Diagnostische Präzision der komponentenbasierten vs. der extraktbasierten In-vitro-Diagnostik von Insektengift-Allergien: Auswirkungen auf das klinische Management.

HINTERGRUND: Die Bestimmung von spezifischen IgE-Antikörpern spielt eine zentrale Bedeutung bei der Diagnostik von Bienen- und Wespengiftallergien. In den letzten Jahren wurden die komponentenbasierte Diagnostik (CRD) eingeführt, die die Bestimmung spezifischer IgE-Antikörper gegen die Allergene Api m 1, Ves v 1, Ves v 5 und Pol d 5 sowie kreuzreaktive Kohlenhydratdeterminanten (CCDs) erlaubt. Hierdurch soll vor allem bei Probanden mit Doppelsensibilisierungen die klinische Relevanz der einzelnen Sensibilisierungen besser beurteilt werden können. PROBANDEN UND METHODIK: Die spezifischen IgE-Antikörper-Bestimmungen an 143 Probanden mit Bienen- und/oder Wespengiftallergie erfolgten mit den extraktbasierten ImmunoCAP®-Allergenen i1 und i3 sowie den ImmunoCAP®-Allergenkomponenten i208-211 und o214 (Api m 1, Ves v 1, Ves v 5, Pol d 5, CCD). Bei Doppelsensibilisierten wurde zusätzlich ein Inhibitionstest durchgeführt. An einem Teilkollektiv der Studienpopulation erfolgten sIgE-Bestimmungen gegen Api m 1, Api m 4, Pol d 5 und Ves v 5 mittels Allergiechip (ISAC®, n  =  44). ERGEBNISSE: Die Sensitivität von Ves v 5 bei isolierten Wespengiftallergikern betrug 78,5 %, gemeinsam mit Ves v 1 stieg diese auf 92,3 %. Die Sensitivität von Api m 1 bei isolierten Bienengiftallergikern betrug 25 %. Die komponentenbasierte Diagnostik und Inhibitionstests bei Doppelsensibilisierten lieferten divergente Ergebnisse. Die CRD mittels ISAC®-Allergiechip erbrachte deutliche Unterschiede vor allem im Hinblick auf die Diagnostik von Api m 1 und CCDs. SCHLUSSFOLGERUNGEN: Die CRD bereichert das diagnostische Spektrum, sofern sie nicht allein sondern zusätzlich zu den etablierten Verfahren eingesetzt wird. Sie sollte neben Ves v 5 stets die Bestimmung der IgE-Antikörper gegen Ves v 1 umfassen.

Diagnostic precision of component-resolved vs. extract-based in vitro diagnosis of hymenoptera venom allergy: effects on clinical management.

BACKGROUND: The measurement of specific IgE (sIgE) antibodies plays a key role in the diagnosis of honeybee and wasp venom allergy. In recent years, component-resolved diagnosis (CRD) has been introduced, which allows for the measurement of sIgE antibodies against Api m 1, Ves v 1, Ves v 5, and Pol d 5, as well as cross-reactive carbohydrate determinants (CCDs). These tests are intended to help determine the clinical relevance of any given sensitization, especially in patients with dual sensitization. PATIENTS AND METHODS: Specific IgE antibody levels were measured in 143 patients with bee and/or wasp venom allergy using the extract-based ImmunoCAP® allergens i1 and i3 as well as the ImmunoCAP® allergen components i208-211 and O214 (Api m 1, Ves v 1, Ves v 5, Pol d 5, CCDs). In patients with dual sensitization, inhibition testing was also performed. In a subgroup of the study population, sIgE to Api m 1, Api m 4, Pol d 5, and Ves v 5 were determined using the ISAC® allergy microarray (n = 44). RESULTS: The sensitivity of Ves v 5 in patients with isolated wasp venom allergy was 78.5 %; in combination with Ves v 1, that figure increased to 92.3 %. The sensitivity of Api m 1 in individuals with isolated bee venom allergy was 25 %. CRD and inhibition testing in individuals with dual sensitization showed divergent results. CRD using the ISAC® allergy microarray showed marked differences, especially with regard to Api m 1 and CCDs. CONCLUSION: Component-resolved tests are a valuable addition to the diagnostic spectrum as long as they are used in combination with established procedures. Apart from Ves v 5, measuring IgE antibodies to Ves v 1 should always be included in the diagnostic workup.

Diagnostic value of the ISAC(®) allergy chip in detecting latex sensitizations.

PURPOSE: Latex allergy can be diagnosed by different test methods such as IgE quantification, Western blot, cellular antigen stimulation test (CAST), and in vivo methods [e.g. skin prick test (SPT)]. Phadia provides two modern methods using recombinant latex allergens: ImmunoCAP(®) and the Immuno Solid-phase Allergen Chip (ISAC(®)), which enables simultaneous determination of specific IgE against five latex allergens. We compared the diagnostic sensitivity of the ISAC(®) test kit and the conventional Hev b 5-spiked ImmunoCAP(®) latex extract. METHODS: Forty sera were sampled from subjects with suspected natural rubber latex (NRL) allergy. These patients had positive SPT to NRL extract, positive NRL Western blots, and positive results in the CAST. All sera were analysed using the ISAC(®) and recombinant NRL ImmunoCAP(®) allergens and compared to the results of 20 negative control sera. RESULTS: Only 22 of the 40 subjects (55 %) showed positivity to at least one latex allergen on the ISAC(®) (sensitivity ISAC(®) 55 %). The sensitivity of the ImmunoCAP(®) latex extract was 70 %. The most frequently detected sensitization was against Hev b 6.01 (n = 12). When the serum samples were tested with all recombinant ImmunoCAP(®) allergens, three additional sensitizations against latex could be detected compared to the ISAC(®). CONCLUSIONS: Microarrays do offer many potential benefits such as elegant simultaneous determination of sensitizations against different NRL allergens with minimal amounts of serum. However, a negative NRL test result should be regarded with caution and at least be confirmed by other in vitro methods.

Development of a standardized testing procedure for assessing the irritation potential of occupational skin cleansers.

BACKGROUND: Frequent skin cleaning fulfils the definition of occupational 'wet work'. Standardized methods are required to assess the irritation potential of workplace cleansers. OBJECTIVES: To develop a standardized procedure for testing the irritation potential of occupational skin cleansers. METHODS: In this single-blind, single-centre trial in 25 healthy volunteers, the irritation potential of five generic reference cleansers was tested by three-times-daily washing with an automated skin cleaning device for 4 days, and quantification of cumulative skin barrier damage was performed by visual scoring, chromametry, transepidermal water loss TEWL, and corneometry. For two cleansers, reproducibility of the irritancy assessment was assessed. Furthermore, the irritation induced by four commercial workplace skin cleansers was studied. RESULTS: Whereas no significant changes were observed for any of the tested cleansers by either visual scoring or chromametry, significant increases in TEWL and significant decreases in stratum corneum hydration were found for all cleansers. Cleansers differed significantly in their irritation potential. On retesting of two cleansers, the first results were confirmed. Among the four commercial cleansers, one that was claimed to be mild was found to be disproportionally irritant. CONCLUSIONS: The presented model for testing cleansing preparations allows a highly controlled, practically relevant and reproducible irritancy assessment of occupational skin cleansers.

Development of a standardized procedure for testing the efficacy of workplace cleansers.

BACKGROUND: Occupational skin cleansing is a risk factor for cumulative irritant dermatitis. Standardized methods are required to assess the cleaning efficacy of occupational skin cleansers, allowing us to choose the optimal cleanser with minimal irritation for specific workplaces. OBJECTIVES: To develop a standardized procedure for testing the cleaning efficacy of occupational skin cleansers. METHODS: In this single-blind, randomized, monocentric clinical trial in 24 healthy volunteers, the cleaning efficacy of generic reference cleansers was assessed against standardized test dirts. The test procedure included standardized dirt application, standardized washing with an automated skin cleaning device, and quantification of the cleaning efficacy by chromametry. For two cleansers, reproducibility of the cleaning efficacy assessment was studied. Furthermore, the performance of two cleansers was compared with that of commercial skin cleansers. RESULTS: Cleaning factors ranged from 12% up to 97% for the respective model dirts. A high reproducibility of the washing and the assessment procedure could be shown. The comparison of the washing efficacies for two reference cleansers with those of commercial cleansers showed partly similar performances, but also a higher cleaning efficacy for some dirts of the reference cleansers. CONCLUSIONS: The testing procedure proved to be applicable to a wide spectrum of cleansers and model dirts.

Trichophyton species of Arthroderma benhamiae - a new infectious agent in dermatology.

In Germany, infections due to the zoophilic dermatophyte Trichophyton (T.) species of Arthroderma benhamiae are being more frequently diagnosed. The source of infection of this emerging pathogen overlaps with that of the zoophilic species T. interdigitale. The most common source are guinea pigs. T. species of Arthroderma benhamiae causes inflammatory dermatophytosis in children and adolescents. In addition to tinea capitis, it may cause both tinea corporis, tinea manus and frequently tinea faciei. In Germany, T. species of Arthroderma benhamiae is a frequent zoophilic dermatophyte, which in regions is probably more frequent than Microsporum canis. The mycological identification of the isolates with their yellow stained colonies is based on their macroscopic and microscopic features. However, some exhibit colony features consistent with those of T. interdigitale. These strains only can be identified unambiguously by means of molecular techniques. Using detection methods such as PCR-ELISA or real-time PCR, the dermatophyte can be identified directly from clinical material. Sequencing of the internal transcribed spacer region (ITS) of the ribosomal DNA has been approved as culture confirmation test for T. species of Arthroderma benhamiae. In addition, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) is useful. Widespread dermatophytosis due to T. species of Arthroderma benhamiae, in particular of tinea capitis, requires oral antifungal agents. Terbinafine is most effective, alternatives are fluconazole and itraconazole.

Standardized dirts for testing the efficacy of workplace cleaning products: validation of their workplace relevance.

BACKGROUND: In order to assess the cleaning efficacy of occupational skin cleansers, standardized test dirts mimicking the spectrum of skin soiling at dirty workplaces are necessary. OBJECTIVES: To validate newly developed standardized test dirts (compliant with the EU Cosmetics Directive) for their occupational relevance. METHODS: In this single-blinded, monocentric questionnaire-based clinical trial, 87 apprentices of three trades (household management; house painting and varnishing; and metal processing) evaluated the cleanability of six standardized test dirts in relation to their workplace dirts. In addition, they judged the similarity of the test dirts to actual dirts encountered in their working environments. RESULTS: Most of the household management participants assessed the hydrophilic model dirt ('mascara'), the lipophilic model dirt ('W/O cream') and a film-forming model dirt ('disperse paint') as best resembling the dirts found at their workplaces. Most of the painters and varnishers judged the filmogenic model dirts ('disperse paint' and 'acrylic paint') as best resembling the dirts found at their workplaces. For the metal workers, the lipophilic and paste-like model dirts were most similar to their workplace dirts. CONCLUSIONS: The spectrum of standardized test dirts developed represents well the dirts encountered at various workplaces. The test dirts may be useful in the development and in vivo efficacy testing of occupational skin cleansers.

Impact of age and heterophilic interference on the basal serum tryptase, a risk indication for anaphylaxis, in 1,092 dermatology patients.

A raised baseline serum tryptase is a risk indicator for anaphylactic reactions, especially in patients with hymenoptera venom allergy. Borderline elevations (> 11.4 µg/l) occur frequently and may necessitate invasive diagnostic procedures to rule out systemic mastocytosis. We retrospectively analysed 1,092 non-mastocytotic patients from our general dermatology clinic with respect to age- and gender-associated effects and investigated the impact of heterophilic antibody interference on the tryptase assay. The results were stratified by gender and five age classes. Sera with raised tryptase (n = 106) were re-tested after pre-incubation with Heterophilic Blocking Tubes (HBT(®), Scantibodies Laboratory; Santee, CA, USA). A significant increase in baseline tryptase was observed with increasing age. Incubation with HBT(®) caused a decline of more than 50% in only one case. In conclusion, older patients showed significantly higher serum tryptase levels and heterophilic interference was of subordinate relevance.

Identification of yeast isolated from dermatological patients by MALDI-TOF mass spectrometry.

Species identification of yeasts is based on biochemical (e.g. API ID 32 C®, bioMérieux) and molecular biological approaches. As an alternative to DNA-dependent methods, mass spectral analysis based identification of micro-organisms has become increasingly recognized. In a number of studies, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been applied for the rapid classification and identification of micro-organisms. In this study, the applicability of MALDI-TOF MS for identifying yeasts isolated from dermatological patients was analysed and compared with the results from the API ID 32 C® system. Furthermore, sequencing the internal transcribed spacer (ITS) regions of the ribosomal DNA was employed as reference method. Candida (C.) albicans was isolated in 41.9% of all cases, C. parapsilosis in 20.3%, C. glabrata in 10.8%, and C. krusei in 6, 8.1%. Rarely isolated yeasts were Candida colliculosa, famata, guilliermondii, lusitaniae, and tropicalis as well as Geotrichum candidum, Rhodotorula mucilaginosa and Trichosporon mucoides. The MALDI TOF results were equal to the results gained by ITS sequence analysis in 94%, whereas API ID 32 C® provided the correct diagnosis in 84.3% (of all cases). This lower identification rate is mostly referable to frequent misidentifications of C. krusei as C. inconspicua/norvegensis,Candida tropicalis, or Geotrichum capitatum. In contrast, all C. krusei strains were correctly identified by MALDI TOF MS. In conclusion, species identification by MALDI-TOF MS was proven to be consistent with ITS sequence analysis; the technique has a resolving power comparatively as high as ITS sequence analysis.

Safety, effectiveness and comparability of professional skin cleansers.

BACKGROUND: There are no widely-accepted methodical specifications with which to objectify cleansing effectiveness and skin compatibility of occuptional skin cleansing products in Europe. Therefore the German Social Insurance Agency (DGUV) initiated a study with the goal to evaluate such products in view of the potency and the safety of hand cleansers. A market analysis was a part of the project. METHODOLOGY: The product descriptions and safety data sheets of 120 products (5-20/manufacturer) of 11 manufacturers were evaluated between 02/2008 and 04/2008. RESULTS: The manufacturers used mainly ingredients of low irritancy. The declaration of the applied ingredients was in the majority of the cases correctly labeled according to the INCI Declaration. Although there was documentation of skin tolerability for most products, the manufacturers used widely differing tests of skin compatibility. Evidences for cleansing effectiveness were not declared or have not been provided. One manufacturer even promised medical effects of its products. There was no uniform general classification of products making it difficult to identify characteristics of cleansers and choose between them. CONCLUSIONS: Presently, there are no commonly accepted criteria to classify products in view of cleansing effectiveness und skin compatibility. Generally accepted criteria and test methods are needed for the evaluation of hand cleansers in order to provide the possibility of transparency and comparability.

Chronotype is associated with psychological well-being depending on the composition of the study sample.

Past studies examining the effect of chronotype and social jetlag on psychological well-being have been inconsistent so far. Here, we recruited participants from the general population and enquired about their natural sleeping behavior, sleep quality, depressive symptoms, and perceived stress. Partial correlations were computed between sleep variables and indicators of psychological well-being, controlling for age and sex. Less sleep during work days was found a good indicator for impairments in psychological well-being. In exploratory follow-up analyses, the same correlations were calculated within groups of early, intermediate, and late chronotype. We observed that the composition of the sample in terms of chronotype influenced whether associations between sleep variables and psychological well-being could be observed, a finding that is advised to be taken into account in future studies.

Sensitization to para-amino compounds in swim fins in a 10-year-old boy.

A ten-year-old boy presented with recurrent eczema on the dorsal of both feet and the thighs. His symptoms became worse when he used racing swim fins. Patch testing included the standard, ointments, preservatives, leather, textile dyes, rubber component, and corticosteroid series of the German Contact Dermatitis Research Group (DKG) as well as the patient's own materials (fin material). At 72 hours positive reactions were observed to blue flipper material, Acid Yellow 36, Bismarck Brown R, and DPPD. DPPD is widely used as an antioxidant in the rubber industries. Therefore, the dermatitis was most probably attributable to DPPD in the blue flipper material, although we were not able to prove this due to lack of cooperation of the manufacturer. Sensitization to Acid Yellow 36 and Bismarck Brown R are interpreted as cross sensitizations, since all substances are para-amino compounds.

The use of ITS DNA sequence analysis and MALDI-TOF mass spectrometry in diagnosing an infection with Fusarium proliferatum.

Although mycoses are among the most common diseases worldwide, infections with Fusarium spp. occur only rarely. Mostly patients suffering from underlying immune deficiency are infected with this mould, resulting in a considerably decreasing prognosis. In immunocompromised patients, cutaneous manifestations are more often associated with Fusarium sp. than with Candida sp. or Aspergillus sp. We describe one patient with acute lymphoblastic leukaemia, who was first treated with chemotherapy after GMALL protocol 07/03. After relapse, the patient was successfully transplanted in second remission with a human leukocyte antigen (HLA)-matched unrelated peripheral blood stem cell graft. Ten months later, the patient died from respiratory insufficiency and recurrence of leukaemia. Previously, Aspergillus antigen was detected in blood. In the latter course, disseminated papules appeared. One of these was examined histologically and mycologically. Conventional cultural diagnostics led to the diagnosis of a fusariosis, further supported by internal transcribed spacer (ITS) sequencing and matrix assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF) mass spectrometry, both determining the isolated strain as Fusarium proliferatum, which is a very infrequent pathogen within this genus. Our investigations underline the potential of MALDI-TOF MS based identification of Fusarium species as an innovative, time and cost efficient alternative to ITS sequencing.

Teaching interventions in contact dermatitis.

Prevention of occupational contact dermatitis is of utmost significance for both insurers and legislators because it can preserve the individual's ability to work and result in decreased costs for public health. In the last 20 years, many concepts for educational interventions and a multitude of teaching aids have been developed by insurance associations and public institutions from different countries (eg, the United States, Sweden, Denmark, Switzerland, and Germany). For didactic purposes, terms of different levels of prevention have been inaugurated in northern Europe in the context of occupational dermatology. This review presents different educational interventions in the fields of primary, secondary, and tertiary prevention as well as evaluation studies of these measures, especially among health care workers, hairdressers, metalworkers, and bakers' apprentices. Special emphasis is put on the prevention of allergic contact dermatitis.

Dry skin, barrier function, and irritant contact dermatitis in the elderly.

Dry skin is characterized by a decreased lipid content and a delayed reconstitution of the epidermal barrier after skin irritation. These are problems of high relevance in the aged population, especially in the development of irritant contact dermatitis. Asteatotic and perineal irritant dermatitis are the most important subtypes of irritant contact dermatitis in the elderly. This contribution presents a compressed survey on these subtypes and elucidates their relation to an impaired barrier function. Typical irritants affecting aged individuals are explained and compared with irritants that seem to be more significant in younger people. Results of biophysical investigations, such as measurement of transepidermal water loss, are discussed regarding their age-dependence. Transepidermal water loss decreases with age, which was formerly interpreted as an indication of a decreased sensitivity. Today, we know that reconstitution of the epidermal barrier after irritation is delayed once it has been impaired. Reasons are decreased activities of enzymes involved in lipid synthesis and processing, a changed cytokine profile, a reduced acidification of aged skin, and alterations in the function of epidermal stem cells. Owing to these new insights, a reevaluation of the sensitivity of aged skin has to be initiated, especially with regard to occupational dermatology.

Atypical manifestations of tinea corporis.

Tinea corporis classically presents as an erythematous annular plaque with a scaly, centrifugally advancing border. However, sometimes vesicles and pustules are observed. Occasionally, even frank bullae appear secondary to severe inflammation. Diagnostic difficulties arise when atypical manifestations mimic other inflammatory skin diseases, including atopic or seborrheic dermatitis, subacute cutaneous lupus erythematosus, or vesicular diseases. We report five cases of atypical tinea corporis, where the initial clinical diagnosis was different from dermatophytosis. The differential diagnoses and the diagnostic difficulties related to atypical manifestations of fungal infections are discussed. Moreover, our cases emphasise the importance of conventional histological examination, which enables a fast, correct diagnosis.

Widespread tinea corporis caused by Trichophyton rubrum with non-typical cultural characteristics--diagnosis via PCR.

From the epidemiological point of view, Trichophyton rubrum is the most important dermatophyte in the Western world. Mostly, the lesions of fungal infections of the skin are restricted and circumscribed. Immunodeficiency, diabetes or treatment with steroids, however, favour widespread disease involving the entire integument. In the present study, we describe a patient without immunodeficiency or diabetes presenting with widespread tinea corporis caused by T. rubrum. The fungal isolate showed atypical morphological features. Moreover, the typical metabolic features (negative urease test, production of red pigment on potato dextrose agar) were expressed with delay after 6 weeks of cultivation. Thus, sequencing of the internal transcribed spacer of the ribosomal DNA was applied to identify the fungal isolate and led us to the correct diagnosis, before conventional mycological methods were successful.