Choroidal Thickness in healthy eyes measured by Widefield Optical Coherence Tomography.

INTRODUCTION: The choroid and its role in posterior segment pathology has become an increasing subject of study. The objective of the present study was to analyze choroidal thickness (CT) in healthy eyes by widefield (WF) optical coherence tomography (OCT) up to the periphery and to compare the reliability of manual versus automatic measurement. METHODS: Cross-sectional and non-interventional study conducted on 191 healthy eyes of 101 patients. All patients were scanned by using WF-OCT (Xephilio WF-OCT S1; Canon Corp, Tokyo, Japan). CT was measured in 2000 µm intervals automatically using the built-in software and manually by two masked observers. All analyses were performed using the IBM-PSSS statistical software program (IBM-SPSS, v. 28.0.0.0, Chicago, IL, USA). RESULTS: CT was measured in 100% of the sample. The mean age of the study cohort was 39.05±19.06 years old. Mean subfoveal (SF)CT measured automatically was 343.67±84.18 µm and manually was 336.55±75.57 µm. The thickest point was located 2000 µm from the fovea in the superior sector in 62.83% of the subjects. According to age distribution, mean CT became significantly thinner from 40 years of age. When comparing automatic and manually measuring, the intraclass correlation coefficient was excellent (p<0.01) in all quadrants. Moreover, manual measurement interobserver agreement was excellent in all quadrants (p<0.01). CONCLUSION: The automatic system is valid and serves as the basis of choroid measurement. In more than 50% of the healthy subjects, superior CT is thicker than subfoveolar CT and mean CT became significantly thinner from 40 years of age.

Could alternative pathways for carotenoid transformation affect colour production efficiency? A correlative study in wild birds.

In many vertebrates, dietary yellow carotenoids are enzymatically transformed into 4C-ketocarotenoid pigments, leading to conspicuous red colourations. These colourations may evolve as signals of individual quality under sexual selection. To evolve as signals, they must transmit reliable information benefiting both the receiver and the signaler. Some argue that the reliability of 4C-ketocarotenoid-based colourations is ensured by the tight link between individual quality and mitochondrial metabolism, which is supposedly involved in transforming yellow carotenoids. We studied how a range of carotenoids covary in the feathers and blood plasma of a large number (n > 140) of wild male common crossbills (Loxia curvirostra). Plumage redness was mainly due to 3-hydroxy-echinenone (3HOE). Two other, less abundant, red 4C-ketocarotenoids (astaxanthin and canthaxanthin) could have contributed to feather colour as they are redder pigments. This was demonstrated for astaxanthin but not canthaxanthin, whose feather levels were clearly uncorrelated to colouration. Moreover, moulting crossbills carried more 3HOE and astaxanthin in blood than non-moulting ones, whereas canthaxanthin did not differ. Canthaxanthin and 3HOE can be formed from echinenone, a probable product of dietary ß-carotene ketolation. Echinenone could thus be ketolated or hydroxylated to produce canthaxanthin or 3HOE, respectively. In moulting birds, 3HOE blood levels positively correlated to astaxanthin, its product, but negatively to canthaxanthin levels. Redder crossbills also had lower plasma canthaxanthin values. A decrease in hydroxylation relative to ketolation could explain canthaxanthin production. We hypothesize that red colouration could indicate birds' ability to avoid inefficient deviations within the complex enzymatic pathways.

Bigger or long-winged male common crossbills exhibit redder carotenoid-based plumage coloration.

Carotenoid-based ornaments are often considered reliable (honest) individual condition signals because their expression implies physiological costs unaffordable for low-quality animals (handicap signals). Recently, it has been suggested that efficient cell respiration is mandatory for producing red ketocarotenoids from dietary yellow carotenoids. This implies that red colorations should be entirely unfalsifiable and independent of expression costs (index signals). In a precedent study, male common crossbills, Loxia curvirostra, showing a red plumage reported higher apparent survival than those showing yellowish-orange colors. The plumage redness in this species is due to ketocarotenoid accumulation in feathers. Here, we correlated the male plumage redness (a 4-level visual score: yellow, patchy, orange, and red) and the body morphology in more than 1,000 adult crossbills captured in 3 Iberian localities to infer the mechanisms responsible for color evolution. A principal component analysis summarized morphometry of 10 variables (beak, wing, tarsus length, etc.). The overall body size (PC1) and the length of flight feathers regarding body size (PC3) showed significant positive relationships with plumage redness. Plumage redness was barely correlated with bill shape measures, suggesting no constraint in acquiring carotenoids from pine cones. However, large body sizes or proportionally long flying feathers could help carotenoid acquisition via social competition or increased foraging ranges. Proportionally longer flight feathers might also be associated with a specific cell respiration profile that would simultaneously favor flying capacities and enzymatic transformations needed for ketocarotenoid synthesis. Such a phenotypic profile would agree with the hypothesis of ketocarotenoid-based colors acting as individual quality index signals.

Wild common crossbills produce redder body feathers when their wings are clipped.

BACKGROUND: The animal signaling theory posits that conspicuous colorations exhibited by many animals have evolved as reliable signals of individual quality. Red carotenoid-based ornaments may depend on enzymatic transformations (oxidation) of dietary yellow carotenoids, which could occur in the inner mitochondrial membrane (IMM). Thus, carotenoid ketolation and cell respiration could share the same biochemical pathways. Accordingly, the level of trait expression (redness) would directly reveal the efficiency of individuals' metabolism and, hence, the bearer quality in an unfalsifiable way. Different avian studies have described that the flying effort may induce oxidative stress. A redox metabolism modified during the flight could thus influence the carotenoid conversion rate and, ultimately, animal coloration. Here, we aimed to infer the link between red carotenoid-based ornament expression and flight metabolism by increasing flying effort in wild male common crossbills Loxia curvirostra (Linnaeus). In this order, 295 adult males were captured with mist nets in an Iberian population during winter. Approximately half of the birds were experimentally handicapped through wing feather clipping to increase their flying effort, the other half being used as a control group. To stimulate the plumage regrown of a small surface during a short time-lapse, we also plucked the rump feathers from all the birds. RESULTS: A fraction of the birds with fully grown rump feathers (34 individuals) could be recaptured during the subsequent weeks. We did not detect any significant bias in recovery rates and morphological variables in this reduced subsample. However, among recaptured birds, individuals with experimentally impaired flying capacity showed body mass loss, whereas controls showed a trend to increase their weight. Moreover, clipped males showed redder feathers in the newly regrown rump area compared to controls. CONCLUSIONS: The results suggest that wing-clipped individuals could have endured higher energy expenditure as they lost body mass. Despite the small sample size, the difference in plumage redness between the two experimental groups would support the hypothesis that the flying metabolism may influence the redox enzymatic reactions required for converting yellow dietary carotenoids to red ketocarotenoids.

Testing the shared-pathway hypothesis in the carotenoid-based coloration of red crossbills.

The mechanisms involved in the production of red carotenoid-based ornaments of vertebrates are still poorly understood. These colorations often depend on enzymatic transformations (ketolation) of dietary yellow carotenoids, which could occur in the inner mitochondrial membrane (IMM). Thus, carotenoid ketolation and cell respiration could share biochemical pathways, favoring the evolution of ketocarotenoid-based ornaments as reliable indices of individual quality under sexual selection. Captive male red crossbills (Loxia curvirostra Linnaeus) were exposed to redox-active compounds designed to penetrate and act in the IMM: an ubiquinone (mitoQ) or a superoxide dismutase mimetic (mitoTEMPO). MitoQ can act as an antioxidant but also distort the IMM structure, increasing mitochondrial free radical production. MitoQ decreased yellow carotenoids and tocopherol levels in blood, perhaps by being consumed as antioxidants. Contrarily, mitoTEMPO-treated birds rose circulating levels of the second most abundant ketocarotenoid in crossbills (i.e., canthaxanthin). It also increased feather total red ketocarotenoid concentration and redness, but only among those birds exhibiting a redder plumage at the start of the study, that is, supposedly high-quality individuals. The fact that mitoTEMPO effects depended on original plumage color suggests that the red-ketocarotenoid-based ornaments indicate individual quality as mitochondrial function efficiency. The findings would thus support the shared pathway hypothesis.