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Erythema is a hallmark skin response to excessive ultraviolet radiation (UVR) and is associated with cutaneous inflammation. Both are mediated by inflammatory mediators including nitric oxide (NO), prostaglandin E2 (PGE2) and chemoattractants such as 12-hydroxyeicosatetraenoic acid (12-HETE) leading to vasodilation and increased leukocyte infiltration. The erythematous response is more pronounced in individuals with low basal melanin levels or who fail to respond to UVR with a robust up-regulation of melanogenesis. While melanin production is a key function of melanocytes, these cells can also produce NO and PGE2, and are located in close proximity to the dermal vasculature. It has been hypothesized that melanocytes with poor melanogenic capacity may participate in the inflammatory response to UVR. The aim of this project was to investigate the inflammatory response in the skin of individuals with either skin phototype (SPT) 1 or 4 to UVR. Sixteen normal healthy individuals were selected for study (8 SPT-1 & 8 SPT-4). Buttock skin was investigated by immunohistochemistry for leukocyte subtypes, eicosanoid producing enzymes and NO synthases under basal and UVR-stimulated conditions. In addition primary cultures of epidermal melanocytes (EM) were established from 16 individuals (8 SPT-1 & 8 SPT-4) and assessed for the presence of eicosanoid-producing enzymes, melanogenic enzymes and NO synthases, by immunocytochemistry, Polymerase Chain Reaction and Western Blotting and for the production of the main pro-inflammatory eicosanoid PGE2 by ELISA and Mass Spectrometry. Moreover, the fatty acid composition of cultured melanocytes was assessed by Gas Chromatography. Results showed that individuals with SPT-1 had significantly greater neutrophil infiltration into the epidermis than those with SPT-4 at 24 hrs post-UVR. Moreover, CD3+ lymphocyte infiltration into the dermis was significantly greater in individuals with SPT-4 than those with SPT-1 at 24 and 72 hrs post-UVR. NOS-1, NOS-3, 12-LOX and COX-2 expression were significantly increased in SPT-1 skin, while NOS-2 and 15-LOX were significantly increased in SPT-4 skin. As 12-LOX and COX-2 products are chemoattractive (for neutrophils) and pro-inflammatory respectively these data could explain the greater observed neutrophil infiltration in SPT-1. The 15-LOX product (15-HETE) is anti-inflammatory and may suggest that 15-LOX up-regulation in SPT-4 skin may aid resolution of the sunburn response, which in part may be mediated by CD3+ lymphocytes and a class-switch in eicosanoid production from COX to LOX products. Melanocyte primary cultures surprisingly showed that SPT was not correlated with melanin content or melanogenic enzyme expression/activity suggesting that all melanocytes in vitro contained the necessary cellular machinery to produce melanin. This finding may reflect also their equal treatment under these enriched culture conditions, which may or may not be available to these cells in situ. Moreover, all melanocytes expressed the necessary machinery (PLA2, COX-1, cPGES) to produce PGE2. However, only some cultures did so at baseline and in response to UVR, and this was not correlated with SPT. A positive correlation was found however between expression level of dopachrome tautomerase (DCT) and protection against PGE2 production in response to UVR, which may suggest a novel role for DCT unrelated to melanogenesis. In summary this research project has generated data that highlights differences between the skin of individuals with SPT-1 and those with SPT-4, and may provide evidence that the keratinocyte partner contributes significantly to the SPT-associated response. This research may also suggest DCT as a novel therapeutic target to protect EM from participation in the UVR-associated inflammatory response in skin. / Wellcome Trust

Bakgrund: Inom ambulanssjukvård i Sverige, likt inom övrig sjukvård, ökar förekomsten av patienter med melaninrik hud. Lagkrav om individanpassad och jämlik vård finns, samtidigt påvisas brister i undervisning om melaninrik hud för sjuksköterskestudenter. I ambulanssjukvård tas ofta snabba beslut grundade på initiala bedömningar, och kunskap om hudbedömning är av vikt för sjuksköterskans arbetsdiagnos eftersom tecken på akut sjukdom snabbt kan avslöjas genom manifestationer på huden. Syfte: Att undersöka sjuksköterskors erfarenheter av att bedöma patienter med melaninrik hud i ambulanssjukvård. Metod: En kvalitativ studie med induktiv ansats där semistrukturerade intervjuer använts som datainsamlingsmetod. Urvalsgruppen bestod av tio sjuksköterskor med yrkeserfarenhet från ambulansverksamhet. Data har analyserats genom kvalitativ innehållsanalys enligt Graneheim och Lundman (2004). Resultat: Sjuksköterskor känner osäkerhet i bedömningen av patienter med melaninrik hud på grund av utbildningsbrister och visuella utmaningar. Detta understryker behovet av förbättrad utbildning för jämlik prehospital omvårdnad. Konklusion: Omvårdnadsutbildningen för sjuksköterskor behöver införliva fler resurser för bedömning av melaninrik hud. Prioritering av varierande simuleringsfall och bilder är nödvändigt för jämlik vård. / Background: In ambulance healthcare in Sweden, as in other healthcare, the incidence of patients with melanin-rich skin is increasing. Legal requirements for individualized and equal care exist, at the same time deficiencies are demonstrated in teaching about melanin-rich skin for nursing students. In ambulance care, quick decisions are often made based on initial assessments, and knowledge of skin assessment is important to the nurse's working diagnosis because signs of acute illness can be quickly revealed by manifestations on the skin. Aim: To investigate nurses' experiences of assessing patients with melanin-rich skin in ambulance care. Method: A qualitative study with an inductive approach where semi-structured interviews were used as data collection method. The sample group consisted of ten nurses with professional experience from ambulance operations. The data has been analyzed through qualitative content analysis according to Graneheim and Lundman (2004). Findings: Nurses experience uncertainty in assessing patients with melanin-rich skin due to educational gaps and visual challenges. This emphasizes the need for enhanced education to ensure equitable prehospital care. Conclusion: Nursing education for nurses needs to incorporate more resources for the assessment of melanin-rich skin. Prioritizing diverse simulation cases and images is essential for equitable care.

No / Patients with acute vitiligo have low epidermal catalase expression/activities and accumulate 10 -3 M H 2O 2. One consequence of this severe oxidative stress is an altered calcium homeostasis in epidermal keratinocytes and melanocytes. Here, we show decreased epidermal calmodulin expression in acute vitiligo. Since 10 -3M H 2O 2 oxidises methionine and tryptophan residues in proteins, we examined calcium binding to calmodulin in the presence and absence of H 2O 2 utilising 45calcium. The results showed that all four calcium atoms exchanged per molecule of calmodulin. Since oxidised calmodulin looses its ability to activate calcium ATPase, enzyme activities were followed in full skin biopsies from lesional skin of patients with acute vitiligo (n = 6) and healthy controls (n = 6). The results yielded a 4-fold decrease of ATPase activities in the patients. Computer simulation of native and oxidised calmodulin confirmed the loss of all four calcium ions from their specific EF-hand domains. Taken together H 2O 2-mediated oxidation affects calcium binding in calmodulin leading to perturbed calcium homeostasis and perturbed L-phenylalanine-uptake in the epidermis of acute vitiligo.

Rainbow trout (Oncorhynchus mykiss) are introduced in large parts of the world and are commonly farmed for consumption as well as a valued sport fish. Many species of salmonids show large intraspecific variation in pigmentation, which has been shown to correlate with stress response and several other behavioral traits. In this study, the behavior and stress response of rainbow trout linked to pigmentation is investigated, depending on previous sport fishing experience. Through data from a previous study on rainbow trout in semi-natural ponds, locomotor activity was measured as a proxy of stress, under three different sport fishing treatments. From previous photos, the pigment spots of each rainbow trout are counted to test the correlation with locomotor activity. In addition, it is tested whether different previous experience of sport fishing means a higher stress response when re-exposed to fishing. No significant relationship between pigment spots and locomotor activity could be obtained for the three treatment groups. However, significant differences in locomotor activity due to angling experience between treatment groups upon re-exposure to fishing were found, with fish that were inexperienced in angling having higher locomotor activity compared to previously caught fish. Sport fishing and catch-and-release had the effect of reducing locomotor activity in rainbow trout, which can be interpreted as fishing could both stress them and trigger the fish's feeding response, depending on previous experience. Fishing is believed to create a passivation due to the negative association of being caught, while fish not previously caught were activated by sport fishing. The study's missing correlation between pigment spots and stress has been both confirmed and denied in other studies and may depend on the origin and the degree of domestication, which means that the correlation within other species of salmonids or fish with different origins would be relevant to investigate. / Regnbåge (Oncorhynchus mykiss) förekommer introducerade i stora delar av världen och är vanliga att odlas för konsumtion samt en uppskattad sportfisk. Många arter av salmonider visar stor intraspecifik variation i pigmentering, vilket visats korrelera med stressrespons och flera andra beteendemässiga karaktärsdrag. I denna studie undersöks regnbågarnas beteende och stressrespons kopplat till pigmentering, beroende på tidigare erfarenhet av sportfiske. Genom data från en tidigare studie på regnbåge i semi-naturliga dammar mäts rörelseaktivitet som indirektmått på stress, under tre olika sportfiskebehandlingar. Från tidigare foton räknas varje regnbåges pigmentfläckar för att testa korrelationen med rörelseaktivitet. Därtill testas om olika tidigare erfarenhet av sportfiske, innebär högre stressrespons vid återexponering för fiske. Inget signifikant samband mellan pigmentfläckar och rörelseaktivitet kunde erhållas för de tre behandlingsgrupperna. Signifikanta skillnader i rörelseaktivitet på grund av erfarenheten av sportfiske mellan behandlingsgrupperna vid återexponering för fiske fanns dock, där fisk som var oerfaren sportfiske hade högre rörelseaktivitet jämfört med fisk som fångats tidigare. Sportfiske och catch-and-release hade effekten att minska rörelseaktiviteten hos regnbågar, vilket kan tolkas som att fisket både kunde stressa och trigga fiskens födorespons, beroende på tidigare erfarenhet. Fisket tros skapa en passivisering på grund av den negativa associationen att bli fångad, medan fisk som inte fångats tidigare aktiverades av sportfisket. Studiens uteblivna samband mellan pigmentfläckar och stress har både bekräftats och dementerats i andra studier och kan bero på ursprung och graden av domesticering, vilket gör att sambandet inom andra arter av salmonider eller fisk med olika ursprung vore aktuellt att undersöka.

Le phytomanagement est une méthode de gestion de sites pollués basée sur la capacité des plantes à séquestrer les polluants. L’utilisation de symbiotes fongiques peut représenter un atout afin d’augmenter la production de biomasse. Les endophytes bruns septés (DSE) pourraient stimuler la croissance végétale et augmenter leur protection contre les éléments-traces-métalliques (ETM). Cette thèse se déroule dans le cadre du projet LORVER et vise à comprendre l’interaction « plantes-DSE-sols pollués ». A ces fins, des champignons DSE isolés de racines de peupliers issues de sols pollués par des ETM ont été caractérisés. Ils appartiennent aux genres Leptodontidium, Phialophora, Cadophora et Phialocephala. Trois de ces souches ont ensuite été inoculées à des plantes en présence d’ETM. Dans certains cas, la croissance végétale a été stimulée et une augmentation de la teneur en chlorophylles, en P et en K, ou une réduction de la teneur en Cd dans les parties aériennes ont été mises en évidence. Puis, la souche la plus efficace sur la croissance végétale, Cadophora sp. Fe06, a été utilisée dans un double inoculum avec une souche endomycorhizienne, chez du ray-grass en présence d’ETM. Cette association diminue la teneur en Cd dans les feuilles. Enfin, les mécanismes impliqués dans la tolérance aux ETM chez les DSE ont été examinés via le criblage d’une banque de mutants d’insertions aléatoires, et via l’étude du rôle de la mélanine. Ce travail met en évidence que l’utilisation de DSE, de part leur capacité à coloniser de nombreuses plantes dans différents sols et à stimuler leur croissance, pourrait représenter un atout considérable pour des opérations de phytomanagement / Phytomanagement is a handling method of contaminated sites based on the ability of plants to sequester pollutants. Fungi could favor plant growth in contaminated sites but were far less studied in the context of phytomanagement. Dark septate endophytes (DSE) were reported to improve plant tolerance against metallic trace elements (TE). The present work is part of the LORVER project and aims to understand the “plant-DSE-TE”. DSE were isolated from poplar roots growing on TE-contaminated soils and characterized. They belong to Leptodontidium, Phialophora, Cadophora and Phialocephala genera. Three strains were inoculated to birch and poplar in TE-contaminated soils. Leptodontidium sp. improved plant growth with an increase in chlorophyll, P and K concentrations, whereas Cadophora sp. decreased Cd concentration in shoots of birch. Then, Cadophora sp. Fe06, was used with an endomycorrhizal fungus, to co-inoculate ryegrass in a TE-polluted soil. The co-inoculation leads to the decrease of Cd concentration in shoots. Finally, the mechanisms involved in TE-tolerance by DSE were studied through the screening of a library of Leptondontidium sp. insertional random-mutants and an evaluation of the role of melanin against TE stress. The present work highlights the complexity of the interactions between DSE and plants under TE stress. These interactions were influenced by different parameters including plant species, DSE strains, and soil properties. Nevertheless, the ability of DSE to colonize a broad spectrum of plants in different soils and to promote plant growth, suggest that DSE could represent a substantial benefit for the fungus-assisted phytomanagement of polluted soils

Estudos hodológicos sugerem um dimorfismo sexual das projeções da área incerto-hipotalâmica (IHy), com áreas relevantes para o controle reprodutivo mais densamente inervadas em fêmeas. Estudos funcionais mostraram que o RNAm do precursor do hormônio concentrador de melanina (ppMCH) varia apenas na IHy, durante o ciclo estral e é reduzido em ratas em anestro induzido por restrição alimentar (RA). Nosso objetivo foi analisar em camundongos fêmeas, as alterações na morfologia de útero e ovários e na expressão do RNAm do ppMCH da IHy ocasionadas pelo anestro induzido por RA. Para isso, camundongos fêmeas C57BL/6 foram distribuídas em controle (C), restrito (R; ração = 70% do C) e realimentado (RR). Os animais R foram perfundidos após 4, 8 e 12 dias para coleta de encéfalo, útero e ovários. Houve um aumento no número de folículos antrais nos animais R12 comparados ao R4 e redução da área uterina nos animais R8 com recuperação dos RR. Não foi possível detectar alterações na expressão do RNAm do ppMCH na IHy. / Hodological studies suggest a sexual dimorphism of the incerto-hypothalamic area (IHy) projections since relevant areas to the reproductive control are more densely innervated in females. Functional studies have shown that the MCH precursor mRNA (ppMCH) varies only in the IHy during the estrous cycle of an intact rat and is decreased in anestrus rats induced by food restriction. We aim to analyze, in female mice, the changes in uterine and ovarian morphology and expression of IHy ppMCH mRNA in anestrus induced by food restriction. For this, C57BL/6 female mice were distributed in control (C), food restricted (FR) and refed (RR). FR were perfused after 4, 8 and 12 days to collect the brain, uterus and ovaries. Our results show an increased number of antral follicles of R12 compared to R4 and a reduction in the uterine area of R8 with restoration in RR. It was not possible to detect changes in ppMCH mRNA expression in IHy.

AbstractIn the field of Biomedical Optics measurement of tissue optical properties, like absorption, scattering, and reduced scattering coefficient, has gained importance for therapeutic and diagnostic applications. Accuracy in determining the optical properties is of vital importance to quantitatively determine chromophores in tissue.There are different techniques used to quantify tissue chromophores. Reflectance spectroscopy is one of the most common methods to rapidly and accurately characterize the blood amount and oxygen saturation in the microcirculation. With a hyper spectral imaging (HSI) device it is possible to capture images with spectral information that depends both on tissue absorption and scattering. To analyze this data software that accounts for both absorption and scattering event needs to be developed.In this thesis work an HSI algorithm, capable of assessing tissue oxygenation while accounting for both tissue absorption and scattering, is developed. The complete imaging system comprises: a light source, a liquid crystal tunable filter (LCTF), a camera lens, a CCD camera, control units and power supply for light source and filter, and a computer.This work also presents a Graphic processing Unit (GPU) implementation of the developed HSI algorithm, which is found computationally demanding. It is found that the GPU implementation outperforms the Matlab “lsqnonneg” function by the order of 5-7X.At the end, the HSI system and the developed algorithm is evaluated in two experiments. In the first experiment the concentration of chromophores is assessed while occluding the finger tip. In the second experiment the skin is provoked by UV light while checking for Erythema development by analyzing the oxyhemoglobin image at different point of time. In this experiment the melanin concentration change is also checked at different point of time from exposure.It is found that the result matches the theory in the time dependent change of oxyhemoglobin and deoxyhemoglobin. However, the result of melanin does not correspond to the theoretically expected result.

Secondary sexual traits play an irreplaceable role in the reproduction of a range of animals and are used as quality and fitness sensors during pairing of individuals. Expression of these traits, ornamentes, can correlated with an individual's personal and behavioral strategies. In the case of melanin ornaments, not only the relationships with personal individuals are considered, but there is the possible connections with stress resistance and the level of stress responses too. This hypothesis is based on the pleiotropic effect of the melanocortin system, which can be used during melanogenesis but also in the production of hormones that contribute to range of stress responses. The aim of this work was to clarify the relationship between an individual's ornaments, his stress response and individuality in the barn swallows (Hirundo rustica rustica). In this work the relationship between selected ornaments and the stress reaction of the organism, stressed glucose levels measured 15 minutes after a stress stimulus, was tested. Both sexes showed a correlation between area of white tail spots and stress response. Only for males a relationship with the length of outermost tail feathers was found and a correlation with the color saturation of feathers on the throat was shown for females. The second part of...

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