Inherent immunological parameters in kutum, Rutilus frisii, larvae obtained from the adult females with orange and green eggs.

Kutum (Rutilus frisii) displays different egg colours during the spawning season, mostly due to the presence of carotenoid pigments. In this study, the authors investigated the relationship between the egg colour and muscle lipid of adult female kutum and the correlation between egg carotenoid content and the immune parameters of larvae. The results from this study highlighted the positive influence of egg carotenoid on post-fertilization stages, such as elevating the innate immune parameters in larvae.

Plasticity and divergence in ultraviolet reflecting structures on Dogface butterfly wings.

The vast diversity of animal coloration is generated through a combination of pigment and structural colors. These colors can greatly influence the fitness and life history of an organism. Butterflies and their wing colors are an excellent model to study how these colors can impact the development and success of an organism. In this study, we explore species differences in structurally-based ultraviolet coloration in the Zerene butterfly. We show clear species differences in ultraviolet (UV) pattern and reflectance spectra. By varying larval diet, we show evidence for developmental plasticity in the structure and organization of UV reflecting scales in Zerene cesonia. We further show that feeding the larval host plant of Zerene eurydice to Z. cesonia does not result in greater similarity in scale structure or UV coloration to the sister species. These results not only demonstrate a connection between plasticity in a male ornamentation, UV wing pattern, and larval resource acquisition, but also identify candidate structural and organizational changes in wing scales responsible for the trait variation.

Efeito da pigmentação extrínseca nas cerâmicas à base de dissilicato de lítio submetidas aos processos de desgaste e biodegradação nas propriedades mecânicas e aderência bacteriana / Effect of extrinsic pigmentation on lithium disilicate-based ceramics subjected to wear and biodegradation processes on mechanical properties and bacterial adherence

O objetivo deste estudo foi avaliar se há alteração no comportamento mecânico e na aderência microbiológica da cerâmica à base de dissilicato de lítio com a técnica de pigmentação extrínseca aplicada, após ser submetida a diferentes condições experimentais. Foram confeccionadas 160 amostras, divididas em grupos com e sem pigmentação (n=80). Destes, cada grupo foi subdividido em Controle, Desgate, Biodegradação e Desgaste com Biodegradação (n=20).15 amostras de cada subgrupo foram submetidas ao teste de resistência à flexão e 5 para o teste de aderência microbiológica. As amostras passaram anteriormente por testes complementares para caracterização da superfície (rugosidade, perfilometria volumétrica e microscopia eletrônica de varredura (MEV)). Os resultados foram submetidos a análise estatística descritiva (média e desvio padrão) e inferencial, mediante o teste paramétrico de análise de variância (ANOVA) dois fatores e teste de Tukey ( = 0,05). O fator pigmentação extrínseca influenciou negativamente no comportamento mecânico da cerâmica, apresentando significância estatística (p = 0,000), assim como a interação entre o tipo de condição experimental e a pigmentação (p = 0,020). Entretanto, na aderência microbiológica, foi a condição experimental que influenciou negativamente no comportamento microbiológico (p = 0,000), assim como a interação entre a condição experimental e a pigmentação (p = 0,000). Nas análises complementares, observou-se que a interação entre os fatores aumentou a rugosidade superfial (p = 0,000) e aumentou o volume perdido pelo desgatse (p = 0,040). As microscopias da superficie mostram as características de cada grupo, mostrando as diferenças entre as condições experimentais com e sem pigmentação extrínseca. E as microscopias da aderência microbiológica ilustram e confirmam os resultados obtidos no teste estatístico. Concluiu-se que a pigmentação extrínseca altera as propriedades mecânicas da cerâmica de dissilicato de lítio, reduzindo a resistência à flexão e aumentando a rugosidade superficial e o desgaste. Porém, a aderência microbiológica foi aumentada pela condição experimental. Entretanto, a interação entre os fatores contribuiu para esse aumento e para agravar a alteração nas propriedades mecânicas(AU)

The objective of this study was to evaluate the mechanical and microbiological behavior of the ceramics based on lithium disilicate with extrinsic characterization. For this, 160 discs were made, divided into two large groups, with extrinsec characterization and without, after which each was divided into four groups (n = 20): Control, Wear, Biodegradation and Biodegradation with Wear. Fifteen samples from each group were submitted to the flexural strength test and 5 submitted to the microbiological adherence test. Prior to the destructive test of flexural strength, the representative samples of each group underwent complementary tests for surface characterization. The results were submitted to descriptive statistical analysis (mean and standard deviation) and inferential, using the parametric analysis of variance (ANOVA) two way and Tukey test ( = 0,05). The extrinsec characterization factor influenced the mechanical behavior of the ceramic, presenting statistical significance (p = 0.000), as well as the interaction between the type of experimental condition and the extrinsec characterization (p = 0.020). However, in the microbiological adherence, it was the experimental condition that influenced the microbiological behavior (p = 0.000), as well as the interaction between the experimental condition and the extrinsec characterization (p = 0.000). It was concluded that the makeup influenced the mechanical behavior of the ceramic, and the experimental condition influenced the microbiological adherence. The interaction between the factors influenced both the mechanical behavior and the microbiological adherence(AU)

Stepwise evolution of floral pigmentation predicted by biochemical pathway structure.

Developmental pathways play a major role in influencing the distribution of naturally occurring phenotypes. For example, pathway structure and regulation could make some phenotypes inaccessible or restrict the routes through which phenotypes evolve. In this study, we examine floral anthocyanin pigments across the Solanaceae family and test whether patterns of phenotypic variation are consistent with predicted constraints based on the structure of the flavonoid biosynthetic pathway. We find that anthocyanin evolution occurs in a stepwise manner whereby transitions between the production of red mono hydroxylated pelargonidin pigments and blue trihydroxylated delphinidin pigments first passes through an intermediate step of producing purple dihydroxylated cyanidin pigments. Although the transitions between these three pigment types differ in frequency, we infer that these shifts are often reversible, suggesting that the functionality of the underlying biochemical pathway is generally conserved. Furthermore, our study finds that some pigment combinations are never observed, pointing to additional constraints on naturally occurring phenotypes. Overall, our findings provide insights into how the structure of an angiosperm-wide biochemical pathway has shaped macroevolutionary variation in floral pigmentation.

No evidence that carotenoid pigments boost either immune or antioxidant defenses in a songbird.

Dietary carotenoids have been proposed to boost immune system and antioxidant functions in vertebrate animals, but studies aimed at testing these physiological functions of carotenoids have often failed to find support. Here we subject yellow canaries (Serinus canaria), which possess high levels of carotenoids in their tissue, and white recessive canaries, which possess a knockdown mutation that results in very low levels of tissue carotenoids, to oxidative and pathogen challenges. Across diverse measures of physiological performance, we detect no differences between carotenoid-rich yellow and carotenoid-deficient white canaries. These results add further challenge to the assumption that carotenoids are directly involved in supporting physiological function in vertebrate animals. While some dietary carotenoids provide indirect benefits as retinoid precursors, our observations suggest that carotenoids themselves may play little to no direct role in key physiological processes in birds.

Involvement of Different Genes Expressions during Immunological and Inflammatory Responses in Vitiligo.

Vitiligo is a condition of the skin distinguished by hypo-pigmentation. Etiology of this disorder is unknown, and several theories and mechanisms have been hypothesized. The inflammatory response in vitiligo is thought to be mediated by polymorphism in genes such as FOXP3, ACE, APE, GSTP1, TLR, SOD, CTLA-4, TAP/LMP gene cluster, etc. Theories including reactive oxygen species model, Nrf2-antioxidant response element (ARE) pathway, WNT pathway, tyrosinase activity, biochemical, molecular, and cellular alterations have been hypothesized to explain vitiligo pathogenesis. Melanosomal proteins are involved in antigen processing. The antigens are expressed to the T-cells in the form of peptides with HLA class II molecules. T-cells are activated in response to the discharge of co-stimulatory molecules such as LFA-3 as well as ICAM-1. An adaptive immune response is thus elicited, and the melanocytes eventually die or start malfunctioning and the skin undergoes hypo-pigmentation. IFN-γ is known to be a melanocyte inhibitor of paracrine origin; it is clearly involved in the early onset of symptoms of vitiligo disease. The surge in the IFN-γ levels mediates augmented expression of ICAM-1 molecule on the melanocytes, thereby establishing cytokine-mediated destruction of melanocytes. Mainly, mediators released by melanocytes and the functionality of keratinocytes decrease the disease activity. Such mediators include ET-1 as well as SCF, increase the pigmentation particularly when a patient is given with the UVB treatment. By scavenging ROS and screening UV radiation, melanin limits the damage caused to the cutaneous cells by UV radiation. Various immune responses play important roles in vitiligo.

The effect of growth rate and ageing on colour variation of European pond turtles.

Many chelonians have colourful dots, patches and stripes throughout their body that are made up, at least in part, of carotenoids. Therefore, turtles are very suitable models to study the evolution and functionality of carotenoid-based colouration. Recent studies suggested a close link between colouration and immune system in these taxa. However, more research is needed to understand the role of these colourful stripes and patches in turtle visual signalling. The purpose of this study was to explore the relationship between growth rate and colouration in European pond turtles. In particular, we wanted to answer the question of whether there is a trade-off between growth and colour expression. We also aimed to explore the effect of body size and age on colour variation. Turtles from a reintroduction-breeding program were recaptured, weighed and measured over an 8-year period to estimate their growth rates and age. We also measured with a spectrometer the reflectance of colour patches in two different body parts: shell and forelimb. We found that turtles with a faster growth rate had brighter limb stripes independently of their age. On the other hand, shell colouration was related to body size with larger turtles having brighter shell stripes and higher values of carotenoid chroma. Our results suggest that fast-growers may afford to express intense colourful limb stripes likely due to their higher intake of carotenoids that would modulate both growth and colour expression. However, shell colouration was related to body size probably due to ontogenetic differences in the diet, as juveniles are strictly carnivorous while adults are omnivorous. Alternatively, shell colouration might be involved in crypsis as the shell is visually exposed to predators.

Immune challenges and visual signalling in tree frogs.

In animals, mate-choice is often based on sexual signals that carry information and help the receiver make the best choice to improve the receiver's fitness. Orange visual sexual signals have been hypothesised to carry immune information because they are often due to carotenoid pigments which are also involved in immunity response. Although many studies have focused on the direct relationships between coloration and immunocompetence, few studies have simultaneously studied immunocompetent response and coloration variation after an immune challenge. We tested this hypothesis on starved and ad libitum-fed males of the European tree frog Hyla arborea. Our results show that male coloration is not a reliable indicator of its immune response capacity in this species. However, after an immune challenge induced by a PHA (Phaseolus vulgaris phytohaemagglutinin) injection, starved males presented a significant coloration loss and this alteration was related to the immune response intensity. Taken together, these results suggest that the brighter (lighter) coloration may be used as a cue by female to exclude males with a recent immune challenge, due to diseases or parasites for example.

Population-Specific Covariation between Immune Function and Color of Nesting Male Threespine Stickleback.

Multiple biological processes can generate sexual selection on male visual signals such as color. For example, females may prefer colorful males because those males are more readily detected (perceptual bias), or because male color conveys information about male quality and associated direct or indirect benefits to females. For example, male threespine stickleback often exhibit red throat coloration, which females prefer both because red is more visible in certain environments, and red color is correlated with male immune function and parasite load. However, not all light environments favor red nuptial coloration: more tannin-stained water tends to favor the evolution of a melanic male phenotype. Do such population differences in stickleback male color, driven by divergent light environments, lead to changes in the relationship between color and immunity? Here, we show that, within stickleback populations, multiple components of male color (brightness and hue of four body parts) are correlated with multiple immune variables (ROS production, phagocytosis rates, and lymphocyte:leukocyte ratios). Some of these color-immune associations persist across stickleback populations with very different male color patterns, whereas other color-immune associations are population-specific. Overall, lakes with red males exhibit stronger color-immune covariance while melanic male populations exhibit weak if any color-immune associations. Our finding that color-immunity relationships are labile implies that any evolution of male color traits (e.g., due to female perceptual bias in a given light environment), can alter the utility of color as an indicator of male quality.

Vitiligo.

Vitiligo is an acquired depigmenting disorder that affects 0.5% to 2% of the world population. Three different forms are classified according to the distribution of lesions; namely non-segmental, segmental and mixed vitiligo. Vitiligo is associated with polymorphisms in genes involved in the immune response and in melanogenesis. However, environmental factors are required for the development of manifest disease. In general, the diagnosis is clinical and no laboratory tests or biopsies are required. Metabolic alterations are central to current concepts in pathophysiology. They induce an increased generation of reactive oxygen species and susceptibility to mild exogenous stimuli in the epidermis. This produces a senescent phenotype of skin cells, leads to the release of innate immune molecules, which trigger autoimmunity, and ultimately causes dysfunction and death of melanocytes. Clinical management aims to halt depigmentation, and to either repigment or depigment the skin, depending on the extent of disease. New therapeutic approaches include stimulation of melanocyte differentiation and proliferation through α-melanocyte-stimulating hormone analogues and through epidermal stem cell engineering. Several questions remain unsolved, including the connection between melanocyte depletion and stem cell exhaustion, the underlying degenerative mechanisms and the biological mediators of cell death. Overall, vitiligo is an excellent model for studying degenerative and autoimmune processes and for testing novel approaches in regenerative medicine. For an illustrated summary of this Primer, visit: http://go.nature.com/vIhFSC.

Honest sexual signaling in turtles: experimental evidence of a trade-off between immune response and coloration in red-eared sliders Trachemys scripta elegans.

Sexual signals can be evolutionarily stable if they are honest and condition dependent or costly to the signaler. One possible cost is the existence of a trade-off between maintaining the immune system and the elaboration of ornaments. This hypothesis has been experimentally tested in some groups of animals but not in others such as turtles. We experimentally challenged the immune system of female red-eared sliders Trachemys scripta elegans, with a bacterial antigen (lipopolysaccharide (LPS)) without pathogenic effects to explore whether the immune activation affected visual colorful ornaments of the head. The LPS injection altered the reflectance patterns of color ornaments. In comparison to the control animals, the yellow chin stripes of injected animals exhibited (1) reduced brightness, (2) lower long wavelength (>470 nm) reflectance, and (3) lower values for carotenoid chroma. The postorbital patches of injected individuals also showed reduced very long wavelength (>570 nm) reflectance but did not change in carotenoid chroma. Thus, experimental turtles showed darker and less "yellowish" chin stripes and less "reddish" postorbital patches at the end of the experiment, whereas control turtles did not change their coloration. This is the first experimental evidence supporting the existence of a trade-off between the immune system and the expression of visual ornaments in turtles. We suggest that this trade-off may allow turtles to honestly signal individual quality via characteristics of coloration, which may have an important role in intersexual selection processes.

Shell colouration and parasite tolerance in two helicoid snail species.

The polymorphism of shell colouration in helicoid snails is a well-known phenomenon attributed to different factors such as predation and climatic effects. Another aspect contributing to this polymorphism could be the interplay of melanin production and phenoloxidase-related immunity. Therefore, in this study we aimed at answering the questions whether there is a differential sensitivity of different snail shell colour morphs to nematode infection, and whether this can be related to differences in phenoloxidase (PO) activity levels using the two helicoid, polymorphic snail species Cepaea hortensis and Cernuella virgata. Snails of both species were artificially infected with the parasitic nematode Phasmarhabditis hermaphrodita, and analysed for mortality and PO activity levels. We found C. virgata to be more severely affected by P. hermaphrodita infection than C. hortensis, and the dark C. virgata morphs to be more resistant to lethal effects of this infection than pale morphs. However, these differences in sensitivity to the parasite could not clearly be related to different PO activity levels.

Sexually dichromatic coloration reflects size and immunocompetence in female Spanish terrapins, Mauremys leprosa.

Many studies have shown the importance of colorful ornamentation in mate choosiness or intrasexual conflict. However, research on color ornaments has focused mainly on birds, lizards or fish, but remains practically unknown in other animal groups such as turtles. In addition, female ornaments and their relation with sexual selection also remain almost unknown. Here, we measured the coloration of the shell and the limb stripes of male and female Spanish terrapins Mauremys leprosa and explored the existence of sexual dichromatism and the relation of color characteristics with body size and health state estimated from the immune response to the injection of an antigen (phytohaemagglutinin test). Our results showed that shell coloration, which could be constrained by natural selection to be cryptic, changed with body size, but did not differ between sexes. In contrast, females had brighter and less ultraviolet-saturated and more orange-saturated limb stripes than males. In females, interindividual variation in limb stripe coloration was related with body size and immune response suggesting that this coloration may inform honestly about multiple traits that could be important in sexual selection. In contrast, coloration of limb stripes of males was duller than in females, and was not related with any trait suggesting that coloration is not important in sexual selection for males.

Multivariate heredity of melanin-based coloration, body mass and immunity.

The genetic covariation among different traits may cause the appearance of correlated response to selection on multivariate phenotypes. Genes responsible for the expression of melanin-based color traits are also involved in other important physiological functions such as immunity and metabolism by pleiotropy, suggesting the possibility of multivariate evolution. However, little is known about the relationship between melanin coloration and these functions at the additive genetic level in wild vertebrates. From a multivariate perspective, we simultaneously explored inheritance and selection of melanin coloration, body mass and phytohemagglutinin (PHA)-mediated immune response by using long-term data over an 18-year period collected in a wild population of the common kestrel Falco tinnunculus. Pedigree-based quantitative genetic analyses showed negative genetic covariance between melanin-based coloration and body mass in male adults and positive genetic covariance between body mass and PHA-mediated immune response in fledglings as predicted by pleiotropic effects of melanocortin receptor activity. Multiple selection analyses showed an increased fitness in male adults with intermediate phenotypic values for melanin color and body mass. In male fledglings, there was evidence for a disruptive selection on rump gray color, but a stabilizing selection on PHA-mediated immune response. Our results provide an insight into the evolution of multivariate traits genetically related with melanin-based coloration. The differences in multivariate inheritance and selection between male and female kestrels might have resulted in sexual dimorphism in size and color. When pleiotropic effects are present, coloration can evolve through a complex pathway involving correlated response to selection on multivariate traits.

Dermal mesenchymal stem cells (DMSCs) inhibit skin-homing CD8+ T cell activity, a determining factor of vitiligo patients' autologous melanocytes transplantation efficiency.

We here investigated the efficiency of autologous melanocyte transplantation of 23 vitiligo patients by focusing on perilesional skin homing CD8+ T lymphocytes, and studied the potential effect of dermal mesenchymal stem cells (DMSCs) on CD8+ T cell activities in vitro. Out of 23 patients with the autologous melanocyte transplantation, 12 patients (52.17%) had an excellent re-pigmentation, 6 patients (26.09%) had a good re-pigmentation, 5 patients (21.74%) had a fair or poor re-pigmentation. CD8+ T cells infiltrating was observed in the perilesional vitiligo area of all patients. Importantly, the efficiency of the transplantation was closely associated with skin-homing CD8+ T cell activities. The patients with high number of perilesional CD8+ T cells or high level of cytokines/chemokines were associated with poor re-pigmentation efficiency. For in-vitro experiments, we successfully isolated and characterized human DMSCs and skin-homing CD8+ T cells. We established DMSCs and CD8+ T cell co-culture system, where DMSCs possessed significant inhibitory effects against skin homing CD8+ T lymphocytes. DMSCs inhibited CD8+ T cells proliferation, induced them apoptosis and regulated their cytokines/chemokines production. Our results suggest that vitiligo patients' autologous melanocytes transplantation efficiency might be predicted by perilesional skin-homing CD8+ T cell activities, and DMSCs might be used as auxiliary agent to improve transplantation efficacy.

Tail loss compromises immunity in the many-lined skink, Eutropis multifasciata.

Tail autotomy incurs energetic costs, and thus, a trade-off in resource allocation may lead to compromised immunity in lizards. We tested the hypothesis that tailless lizards will favor constitutive innate immunity responses over an energetically costly inflammatory response. The influence of fasting and colorful ornamentation was also investigated. We experimentally induced tail autotomy in the lizard Eutropis multifasciata and found that inflammation was suppressed by tail loss, but not further affected by fasting; the suppressive effect of colorful ornamentation was manifested only in males, but not in females. Constitutive innate immunity was not affected by any of these factors. As expected, only costly inflammation was compromised, and a less expensive constitutive innate immunity might be favored as a competent first-line defense during energetically demanding periods. After considering conventional trade-offs among tail regeneration and reproduction, further extending these studies to incorporate disease risk and how this influences escape responses to predators and future reproduction would make worthwhile studies.

Biolistic DNA vaccination against melanoma.

We describe here the use of particle-mediated gene transfer for the induction of immune responses against melanoma antigens in murine tumor models using the melanocyte differentiation antigen tyrosinase-related protein 2 (TRP2) as an antigen in a murine B16 melanoma model. We have utilized marker genes such as ß-galactosidase (ßgal) and EGFP, which can be readily detected, as control antigens to establish the gene delivery and to detect antigen-specific humoral and cellular immune responses. After biolistic DNA vaccination with plasmids encoding the TRP2 gene we observed the induction of TRP2-specific T-cells and antibodies associated with vitiligo-like fur depigmentation and tumor immunity against B16 melanoma cells. Here we describe the preparation of cartridges with DNA-coated gold beads and the in vivo gene transfer into skin using the Helios Gene Gun system. We also describe protocols for the measurement of humoral and cellular immune responses against the melanocyte differentiation antigen TRP2. These protocols can subsequently be adapted to other antigens.

What does carotenoid-dependent coloration tell? Plasma carotenoid level signals immunocompetence and oxidative stress state in birds-A meta-analysis.

Mechanisms maintaining honesty of sexual signals are far from resolved, limiting our understanding of sexual selection and potential important parts of physiology. Carotenoid pigmented visual signals are among the most extensively studied sexual displays, but evidence regarding hypotheses on how carotenoids ensure signal honesty is mixed. Using a phylogenetically controlled meta-analysis of 357 effect sizes across 88 different species of birds, we tested two prominent hypotheses in the field: that carotenoid-dependent coloration signals i) immunocompetence and/or ii) oxidative stress state. Separate meta-analyses were performed for the relationships of trait coloration and circulating carotenoid level with different measures of immunocompetence and oxidative stress state. For immunocompetence we find that carotenoid levels (r = 0.20) and trait color intensity (r = 0.17) are significantly positively related to PHA response. Additionally we find that carotenoids are significantly positively related to antioxidant capacity (r = 0.10), but not significantly related to oxidative damage (r = -0.02). Thus our analyses provide support for both hypotheses, in that at least for some aspects of immunity and oxidative stress state the predicted correlations were found. Furthermore, we tested for differences in effect size between experimental and observational studies; a larger effect in observational studies would indicate that co-variation might not be causal. However, we detected no significant difference, suggesting that the relationships we found are causal. The overall effect sizes we report are modest and we discuss potential factors contributing to this, including differences between species. We suggest complementary mechanisms maintaining honesty rather than the involvement of carotenoids in immune function and oxidative stress and suggest experiments on how to test these.

One trait, many signals: different information on male quality is enclosed within the same trait in a blenny fish.

Colourful ornaments are traditionally evaluated as one trait. However, they could consist of several components, such as total size, colour intensity and extension, each possibly bearing its own message about one aspect of quality. Males of the blenny Salaria pavo exhibit a colourful head crest and solely care for eggs. During the breeding season, the head crest shows a yellow colouration, the intensity and relative extension of which are independent of crest size. Here, we show that: (1) carotenoids are responsible for the head crest yellow patch; (2) activating the immune system by injecting the bacterial antigen lipopolysaccharides affects both the intensity and extent of the yellow colouration; and (3) females assess males on the basis of colour patch expression. However, the response of the yellow patch to the immune challenge was dependent on head crest size. Indeed, males with a larger head crest reacted better to the simulated infection, sustaining a level of yellow patch close to pre-challenge size.