Elevated oxidative stress in pied flycatcher nestlings of eumelanic foster fathers under low rearing temperatures.

Striking variation in melanin coloration within natural populations is likely due to the different fitness outcomes of alternative phenotypes in varying environmental conditions. There are two types of melanin: eumelanins yield blackish hues, whereas pheomelanins yield reddish hues. The production of eumelanins requires low levels of glutathione (GSH), which is the most important intracellular antioxidant, whereas the production of pheomelanins requires high levels of GSH. We investigated the oxidative status of male pied flycatchers (Ficedula hypoleuca) with different degrees of melanin coloration under different temperatures during the nestling period. Moreover, we assessed the oxidative status of offspring in relation to their biological or foster father's melanin coloration and ambient temperature. To separate offspring genotype effects and paternal effects in different temperatures, we used a partial cross-foster design. The temperature differently affected the oxidative status of differently colored male pied flycatchers and their foster offspring. When the weather was relatively cold, black males had higher glutathione S-transferase levels compared with brown males, indicating enhanced stress in black males. Foster offspring of black males had a lower ratio between reduced and oxidized GSH followed by higher total amount of GSH than foster offspring of brown males. Thus, foster offspring of black males seem to suffer from oxidative stress under relatively cold weather compared with those of brown males, and vice versa under relatively warm weather. Although differently colored males experienced changes in their oxidative status under different temperatures, the link between paternal melanin coloration and offspring oxidative stress appears to be environmentally induced.

Carry-over effects of conditions at the wintering grounds on breeding plumage signals in a migratory bird: roles of phenotypic plasticity and selection.

To understand the consequences of ever-changing environment on the dynamics of phenotypic traits, distinguishing between selection processes and individual plasticity is crucial. We examined individual consistency/plasticity in several male secondary sexual traits expressed during the breeding season (white wing and forehead patch size, UV reflectance of white wing patch and dorsal melanin coloration) in a migratory pied flycatcher (Ficedula hypoleuca) population over an 11-year period. Furthermore, we studied carry-over effects of three environmental variables (NAO, a climatic index; NDVI, a vegetation index; and rainfall) at the wintering grounds (during prebreeding moult) on the expression of these breeding plumage traits of pied flycatcher males at individual and population levels. Whereas NAO correlates negatively with moisture in West Africa, NDVI correlates positively with primary production. Forehead patch size and melanin coloration were highly consistent within individuals among years, whereas the consistency of the other two traits was moderate. Wing patch size decreased with higher NAO and increased with higher rainfall and NDVI at the individual level. Interestingly, small-patched males suffered lower survival during high NAO winters than large-patched males, and vice versa during low NAO winters. These counteracting processes meant that the individual-level change was masked at the population level where no relationship was found. Our results provide a good example of how variation in the phenotypic composition of a natural population can be a result of both environment-dependent individual plasticity and short-term microevolution. Moreover, when plasticity and viability selection operate simultaneously, their impacts on population composition may not be evident.

Sympatric divergence and clinal variation in multiple coloration traits of Ficedula flycatchers.

Geographic variation in phenotypes plays a key role in fundamental evolutionary processes such as local adaptation, population differentiation and speciation, but the selective forces behind it are rarely known. We found support for the hypothesis that geographic variation in plumage traits of the pied flycatcher Ficedula hypoleuca is explained by character displacement with the collared flycatcher Ficedula albicollis in the contact zone. The plumage traits of the pied flycatcher differed strongly from the more conspicuous collared flycatcher in a sympatric area but increased in conspicuousness with increasing distance to there. Phenotypic differentiation (PST ) was higher than that in neutral genetic markers (FST ), and the effect of geographic distance remained when statistically controlling for neutral genetic differentiation. This suggests that a cline created by character displacement and gene flow explains phenotypic variation across the distribution of this species. The different plumage traits of the pied flycatcher are strongly to moderately correlated, indicating that they evolve non-independently from each other. The flycatchers provide an example of plumage patterns diverging in two species that differ in several aspects of appearance. The divergence in sympatry and convergence in allopatry in these birds provide a possibility to study the evolutionary mechanisms behind the highly divergent avian plumage patterns.

Heterospecific female mimicry in Ficedula flycatchers.

Mimicry is a widespread phenomenon. Vertebrate visual mimicry often operates in an intraspecific sexual context, with some males resembling conspecific females. Pied flycatcher (Ficedula hypoleuca) dorsal plumage varies from the ancestral black to female-like brown. Experimental studies have shown that conspecific and heterospecific (collared flycatcher, F. albicollis) individuals of both sexes respond, at least initially, to brown individuals as if they were female. We quantified the perceptual and biochemical differences between brown feathers and found that brown pied flycatcher males are indistinguishable from heterospecific, but not from conspecific, females in both aspects. To our knowledge, this is the first evidence of a visual mimetic signalling system in a sexual context where the model is heterospecific to the mimic. By only mimicking heterospecific females, brown pied flycatcher males can establish territories next to the more dominant collared flycatcher in sympatry, suffer less aggression by darker conspecifics in allopatry and preserve within-species sexual recognition throughout the breeding range. A closer look at the evolutionary history and ecology of these two species illustrates how such a mimetic system can evolve. Although likely rare, this phenomenon might not be unique to Ficedula flycatchers.

Fecal calcium levels of bird nestlings as a potential indicator of species-specific metal sensitivity.

Sensitivity of bird species to environmental metal pollution varies but there is currently no general framework to predict species-specific sensitivity. Such information would be valuable from a conservation point-of-view. Calcium (Ca) has antagonistic effects on metal toxicity and studies with some common model species show that low dietary and circulating calcium (Ca) levels indicate higher sensitivity to harmful effects of toxic metals. Here we measured fecal Ca and five other macroelement (potassium K, magnesium Mg, sodium Na, phosphorus P, sulphur S) concentrations as proxies for dietary levels in 66 bird species to better understand their interspecific variation and potential use as an indicator of metal sensitivity in a wider range of species (the main analyses include 39 species). We found marked interspecific differences in fecal Ca concentration, which correlated positively with Mg and negatively with Na, P and S levels. Lowest Ca concentrations were found in insectivorous species and especially aerial foragers, such as swifts (Apodidae) and swallows (Hirundinidae). Instead, ground foraging species like starlings (Sturnidae), sparrows (Passeridae), cranes (Gruidae) and larks (Alaudidae) showed relatively high fecal Ca levels. Independent of phylogeny, insectivorous diet and aerial foraging seem to indicate low Ca levels and potential sensitivity to toxic metals. Our results, together with information published on fecal Ca levels and toxic metal impacts, suggest that fecal Ca levels are a promising new tool to evaluate potential metal-sensitivity of birds, and we encourage gathering such information in other bird species. Information on the effects of metals on breeding parameters in a wider range of bird species would also help in ranking species by their sensitivity to metal pollution.

Fluctuating selection and immigration as determinants of the phenotypic composition of a population.

It is important to identify the factors that affect the evolutionary potential of populations to respond to environmental changes. Such processes are for example the ones affecting the amount of heritable phenotypic variation in a population. We examined factors explaining the wide phenotypic variation in the genetically determined black-brown dorsal colouration of male pied flycatchers (Ficedula hypoleuca) during a period of >50 years in a northern European breeding population. We demonstrate that the temperature-dependent relative breeding success of brown males predicts the inter-annual change in the proportion of the brown male phenotype. The proportion of brown males also appears to reflect immigration from Central Europe, where the brown type prevails due to local selection pressure. Warm springs in northern Central Europe had a positive effect on the proportion of the brown phenotype in the north in the early part of the study period, which suggests prolonged migration in favourable conditions. However, the association between warm springs and a high proportion of brown males has weakened from the 1950s to the present, which may explain why the proportion of the brown males in our study area decreased by a third during the period 1954 to 2008. This is likely a result of decreasing population size in Central Europe. These results demonstrate that temporal variation in environmental conditions is maintaining variation in the pied flycatcher male phenotype. They also indicate that climate warming has the potential to change the population composition both through temperature-dependent selection and environmental factors affecting long-distance immigration.

Candidate genes for colour and vision exhibit signals of selection across the pied flycatcher (Ficedula hypoleuca) breeding range.

The role of natural selection in shaping adaptive trait differentiation in natural populations has long been recognized. Determining its molecular basis, however, remains a challenge. Here, we search for signals of selection in candidate genes for colour and its perception in a passerine bird. Pied flycatcher plumage varies geographically in both its structural and pigment-based properties. Both characteristics appear to be shaped by selection. A single-locus outlier test revealed 2 of 14 loci to show significantly elevated signals of divergence. The first of these, the follistatin gene, is expressed in the developing feather bud and is found in pathways with genes that determine the structure of feathers and may thus be important in generating variation in structural colouration. The second is a gene potentially underlying the ability to detect this variation: SWS1 opsin. These two loci were most differentiated in two Spanish pied flycatcher populations, which are also among the populations that have the highest UV reflectance. The follistatin and SWS1 opsin genes thus provide strong candidates for future investigations on the molecular basis of adaptively significant traits and their co-evolution.

Reduced Number of Pediatric Orthopedic Trauma Requiring Operative Treatment during COVID-19 Restrictions: A Nationwide Cohort Study.

BACKGROUND AND AIMS: The coronavirus outbreak significantly changed the need of healthcare services. We hypothesized that the COVID-19 pandemic decreased the frequency of pediatric fracture operations. We also hypothesized that the frequency of emergency pediatric surgical operations decreased as well, as a result of patient-related reasons, such as neglecting or underestimating the symptoms, to avoid hospital admission. MATERIALS AND METHODS: Nationwide data were individually collected and analyzed in all five tertiary pediatric surgical/trauma centers in Finland. Operations related to fractures, appendicitis, and acute scrotum in children aged above 16 years between March 1 and May 31 from 2017 to 2020 were identified. The monthly frequencies of operations and type of traumas were compared between prepandemic 3 years and 2020. RESULTS: Altogether, 1755 patients were identified in five tertiary hospitals who had an emergency operation during the investigation period. There was a significant decrease (31%, p = 0.03) in trauma operations. It was mostly due to reduction in lower limb trauma operations (32%, p = 0.006). Daycare, school, and organized sports-related injuries decreased significantly during the pandemic. These reductions were observed in March and in April. The frequencies of appendectomies and scrotal explorations remained constant. CONCLUSION: According to the postulation, a great decrease in the need of trauma operations was observed during the peak of COVID-19 pandemic. In the future, in case similar public restrictions are ordered, the spared resources could be deployed to other clinical areas. However, the need of pediatric surgical emergencies held stable during the COVID-19 restrictions.

Melanin coloration has temperature-dependent effects on breeding performance that may maintain phenotypic variation in a passerine bird.

Fluctuating selection pressure may maintain phenotypic variation because of different types of individuals being adapted to different environmental conditions. We show that the extensive variation in the coloration of male pied flycatchers (Ficedula hypoleuca) can be maintained through differences in the reproductive success of male phenotypes under different conditions. The effects of weather conditions on the relative success of different male phenotypes varied between different phases of breeding. The reproductive output of black males was the highest when it was cold during egg-laying but warm during the nestling period, whereas the fledgling production of brown males was highest when it was continuously warm. In addition, male forehead and wing patch sizes had context-dependent effects on timing of breeding and nestling mortality, respectively. These results indicate that environmental heterogeneity plays a role in maintaining phenotypic variation. As melanin-based coloration is heritable, climate change may alter phenotype frequencies depending on the patterns of warming.

White-tailed eagle (Haliaeetus albicilla) and great cormorant (Phalacrocorax carbo) nestlings as spatial sentinels of Baltic acidic sulphate soil associated metal contamination.

Sulphate soils, characterised by low pH conditions, are found worldwide, and are potentially large sources of metal contamination, often exceeding industrial emissions. Metal leaching from sulphate soils has been shown to be harmful to aquatic organisms, but the cascading effect on exposure in apex avian predators has not been studied earlier. With the present study we aimed at evaluating the potential of white-tailed eagle (Haliaeetus albicilla) and great cormorant (Phalacrocorax carbo) nestlings, collected from nests located either in sulphate soil or control areas, for monitoring spatial contaminant trends of metals typically associated with sulphate soils. In blood of white-tailed eagles, the concentrations of aluminium and cobalt were significantly higher in sulphate soil areas. In blood of great cormorants, the concentrations of copper and manganese were so, while the concentration of zinc was found to be lower. Also, we observed an interaction between the latitude and soil type in cobalt and lithium concentrations of great cormorants, showing that concentrations in the sulphate soil associated nestlings rose more steeply towards the north than in the control group. Latitudinal trends of higher concentrations in the south were found in cadmium, manganese, and copper of white-tailed eagle nestlings, while thallium of white-tailed eagle nestlings, and thallium and zinc of great cormorant nestlings showed a latitudinal trend of higher concentrations in the north. Concentrations of several metals correlated positively within a species indicating covariation in metal exposure. Generally, the metal concentrations in both species were similar to levels reported to be below toxicity thresholds in other species. These results indicate, that white-tailed eagle and great cormorant nestling metal burdens may indicate environmental contamination from acidic sulphate soil runoff, and that they may act as indicators of latitudinal gradient identifying different contamination sources.

Population fragmentation leads to isolation by distance but not genetic impoverishment in the philopatric Lesser Kestrel: a comparison with the widespread and sympatric Eurasian Kestrel.

Population fragmentation is a widespread phenomenon usually associated with human activity. As a result of habitat transformation, the philopatric and steppe-specialist Lesser Kestrel Falco naumanni underwent a severe population decline during the last century that increased population fragmentation throughout its breeding range. In contrast, the ubiquitous Eurasian Kestrel Falco tinnunculus did not suffer such adverse effects, its breeding range still remaining rather continuous. Using microsatellites, we tested the effects of population fragmentation on large-scale spatial patterns of genetic differentiation and diversity by comparing these two sympatric and phylogenetically related species. Our results suggest that habitat fragmentation has increased genetic differentiation between Lesser Kestrel populations, following an isolation-by-distance pattern, while the population of Eurasian Kestrels is panmictic. Contrary to expectations, we did not detect significant evidence of reduced genetic variation or increased inbreeding in Lesser Kestrels. Although this study reports genetic differentiation in a species that has potential for long-distance dispersal but philopatry-limited gene flow, large enough effective population sizes and migration may have been sufficient to mitigate genetic depauperation. A serious reduction of genetic diversity in Lesser Kestrels would, therefore, only be expected after severe population bottlenecks following extreme geographic isolation.

One in Three of Pediatric Tibia Shaft Fractures is Currently Treated Operatively: A 6-Year Epidemiological Study in two University Hospitals in Finland Treatment of Pediatric Tibia Shaft Fractures.

BACKGROUND: Internal fixation of pediatric tibia shaft fractures has become increasingly popular despite the fact that non-operative treatment results in satisfactory outcome with few exceptions. Indications for surgery and benefits of internal fixation are however still debatable. MATERIALS AND METHODS: All 296 less than 16-year-old patients treated for a tibia shaft fracture between 2010 and 2015 in two of the five university hospitals in Finland were included in the study. Patient data were analyzed in three treatment groups: cast immobilization in emergency department, manipulation under anesthesia, and operative treatment. Incidence of operative treatment of pediatric tibia shaft fractures was calculated in the cities of Helsinki and Kuopio. RESULTS: A total of 143 (47.3%) children's tibia shaft fractures were treated with casting in emergency department, 71 (22.3%) with manipulation under anesthesia, and 82 (30.4%) with surgery. Mean age of the patients in these treatment groups was 6.2, 8.7, and 12.7 years. Fibula was intact in 89%, 51%, and 27% of the patients, respectively. All 6 patients with multiple fractures and 16 of 18 patients with open fractures were treated operatively. In eight patients, primary non-operative treatment was converted to internal fixation. Operatively treated patients with isolated closed fractures were more likely to have a fibula fracture (46/66 vs 52/214, p < 0.001), be older in age (13.08 ± 2.4 vs 6.4 ± 3.7, p < 0.001), and have more primary angulation (6.9 ± 5.8 vs 0.48 ± 3.1, p < 0.001). Re-operations were done to eight and corrective osteotomy to two operatively treated children. CONCLUSION: Operative treatment of a pediatric tibia shaft fracture is currently nearly a rule in patients with open or multiple fractures. Surgical treatment of closed tibia shaft fractures is based on surgeon's personal preference, type of fracture and age of the patient.

FLIM reveals alternative EV-mediated cellular up-take pathways of paclitaxel.

In response to physiological and artificial stimuli, cells generate nano-scale extracellular vesicles (EVs) by encapsulating biomolecules in plasma membrane-derived phospholipid envelopes. These vesicles are released to bodily fluids, hence acting as powerful endogenous mediators in intercellular signaling. EVs provide a compelling alternative for biomarker discovery and targeted drug delivery, but their kinetics and dynamics while interacting with living cells are poorly understood. Here we introduce a novel method, fluorescence lifetime imaging microscopy (FLIM) to investigate these interaction attributes. By FLIM, we show distinct cellular uptake mechanisms of different EV subtypes, exosomes and microvesicles, loaded with anti-cancer agent, paclitaxel. We demonstrate differences in intracellular behavior and drug release profiles of paclitaxel-containing EVs. Exosomes seem to deliver the drug mostly by endocytosis while microvesicles enter the cells by both endocytosis and fusion with cell membrane. This research offers a new real-time method to investigate EV kinetics with living cells, and it is a potential advancement to complement the existing techniques. The findings of this study improve the current knowledge in exploiting EVs as next-generation targeted drug delivery systems.

Comparison of liposomal drug formulations for transdermal iontophoretic drug delivery.

This study was aimed to evaluate the in vitro transdermal direct/pulsed current iontophoretic delivery of an amphiphilic model compound from various lipid vesicle-encapsulated formulations compared to free-drug formulation. Conventional, pegylated, ultradeformable liposomes (transfersomes) and ethosomes loaded with a negatively charged drug diclofenac sodium (DS) were prepared and characterized. All the liposomes possessed an average size of ≈100-150nm and negative zeta potential. No changes in colloidal stability were detected after 8h incubation of any vesicle formulation under constant or pulsed iontophoretic current. DS was released from all the liposome formulations with a similar, limited rate (≈50% in 24h), leading therefore to significantly lower transdermal fluxes across full-thickness porcine skin compared to the respective free drug formulation. From the tested lipid vesicle formulations, the transfersomes resulted in the highest passive flux and the ethosomes in the highest iontophoretic flux under direct constant current treatment. Higher negative surface charge of the vesicle led to better transport efficiency due to the higher mobility of the drug carrier under electric field. Pulsed current iontophoresis had no advantage over constant current treatment in combination with any type of lipid vesicular nanocarriers, in contrast to what has been described earlier with drug-loaded polymeric nanocarriers.

Transdermal iontophoresis of flufenamic acid loaded PLGA nanoparticles.

The objective of this study was to test in vitro a drug delivery system that combines nanoencapsulation and iontophoresis for the transdermal delivery of lipophilic model drug using poly(lactic-co-glycolic acid) (PLGA) as the carrier polymer. Negatively charged fluorescent nanoparticles loaded with negatively charged flufenamic acid were prepared. The colloidal properties of the particles were stable under iontophoretic current (constant, pulsed and alternating) profiles and in contact with skin barrier. The release of the drug from the particles was not affected by iontophoresis and remained always limited (≈50%), leading to significantly lower transdermal fluxes across human epidermis and full thickness porcine skin compared to respective free drug formulation. From nanoparticles, pulsed current profile resulted in comparable or higher fluxes compared to constant current profile although fluorescence imaging was not able to confirm deeper distribution of nanoparticles in skin. Based on our results, there is no clear advantage with respect to drug permeation from nanoencapsulating flufenamic acid into PLGA nanoparticles compared to free drug formulation, either in passive or iontophoretic delivery regimens. However, pulsed current iontophoresis could be an effective alternative instead of traditional constant current iontophoresis to enhance transdermal permeation of drugs from nanoencapsulated formulations.

Influence of exteroceptive contextual conditions upon internal drug stimulus control.

Rats were trained in a spatial T-maze discrimination either in a drugged (D = pentobarbital, 17.5 mg/kg) or in a non-drugged (N = saline) state (drug discrimination learning). Either of two external discriminative stimulus sets (light vs complete darkness) was consistently associated with the D or N state. When tested in the presence of the external stimulus previously associated with training in the D state, the animals made more drug-appropriate choices when tested with low pentobarbital doses as compared to testing in the external stimulus condition previously associated with the N state. This was reflected both in the ED50 values and the slopes of the dose-generalization gradients. The gradients of the controls were intermediate to those of the experimental rats. The present data suggest a new approach for studying interactions between controlling features in environmental events and the internal state.

Use of children's oral health services in Finland with special reference to heavy consumption.

The consumption of oral health services among 4-16-yr-old children in Finland is described, including an analysis of heavy consumption. Data representative of all Finnish children were collected for 4518 children from municipal health centers and homes. The response rate was 85%. Children with orthodontic visits during the year were excluded from the analyses. Of the preschoolchildren 12% and of the school-age children 4% did not use any oral health services during the studied year. Non-users did not differ from the others in terms of socioeconomic status, distance to a health center or type of residential area. Public oral health services were used very unevenly. Half of the children consumed only one-fifth of the services and the next one-third used about one-third of the services. The heavy consumers, about one-fifth of the total, used as much as half of all the services. Among preschoolchildren, the heavy consumers were mainly from lower socioeconomic groups. In contrast, no clear predictive factors were found for heavy consumption among the schoolchildren. Heavy consumers had more both treated and untreated caries than the other children did. Their treatment was mainly restorative; at least half of them did not receive adequate preventive care. Finnish public oral health care clearly should make more efforts to ensure that heavy consumers of dental services do not remain so.

Continuous noise, intermittent noise, and annoyance.

In a within-subjects design, 18 subjects listened to white noise, the intensity of which was controlled by themselves. They were instructed to choose the intensity they experienced as "clearly annoying." Three kinds of white noise were used: continuous, intermittent regular, and intermittent irregular. In the two intermittent conditions, the proportion of time with the noise on was 50%. The duration of on- and off-periods in the regular condition was 1.15 sec.; in the irregular condition, it varied between 0.25 and 1.65 sec. The subjects chose their "clearly annoying" level three times for each noise type. The results showed that the mean level chosen was 83.9 dB for continuous, 90 dB for intermittent regular, and 89.6 dB for intermittent irregular noise. Pairwise comparisons indicated a significant difference between continuous noise and each of the two intermittent noises while there was no difference between the two intermittent noises. The results are interpreted as indicating that noise-induced annoyance may be a function of the over-all amount of noise rather than the mere presence or absence of intermittency, at least when no concurrent demanding task is performed and when the required annoyance level is set by the subjects themselves. The results further showed that the intensity chosen by the subjects correlated negatively with scores from Weinstein's Noise Sensitivity Scale while the intensity chosen was unrelated to extraversion or neuroticism scores as measured by Eysenck Personality Inventory.

A repetitive reasoning task based on simple spatial relations.

This note describes a task that requires simple reasoning and can be used, e.g., in research on noise. The task includes a series of items of approximately equal difficulty. Each item consists of five geometric figures and a sentence that claims to describe spatial relations between the figures. The subject must decide whether the description is true. The task is short (3 1/2 min.) but can be extended at will.

Coverage of public oral health services for young children in Finland.

In the Finnish programme of public oral health care for preschool children, every child is eligible for free care. The coverage of the programme has been high, about 85%. The aim of this study was to determine, whether the nonparticipants of the programme actually do have equality in making the choice to participate. The barriers to care were identified. All 4- to 6-year-old children who were registered administratively as nonparticipants in oral health care in 1981 and their randomly selected controls were studied in the city of Lahti. Detailed analysis of nonparticipation showed that some children had been misclassified as nonparticipants in the administrative statistics, indicating the national coverage of the programme to be higher than reported. Based on this finding, suggestions are made for increasing reliability of official statistics. Multiple logistic regression analysis for 36 factors resulted in three main barriers to utilization. Children of single-parent families in which the parent had a low level of education and difficulties to take time off work had the highest risk of nonutilization and thus had little choice to participate. Children with fear to dental care, regardless of any characteristic of the family or parents, were at increased risk of nonparticipation. Not having received the invitation to an examination also increased nonutilization, regardless of any factors related to family, parents or child. It was concluded, however, that these barriers to utilization could be manipulated by the system of public oral health care.