Structural Elucidation and Relative Quantification of Sodium- and Potassium-Cationized Phosphatidylcholine Regioisomers Directly from Tissue Using Electron Induced Dissociation.

Comprehensive structural characterization of phosphatidylcholines (PCs) is essential to understanding their biological functions and roles in metabolism. Electron induced dissociation (EID) of protonated PCs directly generated from biological tissues has previously been shown to provide in-depth structural information on the lipid headgroup, regiosiomerism of fatty acyl tails and double bond positions. Although phosphatidylcholine ions formed via alkali metal cationization (i.e., [M + Na]+ and [M + K]+) are commonly generated during matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry experiments, the gas-phase ion chemistry behavior of EID on sodium- and potassium-cationized phosphatidylcholine ion types has not been studied for ions generated directly from tissue. Herein, we demonstrate EID on [M + Na]+ and [M + K]+ ion types in a MALDI imaging mass spectrometry workflow for lipid structural characterization. Briefly, near-complete structural information can be obtained upon EID of sodium- and potassium-cationized PCs, including diagnostic fragmentation of the lipid headgroup as well as identification of fatty acyl chain positions and double bond position. EID of cationized lipids generates sn-specific glycerol backbone cleavages as well as a favorable combined loss of sn-2 fatty acid with choline over sn-1, allowing for facile differentiation and relative quantification of PC regioisomers. Moreover, relative quantification of sn-positional isomers from biological tissue reveals that the relative percentages of sodium- and potassium-cationized sn-positional isomers varies significantly in different regions of rat brain tissue.

Full-body blue light irradiation as treatment for atopic dermatitis: a randomized sham-controlled clinical trial (AD-Blue).

BACKGROUND: Visible blue light (wavelength 400-495 nm) is a promising new treatment option for both psoriasis and atopic dermatitis (AD). Whilst previous clinical trials featured various devices and blue light at a variety of wavelengths, none of these interventions were challenged in objective clinical criteria. PATIENTS AND METHODS: Eighty-seven patients diagnosed with AD were enrolled in AD-Blue, an international, prospective, double-blinded, three-armed (415 nm vs. 450 nm vs. sham control), randomized trial designed to investigate the safety and efficacy of prototype full-body blue light devices. RESULTS: Full-body irradiation with 450 nm blue light but not 415 nm had a significant impact on itch (Itch-VAS, -1.6 ± 2.3; p  =  0.023 vs. sham irradiation). PO-SCORAD values also decreased significantly in response to irradiation at 415 nm (-11.5 ± 18.4; p = 0.028 vs. sham irradiation). None of the other outcome measures (EASI, SCORAD, IGA, DLQI) changed significantly. No safety signals were observed. Evaluation of skin transcriptomes, cytokine levels in serum, and ELISpots from peripheral blood mononuclear cells isolated from a subset of patients revealed moderate decreases in IL-31 in response to irradiation with blue light. CONCLUSIONS: Despite its favorable safety profile and moderate reductions in itch and IL-31 levels, full-body blue light irradiation did not lead to an amelioration of any of the objective measures of AD.

Prospective, Randomized Study on the Efficacy and Safety of Local UV-Free Blue Light Treatment of Eczema.

BACKGROUND: Blue light was shown to reduce the activation of T cells and modulate cytokine release in vitro. Therefore, we investigated the efficacy of blue light in the treatment of eczema. METHODS: A sample of 21 patients with mild to moderate eczema were locally treated with blue LED light (light-emitting diode, emission maximum: 453 nm). They received light treatment 3 times per week for 4 weeks. A contralateral control lesion remained untreated. RESULTS: A total of 20 patients completed the trial with a compliance rate of 100%. The blue light treatment was safe with no adverse events and no side effects. The primary end point change from baseline in the mean sum score of the local Eczema Severity Index (local ESI) was more pronounced for the treated area than for the control area (-1.9 ± 2.02 vs. -1.3 ± 2.24). The treatment difference was statistically significant (p = 0.0152, paired t test, two-sided). CONCLUSION: In this study UV-free blue light was safe and effective in the reduction of eczema lesions.

Prospective Randomized Long-Term Study on the Efficacy and Safety of UV-Free Blue Light for Treating Mild Psoriasis Vulgaris.

BACKGROUND: Blue light irradiation reduces the proliferation of keratinocytes and modulates T-cell immune response in vitro and has been shown to reduce the severity of psoriasis vulgaris (Pv) in two clinical trials. OBJECTIVE: Evaluation of safety and efficacy of long-term UV-free blue light treatment at home for mild Pv. METHODS: Forty-seven patients with mild Pv were randomized for receiving high-intensity blue light treatment (HI: 453 nm LED, 200 mW/cm(2), n = 24) and low-intensity treatment (LI: 453 nm LED, 100 mW/cm(2), n = 23) of one Pv plaque for 12 weeks. A contralateral control plaque remained untreated. RESULTS: Patient compliance and satisfaction were high. The primary endpoint, change from baseline (CfB) of the Local Psoriasis Severity Index, revealed a significant improvement of the target compared to the control plaques (ΔCfB for the HI group: -0.92 ± 1.10, p = 0.0005; for the LI group: -0.74 ± 1.18, p = 0.0064). CONCLUSION: UV-free blue light home treatment is safe and improves Pv plaques.

Blue light inhibits transforming growth factor-ß1-induced myofibroblast differentiation of human dermal fibroblasts.

Transforming growth factor-ß1 (TGF-ß1) is the major promoter of phenotypic shift between fibroblasts and myofibroblasts accompanied by the expression and incorporation of α-smooth muscle actin (α-SMA). This differentiation is crucial during normal wound healing and wound closure; however, myofibroblasts are considered as the main effecter cell type in fibrosis, for example in scleroderma and hypertrophic scarring. As blue light has exerted antiprolific and toxic effects in several cell types, we investigated whether blue light irradiations with a light-emitting diode array (420 nm) were able to affect proliferation and differentiation of human dermal fibroblasts (HDF). We found that repeated irradiation with non-toxic doses significantly inhibits TGF-ß1-induced differentiation of HDF into myofibroblasts shown by α-SMA immunocytochemistry and Western blotting. Additionally, used doses reduced proliferation and myofibroblast contractibility measured by resazurin and collagen gel contraction assays. It could be demonstrated that blue light mediates cell toxicity by oxidative stress due to the generation of singlet oxygen. We postulate that irradiations at non-toxic doses induce low-level oxidative stress and energy-consuming cellular responses, which both may effect proliferation stop and interfere with myofibroblast differentiation. Thus, targeting differentiation, proliferation and activity of myofibroblasts by blue light may represent a useful strategy to prevent or reduce pathological fibrotic conditions.

Insights into blue light accelerated tooth whitening.

OBJECTIVE: To test the hypotheses that blue light accelerates whitening through either (1) direct photobleaching or (2) photon-assisted oxidation using sequential longitudinal bleaching. METHODS: Thirty extracted human tooth samples having natural life accumulated color were divided over five groups: A. 9h light + 10h 6% H2O2 gel + 6h light & 6% H2O2 combined; B. 9h 6% H2O2 gel + 10h light + 6h light & 6% H2O2 combined; C. 11 h light & 6% H2O2 combined; D. 8.45h 25 %H2O2 gel + 10h of light only + 6h light & 25% H2O2 combined E. 10.45 h light & 25 %H2O2 combined. Blue light (456nm) was used at 190 mW/cm2. Color change (ΔE) was measured over time, and reported after 48h color stabilization. RESULTS: Groups A, B and D reached saturation in the first phase (at 9h) at a ΔE of 4.3 ± 0.7, 4.9 ± 1.3 and 10.9 ± 2.2, respectively. Groups C and E achieved in the same time a significantly higher ΔE of 14.2 ± 1.7 and 15.6 ± 1.9, respectively. Subsequently adding the opposite single modality to groups A, B and D for 10h did reach an end stage at 8.1 ± 1.3, 8.8 ± 1.8 and 10.8 ± 1.4 ΔE, respectively. The final 6h treatment combining light and H2O2 showed in these groups a statistically significant step in ΔE reaching 12.9 ± 1.4, 10.7 ± 2.5 and 15.3 ± 1.7, respectively. CONCLUSIONS: Blue light significantly increases bleaching rate and final achievable ΔE.This sequential whitening study provides a first indication that this enhanced bleaching is the result of the hypothesized light mechanisms acting in parallel to hydrogen peroxide bleaching. CLINICAL SIGNIFICANCE: This study shows that blue light can accelerate whitening, within the limits of an in-vitro model. The findings help the clinician explain to their patients that in light accelerated whitening the light not merely accelerates the bleaching process, but that it attacks more stain compounds than peroxide alone does.

A new method for sustained generation of ultra-pure nitric oxide-containing gas mixtures via controlled UVA-photolysis of nitrite solutions.

Exogenous gaseous nitric oxide (gNO) is an FDA approved drug for treatment of a variety of human pathologies like Persistent Pulmonary Hypertension in neonates and premature babies, skin lesions and fungal dermatophyte infections. Substantial disadvantages of current gNO-based therapies are the high therapy costs, high storage costs of the gas cylinders, and the rapid contamination of compressed NO gases with various decomposition products. Here we describe a new, very simple, and inexpensive photolytic generator of uncontaminated NO-containing gas mixtures at therapeutic concentrations. The new method bases on UVA-induced and redox-assisted decomposition of nitrite ions in aqueous solutions. NO formation via UVA-induced photolysis of nitrite is accompanied by an OH radical-dependent production of NO(2) that beside its toxic character additionally strongly reduces the NO yield by consuming NO in its reaction to N(2)O(3). During the UVA-induced photodecomposition process both, inhibition of NO(2) formation or NO(2) depletion by antioxidants hinders the NO-consuming reaction with NO(2) and ensured a maximal purity and maximal yield of NO-containing gas mixtures. Therefore, NO-containing gas mixtures generated by the described method are suitable for medical applications like inhalation or gassing of chronic non-healing wounds. Control of temperature, UVA intensity and composition of the reaction mixture allows facile control over the final NO level in the carrier gas over a wide concentration range. We demonstrate the sustained and stable release of NO over a wide dynamic range (10-5000 ppm NO) for many hours. The method avoids contamination-prone long time storage of NO gas. As such, it appears particularly relevant for applications involving the additional presence of oxygen (e.g. inhalation).

Characterization of disease-specific cellular abundance profiles of chronic inflammatory skin conditions from deconvolution of biopsy samples.

BACKGROUND: Psoriasis and atopic dermatitis are two inflammatory skin diseases with a high prevalence and a significant burden on the patients. Underlying molecular mechanisms include chronic inflammation and abnormal proliferation. However, the cell types contributing to these molecular mechanisms are much less understood. Recently, deconvolution methodologies have allowed the digital quantification of cell types in bulk tissue based on mRNA expression data from biopsies. Using these methods to study the cellular composition of the skin enables the rapid enumeration of multiple cell types, providing insight into the numerical changes of cell types associated with chronic inflammatory skin conditions. Here, we use deconvolution to enumerate the cellular composition of the skin and estimate changes related to onset, progress, and treatment of these skin diseases. METHODS: A novel signature matrix, i.e. DerM22, containing expression data from 22 reference cell types, is used, in combination with the CIBERSORT algorithm, to identify and quantify the cellular subsets within whole skin biopsy samples. We apply the approach to public microarray mRNA expression data from the skin layers and 648 samples from healthy subjects and patients with psoriasis or atopic dermatitis. The methodology is validated by comparison to experimental results from flow cytometry and immunohistochemistry studies, and the deconvolution of independent data from isolated cell types. RESULTS: We derived the relative abundance of cell types from healthy, lesional, and non-lesional skin and observed a marked increase in the abundance of keratinocytes and leukocytes in the lesions of both inflammatory dermatological conditions. The relative fraction of these cells varied from healthy to diseased skin and from non-lesional to lesional skin. We show that changes in the relative abundance of skin-related cell types can be used to distinguish between mild and severe cases of psoriasis and atopic dermatitis, and trace the effect of treatment. CONCLUSIONS: Our analysis demonstrates the value of this new resource in interpreting skin-derived transcriptomics data by enabling the direct quantification of cell types in a skin sample and the characterization of pathological changes in tissue composition.

Treatment of Atopic Dermatitis Using a Full-Body Blue Light Device (AD-Blue): Protocol of a Randomized Controlled Trial.

BACKGROUND: Irradiation with visible blue light (wavelength 400-495 nm) is a promising, effective, and safe new treatment option for chronic inflammatory skin diseases such as psoriasis and atopic dermatitis. OBJECTIVE: We will perform a multicenter, placebo-controlled, double-blinded, 3-armed, prospective, randomized controlled trial to investigate the efficacy and safety of full-body blue light devices (wavelengths: 415 nm and 450 nm) compared with that of placebo irradiation for the treatment of atopic dermatitis. METHODS: We are planning to enroll a total of 150 patients at the University hospitals in Göttingen (Germany), Marburg (Germany), and Geneva (Switzerland). RESULTS: The trial was approved by the lead ethics committee of the medical faculty of the University of Göttingen (21/11/16). Further approvals were obtained from local and federal authorities (ethics committee Marburg, Cantonal Commission for Research Ethics Geneva, Suisse Medic, and Bundesinstitut für Arzneimittel und Medizinprodukte). CONCLUSIONS: We will disseminate the results in a peer-reviewed journal. TRIAL REGISTRATION: ClinicalTrials.gov NCT03085303; https://clinicaltrials.gov/ct2/show/NCT03085303 (Archived by WebCite at http://www.webcitation.org/73ucqkkA1). INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/11911.

Blue light exposure decreases systolic blood pressure, arterial stiffness, and improves endothelial function in humans.

AIMS: Previous studies have shown that ultraviolet light can lead to the release of nitric oxide from the skin and decrease blood pressure. In contrast to visible light the local application of ultraviolet light bears a cancerogenic risk. Here, we investigated whether whole body exposure to visible blue light can also decrease blood pressure and increase endothelial function in healthy subjects. METHODS: In a randomised crossover study, 14 healthy male subjects were exposed on 2 days to monochromatic blue light or blue light with a filter foil (control light) over 30 minutes. We measured blood pressure (primary endpoint), heart rate, forearm vascular resistance, forearm blood flow, endothelial function (flow-mediated dilation), pulse wave velocity and plasma nitric oxide species, nitrite and nitroso compounds (secondary endpoints) during and up to 2 hours after exposure. RESULTS: Blue light exposure significantly decreased systolic blood pressure and increased heart rate as compared to control. In parallel, blue light significantly increased forearm blood flow, flow-mediated dilation, circulating nitric oxide species and nitroso compounds while it decreased forearm vascular resistance and pulse wave velocity. CONCLUSION: Whole body irradiation with visible blue light at real world doses improves blood pressure, endothelial function and arterial stiffness by nitric oxide released from photolabile intracutanous nitric oxide metabolites into circulating blood.

A Dynamic Model for Prediction of Psoriasis Management by Blue Light Irradiation.

Clinical investigations prove that blue light irradiation reduces the severity of psoriasis vulgaris. Nevertheless, the mechanisms involved in the management of this condition remain poorly defined. Despite the encouraging results of the clinical studies, no clear guidelines are specified in the literature for the irradiation scheme regime of blue light-based therapy for psoriasis. We investigated the underlying mechanism of blue light irradiation of psoriatic skin, and tested the hypothesis that regulation of proliferation is a key process. We implemented a mechanistic model of cellular epidermal dynamics to analyze whether a temporary decrease of keratinocytes hyper-proliferation can explain the outcome of phototherapy with blue light. Our results suggest that the main effect of blue light on keratinocytes impacts the proliferative cells. They show that the decrease in the keratinocytes proliferative capacity is sufficient to induce a transient decrease in the severity of psoriasis. To study the impact of the therapeutic regime on the efficacy of psoriasis treatment, we performed simulations for different combinations of the treatment parameters, i.e., length of treatment, fluence (also referred to as dose), and intensity. These simulations indicate that high efficacy is achieved by regimes with long duration and high fluence levels, regardless of the chosen intensity. Our modeling approach constitutes a framework for testing diverse hypotheses on the underlying mechanism of blue light-based phototherapy, and for designing effective strategies for the treatment of psoriasis.

Mechanism and biological relevance of blue-light (420-453 nm)-induced nonenzymatic nitric oxide generation from photolabile nitric oxide derivates in human skin in vitro and in vivo.

Human skin contains photolabile nitric oxide (NO) derivates such as nitrite and S-nitrosothiols, which upon UVA radiation decompose under high-output NO formation and exert NO-specific biological responses such as increased local blood flow or reduced blood pressure. To avoid the injurious effects of UVA radiation, we here investigated the mechanism and biological relevance of blue-light (420-453 nm)-induced nonenzymatic NO generation from photolabile nitric oxide derivates in human skin in vitro and in vivo. As quantified by chemiluminescence detection (CLD), at physiological pH blue light at 420 or 453 nm induced a significant NO formation from S-nitrosoalbumin and also from aqueous nitrite solutions by a to-date not entirely identified Cu(1+)-dependent mechanism. As detected by electron paramagnetic resonance spectrometry in vitro with human skin specimens, blue light irradiation significantly increased the intradermal levels of free NO. As detected by CLD in vivo in healthy volunteers, irradiation of human skin with blue light induced a significant emanation of NO from the irradiated skin area as well as a significant translocation of NO from the skin surface into the underlying tissue. In parallel, blue light irradiation caused a rapid and significant rise in local cutaneous blood flow as detected noninvasively by using micro-light-guide spectrophotometry. Irradiation of human skin with moderate doses of blue light caused a significant increase in enzyme-independent cutaneous NO formation as well as NO-dependent local biological responses, i.e., increased blood flow. The effects were attributed to blue-light-induced release of NO from cutaneous photolabile NO derivates. Thus, in contrast to UVA, blue-light-induced NO generation might be therapeutically used in the treatment of systemic and local hemodynamic disorders that are based on impaired physiological NO production or bioavailability.

Effects of blue light irradiation on human dermal fibroblasts.

Previous studies have reported that separately from UV-radiation also blue light influences cellular physiology in different cell types. However, little is known about the blue light action spectrum. The purpose of this study was to investigate effects of blue light at distinct wavelengths (410, 420, 453, 480 nm) emitted by well defined light-emitting-diodes on viability, proliferation and antioxidative capacity of human dermal fibroblasts. We found that irradiation with blue light (410, 420 nm) led to intracellular oxidative stress and toxic effects in a dose and wavelength dependent manner. No toxicity was observed using light at 453 nm and 480 nm. Furthermore, blue light (410, 420, 453 nm) at low doses reduced the antioxidative capacity of fibroblasts. At non-toxic doses, irradiations at 410, 420 and 453 nm reduced proliferation indicating a higher susceptibility of proliferating fibroblasts to blue light. Our results show that blue light at different wavelengths may induce varying degrees of intracellular oxidative stress with different physiological outcome, which could contribute to premature skin photoaging. On the other hand, the use of blue light due to its antiproliferative and toxic properties may represent a new approach in treatment and prevention of keloids, hypertrophic scars and fibrotic skin diseases.

Revealing the neural response to imperceptible peripheral flicker with machine learning.

Lighting in modern-day devices is often discrete. The sharp onsets and offsets of light are known to induce a steady-state visually evoked potential (SSVEP) in the electroencephalogram (EEG) at low frequencies. However, it is not well-known how the brain processes visual flicker at the threshold of conscious perception and beyond. To shed more light on this, we ran an EEG study in which we asked participants (N=6) to discriminate on a behavioral level between visual stimuli in which they perceived flicker and those that they perceived as constant wave light. We found that high frequency flicker which is not perceived consciously anymore still elicits a neural response in the corresponding frequency band of EEG, con-tralateral to the stimulated hemifield. The main contribution of this paper is to show the benefit of machine learning techniques for investigating this effect of subconscious processing: Common Spatial Pattern (CSP) filtering in combination with classification based on Linear Discriminant Analysis (LDA) could be used to reveal the effect for additional participants and stimuli, with high statistical significance. We conclude that machine learning techniques are a valuable extension of conventional neurophysiological analysis that can substantially boost the sensitivity to subconscious effects, such as the processing of imperceptible flicker.

Blue-light irradiation regulates proliferation and differentiation in human skin cells.

Sunlight influences the physiology of the human skin in beneficial as well as harmful ways, as has been shown for UV light. However, little is known about the effects of other wavelengths of solar irradiation. In this study we irradiated human keratinocytes and skin-derived endothelial cells with light-emitting-diode devices of distinct wavelengths to study the effects on cell physiology. We found that light at wavelengths of 632-940 nm has no effect, but irradiation with blue light at 412-426 nm exerts toxic effects at high fluences. Light at 453 nm is nontoxic up to a fluence of 500 J/cm(2). At nontoxic fluences, blue light reduces proliferation dose dependently by up to 50%, which is attributable to differentiation induction as shown by an increase of differentiation markers. Experiments with BSA demonstrate that blue-light irradiation up to 453 nm photolytically generates nitric oxide (NO) from nitrosated proteins, which is known to initiate differentiation in skin cells. Our data provide evidence for a molecular mechanism by which blue light may be effective in treating hyperproliferative skin conditions by reducing proliferation due to the induction of differentiation. We observed a photolytic release of NO from nitrosated proteins, indicating that they are light acceptors and signal transducers up to a wavelength of 453 nm.

The osteoblast mechano-receptor, microgravity perception and thermodynamics.

Mechano-sensing in cells is tightly obliged with changes in intracellular free calcium (IFC), regulation of specific genes and activation of specific second messenger systems. To investigate whether single non-professional cells like osteoblasts can detect microgravity through the mechano-sensor, measurements on a sub-orbital rocket and parabolic flights observing the IFC and gene expression were performed. We find that microgravity did neither effect IFC nor gene expression. Thermal and mechanical noise within cells is too high in relation to the change of force due to the change from gravity to microgravity. Complementary force measurements have shown that cells exert high forces on the substrate and that these high forces have to be applied for activation.