Access to bacteriophage therapy: discouraging experiences from the human cell and tissue legal framework.

Cultures of human epithelial cells (keratinocytes) are used as an additional surgical tool to treat critically burnt patients. Initially, the production environment of keratinocyte grafts was regulated exclusively by national regulations. In 2004, the European Tissues and Cells Directive 2004/23/EC (transposed into Belgian Law) imposed requirements that resulted in increased production costs and no significant increase in quality and/or safety. In 2007, Europe published Regulation (EC) No. 1394/2007 on Advanced Therapy Medicinal Products. Overnight, cultured keratinocytes became (arguably) 'Advanced' Therapy Medicinal Products to be produced as human medicinal products. The practical impact of these amendments was (and still is) considerable. A similar development appears imminent in bacteriophage therapy. Bacteriophages are bacterial viruses that can be used for tackling the problem of bacterial resistance development to antibiotics. Therapeutic natural bacteriophages have been in clinical use for almost 100 years. Regulators today are framing the (re-)introduction of (natural) bacteriophage therapy into 'modern western' medicine as biological medicinal products, also subject to stringent regulatory medicinal products requirements. In this paper, we look back on a century of bacteriophage therapy to make the case that therapeutic natural bacteriophages should not be classified under the medicinal product regulatory frames as they exist today. It is our call to authorities to not repeat the mistake of the past.

LEKTI-1 in sickness and in health.

The stratum corneum (SC) is a biosensor that mediates responses to a variety of exogenous insults through various signalling mechanisms, including the activation of SC serine proteases (SP) kallikrein cascade. The SPINK5 gene encodes an SP inhibitor, the lympho-epithelial-Kazal-type-1 inhibitor (LEKTI-1), which in turn will buffer the excess of SP cascade initiation, key in the maintenance of permeability barrier homeostasis. We demonstrate that LEKTI processing can occur within the SC after secretion from stratum granulosum keratinocytes at least partially by klk7, an SC-specific chymotryptic SP. Unlike the recently described LEKTI-2, neither recombinant full-length LEKTI-1 nor recombinant LEKTI-1 fragments exhibit antimicrobial activity. Finally, we discuss the pathophysiological implications of LEKTI-1 in skin biology as well as its contribution to the pathogenesis of Netherton Syndrome and its potential involvement in atopic dermatitis.

Alexithymia in patients with alopecia areata: educational background much more important than traumatic events.

BACKGROUND: Recent studies suggest a higher prevalence of alexithymia in patients with alopecia areata (AA). Some authors link alexithymia with the presence of early traumatic events, such as dysfunctional parent-child relationships. However, until today, no studies have been carried out on the association of alexithymia and early traumatic events in AA patients. OBJECTIVE: The primary aim of this study was to explore if an association exists between the presence of traumatic childhood experiences and alexithymia in AA patients. A secondary aim was to confirm earlier observations indicating that the occurrence and/or degree of alexithymia is higher in patients with AA compared with individuals from the general population. METHODS: We enrolled 90 patients with AA. Data on alexithymia and traumatic events were collected with two self-report questionnaires: the Toronto Alexithymia Scale-20 and the Traumatic Experiences Checklist. These data were compared with data obtained from control patients without AA randomly selected from patients presenting for dermatological surgery. RESULTS: In adult AA patients, we found no evidence for a significant association between Toronto Alexithymia Scale (TAS) scores and emotional neglect or childhood traumatic experiences. We found a significant association with educational level, higher levels of education being associated with lower TAS-20 scores (P = 0.002). The mean TAS-20 score of 51.22 (SD 11.90) in our adult AA patient group was significantly higher compared with control patients from the same setting (44.00, SD 10.33, P < 0.001). CONCLUSION: In adult AA patients, higher levels of education are significantly associated with lower alexithymia scores. Somewhat unexpectedly, we found no association between alexithymia score and emotional neglect or childhood traumatic experiences. Our results also confirm that alexithymia scores are significantly higher in adult patient with AA compared with control patients.

Seasonal effects on the nasolabial skin condition.

In the present work, nasolabial skin condition and the influence of seasonal changes during autumn and winter were studied in 16 healthy female volunteers. Apart from visual scoring of erythema and skin scaliness, transepidermal water loss (TEWL), skin hydration, apparent skin pH, skin colour and skin desquamation were biophysically measured. The study results showed that nasolabial TEWL was significantly higher during wintertime than in autumn. Also skin colour measurements and squamometry scorings revealed higher values, indicating a more reddish and scaly nasolabial skin during winter compared to autumn. Results from tape stripping and skin surface lipid analysis by high-performance thin-layer chromatography demonstrated significant differences for triglycerides and cholesterol esters, indicating a functionally inferior hydrolipidic layer during the winter season.

Trichothiodystrophy-like hair abnormalities in a child with keratitis ichthyosis deafness syndrome.

Keratitis ichthyosis deafness syndrome is a rare congenital ectodermal disorder. It appears to be genetically heterogeneous and may be caused by mutations in the connexin 26 (Cx26) gene (GJB2) or in the connexin 30 gene. It is characterized by the association of ichthyosis-like skin lesions, hearing loss, and vascularizing keratitis. We report the clinical and molecular findings in a 5-year-old girl with keratitis ichthyosis deafness syndrome. DNA sequencing in our patient revealed a p.Ser17Phe mutation in GJB2. Besides the typical clinical features of keratitis ichthyosis deafness syndrome, a peculiar intriguing finding not previously described in the literature in this condition was that polarizing light microscopy of the scalp hair in our patient revealed striking bright and dark bands as seen in trichothiodystrophy. Amino acid analysis of the hair sample also disclosed a reduced cysteine index. We emphasize that it would be of great benefit to examine hair shafts in other patients with keratitis ichthyosis deafness syndrome for trichothiodystrophy-like abnormalities.

Oral R115866 in the treatment of moderate to severe facial acne vulgaris: an exploratory study.

BACKGROUND: R115866 (Rambazole; Barrier Therapeutics NV, Geel, Belgium), a new-generation retinoic acid metabolism-blocking agent, is a nonretinoid compound enhancing intracellularly the endogenous levels of all-trans-retinoic acid by blocking its catabolism. By virtue of this property, and the proven positive effects of retinoids in the treatment of acne, R115866 could potentially be a useful drug for acne. OBJECTIVES: To explore the efficacy, safety and tolerability of systemic R115866 in male patients with moderate to severe facial acne vulgaris (at least 15 papules and/or pustules and at least two nodulocystic lesions). METHODS: In this exploratory trial, 17 patients were treated with oral R115866 1 mg once daily for 12 weeks, followed by a 4-week treatment-free period. RESULTS: At the end of treatment (week 12, n = 16) a mean reduction in inflammatory lesion count of 77.4% (P < 0.001), in noninflammatory lesion count of 58.3% (P < 0.001) and in total lesion count of 76.0% (P < 0.001) was observed as compared with baseline. All lesion counts were significantly reduced from week 4 onwards. Mild side-effects were reported occasionally. CONCLUSIONS: The current data indicate that treatment with oral R115866 1 mg once daily for 12 weeks in patients with moderate to severe facial acne vulgaris is efficacious and well tolerated and merits further investigation.

Distinct phenotypic changes between the superficial and deep component of giant congenital melanocytic naevi: a rationale for curettage.

BACKGROUND: Giant congenital melanocytic naevi (GCMN) convey a 14-fold increased melanoma risk. In contrast, medium congenital melanocytic naevi (MCMN) are rarely associated with malignant transformation. Management of patients with GCMN is challenging and there is no consensus on the most appropriate strategy for treating these patients. OBJECTIVES: To provide a rationale for performing curettage of GCMN in the neonatal period in order to reduce the risk of malignant transformation to melanoma. METHODS: Twenty-six infants with GCMN who underwent biopsies before excisional surgery (n = 7) or curettage (n = 19) during the past 14 years (Academic Hospital, Vrije Universiteit Brussel) and 10 MCMN patients who underwent excision biopsies (Radboud University Nijmegen Medical Centre) were included in this study. Using these biopsies, we performed genetic and detailed immunohistochemical evaluations of changes that are associated with malignant transformation. Variables of interest included melanoma-associated BRAF mutations, proliferative activity, vascularity, cellular context and extracellular matrix architecture. RESULTS: GCMN and MCMN did not show oncogenic BRAF mutation and displayed similar features with respect to the amount of nonmelanocytic cells within the naevus and matrix architecture. Naevus cells in the superficial component of the GCMN, however, were more proliferative, and this component was more vascular compared with its deep component and with MCMN. In this study, none of the 19 newborn patients who underwent curettage developed a melanoma within a mean follow-up time of 7 years. CONCLUSIONS: The data presented here support the idea that curettage of GCMN in neonates has the potential for lowering the risk of developing cutaneous melanoma by not only obtaining an important numerical reduction of naevus cells but also removing the 'active' melanocytes.

Analysis of epidermal lipids of the healthy human skin: factors affecting the design of a control population.

The intervariability of studies on the lipids of human epidermis and stratum corneum is high because of the different origin of the skin samples and the variety of extraction methods used. In the present work, a high-performance thin-layer chromatographic technique has been used to study the parameters age, sex, and anatomical site for their effects on the lipid profiles recovered from healthy epidermal skin biopsy specimens. It was found that sex-related differences were seen at the level of the total ceramide concentration. Observed decreases in lipid concentration, due to ageing, depended on the anatomical site. Therefore, these variables should be controlled in a reproducible and standardized way in order to be able to study the direct relationship between skin condition and barrier lipid composition. Only when this relation is established, results of topical treatment can be scientifically evaluated.

High prevalence of foot diseases in Europe: results of the Achilles Project.

OBJECTIVE: To provide an insight into the prevalence of foot disease in Europe, and to include an assessment of the prevalence of predisposing factors and their correlation with foot disease. DESIGN: Large population-based survey conducted in 16 European countries. SETTING: The project consisted of two parts (study I and study II), in which all patients presenting to general practitioners and dermatologists over a defined time period were invited to participate. Patients. In study I, 70,497 patients presenting to dermatologists or general practitioners were recruited, and in study II 19,588 patients presenting to dermatologists were recruited. MAIN OUTCOME MEASURE: The feet of all participants were examined for signs of foot disease. The assessors also recorded relevant details such as the age and sex of patients, and the presence of predisposing factors for foot disease. In addition, patients in study II were offered a free mycological examination of the toenails and skin on the feet. RESULTS: In study I, 57.0% of patients had at least one foot disease. In study II, 61.3% had at least one foot disease. The proportions of patients with fungal foot disease and non-fungal foot disease in study I were 34.9% and 38.4%, respectively, and in study II were 40.6% and 41.7%, respectively. Orthopedic conditions and metatarsal corns were the most frequently reported non-fungal foot diseases, and onychomycosis and tinea pedis were the most frequently observed fungal infections. CONCLUSIONS: This large-scale survey suggests that the prevalence of fungal and non-fungal foot disease is higher than previously estimated.

The treatment of basal skin carcinomas in two sisters with xeroderma pigmentosum.

Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder that causes defects in the DNA repair system. It is characterized by a marked sensitivity to sunlight, and sufferers develop serious sunburns with onset of poikilodermia in the light-exposed skin. Squamous cell carcinomas, basal cell carcinomas (BCCs) and malignant melanomas appear in childhood. Two sisters with XP presented with previously treated facial BCCs. They were treated with imiquimod 5% cream three times weekly, one for 6 weeks and the other for 10 weeks. Although both sisters temporarily discontinued treatment due to severe erythema and erosion, successful long-term clearance was observed with no recurrences in both cases.

Repair of acetone- and sodium lauryl sulphate-damaged human skin barrier function using topically applied emulsions containing barrier lipids.

BACKGROUND: It is generally acknowledged that well-formulated moisturizing skin care products can restore disturbed barrier function that can be assessed by transepidermal water loss (TEWL) measurements. When ceramides and/or other barrier lipids are incorporated, it is, however, not always clearly demonstrated which ingredients of the formulation exert the beneficial effects. OBJECTIVES: In this study the effects of topically applied ceramide-containing mixtures on the barrier repair of sodium lauryl sulphate (SLS)- and acetone-induced skin damage have been studied in human volunteers. TEWL and stratum corneum hydration measurements were carried out. The emulsions applied contained either a mixture of two types of ceramides, CerIII and CerIIIB (emulsion 1) or a complete mixture of ceramides III, IIIB and VI together with phytosphingosine, cholesterol and the free fatty acid linoleic acid (emulsion 2). RESULTS: After SLS damage, it was observed that barrier recovery was significantly accelerated by topical application (14 days, 2 x/d) of emulsion 2 compared with the results obtained with emulsion 1. Corneometrical results were not relevant due to the occurrence of scaly fissured skin, failing to provide a good skin/probe contact. Although no effect on TEWL could be observed, the improvement of skin hydration after acetone treatment and a single application of the emulsions, was significantly more positive for emulsion 2 than for emulsion 1. CONCLUSIONS: The investigative methods used in this study show that ceramides combined with other skin lipids can improve barrier repair after damage.

Efficacy of topical corticosteroids in nickel-induced contact allergy.

In this study we used the nickel contact allergy patch (CAP) test to investigate the effect of topical corticosteroids on allergic contact dermatitis (ACD). On day 1, three CAP tests were applied for 48 h on the forearms of 20 female volunteers with a known nickel ACD. CAP of the right forearm contained 5% nickel, and of the left forearm physiological saline. Clinical scoring, transepidermal water loss and skin hydration were measured on day 1 before CAP application, on day 4 (0, 2 and 6 h) after ACD and from days 5 to 8 (0 h). A topical corticosteroid and its vehicle were applied twice daily starting from day 4 on two ACD sites. Transepidermal water loss values were significantly decreased on the topical-corticosteroid-treated sites in the early phase of ACD (day 4, 6 h after the first application) while clinical efficacy showed significant improvement on days 7 and 8. The vehicle was found to improve skin hydration only on day 8. In conclusion the topical corticosteroid improved the skin barrier function in the early inflammatory phase of ACD (day 4, 6 h). The lack of improvement in transepidermal water loss in the later phase of ACD might be accounted for by the secondary effects of the corticosteroid on proliferation and differentiation of keratinocytes.

Squamometry as a screening method for the evaluation of hydrating products.

BACKGROUND/AIMS: Squamometry is a combination of sampling corneocytes by adhesive coated discs followed by colour measurements after staining the cells. In this study, the correlation between stratum corneum (SC) hydration and scaling was investigated using capacitance measurements and squamometry, respectively. METHODS: Stratum corneum hydration and assessment of barrier function by transepidermal water loss (TEWL) measurements were carried out on different sites of left and right volar forearm skin of female volunteers (n = 13; 24 +/- 3 years). D-Squame samples were taken on the same test spots. RESULTS: Visual evaluation of the coloured samples by light microscopy and the development of a four-point-scale scoring system was found to be necessary to detect and minimise overestimation of chroma C* values. Capacitance measurements revealed neither significant differences between corresponding sites on left and right forearms no between different skin areas on the same forearm. Squamometric measurements, on the contrary, did not show any symmetry between corresponding test sites on both forearms or between different sites on one forearm. No correlation could be found between squamometric measurements and SC hydration values obtained at the same test sites. No skin barrier function impairment lays at the origin of this observation since TEWL values were found to be similar at all test sites in comparison to control skin. In a randomised single blind study, hydration and TEWL showed a significant improvement of 25% and 15%, respectively, after a 14-day application period of a moisturising cream. Although visual scoring of the coloured samples of both treated and untreated test spots revealed a good correlation with chroma C* values, the quantitative results found with squamometry were very doubtful. CONCLUSION: Squamometry with visual scoring can be proposed as a screening technique for SC hydration rather than a quantitative method to appreciate skin moisturisation.

Combination therapy improves the recovery of the skin barrier function: an experimental model using a contact allergy patch test combined with TEWL measurements.

BACKGROUND: Nickel (Ni) allergic contact dermatitis (ACD) alters the skin barrier. OBJECTIVE: Our aim was to compare the efficacy of combination therapies on ACD, using a topical corticosteroid and a corneotherapy agent (barrier cream), with that of a single therapy with corticosteroids. METHODS: On day 1, 3 Ni test patches were applied on each forearm of 14 Ni-patch-test-positive females. Four contained 5% Ni and 2 physiological saline. Either topical corticosteroid or barrier cream were matched with the combination of both products on 3 of the 4 Ni ACD. The fourth was not treated. Clinical scoring, transepidermal water loss (TEWL) and stratum corneum (SC) capacitance were measured before (day 1) and after (days 4-8) ACD. RESULTS: The combination therapy showed a significant decrease in TEWL values and an increase in SC capacitance. CONCLUSION: Combining a topical corticosteroid with corneotherapy agents prevents the delay in the healing process of skin barrier disruption due to ACD.

Beneficial effects of a skin tolerance-tested moisturizing cream on the barrier function in experimentally-elicited irritant and allergic contact dermatitis.

In experimentally-induced irritant (ICD) and allergic (ACD) contact dermatitis, an oil-in-water (o/w) cream was applied to investigate its effects on a disturbed barrier function compared to untreated physiological barrier repair. Transepidermal water loss (TEWL) measurements were performed. Before the start of the experiments, the skin tolerance of the cream was examined, revealing the non-irritating characteristics of the ingredients and the absence of any contact allergic patch test reaction. In the ICD study, sodium lauryl sulfate (SLS) patches were applied to the forearms of young female volunteers. Consequently, it was observed that repeated cream application (14 days, 2x/day) significantly improved the TEWL of SLS-damaged skin, leading to a complete recovery on day 15. In the ACD study, disruption of skin barrier function was obtained by a nickel-mediated contact allergy patch (CAP) test. The cream was then applied 2x/day for 4 consecutive days. Assessment of TEWL clearly showed that recovery of the disrupted skin significantly improved after cream application in comparison to untreated barrier repair.

Claim substantiation and efficiency of hydrating body lotions and protective creams.

In the present work a practical claim substantiation study is shown by the example of 5 commercially available body lotions. Their efficacy with respect to effects on transepidermal water loss (TEWL) and stratum corneum (SC) hydration of ageing skin has been examined. Results were obtained after single and repeated application (14 days, 2 x a day). The best performing product was then selected and further tested for its potential effects on sodium lauryl sulfate (SLS)-damaged skin. This was done in a younger population and the recovery of the impaired barrier function was followed by TEWL measurements. The selected body lotion had a high efficacy, improving both the TEWL and SC hydration of ageing skin by more than 30%. When applied to SLS-damaged skin, the product was able to improve skin barrier repair in comparison with physiological barrier repair. The results of this study show that a combination of non-invasive objective measurements can be used to substantiate product claims. Claims can be made with respect to protective and preventive properties of products, but also as to effectiveness of topical skin treatment in the case of abnormal barrier function or barrier restoration.