Background: Collagen cross-linking of the sclera is a promising approach to strengthen scleral rigidity and thus to inhibit eye growth in progressive myopia. Additionally, cross-linking might inhibit degrading processes in idiopathic melting or in ocular inflammatory diseases of the sclera. Different cross-linking treatments were tested to increase resistance to enzymatic degradation of the rabbit sclera. Methods: Scleral patches from rabbit eyes were cross-linked using paraformaldehyde, glutaraldehyde or riboflavin combined with UV-A-light or with blue light. The patches were incubated with collagenase I (MMP1) for various durations up to 24 h to elucidate differences in scleral resistance to enzymatic degradation. Degraded protein components in the supernatant were detected and quantified using measurements of Fluoraldehyde o-Phthaldialdehyde (OPA) fluorescence . Results: All cross-linking methods reduced the enzymatic degradation of rabbit scleral tissue by MMP1. Incubation with glutaraldehyde (1%) and paraformaldehyde (4%) caused nearly a complete inhibition of enzymatic degradation (down to 7% ± 2.8 of digested protein compared to control). Cross-linking with riboflavin/UV-A-light reduced the degradation by MMP1 to 62% ± 12.7 after 24 h. Cross-linking with riboflavin/blue light reduced the degradation by MMP1 to 77% ± 13.5 after 24 h. No significant differences could be detected comparing different light intensities, light exposure times or riboflavin concentrations. Conclusions: The application of all cross-linking methods increased the resistance of rabbit scleral tissue to MMP1-degradation. Especially, gentle cross-linking with riboflavin and UV-A or blue light might be a clinical approach in future.

No description available.

Light and optical techniques have made a profound impact on modern medicine both in diagnostics and in therapy. Therapeutic action of light is based on photomechanical, photothermal, photochemical and photobiological interactions, depending on the wavelength, power density, exposure time and optical properties of tissue and cells. Last decade experienced a growing rise of commercial devices for management of hair growth, where all of them are based on low levels of light resulting into photobiological, non-thermal interaction of photons with cells, a process that recently has received an official term ‘photobiomodulation’. However, the design and analysis of the reported clinical studies are highly debated in a wider scientific community. The picture is further complicated by a virtual lack of proof about the exact molecular targets that mediate the physiological response of skin and hair follicles (HF) to low levels of light. The goal of this project was to investigate the expression of light-sensitive receptors in the human HF and to study the impact of UV-free blue light on hair growth ex vivo. The expression of Cryptochromes 1 and 2 (CRY1, 2), Opsin 2 and 3 (OPN2 and OPN3), but not other Opsins 1, 4 and 5 was detected in the distinct compartments of skin and anagen HF. Evaluation of the physiological role of detected light-sensitive receptors on hair growth was performed by the modulation of photoreceptors activity in HF ex vivo model. HFs treated with KL001, a stabilizer of CRY1 protein that lengthens the circadian period, delayed HF anagen-catagen transition; while silencing of CRY1 induced premature catagen development accompanied by reduced cell proliferation. Silencing of CRY1 in the HF outer root sheath (ORS) cells in vitro caused downregulation of ii genes involved in the control of proliferation; including the cyclin dependent kinase 6 (CDK6). OPN3 also had a positive effect on metabolic activity and proliferation of the ORS cells in vitro. OPN3 silencing resulted in the altered expression of genes involved in the control of proliferation and apoptosis. Investigated CRY1, OPN2 and 3 greatly absorb in the blue to green-region of the visible spectrum. This led us to investigate the effect of blue light on HF growth. Daily treatment with blue light (453 nm, 3.2 J/cm2, 16 nm full width half maximum) prolonged anagen phase in HF ex vivo that was associated with sustained proliferation. In addition, blue light (3.2 J/cm2) significantly stimulated proliferation of ORS cells in vitro. This effect was abrogated by silencing of OPN3. To summarize, CRY 1, OPN 2 and OPN 3 are expressed in the distinct compartments of the HF, including HF stem cells. Blue light (453 nm) at low radiant exposure exerts a positive effect on hair growth ex vivo, potentially via interaction with OPN3. The further research should be conducted to decipher interactions between blue light and the investigated receptors in the HFs. In addition, the beneficial effect of blue light at low radiant exposure on hair growth raises a possibility of increasing therapeutic efficacy when combined with topical chemistry used for management of hair growth.

Indiana University-Purdue University Indianapolis (IUPUI) / The phototropin photoreceptors are important regulators of plant growth and development and can therefore affect the photosynthetic activity of plants. Phototropin1 and Phototropin2 are versatile protein kinases that become activated when exposed to blue light. Their photobiological actions are best understood in the model plant Arabidopsis thaliana, where they are known to trigger several responses to blue light, one of which is phototropism, the bending of plant organs towards light. Additionally, phot1 and phot2 drive stomatal opening, chloroplast arrangement in leaf cells, leaf expansion, and leaf orientation. The phot1-specific response is rapid inhibition of hypocotyl growth, leaf positioning and mRNA stability whereas phot2 mediates the chloroplast avoidance response to high light. These responses impact a plant’s ability to capture light for photosynthesis, therefore the phototropins play important roles in optimizing a plant’s photosynthetic activity. Soybean (Glycine max) is a very important crop plant in Indiana known for its nutritional versatility and is also utilized for biodiesel production.In spite of soybean being a key crop, there is currently no information about the functionality of soybean phototropins. Also, being a legume, soybean has many structural and functional features that are not present in Arabidopsis. Interestingly, PsPHOT1A (a photoreceptor from garden pea) was found to be a functional phototropin as it was able to complement the phot1 mutation in Arabidopsis. The roles of these proteins in soybean will be elucidated based on the hypothesis that soybean phototropins play essential roles in regulating photosynthetic activity as do the Arabidopsis phototropins. To date, five soybean phototropins, 3 PHOT1s and 2 PHOT2s, are believed to exist. These GmPHOT protein coding regions were amplified by RT-PCR and cloned into pCR8/TOPO or pENTR-D/TOPO vectors via TOPO cloning to utilize Gateway cloning technology to create plant transformation constructs subsequently. The cloned GmPHOT cDNAs from each of the 5 GmPHOTs were sequenced and compared to the GmPHOT sequences from the Phytozome database to assess the accuracy of the gene models. The gene models of all the GmPHOTs were found to be accurate except that of GmPHOT1B-2. The high level of sequence identity between the GmPHOTs and AtPHOTs and the conservation of LOV domains and catalytic domains indicate structural resemblance between them. This suggests that soybean phototropins should encode active photoreceptors. The cloned protein coding regions from soybean were then recombined into a plant expression vector via Gateway technology,which were then used for transformation of Agrobacterium tumefaciens. These plant expression constructs will be utilized in the future to determine the functionality of soybean phototropins in Arabidopsis.

Problématique : Certaines personnes souffrent d'avoir un horaire de sommeil trop tardif qu'elles n'arrivent pas à modifier pour satisfaire les exigences liées à leur emploi ou à leurs études. Ces individus souffrent de privation de sommeil et de somnolence lorsqu'ils doivent se conformer à un horaire socialement acceptable. Malgré sa prévalence importante, l’étiologie d’un horaire trop tardif reste méconnue. Objectif: Évaluer des facteurs pouvant contribuer au maintien d’un horaire de sommeil tardif chez des jeunes adultes. Méthodes : Quatorze jeunes adultes se plaignant d’un horaire de sommeil trop tardif ont été comparés à des sujets appariés qui avaient un horaire de sommeil adapté. L’heure de coucher (HC) était après minuit pour les sujets tardifs et avant minuit pour les sujets adaptés. Les sujets étaient admis au laboratoire 5h avant l’HC et garder en obscurité pour 6h. Ils étaient ensuite exposés à 1,5h de lumière bleue. La mélatonine salivaire et la vigilance subjective étaient mesurées aux 30 minutes. La suppression de mélatonine a été utilisée pour déterminer la sensibilité circadienne à la lumière. Le dim light melatonin onset (DLMO) a été utilisé pour déterminer la phase circadienne. Résultats : Le DLMO survenait plus tard dans le groupe tardif que dans le groupe adapté. Il n’y avait pas de différence de suppression de mélatonine après 1,5h d’exposition à la lumière. Néanmoins, une corrélation entre la sensibilité à la lumière et la phase circadienne a été trouvée dans le groupe tardif. Les sujets tardifs présentaient aussi une augmentation plus lente de la somnolence subjective en soirée. Conclusion : Nos résultats suggèrent qu’une phase circadienne en délai, une augmentation plus lente du besoin de dormir et une sensibilité circadienne à la lumière accrue contribuent à la plainte d’un horaire de sommeil trop tardif. / Problem: Some people suffer from having a delayed sleep schedule that they can’t modify to satisfy school/work requirements. These individuals suffer from sleep deprivation and sleepiness when they have to comply with a socially acceptable schedule. Despite its high prevalence, the etiology of a delayed schedule remains unknown. Objective: This study aims to elucidate factors that might contribute to the maintenance of a delayed sleep schedule in young adults. Methods: Fourteen young adults (8 women) complaining of delayed sleep schedule were compared to matched subjects with an adapted sleep schedule. Habitual bedtime (HB) was after midnight in all delayed subjects and before midnight in all adapted subjects. Subjects were admitted 5h before HB and kept in dim light for 6h. They were then exposed for 1.5h to blue light. Salivary melatonin and subjective sleepiness were assessed every 30 min. Melatonin suppression was used to measure circadian sensitivity to light. Dim light melatonin onset (DLMO) was used to estimate circadian phase. Results: DLMO was later in the delayed than in the adapted group. There was no difference for melatonin suppression over the 1.5h of light exposure. However, in the delayed group, there was a significant correlation between DLMO and melatonin suppression. There was a smaller increase of subjective sleepiness in the delayed subjects than in the adapted subjects before HB. Conclusions: Our results suggest that a delayed circadian phase, a slower build-up of sleep propensity and an enhanced circadian sensitivity to evening light contribute to the complaint of a delayed sleep schedule.

Fluorescence in blue light (FLU) ist ein negativer Feedbackregulator der Chlorophyllbiosynthese, welcher an der Dunkelrepression der 5-Aminolävulinsäure (ALA)-Synthese beteiligt ist. FLU ist Teil eines Komplexes, der die Enzyme umfasst, welche an der Katalyse der finalen Schritte der Chlorophyllbiosynthese beteiligt sind. Drei funktionelle Domänen wurden für das Arabidopsis FLU Protein postuliert: eine Tetratricopeptid-Wiederholungsdomäne (TPR) befindet sich am C-Terminus; eine Transmembrandomäne (TM) ist am N-Terminus lokalisiert; eine Coiled-coil-Domäne (linker) liegt dazwischen. Die TPR-Domäne von FLU Domäne interagiert mit dem C-terminalen Ende der Glutamyl-tRNA Reduktase (GluTR), dem geschwindigkeitsbestimmenden Enzym der ALA-Synthese. Diese Arbeit zur Erweiterung des Wissen über die Funktion von FLU im Licht sowie über die Rolle der funktionellen Domänen von FLU bei der Inaktivierung der ALA-Synthese bei. / Fluorescence in blue light (FLU), a negative feedback regulator of chlorophyll biosynthesis, is involved in dark repression of 5-aminolevulinic acid (ALA) synthesis. FLU is part of a complex comprising the enzymes catalyzing the final steps of chlorophyll synthesis. Three functional domains were proposed in the Arabidopsis FLU protein: a tetratricopeptide repeat (TPR) domain is at the C-terminus; a transmembrane domain (TM) is at the N-terminus; a coiled-coil domain (linker) is in between. The TPR(FLU) domain interacts with the C-terminal end of glutamyl-tRNA reductase (GluTR), the rate-limiting enzyme of ALA synthesis. This thesis contributes to the extended knowledge about the function of FLU in light as well as the role of the structural domains of FLU in the inactivation of ALA synthesis.

This diploma thesis focuses on electrochemical sensors to detect radiation in the blue region of the visible spectrum. As a semiconducting material for the manufacture of the sensor, a suspension of tungsten trioxide was found. The individual elements of the sensor were applied by pad printing techniques or screen printing. Using linear voltammetry and chronoamperometry, the magnitude of the generated current was measured and radiometric calibration was performed. The properties of the layers were characterized by profilometry, optical and electron microscopy.

This doctoral thesis called “Photobiological safety of luminaires and light sources” has focused on an optical radiation and its relation to human body. The thesis describes possible consequences on living tissue during excessive exposure to optical radiation. Among others the work deals with the light source, lamp and optical devices evaluation in dependence of photobiological safety. There is created a method of light source evaluation procedure for photobiological safety in accordance with ČSN EN 62471. The meaning of this procedure is based on its hazard calculation from measured values of irradiation to different photometric distances. Validity of procedure is verified by light source measuring, in which visual angle and photometric distance have been changed. There we can also find an application of this procedure in the UV source evaluation, which helps to determine a safe distance away from UV sources not to create any danger. The evaluated UV sources include a UVA luminaire with a dominant wavelength of 365 nm or an arc of an electric welder. The thesis also describes a method of evaluating light sources for blue light hazard for an aphakic and pseudophakic eye. This method can also help to determine the hazard for the eye without lens or with some implanted lens. Furthermore, there was developed the method of the blue light hazard assessment depending on the age of an exposed person. In terms of the work there was also created a computer programme evaluating photobiological safety resulting from the spectral data having been stored in the library programme or from spectroradiometric measurement data. The work also marginally deals with the topic of circadian rhythms, which are closely related to photobiological manifestation in an human body. Above all the work compares light sources for which equivalent illuminances are calculated, consequently light sources have the same effect on suppressing the melatonin hormone production.

The thesis deals with the problems of the photobiological lamp and lamps systems safety. It is divided into theoretical and practical parts. The theoretical part touches the basic knowledge concerning the eye and vision, eye and skin diseases caused by excessive exposure to non-ionizing radiation. And besides that there are described the sources of light causing the possible exposure and further more it includes the methodology of measurement and evaluation of photobiological safety. The practical part investigates a measurement of the spectrum of light sources. The measured data are evaluated according to the methods mentioned in the theoretical part.

No description available.