Summary of Abstracts

NO DIFFERENCE DETECTED IN PFA VALUE DETERMINED USING A 300 WATT SOLAR SIMULATOR VERSUS A 150 WATT SOLAR SIMULATOR.

M Caswell, C Wood, G Roberts, and A Martinez. Consumer Product Testing Company, Inc., Fairfield NJ.

he objective of the clinical trial was to ascertain if the Protection Factor of UVA [PFA] determined using the two different solar simulators were comparable. The PFA of the 5% 4-tert-butyl -4’-methoxydibenzoylmethane and 3% 2-ethylhexyl-p-methoxycinnamate (JCIA) standard (PFA = 3.75 ± 1.01) and the 7% padimate O and 3% oxybenzone standard (PFA = 3.2 ± 0.50) were tested using a 300 watt xenon arc solar simulator and a 150 watt xenon arc solar simulator. The solar simulator beams were filtered with UG11 and WG335 filters, were a minimum of 1 cm2, and exhibited less than 20% time-related fluctuation. All solar simulators were calibrated and adjusted to deliver energies within 10% variance. PFAs were determined using the method described in the Japanese Cosmetic Industry Association Technical Bulletin (Measurement standards for protection efficacy, Issued November 21, 1995). Each subject was irradiated to determine their Minimal Persistent Pigmenting Dose [MPPD] and tested on each of the two sunscreen standards for each of the two solar simulators, so each subject had six sites tested.

The subject panel consisted of 7 males and 3 females. Their ages ranged from 20 to 58 years with a mean of 39 years. Of the 10 subjects, 9 were Fitzpatrick skin type III and 1 was type IV. Using the 150 watt xenon arc solar simulator, the PFA value of the JCIA standard was found to be 4.06 ± 0.70 and the PFA value of the 7% padimate O and 3% oxybenzone standard was determined to be 3.28 ± 0.25. Using the 300 watt xenon arc solar simulator, the PFA value of the JCIA standard was found to be 4.06 ±0.70 and the PFA value of the 7% padimate O and 3% oxybenzone standard was determined to be 3.44 ± 0.39.

These results are consistent with their being no significant difference in the resultant PFA value determined using a 150 watt xenon arc solar simulator versus that using a 300 watt xenon arc solar simulator.

IN VIVO EXCITATION-EMISSION MATRIX FLUORESCENCE SPECTROSCOPY OF NORMAL HUMAN SKIN REVEALS BODY SITE SPECIFIC FEATURES.

Soodabeh Zandi, Jianhua Zhao, Haishan Zeng, Florina Feng, and Harvey Lui, Department of Dermatology and Skin Science, University of British Columbia & Vancouver Coastal Health Research Institute; BC Cancer Research Centre, Vancouver, BC.; Kerman University of Medical Sciences, Iran.

Background/Objective: An excitation emission matrix (EEM) is a two-dimensional contour plot that depicts fluorescence intensity as a combined function of the excitation and emission wavelengths for materials or tissues. Major skin fluorophores that can be identified from fluorescence EEM include tryptophan, collagen, elastin, keratin, NADH, and porphyrins. There are very limited data on the variability of skin fluorescence EEM according to different body sites. The objective is to study the cutaneous autofluorescence properties of different body sites skin using EEM spectroscopy.

Methods: We compared skin autofluorescence among 10 anatomic skin sites using a spectrofluorometer equipped with two double-grating monochromators. The sites included the forehead, cheek, nose, neck, palm, thumbnail, dorsal hand, dorsal forearm, medial arm and mid-back. The excitation and emission wavelengths were 260 - 450 nm and 300 - 700 nm with 5 nm steps,

Results: Overall fluorescence emission differed amongst the body sites (p<0.0001, Friedman test). Fluorescence was highest from the palm, thumbnail and mid-back; the lowest fluorescence was demonstrated from the dorsal hand, dorsal forearm, cheek, neck, and forehead; the nose and arm showed intermediate levels of fluorescence. There were two fluorescence peaks present at all skin sites. One was at 290~300/340~350 nm (excitation/emission) and probably due to tryptophan; the second peak at 365~385/455~470 nm (excitation/emission) may be related to collagen and/or elastin fluorescence. The neck, forehead, nose and cheek also showed a distinct fluorescence peak at 405~450/610~625 nm (excitation/emission) that was likely related to porphyrins.

Conclusions: The fluorescence of the skin varies significantly across different body sites not only in terms of overall emission, but also with respect to the relative amount of fluorophores. These topographic differences need to be accounted for when interpreting fluorescence spectra of the skin in normal and diseased states.

EXPOSURES OF MICROSCOPIC SKIN SITES TO UV RADIATION PROVIDE NEW INSIGHTS ON THE RELATIONSHIP OF CHALLENGE TO RESPONSE.

Seo I., Bargo P.R., Chu., Kollias N. Johnson and Johnson Consumer Products Companies, Skillman, NJ

The traditional exposure area for a phototest (a test to determine the sensitivity of skin to ultraviolet radiation) has been between 1 cm2 and 1 in2. There have been reports that the apparent sensitivity of the skin, expressed as erythema, decreases for areas of the order of 1 mm2 and smaller. In this study we investigated the responses of human skin to solar simulated radiation with a beam diameter of 200 SEQ CHAPTER \h \r 1µm (0.03 mm2).

Twelve human subjects were exposed to solar simulated radiation on their dorsal upper arm or on their lower back with a series of doses in increments of 20% in order to determine the threshold dose to induce a minimal perceptible erythema response (MED). Each dose was delivered with a liquid light guide of 8 mm diameter and with quartz optical fibers of 200 μm diameter. The resulting skin responses were evaluated visually for the 8 mm diameter spot and with a video microscope for the 200 μm spots. Both types of sites were investigated with a Reflectance Confocal Microscope.

The threshold dose to elicit a minimal perceptible erythema was found to be greater for the microscopic exposures as was earlier determined. The erythema response to the microscopic beam was always diffuse with no clear boundaries for all given doses extending to several times the exposed site diameter for doses >3MED. The pigment responses, at 5-7 days after exposure, were always confined within the irradiated sites and upon microscopic inspection showed a non uniform distribution of pigment that was modulated by the undulation of the stratum corneum. The skin was found to return to normal appearance after the microscopic challenge within two weeks following exposure while changes in appearance for the larger areas persisted for several weeks to months. This new modality of testing provides the possibility to study skin at the microscopic level with a rapid recovery following challenge.

OMEPRAZOLE (PROTON PUMP INHIBITOR) INHIBITS MELANIN SYNTHESIS ACCOMPANIED BY A DECREASED PH IN THE ACIDIC CELL COMPARTMENT.

Yoko Niki,1 Hideya Ando,1,2 Mary S. Matsui,3 Daniel B. Yarosh,3 and Masamitsu Ichihashi1,2
1 Kobe Skin Research Institute, Kobe, Japan
2 Doshisha University, Skin Aging and Photo-aging Research Center, Kyoto, Japan
3 Estée Lauder Companies Inc., NY, USA

Proton pump inhibitors (PPIs), which act by irreversibly blocking the H+/K+ ATPase of the gastric parietal cells, are used in the treatment of peptic ulcer disease and gastro-esophageal reflux disease. During a screening test using B16F10 mouse melanoma cells, we found that PPIs reduce melanogenesis. Among 14 different PPIs, omeprazole (OPZ) was the most potent at reducing cellular melanin content with low cytotoxicity. OPZ also inhibited melanin synthesis in a reconstructed human skin model. Although OPZ slightly decreased tyrosinase DOPA oxidase activity of mushroom and B16F10 cell extract in vitro, in situ tyrosine hydroxylase activity of tyrosinase was decreased significantly in OPZ-treated B16F10 cells. We also examined the effects of OPZ on mRNA and protein levels of melanin synthesis-related proteins, including tyrosinase, tyrosinase related protein-2, Mitf and Pmel17 using B16F10 cells. OPZ did not inhibit transcription at non-cytotoxic concentrations. However, western-blotting revealed that treatment of B16F10 with 50 M OPZ for 72h decreased tyrosinase protein level to approximately 50% that of control cells. Other research groups have showed that the pH of cell organelles, such as ER, Golgi and melanosomes may have an important regulatory role in melanogenesis, but it has not been clear whether an increase or decrease in pH favors melanogenesis. We therefore investigated the effect of OPZ on melanocyte pH using an acidic probe, DAMP. DAMP- treated B16F10 cells demonstrated lower pH in the acidic compartment within 4h of exposure to OPZ. These results suggest that OPZ may inhibit melanin synthesis via a decrease in melanosomal pH, leading to the inhibition of tyrosinase activity, increased tyrosinase degradation, and/or to the suppression of tyrosinase trafficking.

PROTECTIVE EFFECTS OF β-CAROTENE AND MELANIN AGAINST PROTOPORPHYRINE IX-INDUCED PHOTOTOXICITY IN THE PHOTO HEN’S EGG TEST.

Bafteh P, Siegesmund M, Hanneken S, Neumann NJ, Dept. of Dermatology, Heinrich-Heine University Duesseldorf, Germany.

The aim of our study was to evaluate the photoprotective potential of melanin and β-carotene against protoporphyrine IX-induced phototoxicity via photo hen’s egg test (PHET). In three independent test groups, the yolk sac blood vessel system (YS) of 12 eggs (one test group) was exposed to protoporphyrine IX (0.1mM) and afterwards immediately irradiated with 5 J/cm² UVA. Two of these test groups were employed, applying additionally 1 ml of 0.1mM melanin (M) or 0.1mM β-carotene (βC) to detect their supposed photoprotective capacity. Moreover, three control groups were exposed to physiological saline, M or βC without subsequent irradiation to exclude plain toxic reactions. Over a test observation period of 24h, the embryo lethality and the morphological changes (membrane discoloration and hemorrhage) were recorded. The test group exposed to protoporphyrine IX and 5 J/cm² UVA showed severe phototoxic damage, as well as a high lethality rate (75%) of the embryos. In contrast, in the control groups which were not irradiated but exposed to physiological saline, M or βC alone, only slight damages and no lethalities were observed. The test groups which were exposed to protoporphyrine IX, UVA and additionally to M or βC revealed remarkably lower lethality rates of 16.7% in case of M and 58% in case of βC. Furthermore, these test groups obviously showed lesser morphologic damages compared with the UVA-irradiated protoporphyrine IX test group. In conclusion, β-carotene revealed only moderate photoprotective effects against protoporphyrine IX-induced phototoxicity. This finding is in good accordance with the moderate efficacy of βC in the treatment of erythropoietic porphyria. In contrast to β-carotene, melanin revealed a remarkable and significant photoprotective potential against protoporphyrine IX-induced phototoxic damage in the PHET. Taking into account that synthetic α-MSH-analoga are able to induce a de novo-synthesis of melanin without any previous UV-irradiation in human skin, α-MSH-analoga might be a new promising therapeutic option for photodermatoses, especially erythropoietic porphyria.

SPECTROSCOPIC AND COLORIMETRIC ANALYSIS OF ACANTHOSIS NIGRICANS IN PATIENTS WITH HYPERINSULINEMIA.

ZU Syed, P Vemulapalli, M Henderson, HW Lim, IH Hamzavi. Department of Dermatology, Henry Ford Health System, Detroit, MI.

Acanthosis nigricans consists of hyperpigmented, velvety plaques that occur in association with hyperinsulinemic and/or insulin-resistant states. It is hypothesized that increased insulin causes hyperproliferation of the skin and thus causes acanthosis nigricans. However, there have been no studies determining the association of actual insulin levels to the severity of acanthosis nigricans. We sought to elucidate any such association using spectroscopic and colorimetric analysis of acanthosis nigricans lesions.

Eight (8) patients with diagnosed acanthosis nigricans were chosen who had high insulin and normal blood glucose levels, indicating an insulin resistant state. On a monthly basis, a diffuse reflectance spectrometer was used to measure melanin (pigmentation), oxyhemoglobin (erythema), and reflectance properties of acanthosis nigricans lesions as well as normal neck skin. A tristimulus colorimeter was used to precisely quantify the color of the lesions in the L*a*b* color space. Patients were also either seen by an endocrinologist for treatment with metformin or given dietary and exercise guidelines. Fasting blood levels of insulin and glucose were taken monthly.

Melanin, L* (a measure of lightness) and specular reflectance were all significantly different in acanthosis nigricans skin as compared to normal skin (p<0.05). Clinically, the acanthosis nigricans lesions did not appear to change over the course of eight (8) months. Similarly, there were no significant shifts in the measured melanin values or the darkness of the lesions (as measured by the L* parameter). Insulin levels varied significantly from month to month, but did not correlate to changes in either melanin or L* values.

The colorimeter and diffuse reflectance spectrometer appear to be useful tools in the diagnosis and investigation of acanthosis nigricans lesions. Further study is required to elucidate the correlation between insulin levels and changes in pigmentation measured by these methods.

RANDOMIZED CONTROLLED TRIAL COMPARING EFFICACY OF ANTIBIOTIC THERAPY ALONE VERSUS ANTIBIOTIC THERAPY IN CONJUNCTION WITH TRIPLE PULSE THERAPY USING NDYAG LASER IN THE TREATMENT OF HIDRADENITIS SUPPURATIVA.

ZU Syed, P Vemulapalli, M Henderson, IH Hamzavi. Department of Dermatology, Henry Ford Health System, Detroit, MI

Hidradenitis suppurativa is a chronic disease of apocrine-gland bearing skin causing inflammation, suppuration and scarring. There are limited studies on the efficacy of treatment for hidradenitis suppurativa. We sought to determine the efficacy of triple pulsed NdYag laser therapy in conjunction with antibiotics in comparison to antibiotics alone in the treatment of this disease.

In the first phase of the study, 12 patients diagnosed with Hurley Stage II hidradenitis suppurativa were randomized to one of two groups: antibiotic only or antibiotic plus laser. The antibiotic only group received 300 mg of clindamycin and 300 mg of rifampin by mouth twice daily for three months. The antibiotic plus laser group received the same combination of antibiotics for two weeks, in addition to monthly laser treatment with the NdYag laser for 3 months. Primary outcome measure was the Hidradenitis Suppurativa Lesion Area Score Index (HS-LASI; a modification of the Sartorius scale).

Six (6) patients were placed in the antibiotic only group, and six (6) were placed in the antibiotic plus laser group. Preliminary results show that over the first three months of treatment, the HS-LASI decreased on average by 22.1% (95% CI: 4.2% to -48.4%) in the antibiotic only group, compared to a decrease of 45.9% (95% CI: -27.5% to -64.2%) in the antibiotic plus laser group (p<0.18). Although the results did not reach statistical significance, all patients in the antibiotic plus laser group showed a decrease in the severity score, while one patient in the antibiotic only group worsened over the three months. The lack of significance is likely due to sample size which should be addressed as the next phase of studies start. These results show a benefit to using NdYag laser treatment in conjunction with antibiotics for the treatment of Stage II hidradenitis suppurativa in efficacy and reducing the need for systemic antibiotics.

COMPARISON OF LONGITUDINAL SUNBURN RATES IN THE CANADIAN POPULATION USING THE NATIONAL POPULATION HEALTH SURVEY.

Kalia S, Lui H. Photomedicine Institute, Department of Dermatology and Skin Science; Vancouver Coastal Health and University of British Columbia; Vancouver, BC.

Skin cancer, the most common type of cancer continues to be a major concern with increasing numbers of skin cancers being detected. A variety of campaigns have targeted the Canadian population to educate the public about the importance of reducing sun exposure. Although studies have shown that these campaigns have increased the knowledge of the importance of sun protection and sun avoidance, limited studies have evaluated the effect on behavior. In particular, within Canada there is a lack of data on the long term effects of educational campaigns at a national level. The National Population Health Survey (NPHS) conducted by Statistics Canada has followed the same cohort of people across Canada over several years, targeting over 98% of the Canadian population. Different health practices and outcomes have been collected on this cohort of people. The NPHS has evaluated different sun protection behaviors, such as the presence or absence of sunburn(s) within the last twelve months. This study compared the rates of sunburn in 2006-2007 to 2000-2001. Correlations with demographic variables were computed. Simple logistic regression analysis was performed using Statistical Analysis Software (SAS) version 9.2, and those variables that were statistically significant were included in multiple logistic regression analysis. The response rates for the NPHS have varied over the years from 80-85%. In 2000-2001, 2914 of 11290 individuals experienced a sunburn within the past twelve months (25.8%), compared to 3123 of 12160 (25.7%) in 2006-2007. These changes show that minimal change has been observed with sunburn rates in the Canadian population. The trend shown in this study may indicate that educational campaigns have not yet been successful in changing sun exposure behavior patterns. Similar trends have been found in another national survey, the Canadian Community Health Survey, however further studies are needed to confirm these findings

“STOP AND GO” NB UVB PHOTOTHERAPY PROTOCOL EFFECTIVENESS VERSUS CONTINOUS NB UVB PROTOCOL IN THE TREATMENT OF PATIENTS WITH VITILIGO: A RANDOMIZED CONTROLLED STUDY.

Pacifico A, Iacovelli P, Paro Vidolin A, Leone G. Phototherapy Unit, S. Gallicano Institute, IRCCS, Rome, Italy

Vitiligo is a common skin disease characterized by loss of normal melanin pigments in the skin and its pathogenesis is still unclear. Treatment modalities include PUVA, NB UVB phototherapy, now considered as the “gold standard”, topical and systemic steroids, topical calcineurin inhibitors, topical vitamin D analogues either in monotherapy or in association with phototherapy, and surgical treatment.. Conventional NB UVB still remains the best treatment option for diffuse vitiligo.

Leaving aside short term side effects such as sunrush/erythema and sunburn, the main problem with phototherapy is represented by the cumulative long term effects of UVR. These can cause premature ageing of the skin and the appearance of skin cancer. Therefore the aim of our study was to compare the effectiveness of two different phototherapy protocols; a first protocol included a non-stop treatment for 6 months; a second protocol included periods of interruption during the treatment in order to investigate if the interruption could make the UVR more effective on the melanocytes, and consequently reduce long term damage caused by phototherapy. A total of 29 patients were enrolled in this study. 13 patients were randomly selected to receive the “stop and go” protocol (Group A) while 14 patients were randomly selected to receive a classic NB UVB phototherapy protocol (Group B). Patients were treated twice a week. Our results indicated that 6 patients belonging to group A obtained an excellent degree of repigmentation versus 4 patients of group B and that 7 patients of group A reached a good degree of repigmentation versus 9 patients of group B. Our preliminary data suggest that an interruption of phototherapy followed by a new NB UVB phototherapy course might increase the responsivity of patients’ melanocytes to UV stimulation. Controlled studies on a larger patients’ population are required in order to confirm our observations as well as to optimize the NB UVB irradiation protocol.

PHOTOTHERAPY TRENDS IN DERMATOLOGY.

Dabade TS1, Davis SA1, Feldman SR1,2,3. Center for Dermatology Research, Departments of 1Dermatology, 2Pathology and 3Public Health Sciences; Wake Forest University School of Medicine; Winston-Salem, NC

Background: Phototherapy is an efficacious outpatient treatment for many dermatologic disorders including psoriasis. Interest in its use continues to grow with investigations for varying purposes. Objective: Characterize the use of phototherapy over time by dermatologists in the outpatient setting. Methods: We analyzed the National Ambulatory Medical Care Survey data to estimate the number of outpatient phototherapy visits to non-federal physicians from 1997 – 2008. Results: There were an estimated 1.5 million outpatient visits for phototherapy during this time period of which 85% were administered by dermatologists. Among patients visiting dermatologists, the most common conditions treated were psoriasis (47%), dermatitis NOS (15%), and actinic keratosis (8%). Patients’ were 49% female and most commonly White (82%) or African American (10%). Regression analysis of the number of visits for the use of phototherapy from 1997 – 2008 showed an increasing trend for all diagnoses cumulatively (p=0.01), but for psoriasis or actinic keratosis specifically there was no significant change (p= 0.82, 0.10, respectively). Recent years showed phototherapy use in less common diagnoses, such as pityriasis rosea, dyschromia, sebaceous disease NOS, or pruritus NOS. Payment sources used for phototherapy visits were private insurance (47%), Medicare (29%), and self-pay (17%). Conclusions: The frequency of phototherapy in dermatology has remained constant for its most common uses over the last decade, including psoriasis; however, its use has increased in a minority of diagnoses. Alternative treatments, such as home phototherapy, biologics, or imiquimod may be contributing to this steady state. Out of pocket payments may be limiting phototherapy use resulting in treatments that are overall more expensive.

INTENSE PULSED LIGHT: A NEW STAR ON THE HORIZON OF PDT LIGHT SOURCES?

Philipp Babilas, Stephan Schreml, Michael Landthaler, Rolf-Markus Szeimies Affiliation: Department of Dermatology, University Hospital, University of Regensburg, Regensburg, Germany

(a) Statement of purpose: To evaluate the painfulness and efficacy of intense pulsed light (IPL), a prospective, randomized, controlled split-face study was performed.

(b) Description of design or methods: Topical MAL-PDT was conducted in 25 patients with AK (n=238) that were suitable for two-side comparison. After incubation with MAL, irradiation was performed with a light emitting diode (LED) (50 mW cm-2; 37 J cm-2) vs. IPL (80 J cm-2, double pulsed, 40 J cm-2, pulse train of 15 impulses each with a duration of 5 ms, 610-950 nm filtered hand piece) followed by re-evaluation up to 3 months.

(c) Summary of results: The painfulness during and after therapy was significantly lower with IPL irradiation [t(df=24)=4.42, p<0.001]. The overall infiltration and keratoses score three months after treatment was 0.86±0.71 (LED-system) vs. 1.05±0.74 (IPL) (no statistically significant difference; p=0.292). Patient satisfaction following both treatment modalities did not significantly vary during the 3 months follow-up (p=0.425).

(d) Statement of conclusions: IPL use for MAL-PDT is an efficient alternative for the treatment of AK that results in complete remission and cosmesis equivalent to LED irradiation but causes significantly less pain.

SKIN MELANOMA INCIDENCE CORRELATES STRONGER WITH GEOMAGNETIC THAN WITH GEOGRAPHIC LATITUDE.

Charles R. Chubb, Ferguson, Missouri.

The purpose of this study was to find if the skin melanoma incidence correlation with geomagnetic latitude is stronger than with geographic latitude and how the incidence is changing with time in the presence of large scatter in the incidence values. The method was to enter the values for melanoma of the skin incidences (world age-standardized) for the registries in Cancer Incidence in Five Continents volumes III and IX into computer files. Geographic latitudes and longitudes for the registries were determined and geomagnetic latitudes for the registries were evaluated. Correlation and regression analyses were conducted. The coefficients for cancer incidence correlation with geomagnetic latitude were greater than the coefficients for correlation with geographic latitude. Regression of melanoma of the skin incidence with the square of the geomagnetic latitude values resulted in a significant incidence increase between the values in volume III, center year 1970, and the values in volume IX, center year 2000, for geomagnetic latitudes greater than thirty-one degrees. For geomagnetic latitude fifty degrees, the nominal male regression incidence increase was from 2.4 to 9.7 annual cases /100,000 males. The fact the geomagnetic correlation is stronger than geographical correlation indicates consideration should be given to ionizing radiation, in addition to the many other factors that are increased at high latitudes, as a possible contributing factor for skin melanoma.

USE OF EXTRACORPOREAL PHOTOPHERESIS FOR THE TREATMENT OF BRONCHIOLITIS OBLITERANS SYNDROME AFTER LUNG TRANSPLANTATION.

Robert Knobler, Ulrike Just, and Peter Jaksch
Departments of Dermatology and Transplantation Surgery, Medical Univ. of Vienna, Austria

BACKGROUND: Extracorporeal photopheresis (ECP) has been used since 1995 to treat acute and chronic rejection in lung transplantation recipients. However, data supporting the use of ECP for bronchiolitis obliterans syndrome (BOS) after lung transplantation are limited.

METHODS: We evaluated the efficacy and safety of ECP for progressive BOS at our institution in a prospective manner. Between 10/ 2000, and 6/2010, 47 lung allograft recipients were treated with ECP for progressive BOS. Only patients with a minimum follow up 6 months were included ( n=45). Time from lung transplantation to BOS grade 1 was 1216±898 days (mean±SD)(range 161-4306 days). The interval between initiation of BOS 1 and start of ECP was 255±332 days. Mean number of ECP cycles was 16.

RESULTS: During the 6-month period before the initiation of ECP, the average rate of decline in forced expiratory volume in 1 second (FEV1) was -123.0 ml/month, but the slope decreased to -13 ml/month during the 3-month period after the initiation of ECP and -16ml/month during the 12month period . The mean difference in the rate of decline between this 12-month period and the period before ECP was 42/month (95% CI, 71-130 ml/month; p =0,006). The FEV1 improved in 17% of patients for more than 1 year after the initiation of ECP, 11% showed an improvement for only 3-6 months, FEV1 stabilized in 36% and showed a progressive decline in 36%.

CONCLUSIONS: ECP reduces the rate of decline in lung function associated with progressive BOS. Subgroups of patients responded with an improvement or stabilization of FEV1 for more than 1 year. Future studies are warranted.

EFFICACY AND FEASIBILITY OF COMBINATION OF EXCIMER LASER THERAPY, CLOBETASOL SPRAY, AND CALCITRIOL OINTMENT FOR GENERALIZED PSORIASIS.

Tina Bhutani, Misha Heller, Eric Lee, Kelly Park, John Koo. University of California, San Francisco Department of Dermatology, San Francisco, CA.

Purpose: To ascertain if a combination of the most powerful excimer laser, clobetasol spray, and calcitriol ointment can treat generalized psoriasis faster and more efficaciously than any current regimen available including oral, biologic, and phototherapy options and without exposing the patient to serious systemic risks. Design: This is a 12-week, open-label, pilot trial evaluating the efficacy and safety of the combination of excimer laser therapy with clobetasol spray as the initial treatment of generalized plaque psoriasis, followed by maintenance therapy with topical calcitriol ointment. Regarding the excimer laser treatments, all patients will receive excimer laser treatments with the Photomedex XTRAC® Velocity machine twice weekly for the first 6 weeks. For the remaining 6 weeks, only patients who are still at less than PASI 75 response will continue twice weekly excimer laser treatments. For topicals, the study will be conducted in three distinct periods (A, B, and C), each of 4 weeks duration. During Period A (weeks 1-4), patients will use clobetasol spray twice daily along. During Period B (weeks 5-8), patients will be treated with only topical calcitriol ointment twice daily. During Period C (weeks 9-12), patients will use clobetasol spray twice daily and calcitriol ointment twice daily. Results: Out of 8 subjects recruited, 6 completed 6 weeks and 5 subjects completed all 12 weeks. 6/6 patients who completed first 6 weeks achieved PASI 75. 5/5 patients who completed all 12 weeks achieved PASI 75. Conclusion: With the combination of excimer laser, clobetasol spray, and calcitriol ointment, 100% achieved PASI 75 in 6 weeks. This outstanding efficacy and fast onset of action may reflect the fact that supraerythemogenic protocol was utilized whereby dosimetry reflecting many times the MED was applied only to the lesional skin, making UVB phototherapy much more effective. External treatment of generalized psoriasis with no serious internal risks with significantly better efficacy than current internal options appears within reach.

DOES PIGMENT MELANIN INFLUENCE KELOID FORMATION?

Menter JM, Nokkaew C, Green A, Naqvi H, and Harris-Hooker S, Morehouse School of Medicine, Atlanta GA.

Purpose: Nitric oxide (NO) has been implicated in the formation of keloids. As keloids preferentially occur in Blacks, pigment melanin might be involved. Cuttlefish sepia melanin can scavenge donor - generated NO through a dialysis membrane in vitro. We hypothesize that reactive nitrogen (RNS) and/or reactive oxygen species (ROS) may occur from melanin – mediated NO oxidation.

Design: Purified sepia melanin extracted from cuttlefish was used. NO was generated in vitro with 0.100 mM 2-(N.N Diethylamino) S - nitroso - N - acetylpenicillamine (SNAP) in phosphate buffer. It was assessed by measurement of its fluorescent adduct with 4, 5 - diaminofluorescein (DAF). Peroxynitrite (ONOO -) was generated in vitro via 0.250 mM 3 – morpholino – syndonimine (SIN – 1), and assessed with the selective scavenger 3.3 µM 3 - methyl - 1, 2 - cyclopentanedione (MCP) in solution. We incubated cultured human adult dermal fibroblasts with and without 0.1 mM melanin, and tested for peroxynitrite via Western and immunocytofluorometric assays using a 3- nitrotyrosine antibody. H2O2 was determined via the scopoletin assay.

Results: We detected significant amounts of peroxynitrite in melanin – containing systems, but not in controls. We could detect little or no H2O2 in these systems. Addition of soluble melanin to the fibroblast culture system resulted in formation of 3-nitrotyrosine. Comparison of the test and control systems by immunocytofluorometry and by Western blotting indicated a small but significant amount of 3-nitrotyrosine in the test cultures. At high 3-nitrotyrosine levels, melanin conferred significant protection from tyrosine nitration.

Conclusions: These results indicate melanin’s chemically complex nature where both protective and cytotoxic effects to fibroblasts can be observed in concentration – dependent manner. Peroxynitrite is produced by redox reactions involving melanin, superoxide, and NO. Peroxide may arise from dismutation of superoxide by melanin, but it will be re-scavenged by melanin. Peroxynitrite may therefore play a significant role in keloid formation. Supported by MBRS Grant #GM08248 , RCMI Grant #RR 03034 and DOD Grant # W911 NF- 10- 1- 0448.

HOW WELL DO SUNSCREENS PROTECT AGAINST ULTRAVIOLET A INDUCED FREE RADICALS?

Osterwalder U1, Floesser-Mueller H1, Jung K2, 
1BASF SE (Ludwigshafen, Germany), 2Gematria Testing Lab (Berlin, Germany)

Currently, the efficiency of UVA protection is assessed via the Persistent Pigment Darkening (PPD) method in vivo and in vitro and by several other in vitro methods based on transmission measurement. Contrary to the SPF method all these UVA methods are lacking a relevant biological endpoint. The determination of the Radical Skin Protection Factor (RSF) is an alternative method that refers to the source of UVA induced skin damage. UVA induced free radicals, e.g. Reactive Oxygen Species (ROS) are measured by Electron Spin Resonance (ESR) spectroscopy, ex vivo in irradiated skin explants.

This study was designed to investigate whether the reduction of free radical formation, expressed as RSF is a measure of UVA protection or, vice versa, whether UVA protection is a suitable predictor of protection from free radical formation.

There is an average correlation UVA-PF = 0.33 SPF over all samples. Half of the current market products comply with the European UVA recommendation. Among the half that complies the average relation UVA-PF = 0.50 SPF was found. The RSF was found to be related to the UVA-PF, RSF = (1.0-1.3) UVA-PF, depending on the SPF range of the samples. The deviation from the perfect correlation can be explained by the various influence factors and differences of both methods.

The RSF is able to provide the information on UVA protection which is required in addition to the SPF. On the other hand in vitro or in silico assessment of UVA protection can also serve as a valuable predictor of the RSF. Good UVA protection means good protection against free radicals.