synthesis without affecting cysteine availability in melanocytes. The influence of body condition on the intensity of pheomelanin‐based pigmentation also differed between populations, being positive in the low‐risk population and negative in the high‐risk population. The resulting pigmentation of birds was more intense in the high‐risk population. These findings suggest that birds perceiving high predation risk may limit the use of cysteine for pheomelanin synthesis, which becomes independent of Slc7a11 expression. Some birds may have thus evolved the ability to adjust their pigmentation phenotype to environmental stress.

Birds are hosts for several zoonotic pathogens. Because of their high mobility, especially of longdistance migrants, birds can disperse these pathogens, affecting their distribution and phylogeography. We focused on Borrelia burgdorferi sensu lato, which includes the causative agents of Lyme borreliosis, as an example for tick-borne pathogens, to address the role of birds as propagation hosts of zoonotic agents at a large geographical scale. We collected ticks from passerine birds in 11 European countries. B. burgdorferi s.l. prevalence in Ixodes spp. was 37% and increased with latitude. The fieldfare Turdus pilaris and the blackbird T. merula carried ticks with the highest Borrelia prevalence (92 and 58%, respectively), whereas robin Erithacus rubecula ticks were the least infected (3.8%). Borrelia garinii was the most prevalent genospecies (61%), followed by B. valaisiana (24%), B. afzelii (9%), B. turdi (5%) and B. lusitaniae (0.5%). A novel Borrelia genospecies “Candidatus Borrelia aligera” was also detected. Multilocus sequence typing (MLST) analysis of B. garinii isolates together with the global collection of B. garinii genotypes obtained from the Borrelia MLST public database revealed that: (a) there was little overlap among genotypes from different continents, (b) there was no geographical structuring within Europe, and (c) there was no evident association pattern detectable among B. garinii genotypes from ticks feeding on birds, questing ticks or human isolates. These findings strengthen the hypothesis that the population structure and evolutionary biology of tick-borne pathogens are shaped by their host associations and the movement patterns of these hosts., This study received financial support from Fundação para a Ciência e a Tecnologia by the strategic program of MARE (MARE ‐ UID/MAR/04292/2013) and the fellowship to Ana Cláudia Norte (SFRH/BPD/108197/2015), and the Portuguese National Institute of Health. Raivo Mänd, Tomi Trilar, Tapio Eeva, Tomas Grim and Dieter Heylen were supported by the Estonian Research Council (research grant # IUT34‐8), the Slovenian Research Agency ‐programme “Communities, relations and communications in the ecosystems” (No. P1‐0255), the Academy of Finland (project 265859), the Internal Grant Agency of Palacky University (PrF_2014_018, PrF_2015_018, PrF_2013_018) and the Marie Sklodowska‐Curie Actions (EU‐Horizon 2020, Individual Global Fellowship, project no 799609), respectively.

The incubation stage in avian reproduction could be as costly as the nestling rearing stage. This is particularly true in the case of uniparental incubation, during which both current and future breeding attempts may be compromised. Therefore, the knowledge of the proximate effects that condition the incubation behaviour in free-living bird populations is of great importance in understanding the evolution of avian life history. In this two-year study, we assessed the incubation behaviours of Blue Cyanistes caeruleus and Great Tits Parus major inhabiting the same Mediterranean area in central Spain through the usage of iButton data loggers. It showed that the incubating behaviour of our tit populations resembles that reported in previous studies, but with peculiarities related to living at lower latitudes, i.e. with a relatively low attentiveness and a shorter active day. Both tit species showed very different incubation strategies, with Blue Tits leaving more frequently the nest (Mean ± SE number of off-bouts, Blue Tit = 27.14 ± 0.63, Great Tit = 16.95 ± 0.58) but for shorter periods than Great Tits (off-bout duration, Blue Tit = 8.76 ± 0.22 min, Great Tit = 14.04 ± 0.56 min; on-bout duration, Blue Tit = 22.63 ± 0.60 min, Great Tit = 36.86 ± 0.86 min). Nonetheless, both species provided a similar nest attentiveness, percentage of time of the active day during which the females were actively incubating (Blue Tit = 70.87 ± 0.57%, Great Tit = 70.75 ± 0.83%). Presumably, differences in the cooling rate of clutches, estimated with the iButtons, could be behind the differences in incubation behaviour between species and the greater capacity of Great Tits to adjust their incubation behaviour., Funding was provided by Ministerio de Economía y Competitividad (CLG2013-48001-C2-1P) and Junta de Comunidades de Castilla-La Mancha and European Social Fund (POIC10-0269- 7632). JBE is supported by a doctoral scholarship from the Junta de Comunidades de Castilla-La Mancha-European Social Fund and RB is a part of the JCCM-FSE 2007/2013 postdoctoral program and has a “Juan de la Cierva” post-doctoral contract (JCI-2011-10945).

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41-6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain. Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000-2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively. Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p < 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 279 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (t(MRCA)) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999-2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin. Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics., This work was supported by Plan Nacional de I + D + I and Fondo Europeo de Desarrollo Regional-FEDER (RD16/0025/0040 and PI18/00819) and Fundacion Progreso y Salud, Junta de Andalucia (PI-0550-2017). The study was in part funded by the Hellenic Scientific Society for the Study of AIDS and Sexually Transmitted Diseases.

Spanish Pediatric TB Research Network (pTBred) and the European NontuberculouS MycoBacterial Lymphadenitis in childrEn (ENSeMBLE) Study., [Objectives] To assess the performance of interferon-gamma release assays (IGRAs) in the differential diagnosis between Mycobacterium avium complex (MAC) and tuberculosis (TB) in children affected with subacute/chronic submandibular/cervical lymphadenitis., [Study design] Multicenter observational study comparing children with microbiologically confirmed MAC lymphadenitis from the European NontuberculouS MycoBacterial Lymphadenitis in childrEn study with children with TB lymphadenitis from the Spanish Network for the Study of Pediatric TB database., [Results] Overall, 78 patients with MAC and 34 with TB lymphadenitis were included. Among MAC cases, 44 out of 74 (59.5%) had positive tuberculin skin test (TST) results at the 5-mm cut-off, compared with 32 out of 33 (97%) TB cases (P < .001); at the 10-mm cut-off TST results were positive in 23 out of 74 (31.1%) vs 26 out of 31 (83.9%), respectively (P < .001). IGRA results were positive in only 1 out of 32 (3.1%) patients with MAC who had undergone IGRA testing, compared with 21 out of 23 (91.3%) TB cases (P < .001). Agreement between TST and IGRA results was poor in MAC (23.3%; κ = 0.017), but good in TB cases (95.6%; κ = 0.646). IGRAs had a specificity of 96.9% (95% CI 84.3%-99.8%), positive predictive value of 95.4% (95% CI 78.2%-99.8%), and negative predictive value of 93.9% (95% CI 80.4%-98.9%) for TB lymphadenitis., [Conclusions] In contrast to TST, IGRAs have high specificity, negative predictive value, and positive predictive value for TB lymphadenitis in children with subacute/chronic lymphadenopathy, and consequently can help to discriminate between TB and MAC disease. Therefore, IGRAs are useful tools in the diagnostic work-up of children with lymphadenopathy, particularly when culture and polymerase chain reaction results are negative.

Capsule: The structure of Great Tit Parus major songs is shaped by the acoustic properties of the habitat within the breeding territory of individuals. Aim: To test whether the structure of the habitat influences song structure within a population of Great Tits P. major. Methods: We recorded Great Tit songs from 42 territories on two different days and measured the habitat structure in each territory. We also trapped the males and estimated the breeding density around each territory, so were able to control the analysis by date, breeding density and male characteristics. Results: Song pause length was positively affected by the ground cover, while the song rate and the minimum frequency were negatively affected by the shrub cover. Male size negatively affected the peak frequency of the songs, whereas the age of the males affected the frequency range; older males sang with a broader bandwidth. Conclusion: This study suggests that Great Tits are capable of adjusting their vocalizations in each territory, presumably to enhance transmission owing to vocal plasticity., This study was funded by the [grant number POIC10-0269-7632] (Junta de Comunidades de Castilla-La Mancha-European Social Fund) and the [grant number GCL2010-21933-C02-01] (MINECO).

In patchy forest areas, the size of the forest patch where birds breed has a strong influence on their breeding success. However, the proximate effects contributing to lowering the breeding success in small forest patches remain unclear; and a shortage of crucial resources in those forest patches has been suggested to account in some degree for this failure. With the aim to further investigate this issue, we have monitored the breeding cycle of blue and great tits in three ‘large’ forest patches (ranging between 26.5-29.6 ha) and twelve ‘small’ forest patches (ranging between 1.1-2.1 ha) in a Mediterranean area in central Spain, during three years (2011-2013). We also recorded the nestling diet inside the nest-boxes with the aid of handy-cams. Only males significantly differed between forest patch size categories; being on average younger and with better body condition in small patches for great and blue tits respectively. Reproductive traits did not vary between forest patch size categories, but the body condition of blue tit nestlings and the size of great tit nestlings did, being significantly better and larger respectively in large forest patches. The recruitment rate of blue tit nestlings was also higher in large patches. Regarding nestling diet, blue tits did not differ but great tits did, delivering a larger amount of caterpillars in large forest patches. Most variation in the reproductive traits occurred between years, probably due to annual differences in environmental conditions. This study suggests that food supply could be limiting the breeding success of birds above all in small patches, but also in large patches under particular environmental conditions., Funding was provided by Ministerio de Ciencia e Innovación (CGL2010-21933-C02-01) and Junta de Comunidades de Castilla-La Mancha and European Social Fund (POIC10-0269-7632). ESF and JBE are both supported by a doctoral scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund. RB benefited from the JCCM-FSE 2007/2013 postdoctoral program and from a “Juan de la Cierva” post-doctoral contract (JCI-2011-10945) and ESD enjoyed a pre-doctoral fellowship from Ministerio de Ciencia e Innovación.

Hepatitis E virus (HEV) has emerged as a relevant pathogen for HIV-infected patients. However, there is scarce data on HEV infection in HIV/HCV-coinfected individuals with advanced fibrosis, which seems to increase the risk of HEV infection and worsen the prognosis of liver disease. We aimed to determine the prevalence of anti-HEV antibodies, acute hepatitis E, resolved hepatitis E, and exposure to HEV in HIV/HCV-coinfected patients and to evaluate associations with clinical and epidemiological characteristics. We performed a cross-sectional study on 198 HIV/HCV-coinfected patients, 30 healthy controls and 36 HIV-monoinfected patients. We found a low concordance between techniques used for detection of anti-HEV antibodies (ELISA versus Immunoblot), particularly in HIV/HCV-coinfected patients. HIV/HCV-coinfected patients showed the highest prevalence of IgG against HEV, resolved hepatitis E, and exposure to HEV (19.2%, 17.2%, and 22.2% respectively). However, we did not find any samples positive for HEV-RNA nor significant differences between groups. Moreover, HIV/HCV-coinfected patients with CD4 T-cells

Certain populations of long‐distance migratory birds are suffering declines, which may be attributed to effects of climate change. In this article, we have analysed a long‐term (1991–2015) data set on a pied flycatcher Ficedula hypoleuca population breeding in nest‐boxes in a Mediterranean montane oak forest, exploring the trends in population size due to changes in nestling recruitment, female survival and female immigration. We have related these changes in population parameters to local climate, winter NAO index and to breeding density. During the last 25 yr the population has declined by half, mainly in association with a decrease in nestling mass and structural size which had repercussions on the probability of nestling recruitment to the population. Lower local nestling recruitment in certain years was linked to lower female immigration rate in the same years. On the other hand, the local survival of females remained stable throughout the study period. Laying date and breeding success were negatively affected by local temperatures while breeding, recruitment rate likewise by minimum temperature prior to breeding in April. As minimum April temperatures have increased across the study period, this may have affected recruitment and immigration rates negatively. On the other hand, tarsus length and body mass of nestlings were positively associated with winter NAO index, pointing to more global climatic links. Moreover, there was also a negative temporal trend in body mass of adults, implying increasingly difficult conditions for breeding. Declining recruit production in the study area could be attributed to a mismatch between the timing of arrival and breeding in the population, and the peak of food availability in this area., Data collection was supported by different projects of the Spanish DGICYT, Ministerio de Ciencia e Innovación and at writing by project CGL2013-48193-C3-3-P to JM from Ministerio de Economía y Competitividad (MINECO). SG-B received a contract of the Museo Nacional de Ciencias Naturales-CSIC with charge to project CGL2010-19233-C03-02 by Ministerio de Ciencia e Innovación (MICINN) while writing this manuscript.

Multiple mating to obtain genetic benefits has been championed as the most likely cause of the evolution of polygamy. However, this assumption has been put into question by an increasing number of recent studies, most of which highlight the importance of considering ecological constraints to comprehend variation in extrapair (EP) behaviour. Here, we studied patterns of extrapair paternity (EPP) in the great tit, Parus major, using data from 11 nestbox plots that differed in population size and breeding conditions. Specifically, we analysed EPP rates in relation to socioecological variables that could influence the way individuals encountered one another in space and time, we tested whether adults engaged in EPP with more heterozygous, more compatible or phenotypically superior individuals than their social mates and we analysed whether extrapair offspring (EPO) were phenotypically or genetically superior to within-pair offspring. Our results do not provide support for the genetic benefit hypothesis from either the male or the female perspective. EPO were heavier than their within-pair paternal half-siblings, but there was no significant difference between EPO and their within-pair maternal half-siblings in terms of phenotypic quality. Regarding socioecological factors, we found a negative relationship between breeding synchrony and EPP rates both within and among plots, which suggests that males face a trade-off between mate guarding and obtaining EPP elsewhere. Our results show that most males engaged in EPP after the fertile period of their social female despite having to travel long distances: about half of the detected cases of EPP involved individuals from different woodlands. This study indicates that when and where to engage in EPP seem to be more relevant factors than with whom to do it and highlights the importance of considering spatiotemporal constraints at a landscape scale to achieve a better understanding of variation in EP mating behaviour.

Hypermethylation of the promoter region of tumor suppressor genes is associated with carcinogenesis in lung cancer (LC). Endobronchial ultrasound with needle aspiration (EBUS-NA) is a semi-invasive method for obtaining cell blocks from lymph nodes, which can be used for epigenetic analyses. To establish the relationship between methylation status of p16, DAPK, RASSF1a, APC and CDH13 genes in lymph nodes sampled by EBUS-NA, tumor staging and prognosis. Methylation status of DAPK, p16, RASSF1a, APC and CDH13 genes was assessed in EBUS-NA cell blocks from LC patients and related to stage and survival. Eighty-five consecutive patients [mean age 67 (SD 8)] were included. Methylation of ≥1 gene was found in 43 malignant nodes (67 %). A higher prevalence of RASSF1a methylation was observed in small cell lung cancer patients [9/10 (90 %) vs. 15/53 (28 %); p < 0.001 χ(2) test]. Methylation of APC and/or p16 was related to advanced staging in non-small cell lung cancer (NSCLC) [15/29 (52 %) vs. 6/24 (25 %), p = 0.048, χ(2) test]. Patients with NSCLC showing methylation of APC and/or p16 had also lower 6-month survival (p = 0.019, log rank test), which persisted after adjustment for age and subtyping (HR = 6, 95 % CI [1.8-19.5], p = 0.003, Cox regression). Epigenetic analyses are feasible in EBUS-NA cell blocks and may identify methylation patterns associated with worse prognosis. Methylation of p16 and APC genes in NSCLC patients was associated with advanced staging and lower 6-month survival.

Background and Purpose Abdominal aortic aneurysm (AAA) is a degenerative vascular disease associated with angiogenesis. Bexarotene is a retinoid X receptor (RXR) ligand with anti-angiogenic activity. Statins also exert anti-angiogenic activity and activate PPARs. Because RXR ligands form permissive heterodimers with PPARs and a single anti-angiogenic drug may not be sufficient to combat the wide array of angiogenic factors produced during AAA, we evaluated the effect of combined low doses of bexarotene and rosuvastatin in a mouse model of AAA.

Funding was provided by Ministerio de Ciencia e Innovacion (CGL2010-21933-C02-01) and Junta de Comunidades de Castilla-La Mancha and European Social Fund (POIC10-0269-7632). ESF and JBE are both supported by a doctoral scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund, VGN is supported by a Forschungskredit of the University of Zurich (FK-14-103) and JO is supported by a research contract funded by Severo Ochoa Program (SEV-2012-0262).

Post-nuptial moult and reproductive success were studied in relation to timing of breeding in blue tits, Parus caeruleus, breeding in southern Europe. A group of experimentally delayed pairs was created by removing first clutches, thereby inducing late repeat clutches. Reproductive success and post-nuptial moult of delayed pairs were compared with both control pairs that bred early and unmanipulated late-breeding pairs. Delayed pairs fledged fewer young and with a lower body mass than control pairs. However, the number of fledged young and fledgling mass did not differ between delayed and late-breeding pairs. These results were more consistent with the date hypothesis, and it is concluded that the timing of breeding and reproductive success may be causally related in the blue tit. This study reveals a harmful effect of relaying on female body mass at the end of the nestling period. Therefore, females apparently pay the costs of relaying, since a reduction in body mass during the nestling period may be accompanied by a lowered survival probability. Delayed and late-breeding males often began moulting while still feeding young, but neither control males nor females from the three study groups did so. These results support the view that timing-related energy constraints on breeding may be important causes of a seasonal decline in reproductive success at different latitudes.

Habitat‐specific selection pressures have been widely recognized, but whether selection favours different personality types in different habitats has rarely been evaluated. This study aimed to test whether personality‐related differences in annual reproductive success differed between two populations of blue tits (Cyanistes caeruleus) occupying different Mediterranean habitats (oakwood and pinewood). We measured exploration and parental provisioning behaviours and used a path analysis to ask how the interplay between these two behavioural traits affected reproductive success in each of these two habitats. We found that blue tits breeding in the pinewood were slow‐exploring compared to blue tits breeding in the oakwood, suggesting the occurrence of personality‐related differences in settlement, or behavioural plasticity in response to habitat. Exploration behaviour and feeding rates were positively associated, suggesting that they affect each other or that there is an environmental factor affecting both traits simultaneously. Finally, fast explorers were favoured in the pinewood, while there was no selection acting on exploration behaviour in the oak habitat. These findings emphasize the importance of integrating habitat selection, plasticity and personality in the study of behavioural evolution., The manuscript was written by ESD, YGA and NJD. ESD was funded by a predoctoral scholarship (BES‐2011‐047046) from Ministerio de Ciencia e Innovación (Spain), YGA and JJS by grant CGL2010‐21933‐C02‐01 (Ministerio de Ciencia e Innovación, Spain).

[Capsule]: The nestling diet of Blue Tits Cyanistes caeruleus is influenced by parental effort and habitat type, and consequently has an impact on breeding success., [Aims]: In a three-year study, we compared the nestling diet of Blue Tits in two Mediterranean forests (pinewood and oakwood) and tested its implications for breeding success., [Methods]: Adults were captured at the nest to obtain morphological measurements, and provisioning behaviour was recorded when chicks were 11 days old. Nestling tarsus length and body mass were measured on day 13 after hatching., [Results]: Caterpillars constituted the largest proportion of nestling diet in both habitats, however, higher numbers and biomass of noctuid, as well as higher numbers of tortricid larvae, were provided to nestlings in the pinewood. Furthermore, females provided tortricids more often than males, whereas males supplied more geometrid larvae and spiders. We found a more generalist diet for nestlings raised in the pinewood. Also, a greater number of young fledged when their diet included more tortricids and was more generalist, and Blue Tit nestlings raised on a diet with a higher number of spiders were in better body condition., [Conclusion]: Differences in nestling diet between habitat types contribute to explain variance in breeding performance and therefore demonstrate diverse foraging behaviour strategies among populations., This study was funded by the project CGL2010-21933-C02-01 (to J.J.S) and the FPI predoctoral fellowship BES-2011-047046 (to E.S.D) from the Spanish Ministry of Science and Innovation (Ministerio de Ciencia e Innovación).

[EN] Because extended incubation recesses, where incubating songbirds are away from nests for periods much longer than usual, occur infrequently, they have been treated as outliers in most previous studies and thus overlooked. However, egg temperatures can potentially fall below the physiological zero temperature during extended recesses, potentially affecting developing embryos. As such, evaluating extended recesses in an ecological context and identifying their possible fitness effects are important. With this aim, we used iButton data loggers to monitor the incubation behavior of female Blue Tits (Cyanistes caeruleus) and Great Tits (Parus major) during two breeding seasons in central Spain. We classified incubation recesses as extended if they were more than four times the mean recess duration for each species. Extended incubation recesses occurred more frequently in 2012 when females exhibited poorer body condition. Female Blue Tits had more extended incubation recesses than female Great Tits and, for both species, more extended recesses occurred at the beginning of the breeding season. Both nest attentiveness and average minimum nest temperature decreased when at least one extended recess occurred. Incubation periods averaged 4 d longer for nests where females had at least one extended recess, potentially increasing predation risk and resulting in lower-quality nestlings. Overall, our results suggest that extended recesses may be more common among songbirds than previously thought and that, due to their effects on egg temperatures and attentiveness, they could impose fitness costs. [ES] Debido a que los periodos extendidos o prolongados de receso durante los cuales el ave que incuba se ausenta del nido por un periodo más largo de lo usual son poco frecuentes, éstos han sido tratados como atípicos o han sido ignorados. Sin embargo, la temperatura de los huevos puede disminuir muy por debajo de la temperatura cero fisiológica durante estos periodos prolongados de recesos, lo que potencialmente puede afectar el desarrollo embrionario. Por tal motivo, es importante evaluar los periodos extensivos de receso en un contexto ecológico e identificar su posible efecto en la aptitud. Con este objetivo, utilizamos data-loggers iButton para monitorear la conducta de incubación de hembras de Cyanistes caeruleus y Parus major durante dos épocas reproductivas en España central. Clasificamos como recesos extendidos en la incubación, si estos eran cuatro veces mayores que el periodo promedio de recesos en cada especie. Los periodos extensivos de receso ocurrieron con mayor frecuencia en el 2012, cuando las hembras mostraron una peor condición corporal. Las hembras de Cyanistes caeruleus tuvieron periodos de receso en la incubación más extendidos que las hembras de Parus major y en ambas especies los periodos de receso extendido ocurrieron al principio de la temporada de reproducción. Tanto el periodo de atención del nido como el promedio mínimo de la temperatura dentro del nido disminuyó cuando se presentó por lo menos un periodo extendido de receso. El periodo de incubación fue, en promedio, cuatro días más largo en nidos en los que las hembras tuvieron por lo menos un receso extendido, incrementando potencialmente el riesgo de depredación y resultando en una calidad más baja de los polluelos. En general, nuestros resultados sugieren que los periodos extendidos de receso, pueden ser más comunes en aves cantoras que lo que se pensaba con anterioridad, y que, dado su efecto en la temperatura de los huevos y atención del nido, pudiera acarrear costos asociados con la aptitud reproductiva. Funding was provided by MINECO (CGL2013-48001-C2-1-P, CGL2010-21933-C02-01) and the Junta de Comunidades de Castilla-La Mancha and the European Social Fund (POIC10-0269-7632).

Understanding the importance of host genetic diversity for coping with parasites and infectious diseases is a long-standing goal in evolutionary biology. Here, we study the association between probability of infection by avian malaria (Plasmodium relictum) and individual genetic diversity in three blue tit (Cyanistes caeruleus) populations that strongly differ in prevalence of this parasite. For this purpose, we screened avian malaria infections and genotyped 789 blue tits across 26 microsatellite markers. We used two different arrays of markers: 14 loci classified as neutral and 12 loci classified as putatively functional. We found a significant relationship between probability of infection and host genetic diversity estimated at the subset of neutral markers that was not explained by strong local effects and did not differ among the studied populations. This relationship was not linear, and probability of infection increased up to values of homozygosity by locus (HL) around 0.15, reached a plateau at values of HL from 0.15 to 0.40 and finally declined among a small proportion of highly homozygous individuals (HL > 0.4). We did not find evidence for significant identity disequilibrium, which may have resulted from a low variance of inbreeding in the study populations and/or the small power of our set of markers to detect it. A combination of subtle positive and negative local effects and/or a saturation threshold in the association between probability of infection and host genetic diversity in combination with increased resistance to parasites in highly homozygous individuals may explain the observed negative quadratic relationship. Overall, our study highlights that parasites play an important role in shaping host genetic variation and suggests that the use of large sets of neutral markers may be more appropriate for the study of heterozygosity–fitness correlations., This study was funded by grants 69/2003 (Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia), CGL2010-21933-C02-01 (Ministerio de Ciencia e Innovación) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund). ESF was supported by a predoctoral scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund, and VGN enjoyed a FPI predoctoral scholarship from Ministerio de Ciencia e Innovación-EuropeanSocial Fund. J.O was supported by ‘Juan de la Cierva’ and ‘Severo Ochoa-EBD’ post-doctoral fellowships (Ministerio de Ciencia e Innovación).

Theoretical models on parental care predict that males should decrease their parental effort when paternity is in doubt. Males may use some cues to assess their certainty of paternity, and try to avoid rearing offspring sired by extra-pair males. We have previously reported in a socially monogamous passerine, the blue tit (Cyanistes caeruleus), that males decorate their nests with feathers, and that when this ornament is manipulated, males appear to have suspicions about the presence of an intruder male. Here, we decrease the male’s certainty of paternity through experimental feather supplementation to analyse whether the outcome of our experiment supports the assumptions of the parental care theory. Male C. caeruleus responded to the feather supplementation experiment by reducing their parental investment (feeding frequency and nest defence) in comparison with control males. The occurrence of extra-pair offspring in experimental nests was double than that in controls. This suggests that the manipulation was successful not only in altering males’ perceived paternity, but also, indirectly, the actual paternity. Furthermore, males that gained extra-pair young also had a higher than average probability to lose paternity in their nest, which may imply that male C. caeruleus faced a trade-off between obtaining extra-pair fertilizations and maintaining paternity in their own nest. Overall, this study supports the idea that males are prone to decrease their parental effort when they perceive that the risk of losing paternity is high. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109, 552–561. ADDITIONAL KEYWORDS: Cyanistes caeruleus – extra-pair paternity – feather carrying – nest ornamentation – parental investment.

Two new species of Acalypha, A. neeana and A. machiensis, are described and illustrated based on Bolivian specimens. Comments on the systematics of related species are included.

Breeding mistiming is increasingly frequent in several ecosystems in the face of current climate change. Species belonging to higher trophic levels must employ mechanisms to reduce it. One of these mechanisms is hatching asynchrony, with the eggs in a clutch hatching over a period of several days. Some authors have suggested it to be adaptive when food is unpredictable. However, these birds can also suffer associated costs. We tested whether a species with higher foraging efficiency avoid hatching asynchrony compared to its sister species. We studied hatching asynchrony and nestling provisioning in relation to food availability in sympatric populations of blue and great tits. For the first time, we show that sister species respond to food availability with different strategies. Blue tit feeding rates readily responded to the abundance of their main prey, and also reduced the impact of nestling size hierarchy on mean nestling weight, consequently increasing fledging rate. Our results suggest that levels of hatching asynchrony seem to be influenced by species-specific life history traits, as generalist foragers rely less on it. They also highlight the importance of multi-species approaches when studying the response of organisms to environmental unpredictability.

The study of the factors structuring genetic variation can help to infer the neutral and adaptive processes shaping the demographic and evolutionary trajectories of natural populations. Here, we analyse the role of isolation by distance (IBD), isolation by resistance (IBR, defined by landscape composition) and isolation by environment (IBE, estimated as habitat and elevation dissimilarity) in structuring genetic variation in 25 blue tit (Cyanistes caeruleus) populations. We typed 1385 individuals at 26 microsatellite loci classified into two groups by considering whether they are located into genomic regions that are actively (TL; 12 loci) or not (NTL; 14 loci) transcribed to RNA. Population genetic differentiation was mostly detected using the panel of NTL. Landscape genetic analyses showed a pattern of IBD for all loci and the panel of NTL, but genetic differentiation estimated at TL was only explained by IBR models considering high resistance for natural vegetation and low resistance for agricultural lands. Finally, the absence for IBE suggests a lack of divergent selection pressures associated with differences in habitat and elevation. Overall, our study shows that markers located in different genomic regions can yield contrasting inferences on landscape-level patterns of realized gene flow in natural populations., This study was funded by grants 69/2003(Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia), CGL2010-21933-C02-01(Ministerio de Ciencia e Innovación) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund). ESF and JBE are supported by a PhD scholarship from Junta de Comunidades de Castilla-La Mancha-European Social Fund, VGN is supported by a Forschungskredit of the University of Zurich (FK-14-103), RB is supported by a Juan de la Cierva fellowship (Ministerio de Ciencia e Innovación), ESD is supported by a PhD scholarship from Ministerio de Ciencia e Innovación-European Social Fund (FPIProgram), and JO is supported by research contracts funded by Severo Ochoa (SEV-2012-0262) and Ramón y Cajal (RYC-2013-12501) Programs.

Opportunistic infections continue to be a cause of morbidity and mortality in HIV-infected patients. They often arise because of severe immunosuppression resulting from poor adherence to antiretroviral therapy, failure of antiretroviral therapy, or unawareness of HIV infection by patients whose first clinical manifestation of AIDS is an opportunistic infection. The present article is an executive summary of the document that updates the previous recommendations on the prevention and treatment of opportunistic infections in HIV-infected patients, namely, infections by parasites, fungi, viruses, mycobacteria, and bacteria, as well as imported infections. The article also addresses immune reconstitution inflammatory syndrome. This document is intended for all professionals who work in clinical practice in the field of HIV infection. (C) 2016 Elsevier Espana, S.L.U. and Sociedad Espanola de Enfermedades Infecciosas y Microbiologia Clinica. All rights reserved.

[EN] The use of nestboxes to study secondary cavity nesting avian species has greatly improved the knowledge related to many fields of environmental sciences. The aim of this study has been to compare the breeding performance of Great Tits (Parus major) and Blue Tits (Cyanistes caeruleus) in two types of nestboxes that differ with regard to their shape and thermal properties. Both nestbox types were placed in the same area to avoid confounding factors. There were significant differences between the extreme values of temperature and relative humidity obtained for the two nestbox types, and this was also the case in their daily patterns. Secondary hole-nesting birds appear to prefer breeding in woodcrete nestboxes. Nest predation was, however, significantly greater in woodcrete nestboxes. The nest height was significantly higher in woodcrete nestboxes, as was the danger distance between the nest cup and the entrance hole, which could account for the fact that Great Tits prefer this type of nestbox. The laying date was earlier in woodcrete boxes, but nestbox type did not influence clutch size. However, the eggs of both species were significantly smaller in woodcrete boxes. Breeding success was worse in woodcrete nestboxes. Nestbox type also affected the incubation pattern in both species, and attentiveness was significantly diminished in woodcrete nestboxes. This study reinforces the idea that the type of nestbox used in avian studies is not a trivial issue and may have strong biological effects on avian populations. It is important to take this into account when nestboxes are used as management measures for bird conservation., [GE] Die Benutzung von Nistkästen für Studien über sekundäre Höhlenbrüter hat zur Weiterentwicklung vieler Bereiche der Biowissenschaften beigetragen. Das Ziel der vorliegenden Studie war der Vergleich der Brutleistung der Kohlmeise (Parus major) und der Blaumeise (Cyanistes caeruleus) in zwei Nistkastentypen mit unterschiedlicher Form und thermischen Eigenschaften. Beide Nistkastentypen wurden in derselben Gegend positioniert, um verfälschende Faktoren zu vermeiden. Es wurden signifikante Unterschiede zwischen den maximalen und minimalen Temperaturen sowie zwischen der relativen Luftfeuchtigkeit gemessen, und dies betraf auch deren Tageschwankungen. Beide Arten bevorzugten die Holzbetonnistkästen. Nestraub war aber signifikant höher in den Holzbetonkästen. Die Nesthöhe war in Holzbetonkästen signifikant größer, ebenso die Distanz zwischen Eingang und das Nest. Dies könnte einer der Gründe sein, dass Kohlmeisen diesen Typ von Nistkasten präferieren. Der Legebeginn war in Holzbetonkästen früher; der Nistkastentyp hatte aber keinen Einfluss auf die Gelegegröße. Allerdings waren die Eier beider Arten in den Holzbetonkästen signifikant kleiner und der Bruterfolg war geringer in den Holzbetonkästen. Der Nistkastentyp beeinflusste auch das Bebrütungsmuster beider Arten und die Aufmerksamkeit der Brutvögel am Nest war geringer bei Holzbetonkästen. Diese Studie unterstützt die Annahme, dass der Typ des Nistkastens keine triviale Rolle in ornithologischen Studien spielt und große biologische Einflüsse haben kann. Dies ist zu berücksichtigen, wenn Nistkästen im Vogelschutz eingesetzt werden., Funding was provided by the MINECO (CGL2010-21933-C02-01; CGL2013-48001-C02-1P), the JCCM and the European Social Fund (POIC10-0269-7632).

The extent of inbreeding depression and the magnitude of heterozygosity–fitness correlations (HFC) have been suggested to depend on the environmental context in which they are assayed, but little evidence is available for wild populations. We combine extensive molecular and capture–mark–recapture data from a blue tit (Cyanistes caeruleus) population to (1) analyze the relationship between heterozygosity and probability of interannual adult local recruitment and (2) test whether environmental stress imposed by physiologically suboptimal temperatures and rainfall influence the magnitude of HFC. To address these questions, we used two different arrays of microsatellite markers: 14 loci classified as neutral and 12 loci classified as putatively functional. We found significant relationships between heterozygosity and probability of interannual local recruitment that were most likely explained by variation in genomewide heterozygosity. The strength of the association between heterozygosity and probability of interannual local recruitment was positively associated with annual accumulated precipitation. Annual mean heterozygosity increased over time, which may have resulted from an overall positive selection on heterozygosity over the course of the study period. Finally, neutral and putatively functional loci showed similar trends, but the former had stronger effect sizes and seemed to better reflect genomewide heterozygosity. Overall, our results show that HFC can be context dependent, emphasizing the need to consider the role of environmental heterogeneity as a key factor when exploring the consequences of individual genetic diversity on fitness in natural populations.

A critical review of the Ecuadorian species of Acalypha L. (Euphorbiaceae) is presented; 20 of the 38 previously recognized species are accepted, 9 are considered synonyms and 9 are based on misidentifications. Comprehensive nomenclatural information is supplied and 13 lectotypes are designated. An identification key is also provided.

Over a five-year period we monitored breeding parameters in a colony of Tree Sparrows Passer montanus in central Spain. This multi-brooded species can raise one to three broods per season. In this study, we analyzed inter-brood variation in reproductive success to infer the existence of seasonal patterns. We also explored inter-annual variation of life-history traits, which is of particular interest in a declining species like the Tree Sparrow. In agreement with our expectation of a multi-brooded species, clutch size showed a quadratic relationship with laying date. We found that the inter-brood interval varied significantly depending on the number of breeding attempts, which suggests that the species may engage in time-saving behaviours as reported for other multi-brooded passerines. Nestlings from second broods fledged in better condition than nestlings from first and third broods. Fewer fledglings were produced in third broods than in preceding breeding attempts. The reproductive output of third broods varied to a large extent among years, whereas inter-annual variation was much less marked in first and second broods. Over the years, we also detected a decrease in nestling size (tarsus and wing length), which was consistent among breeding attempts. Finally, the productivity of the colony remained fairly stable over the study period. This pattern contrasts with negative population trends detected by national monitoring programmes. The proximity of wetland habitats in the surroundings of our colony — essential for adequate invertebrates for the chicks — is suggested as a main reason for this difference.

Carotenoid-based coloration in adult birds has been often regarded as an honest signal of individual quality. However, few studies have demonstrated a link between carotenoid display and the quantity or quality of resources provided to the offspring. The present study investigated the expression of a carotenoid-based ornament, the breast plumage yellowness of the blue tit Cyanistes caeruleus, in relation to the level of parental provisioning effort and the amount of carotenoid-rich prey provided to the young. The study was conducted in two forest types (evergreen and deciduous), which also allowed an exploration of the possible existence of habitat effects on the coloration of breeding birds. It was found that plumage colour intensity (carotenoid chroma) correlated positively with nestling provisioning rates of both males and females, supporting the good parent hypothesis. In addition, carotenoid chroma was positively related with the proportion of Lepidoptera larvae brought to the nest in both sexes. Female but not male coloration was positively linked to breeding success (proportion of fledged young). Nestling coloration did not correlate with that of their parents, nor the frequency with which they were fed. Hue and lightness of nestling’s plumage correlated positively with body mass and tarsus length, respectively. The results obtained in the present study indicate that ventral plumage coloration in blue tits may advertise the ingested carotenoids (carotenoid foraging ability) and also their overall parental quality in terms of nestling provisioning rates. This suggests that plumage yellowness can be used as an indicator of foraging ability in this species. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106, 418–429.

The study of the genetic consequences of dispersal is a central topic in evolutionary, conservation and behavioural research. However, few studies have simultaneously considered dispersal movements from marked individuals and contemporary patterns of gene flow. We analysed the link between dispersal behaviour and gene flow in four populations of blue tits with different degrees of connectivity. For this purpose, we monitored four breeding patches and used genotypic and capture–mark–recapture data to study the genetic consequences of dispersal at different spatial scales. Data on natal dispersal movements revealed that both males and females dispersed less than expected under a random pattern of settlement at the two large spatial scales considered: the whole study area and the two main localities. However, natal dispersal distance was lower than expected under random settlement within natal patches in males whereas an opposite pattern was found for females. Accordingly, microsatellite data revealed limited gene flow between the localities studied and an isolation-by-distance pattern of genetic structure that was particularly strong at the large spatial scale (i.e. considering geographically distant breeding patches). Finally, the strong male philopatry was reflected by a stronger genetic structure and a lower admixed ancestry in this sex. Overall, we found evidence that restricted dispersal and fragmentation may have both contributed to reduce interpopulation gene flow at different spatial scales in a forest species and that there is concordance between genetic studies and those based on capture–mark–recapture., During this work V.G.N. and J.O. were supported by respective predoctoral FPI (MEC) and postdoctoral JAE-Doc (CSIC) fellowships. This study was funded by the grants 69/2003 (Organismo Autónomo de Parques Nacionales-Ministerio de Medio Ambiente), GCL2007-61395 (Ministerio de Educación y Ciencia) and POIC10-0269-7632 (Junta de Comunidades de Castilla-La Mancha-European Social Fund).

In many animals, females are fed by males during courtship or incubation (mate-feeding). According to the mate appraisal hypothesis, females may evaluate the parental capacity of males, whereas the pair bond hypothesis suggests that feeding may strengthen the pair bonds with them. Following the nutrition hypothesis, by contrast, females obtain direct nutritional benefits from being fed by males during periods of high-energy expenditure, such as egg formation and incubation. However, there is little support for these hypotheses at an interspecific level. We tested predictions from these hypotheses in a dataset of 170 species of passerine birds. As predicted by the nutrition hypothesis, we found that mate-feeding has evolved more often in species in which the female incubates and builds the nest alone and have noncarnivorous diets. This suggests that mate-feeding is a behavioral strategy that compensates for nutritional limitations of females during breeding, as both incubation and nest building are energetically costly processes, and noncarnivorous diets are deficient in proteins. We also found that incubation feeding has evolved more often in species that place nests at elevated sites, suggesting that these species face low predation risk that allows males to feed females. In the particular case of incubation feeding, we found that species that have evolved this behavior produce larger clutch size and have higher hatching success. Our results support the nutrition hypothesis from an interspecific perspective, suggesting that mate-feeding is a strategy to compensate for nutritional limitations of females during reproduction and that it has fitness consequences.

Plant responses to enhanced ozone levels have been studied in two pairs of evergreen-deciduous species (Pistacia terebinthus vs. P. lentiscus; Viburnum lantana vs. V. tinus) in Open Top Chambers. Ozone induced widespread visible injury, significantly reduced CO(2) assimilation and stomatal conductance (g(s)), impaired Rubisco efficiency and regeneration capacity (V(c,max,)J(max)) and altered fluorescence parameters only in the deciduous species. Differences in stomatal conductance could not explain the observed differences in sensitivity. In control plants, deciduous species showed higher superoxide dismutase (SOD) activity than their evergreen counterparts, suggesting metabolic differences that could make them more prone to redox imbalances. Ozone induced increases in SOD and/or peroxidase activities in all the species, but only evergreens were able to cope with the oxidative stress. The relevancy of these results for the effective ozone flux approach and for the current ozone Critical Levels is also discussed.

A common feature of many birds breeding in seasonal environments is that fitnessrelated parameters such as nestling mass or survival decline as the breeding season progresses. Consequently, there is a tendency for early breeders to have better reproductive performance than individuals breeding later in the season. This variation could be caused by factors associated with the date of laying, such as changing environmental conditions (date hypothesis), or by differences in parental quality between early and late breeders (parent quality hypothesis). To evaluate the relative importance of both hypotheses, we manipulated hatch dates of Blue Tits Cyanistes caeruleus by exchanging clutches with different incubation stages and assessed the impact on nestling mass, nestling diet and provisioning rates. Mean nestling mass declined significantly over the season. This was the combined result of differences in parental quality, which dominated in the early part of the season, and the influence of hatching date (date effect per se), which prevailed later in the season. Nestling diet composition was apparently uninfluenced by the manipulation, suggesting that deteriorating food supplies are the primary reason for the seasonal variation in the nestling diet. Counter to the date hypothesis, delayed parents did not feed their young less than control pairs did, but in fact exhibited higher provisioning rates. Our results suggest that in this population, parental quality seems to constrain post-hatching reproductive performance and such intrinsic limitations may help to explain why certain individuals breed later.

Plant phenology is strongly controlled by climate and has consequently become one of the most reliable bioindicators of ongoing climate change. We used a dataset of more than 200 000 records for six phenological events of 29 perennial plant species monitored from 1943 to 2003 for a comprehensive assessment of plant phenological responses to climate change in the Mediterranean region. Temperature, precipitation and North Atlantic Oscillation (NAO) were studied together during a complete annual cycle before phenological events to determine their relative importance and potential seasonal carry-over effects. Warm and dry springs under a positive phase of NAO advance flowering, leaf unfolding and fruiting dates and lengthen the growing season. Spatial variability of dates (range among sites) was also reduced during warm and dry years, especially for spring events. Climate during previous weeks to phenophases occurrence had the greatest impact on plants, although all events were also affected by climate conditions several months before. Immediate along with delayed climate effects suggest dual triggers in plant phenology. Climatic models accounted for more than 80% of variability in flowering and leaf unfolding dates, and in length of the growing season, but for lower proportions in fruiting and leaf falling. Most part of year-to-year changes in dates was accounted for temperature, while precipitation and NAO accounted for o10% of dates’ variability. In the case of flowering, insect-pollinated species were better modelled by climate than wind-pollinated species. Differences in temporal responses of plant phenology to recent climate change are due to differences in the sensitivity to climate among events and species. Spring events are changing more than autumn events as they are more sensitive to climate and are also undergoing the greatest alterations of climate relative to other seasons. In conclusion, climate change has shifted plant phenology in the Mediterranean region.

The presence of somatic mutations of the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) gene in patients with advanced nonsmall cell lung cancer (NSCLC) correlates with a good response to tyrosine kinase inhibitors. The usefulness of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for the detection of EGFR mutations in cells recovered from malignant mediastinal nodes in patients with NSCLC was assessed. All patients with lung adenocarcinoma or unspecified NSCLC referred for staging with EBUS-TBNA were included. Nodes with a short-axis diameter of >5 mm were sampled, and genomic DNA from metastatic tumour cells was obtained for analysis of exons 19 and 21. The impact of sampling on management was assessed. EGFR gene analysis of the EBUS-TBNA sample was feasible in 26 (72.2%) out of the 36 patients with lymph node metastasis. Somatic mutations of the EGFR gene were detected in tissue obtained through EBUS-TBNA in two (10%) out of 20 patients with metastasic lung adenocarcinoma. Malignant tissue samples obtained by EBUS-TBNA from patients with nodal metastasis of NSCLC are suitable for the detection of EGFR mutations in most cases, and this technique demonstrates mutated neoplastic cells in a tenth of patients with adenocarcinoma.

The aim of this study was to investigate how the potato (Solanum tuberosum) cv Desiree, would respond to ozone exposure under three different manure supplies, in order to test the efficiency of manure as a protector against ozone. Potato plants were exposed in the greenhouse to three ozone levels: charcoal filtered air (F); non-filtered air (NF) and non-filtered air plus ozone (NF+: 57 ppb as 8 h mean for 74 days). Subsequently, the ozone treatments were combined with three fertilisation levels: 0 kg m−2, 3 kg m−2 and 6 kg m−2 of manure. Manure reduced ozone effects on visible symptoms, assimilation, biomass and tuber production. In comparison with the filtered treatment, the ozone treatments significantly reduced commercial tuber production, by 53% (NF) and 65% (NF+) in the 0-kg manure treatment and by 26% (NF) and 44% (NF+) in the 3-kg manure treatment, with the large tubers (>55 mm of diameter) being the most affected. The 6-kg manure dose delayed the onset of visible symptoms and compensated for the decrease in biomass and crop production caused by ambient and enhanced ozone levels. The use of a suitable manure content can be effective as a palliative against oxidative stress in potato crops.

1. We test the consequences, in terms of breeding success and parental effort, of eggshell pigmentation pattern in a hole-nesting bird, the blue tit Cyanistes caeruleus that lays eggs asymmetrically speckled with reddish spots (maculated eggs). 2. We assess the effect of distribution of spots (pigment 'spread') and spot size and pigment intensity (pigment 'darkness') on eggshell physical properties and breeding parameters concerning nestling condition, investment of parents in offspring care and reproductive output in two different habitat types: a deciduous oakwoodland and an evergreen forest. 3. Blue tit clutches with more widely distributed spots showed a thicker eggshell, a shorter incubation period, a lesser amount of mass loss per day and a higher hatching probability than those with spots forming a 'corona' ring. While eggs with larger and darker (more pigment intensity) spots showed a thicker eggshell and a shorter incubation period. In the light of 'signal function hypothesis', these egg traits may reflect female health status and, consequently, this could affect male parental effort. 4. Here we show supports for some of the necessary assumptions of this hypothesis. We found a positive relationship between egg pigment 'spread' and male but not female provisioning rates per day. On the other hand, pigment 'darkness' of blue tits' clutches was positively related to female tarsus length, while pigment 'spread' was positively related to clutch size, male body mass and nestling tarsus length. Our study shows that eggshell pigment 'spread' can be used as an indicator of clutch quality. Further investigations are needed to understand the role of calcium availability as possible causal agent of deviant eggs and its relation to the maculation phenomenon.

Avian life-history studies are often conducted on populations nesting in nestboxes. However, the type of nestbox used is a potentially confounding variable, as each model shows specific characteristics. In recent decades, a new commercial model of artificial nestbox made of woodcrete, a mixture of sawdust and additives, has become popular owing to its better protection against predators compared with the classic wooden design. We assess the effect of nestbox type on the breeding ecology of Tree Sparrows Passer montanus , focusing on their thermal properties, an influential factor in nestbox selection. Occupancy rates and reproductive parameters of Tree Sparrows were compared between birds breeding in woodcrete and wooden nestboxes over 5 years. Woodcrete nestboxes had a higher occupancy rate and birds breeding in them had earlier clutches, a shorter incubation period and more reproductive attempts per season than birds nesting in wooden boxes. Clutch size and nestling condition did not differ between nestbox types, but reproductive success was higher in woodcrete nestboxes. The higher temperature in woodcrete nestboxes (1.5 ° C higher on average) might explain these differences. Such differences may bias results obtained in comparative studies where more than one nestbox type is used.

1. Although the ability to detect chemical cues is widespread in many organisms, it is surprising how little is known about the role of chemical communication in avian life histories. Nowadays, growing evidence suggests that birds can use olfaction in several contexts. However, we still do not know the role of bird olfaction in one of the most important determinants of survival, predator detection. 2. Blue tits, Cyanistes caeruleus L., were exposed to chemical cues of: (i) mustelid (predator), (ii) quail (odorous control); or (iii) water (odourless control) inside the nest-box where they were provisioning 8-day-old nestlings. 3. We show that blue tits were able to detect the chemical cues and showed antipredatory behaviours to cope with the risk of predation. Birds delayed their entry to the nest-box, and they perched on the hole of the nest-box and refused to enter more times when they found predator scent than control scents inside the nest-box. In addition, birds decreased the time spent inside the predator-scented nest-box when feeding nestlings. 4. The discovery of the ability of birds to use chemical cues of predators to accurately assess predation may help to understand many aspects of bird life histories that have been neglected until now.

Remote sensing data have been used in previous studies to assess the effects of winter ecological conditions in Africa on biological parameters recorded in bird populations during the following breeding season in Europe. Based on the results of these studies, we hypothesized that a high productivity of vegetation during the winter and, thus, high resource availability, should advance the arrival of long-distance migrants to the European breeding areas due to enhanced ecological conditions. To test this hypothesis, between 1982 and 2000 we examined the first arrival date to the Iberian Peninsula of five species (White Stork, Cuckoo, Common Swift, Barn Swallow and Nightingale) in relation to several explanatory variables: ecological conditions in their African wintering grounds and passage areas, as reflected by the normalized difference vegetation index (NDVI), temperature and precipitation in their passage areas and the winter North Atlantic Oscillation (NAO). Ecological conditions in the wintering areas were important for White Stork, Cuckoo and Barn Swallow phenology, while both NDVI in passage areas and NAO did not have an effect on any species. Migratory birds arrived earlier after winters with high vegetation productivity in Africa. Temperature in passage areas was important for the later species (i.e. Cuckoo, Common Swift and Nightingale), although in all cases the true relevance of this factor was scarce due to the poor explanatory capacity of the models. These species were recorded in the Iberian Peninsula earlier in the spring of those years with warmer temperatures in passage areas. The nexus between African NDVI and arrival phenology is hypothesized through increases in wintering survival rates and/or the faster acquisition of pre-migratory body condition and progression through sub-Saharan areas.

Five tomato cultivars (Nikita, Ailsa Craig, Moneymaker, UC 82 L and Piedmont) were exposed in open-top chambers to three ozone treatments: charcoal-filtered air (F); non-filtered air (NF); and non-filtered air plus 70 ppb ozone (NF+, 8 h, 133 days). Ozone-specific visible symptoms were recorded for all cultivars in the NF and NF+ treatments. All cultivars showed a reduction of root biomass in the NF+ treatment, root-biomass reductions were also observed in the NF treatment in Nikita (29%) and UC 82 L (33%). Four cultivars from the NF+ treatment showed reductions in aboveground dry biomass (25–50%). The sensitive cultivar Nikita also decreased its aboveground dry biomass (28%) at the end of crop cultivation in the NF treatment. Crop production showed generalized reductions in the total number of ripe and unripe fruits in the NF+ treatment (53–73%), with Nikita presenting a 24% decrease in the total number of ripe fruits in the NF treatment as well. A decreased ripeness rate was detected in all cultivars in NF+ and in three cultivars in the NF treatment (Nikita, Moneymaker and Piedmont). The effect of ozone on total crop production (kg m−2) was more important in the early harvest (50–63%) than in the late harvest. The ozone doses in the NF+ treatment affected fruit quality in all cultivars with the exception of Piedmont. Nikita and Ailsa Craig also showed fruit quality changes in the NF treatment. Nikita seems to be an ozone-sensitive cultivar, UC 82 L and Piedmont are more ozone-tolerant and the other two cultivars are in an intermediate position.