Identification of Seasonal Variations of Antibodies against PR-10-Specific Epitopes Can Be Improved Using Peptide-Phage Display.

BACKGROUND: In pollinosis patients, allergen-specific antibody titers show seasonal variations. Little is known about these variations at the epitope level. OBJECTIVES: We aimed at investigating seasonal variations on the level of allergen epitope recognition in patients with Bet v 1-related food allergy using a peptide phage display approach. METHODS: Serum samples collected over 1 year from 4 patients of the placebo arm of the birch-associated soya allergy immunotherapy trial were included. To identify epitopes from Bet v 1-related food allergens, patient sera were used in peptide phage display experiments. In silico analysis of enriched allergen-related motifs was performed. RESULTS: We identified epitope motifs related to Bet v 1 and its homologs in soya and hazelnut (Gly m 4 and Cor a 1, respectively) that were enriched in accordance with birch and hazel pollen exposure. Within several weeks after the birch pollen season peak, the pattern of identified epitope motifs differed considerably among patients. Data for amino acid preferences in homologous Bet v 1 and Cor a 1 epitope motifs identified for one of the investigated patients suggest changes in concentration or specificity of serum antibodies for the Cor a 1 epitope motif. CONCLUSIONS: Peptide phage display data suggest an impact of birch and hazel pollen exposure on the recognition pattern of Bet v 1-like allergen epitopes. Epitope-oriented analyses could provide deeper, personalized details regarding the allergen epitope recognition influenced by pollen exposure beyond the capability of current methods.

High-Fat Diet Exacerbates Early Psoriatic Skin Inflammation Independent of Obesity: Saturated Fatty Acids as Key Players.

In obesity, hypertrophic adipocytes secrete high amounts of adipocytokines, resulting in low-grade inflammation amplified by infiltrating proinflammatory macrophages, oxidative stress, hypoxia, and lipolysis. These chronic proinflammatory conditions support the development of type II diabetes and cardiovascular diseases, but the mechanisms of obesity-related exacerbation of inflammatory skin disorders like psoriasis are unclear. In this study, we uncovered dietary saturated fatty acids (SFAs) as major risk factors for the amplification of skin inflammation, independent of obesity-related parameters such as fat mass extension, adipocytokine levels, and glucose homeostasis. Correlation analyses in a cohort of psoriasis vulgaris patients showed that free fatty acid serum level was the only obesity-associated parameter affecting disease severity. Studies in mice with high-fat diet-induced obesity with psoriasiform inflammation confirmed this critical role of free fatty acids. An increase of free fatty acids in healthy, lean mice alone was sufficient to induce an exacerbation of psoriasiform inflammation. In particular, saturated fatty acids sensitize myeloid cells to an increased inflammatory response in answer to proinflammatory stimuli, which in turn augments the activation of keratinocytes. Consequently, reduction of nutritional saturated fatty acids alone diminished the psoriatic phenotype in obese mice. Thus, our findings may open new perspectives for adjuvant dietary measures accompanying anti-inflammatory psoriasis therapies in lean and obese patients.

Critical role for syndecan-4 in dendritic cell migration during development of allergic airway inflammation.

Syndecan-4 (SDC4), expressed on dendritic cells (DCs) and activated T cells, plays a crucial role in DC motility and has been shown as a potential target for activated T-cell-driven diseases. In the present study, we investigate the role of SDC4 in the development of T-helper 2 cell-mediated allergic asthma. Using SDC4-deficient mice or an anti-SDC4 antibody we show that the absence or blocking of SDC4 signalling in ovalbumin-sensitized mice results in a reduced asthma phenotype compared with control animals. Most importantly, even established asthma is significantly decreased using the anti-SDC4 antibody. The disturbed SDC4 signalling leads to an impaired motility and directional migration of antigen-presenting DCs and therefore, to a modified sensitization leading to diminished airway inflammation. Our results demonstrate that SDC4 plays an important role in asthma induction and indicate SDC4 as possible target for therapeutic intervention in this disease.

Fractional CO2 laser is as effective as Q-switched ruby laser for the initial treatment of a traumatic tattoo.

BACKGROUND AND OBJECTIVE: Q-switched laser treatments are considered the standard method for removing both regular and traumatic tattoos. Recently, the removal of tattoo ink using ablative fractional lasers has been reported. Ablative fractional CO2 laser and q-switched ruby laser treatments were used in a split-face mode to compare the safety and efficacy of the two types of laser in removing a traumatic tattoo caused by the explosion of a firework. STUDY DESIGN/PATIENTS AND METHODS: A male patient suffering from a traumatic tattoo due to explosive deposits in his entire face was subjected to therapy. A series of eleven treatments were performed. The right side of the face was always treated using an ablative fractional CO2 laser, whereas the left side was treated only using a q-switched ruby laser. RESULTS: After a series of eleven treatments, the patient demonstrated a significant lightening on both sides of his traumatic tattoo, with no clinical difference. After the first six treatments, the patient displayed greater lightening on the right side of his face, whereas after another five treatments, the left side of the patient's face appeared lighter. No side effects were reported. CONCLUSIONS: In the initial stage of removing the traumatic tattoo, the ablative fractional laser treatment appeared to be as effective as the standard ruby laser therapy. However, from the 6th treatment onward, the ruby laser therapy was more effective. Although ablative fractional CO2 lasers have the potential to remove traumatic tattoos, they remain a second-line treatment option.

Local anesthesia with lidocaine and prilocaine, using the tumescent technique, for the radiofrequency ablation of lower extremity varicose veins.

AIM: To demonstrate the possible use of a tumescent solution containing lidocaine and prilocaine for radiofrequency-assisted segmental thermal ablation and to reduce the need for postoperative analgesics. METHODS: Fifty patients (51 limbs) underwent radiofrequency-assisted segmental thermal ablation (VNUS Closure FAST(™) ) of insufficient great and small saphenous veins and phlebectomy. The amount of tumescent fluid injected was noted, and evaluation of pain reported on visual analog scale (VAS) and quantity of the postoperative analgesics used and their side effects were recorded one day and six weeks after the procedure. RESULTS: The average amount of a tumescent fluid injected was 852 ml/patient (250-1470 ml). One day after surgery, 65% of the patients reported no pain (VAS 0), 27% reported a VAS of 1, and 4% reported a VAS of 2 on a scale of 0-10. Overall, 4% of the total patient population used analgesics after the procedure. Six weeks after the procedure, 84% of the patients reported no pain. CONCLUSION: Local anesthesia with lidocaine and prilocaine, and the tumescent technique, were found to be effective and safe modalities for radiofrequency ablation of varicose veins of the lower extremities.

Electrochemotherapy under tumescent local anesthesia for the treatment of cutaneous metastases.

BACKGROUND: The surgical management of cutaneous metastasis (CM) is challenging, particularly in elderly patients, in whom general anesthesia can be difficult because of comorbidity. OBJECTIVES: To test the effectiveness of tumescent local anesthesia (TLA) to achieve adequate anesthesia during the treatment of extensive CM with electrochemotherapy (ECT), previously only performed under general anesthesia. METHODS: We conducted five ECT treatments of CM with intralesional bleomycin under TLA. We examined pain scores before, during, and after treatment; analgesic use; and side effects. The intention of the treatment was palliative in all cases. RESULTS: We treated four patients (ages 75-88) with CM with a mean area of 126 cm(2) (range 12-198 cm(2) ) with 356 mL of TLA per treatment (range 180-450 mL). The ECT treatment under TLA demonstrated that anesthesia was adequate, with moderate pain during and slight pain after the procedure as measured on a visual analog scale (VAS). CONCLUSIONS: In this proof-of-principle study, we demonstrated that ECT can be performed under TLA; TLA might be a useful new anesthesia option for patients treated with ECT.

Update dermatologic laser therapy.

New trends in dermatological laser therapy during the last years are based on new wavelengths, concepts and treatment combinations resulting in a variety of new dermatologic indications. Fractional laser therapy of chronic actinic damage of the skin has been introduced and already represents a standard technique. The concept of fractional non-ablative and ablative laser treatment has been shown to be safe and effective. Also pigmented and vascular skin changes can be treated by this method. New, very promising concepts for laser epilation include linear scanned as well as low fluence laser systems. The first enable very short treatment times for large areas; the latter are the basis for the growing market of laser epilation devices for home use. Nevertheless, the potential of low fluence laser devices for long-term hair reduction has not been tested so far. Furthermore, no data exist on side effects resulting from repetitive application of laser light to melanocytic lesions. Laser lipolysis has been introduced as the latest, minimally invasive way of removing small localised fat deposits. The new procedure may have a great potential for liposculpture; its further development should be thoughtfully observed. The latest innovations for precise ablation are ultra-short pulsed laser systems. Femtosecond lasers avoid thermal damage at the border areas of ablation zones.

Local anesthesia in dermatology.

Local and regional anesthetic procedures are an integral part of daily dermatological practice. Safe and effective analgesia in skin and soft tissues is crucial for otherwise painful diagnostic or therapeutic interventions. Tumescent local anesthesia allows for pain-free interventions that previously had to be done by using general anesthesia. Older patients with multiple co-morbidities are especially suited for local anesthetic procedures, because they may significantly reduce surgical risks. For dermatologists, the knowledge of mode of action and toxicity of local anesthetics, as well as the emergency management of their potential complications, is essential.

Exposure to mycotoxins increases the allergic immune response in a murine asthma model.

RATIONALE: Epidemiological studies have shown that indoor molds are associated with increased prevalence and exacerbation of respiratory symptoms and asthma. Mycotoxins, secondary metabolites of molds, may contribute to these effects. OBJECTIVES: To investigate the adjuvant activity of mycotoxins on allergic airway inflammation. METHODS: Balb/c mice were exposed via the airways to gliotoxin and via the intestine to patulin, sensitized with ovalbumin (OVA), and then analyzed in acute and chronic murine asthma models. In addition, the effect of mycotoxin exposure on dendritic cell (DC) function was investigated using murine bone marrow-derived DCs. MEASUREMENTS AND MAIN RESULTS: Exposure of mice to both mycotoxins enhanced dose-dependently airway hyperreactivity, eosinophilic lung inflammation, and OVA-specific IgE serum levels compared with mice that received only the antigen. These findings correlated with increased Th2 cytokine levels and decreased IFN-gamma production. Long-term mycotoxin exposure exacerbated chronic airway inflammation and airway remodeling. In vitro or in vivo mycotoxin exposure inhibited IL-12 production in maturing DCs and enhanced airway inflammation after adoptive DC transfer into Balb/c mice. Mycotoxin exposure enhanced OVA-induced lung lipid peroxidation and moderately increased isoprostane levels in naive mice. Treatment of mycotoxin-exposed DCs with the antioxidants N-acetylcysteine or glutathione ethyl ester restored IL-12 secretion and pretreatment of exposed mice with N-acetylcysteine prevented the mycotoxin-induced increase of airway inflammation and AHR. CONCLUSIONS: Our results demonstrate that gliotoxin and patulin increase the allergic immune response in mice by modulating the Th1/Th2 balance via direct effects on IL-12 secretion in DCs and by inducing oxidative stress.

Fractional laser skin therapy.

Modern treatment of chronically photodamaged skin provides new, minimally invasive methods for laser intervention, using fractional, non-ablative thermal energy for an induction of dermal remodeling. The clinical efficacy coupled with a minimum "down time" for the patient has directed the development of fractional ablative laser systems. Recently introduced systems are based on CO(2) and heated Er:YAG laser systems. The clinical efficacy has been tested on one prototype each and verified their effects at a microscopic level. Initial reports suggest the results are comparable to those achieved with fractional ablative systems. We review the current possibilities incorporating our personal experience. Systematic investigations of clinical outcomes with various system settings are still needed. The possible combination of ablative and non-ablative fractional technology may also lead to increased efficacy and safety of the procedure.

Microcavity array (MCA)-based biosensor chip for functional drug screening of 3D tissue models.

Multicellular tumour spheroids that mimic a native cellular environment are widely used as model systems for drug testing. To study drug effects on three-dimensional cultures in real-time we designed and fabricated a novel type of sensor chip for fast, non-destructive impedance spectroscopy and extracellular recording. Precultured spheroids are trapped between four gold electrodes. Fifteen individual 100microm deep square microcavities with sizes from 200 to 400microm allow an optimised positioning during the measurement. Although apoptosis was induced in human melanoma spheroids by Camptothecin (CTT), treated cultures did not show disintegration but displayed increased impedance magnitudes compared to controls after 8h resulting from an altered morphology of the outer cells. Contractions in cardiomyocyte spheroids were monitored when the innovative chip was used for recording of extracellular potentials. The silicon-based electrode array is used as an acute test system for the monitoring of any kind of 3D cell cultures. Since no adherence of cells or labelling is necessary the multifunctional sensor chip provides a basis for improved drug development by high content screenings with reduced costs and assay times. Additional improvements for parallel testing of different substances on one chip are presented.

Solar simulator-induced phototoxicity of the furoquinoline alkaloid dictamnine compared to 8-methoxypsoralen and 5-methoxypsoralen.

Dictamnine, a furoquinoline alkaloid of the Rutaceae plant family, has been shown to be mutagenic and phototoxic in bacteria and yeasts. Here, we have investigated the phototoxic effect of dictamnine in human Jurkat T cells and HaCaT keratinocytes. Dictamnine was isolated from the roots of DICTAMNUS ALBA L. and was photoactivated with solar simulated radiation, delivered from a 1000-W xenon arc lamp with a maximal output between 300 - 800 nm. Dictamnine displayed concentration- and light-dependent phototoxic effects in both cell lines. In comparison to the structurally related furocoumarins 5-methoxypsoralen and 8-methoxypsoralen, dictamnine was less phototoxic. Nevertheless, it may play a major role in the elicitation of phytophotodermatitis because of its abundance in plants of the Rutaceae family.

Hyperforin acts as an angiogenesis inhibitor.

Hyperforin is a plant compound from Hypericum perforatum that inhibits tumor cell proliferation in vitro by induction of apoptosis. Here, we report that hyperforin also acts as an angiogenesis inhibitor in vitro and in vivo. In vitro, hyperforin blocked microvessel formation of human dermal microvascular endothelial cells (HDMEC) on a complex extracellular matrix. Furthermore, hyperforin reduced proliferation of HDMEC in a dose-dependent manner, without displaying toxic effects or inducing apoptosis of the cells. To evaluate the antiangiogenic activity of hyperforin in vivo, Wistar rats were subcutaneously injected with MT-450 mammary carcinoma cells and were treated with peritumoral injections of hyperforin or solvent. Hyperforin significantly inhibited tumor growth, induced apoptosis of tumor cells and reduced tumor vascularization, as shown by in situ staining of CD31-positive microvessels in the tumor stroma. These data suggest that, in addition to the induction of tumor cell apoptosis, hyperforin can also suppress angiogenesis by a direct, non-toxic effect on endothelial cells.

Topical treatment of atopic dermatitis with St. John's wort cream--a randomized, placebo controlled, double blind half-side comparison.

BACKGROUND: Recent investigations suggest an anti-inflammatory and antibacterial effect of hyperforin, which is a major constituent of Hypericum perforatum L. (Saint John's wort). OBJECTIVE: In the present half-side comparison study we assessed the efficacy of a cream containing Hypericum: extract standardised to 1.5% hyperforin (verum) in comparison to the corresponding vehicle (placebo) for the treatment of subacute Atopic Dermatitis. The study design was a prospective randomised placebo-controlled double-blind monocentric study. METHODS: In twenty one patients suffering from mild to moderate Atopic Dermatitis (mean SCORAD 44.5) the treatment with verum or placebo was randomly allocated to the left or right site of the body, respectively. The patients were treated twice daily over a period of four weeks. Eighteen patients completed the study. The severity of the skin lesions on the left and right site was determined by means of a modified SCORAD-index (primary endpoint). RESULTS: The intensity of the eczematous lesions improved on both sites of treatment. However, the hypericum-cream was significantly superior to the vehicle at all clinical visits (days 7, 14, 28) (p < 0.05). Skin colonisation with Staphylococcus aureus was reduced by both verum and placebo, showing a trend to better antibacterial activity of the hypericum-cream (p = 0.064). Skin tolerance and cosmetic acceptability was good or excellent with both the hypericum-cream and the vehicle (secondary endpoints). CONCLUSION: Taken together, the present study shows a significant superiority of the hypericum-cream compared to the vehicle in the topical treatment of mild to moderate Atopic Dermatitis. The therapeutic efficacy of the hypericum-cream, however, has to be evaluated in further studies with larger patient cohorts, in comparison to therapeutic standards (i.e. glucocorticoids).

Effect of oral administration ofHypericum perforatum extract (St. John's Wort) on skin erythema and pigmentation induced by UVB, UVA, visible light and solar simulated radiation.

Hypericin from St John's wort (Hypericum perforatum L.) is a photosensitizing agent that may cause a severe photodermatitis when higher amounts of St John's wort are ingested by animals. Although Hypericum extracts are widely used in the treatment of depressive disorders, only a little information on the photosensitizing capacity of St John's wort in humans is available. In the present prospective randomized study we investigated the effect of the Hypericum extract LI 160 on skin sensitivity to ultraviolet B (UVB), ultraviolet A (UVA), visible light (VIS) and solar simulated radiation (SIM). Seventy two volunteers of skin types II and III were included and were divided into six groups, each consisting of 12 volunteers. In the single-dose study the volunteers (n = 48) received 6 or 12 coated tablets (5400 or 10 800 microgram hypericin). In the steady-state study the volunteers (n = 24) received an initial dose of 6 tablets (5400 microgram hypericin), and subsequently 3 x 1 tablets (2700 microgram hypericin) per day for 7 days. Phototesting was performed on the volar forearms prior to medication and 6 h after the last administration of Hypericum extract. The erythema-index and melanin-index were evaluated photometrically using a mexameter. After both single-dose and steady-state administration, no significant influence on the erythema-index or melanin-index could be detected, with the exception of a marginal influence on UVB induced pigmentation (p = 0.0471) in the single-dose study. The results do not provide evidence for a phototoxic potential of the Hypericum extract LI 160 in humans when administered orally in typical clinical doses up to 1800 mg daily. This is in accordance with previous pharmacokinetic studies that found hypericin serum and skin levels after oral ingestion of Hypericum extract always to be lower than the assumed phototoxic hypericin threshold level of 1000 ng/mL.

Phototoxic and apoptosis-inducing capacity of pseudohypericin.

Pseudohypericin (PH) and hypericin (H) are photosensitizing plant pigments from Hypericum perforatum. H displays cytotoxic and apoptosis-inducing effects in neoplastic cell lines. Here, we have assessed the phototoxic and apoptosis-inducing capacity of PH compared to H in a cell culture model with human leukemic lymphoma cells (Jurkat). Treatment with both photoactivated H and PH resulted in a dose-dependent inhibition of cell proliferation, whereas not photoactivated H and PH had no effect at the concentrations tested. The half-maximal inhibitory concentration (IC50) of H was lower (100 ng/mL) as compared to PH (200 ng/mL) (p < 0.05). In an apoptosis assay we found a dose-dependent increase of DNA fragmentation after treatment with both photoactivated H and PH. The cytotoxic potential of PH should be taken into account during systemic therapy with Hypericum extracts, since PH is about two times more abundant than H in Hypericum perforatum.

Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John's wort that acts by induction of apoptosis.

Hyperforin is a plant derived antibiotic from St. John's wort. Here we describe a novel activity of hyperforin, namely its ability to inhibit the growth of tumour cells by induction of apoptosis. Hyperforin inhibited the growth of various human and rat tumour cell lines in vivo, with IC(50) values between 3-15 microM. Treatment of tumour cells with hyperforin resulted in a dose-dependent generation of apoptotic oligonucleosomes, typical DNA-laddering and apoptosis-specific morphological changes. In MT-450 mammary carcinoma cells hyperforin increased the activity of caspase-9 and caspase-3, and hyperforin-mediated apoptosis was blocked by the broad-range caspase inhibitor zVAD.fmk. When added to MT-450 cells, hyperforin, but not paclitaxel, induced a rapid loss of the mitochondrial transmembrane potential Deltapsi(m), and subsequent morphological changes such as homogenization and vacuolization of mitochondria. Monitoring of Deltapsi(m) revealed that the hyperforin-mediated mitochondrial permeability transition can not be prevented by zVAD.fmk. This indicates that mitochondrial permeabilization is a cause rather than a consequence of caspase activation. Moreover, hyperforin was capable of releasing cytochrome c from isolated mitochondria. These findings suggest that hyperforin activates a mitochondria-mediated apoptosis pathway. In vivo, hyperforin inhibited the growth of autologous MT-450 breast carcinoma in immunocompetent Wistar rats to a similar extent as the cytotoxic drug paclitaxel, without any signs of acute toxicity. Owing to the combination of significant antitumour activity, low toxicity in vivo and natural abundance of the compound, hyperforin holds the promise of being an interesting novel antineoplastic agent that deserves further laboratory and in vivo exploration.