Mast cells play a key role in neutrophil recruitment in experimental bullous pemphigoid.

Bullous pemphigoid (BP) is an inflammatory subepidermal blistering disease associated with an IgG autoimmune response to the hemidesmosomal protein BP180. Passive transfer of antibodies to the murine BP180 (mBP180) ectodomain triggers a blistering skin disease in mice that depends on complement activation and neutrophil infiltration and closely mimics human BP. In the present study, we show that mast cells (MCs) play a crucial role in experimental BP. Wild-type mice injected intradermally with pathogenic anti-mBP180 IgG exhibited extensive MC degranulation in skin, which preceded neutrophil infiltration and subsequent subepidermal blistering. In contrast, mice genetically deficient in MCs or MC-sufficient mice pretreated with an inhibitor of MC degranulation failed to develop BP. Further, MC-deficient mice reconstituted in skin with MCs became susceptible to experimental BP. Despite the activation of complement to yield C3a and C5a, in the absence of MCs, accumulation of neutrophils at the injection site was blunted. The lack of response due to MC deficiency was overcome by intradermal administration of a neutrophil chemoattractant, IL-8, or by reconstitution of the injection sites with neutrophils. These findings provide the first direct evidence to our knowledge that MCs play an essential role in neutrophil recruitment during subepidermal blister formation in experimental BP.

Multiparametric image cytometry in mycosis fungoides.

Eighteen cases of early mycosis fungoides were compared with 18 cases of eczematous dermatitis by multiparametric image cytometry. A minimum of 100 lymphocytes was measured in each case. A large number of measurements was acquired for each lymphocyte, characterizing nuclear DNA content, area, shape, and texture. There were significant differences between the two groups, especially in nuclear DNA content and texture. These differences allowed the two groups of nuclei to be distinguished with 78% accuracy. The two groups of lesions were distinguished with 94% accuracy, using neural network analysis.

Image analysis cytometry of dysplastic nevi.

Computerized image analysis was used to assess nuclear atypia in 24 dysplastic nevi (DN), 19 CN (CN), and five thin melanomas. DN were selected for the study using architectural criteria alone. Feulgen-stained, 6-um sections were analyzed with a microTICAS cytometer. At least 100 nuclei were measured in each case. The standard deviation of nuclear area, mean nuclear roundness, standard deviation of nuclear roundness, mean ploidy, and standard deviation of ploidy were found to be significantly greater for DN than for CN. DNA histograms from DN showed an increased fraction above 2N, suggesting that DN are more proliferative than CN. No DN were aneuploid. All melanomas were aneuploid, and differed significantly from DN in mean nuclear area, standard deviation of nuclear area, mean ploidy, and standard deviation of ploidy. There were no significant differences between the junctional and intradermal populations of compound DN in any of the measured parameters, except that the intradermal nuclei were significantly rounder than the junctional nuclei. There were no significant differences between DN from patients with only a single DN and DN from patients with at least two dysplastic nevi.

Cutaneous insulin reaction resembling acanthosis nigricans.

We describe a patient who developed hyperkeratotic, verrucous plaques at sites of repeated insulin injections. One similar case has previously been reported. This phenomenon may be an unusual manifestation of the trophic actions of insulin.

Acantholytic epidermolysis bullosa.

BACKGROUND: We describe a new variant of inherited epidermolysis bullosa and elucidate the clinical, histologic, and ultrastructural features of this condition. OBSERVATIONS: This form of epidermolysis bullosa displays an autosomal dominant inheritance pattern, is characterized by acral bullae, and histologically demonstrates suprabasal clefting with acantholysis. Ultrastructural findings are nonspecific but reminiscent of those observed in benign familial pemphigus. CONCLUSION: Acantholytic epidermolysis bullosa is a rare but distinct clinicopathologic entity that warrants inclusion in the nosologic classification of epidermolysis bullosa.

Mucous gland basement membrane immunofluorescence in cicatricial pemphigoid.

Three patients are described with cicatricial pemphigoid and positive immunofluorescence findings in the basement membrane zone of mucous glands of the pharynx, mouth, and nose. These findings appear to be unique to cicatricial pemphigoid.

Digital dermoscopy.

Digital dermatoscopic images are acquired with digital cameras or video camera and frame grabber combinations. These images can be compressed, transmitted, or archived; combined with clinical anamnestic information for medical record purposes; or attached to body surface diagrams for mole mapping applications. Image analysis software, which might interpret the images to produce a computer-assisted or fully automated diagnosis, is under development.

Techniques for a structural analysis of dermatoscopic imagery.

Techniques were developed for automated detection and characterization of dermatoscopic structures, including the pigment network and brown globules. These techniques incorporate algorithms for grayscale shape extraction based on differential geometry developed by Steger, a snake algorithm, and a modification of the region competition strategy of Zhu and Yuille. A novel approach was developed for global segmentation of pigmented lesions, based on stabilized inverse diffusion equations. Procedures for detection of air bubbles and hairs in dermatoscopic images are also reported.

Design of a high resolution image cytometer with open software architecture.

A relatively inexpensive, non-proprietary high resolution color pathology workstation is described. Hardware consists of a JVC frame capture camera with adjustable resolution up to 4416 x 3456, matching frame grabber and a Unix workstation. Analytic software was developed using Khoros 1.0.5, a powerful and flexible system for the development of image analysis applications that is based on a visual programming language. Applications have been developed for DNA ploidy, quantitative immunohistochemistry and texture and shape analysis. The instrument's software is uniquely extensible and transparent, and has been made publicly available over the Internet.

Image analysis in dermatopathology.

BACKGROUND/AIMS: Image analysis in dermatopathology has been used for DNA ploidy analysis, morphometry, stereology, and quantitative immunohistochemistry. The object is to review image analysis in dermatopathology and evaluate these modalities and their application in pigmented lesion pathology, for elucidation of tumor behaviour and architecture and as an aid in tumor identification and prognostication. CONCLUSION: Image analysis in dermapathology has a huge potential. The techniques are difficult and at present mainly used in specialized centres.

A simple immunochemical technique for distinguishing melanocytes and melanophages in paraffin-embedded tissue.

We describe an immunoperoxidase technique for distinguishing melanocytes and melanophages in paraffin-embedded tissue. Two primary antibodies were used: anti-S100 and HAM-56, a monoclonal antibody directed against macrophages. In a variety of pigmented lesions, HAM-56 labelled melanophages while S100 labelled cells of melanocytic lineage.

Multiparametric image cytometry of nevi and melanomas.

Multiparametric image cytometry was applied to 10 examples each of malignant melanoma and common, Spitz, and dysplastic nevus. DNA index, area, and 21 parameters describing chromatin texture were measured for 50 nuclei in each lesion. Linear discriminant analysis was used to derive discriminant functions based on the measured parameters. The analysis demonstrated that chromatin texture provides more diagnostic information than either DNA index or nuclear area. The discriminant functions allowed 68% of nuclei to be accurately classified among the four groups, and allowed 37 of the 40 lesions to be accurately classified as nevus or melanoma.

A new technique for laser treatment of cutaneous tumors.

The CO2 laser was used to treat three patients with discrete, cutaneous tumors. For each patient, we established the relationship between the width of the lesions and their maximum thickness, and between pulse energy and the depth of the resultant crater. Optimal parameters for therapy were established from these results.

Malignant clear-cell hidradenoma of the toe.

Malignant sweat gland tumors are rare tumors of the extremity. Their insidious growth patterns and often confusing pathological characteristics can cause confusion with more common benign tumors. However, these tumors cannot be neglected because they do have a propensity to metastasize. Presented is a 56-year-old woman with a malignant clear-cell hidradenoma of the foot actually presenting as a benign lesion.

Photopolymerization of composite resin using the argon laser.

Because of the dental profession's increased utilization of light-cured restorative materials, there has been a corresponding increase in research into the light sources used to initiate polymerization. The argon laser is one promising source, as the wavelength of light emitted by this laser is optimal for the initiation of polymerization of composite resins. The literature reflects a strong divergence of opinion about many aspects of the effectiveness of laser curing compared to conventional light curing. Research indicates that the argon laser offers a greater depth and degree of polymerization, less time required and an enhancement of the physical properties of composite resins polymerized. These advantages are offset by reports that the increased polymerization caused by the laser results in increased shrinkage, brittleness and marginal leakage. Dentists interested in the new technology need to monitor ongoing studies.

Immunohistochemical localization of cytokines in nevi.

Eight common nevi and 11 dysplastic nevi were evaluated for the presence of basic fibroblast growth factor, platelet-derived growth factor, transforming growth factor-alpha, interleukin-1-alpha, and interleukin-1-beta by immunohistochemical labelling with highly specific monoclonal antibodies. Basic fibroblast growth factor was abundant in the nevus cells and keratinocytes of nevi. Dysplastic nevus cells on average stained less intensely for basic fibroblast growth factor than did common nevus cells. In both types of nevi, basic fibroblast growth factor was identified in the basement membranes at the dermoepidermal junction and surrounding nevus cell nests and individual nevus cells. Labelling of nevus cells for transforming growth factor-alpha was variable, while there was moderate labelling for platelet-derived growth factor and light labelling for interleukin-1-alpha. Only two nevi, both dysplastic, stained (very faintly) for interleukin-1-beta. It is possible that these cytokines, especially basic fibroblast growth factor, act in autocrine fashion to maintain nevocellular growth and may also contribute to the epidermal hyperplasia and fibrosis frequently observed in nevi.

Caseating cutaneous granulomas in a patient with X-linked infantile hypogammaglobulinemia.

A 34-year-old man with X-linked infantile hypogammaglobulinemia, bronchiectasis, and chronic liver disease had a papular eruption on the trunk and upper extremities. A biopsy specimen revealed caseating granulomas, but special stains, cultures, and electron microscopy failed to reveal an infectious organism. Immunohistochemistry showed that the lymphocytes within the granulomas were almost exclusively of the CD8+ cytotoxic/suppressor T phenotype. Phenotypic analysis of the circulating lymphocytes showed normal numbers of CD4+ (helper/inducer) and CD8+ T cells, whereas B cells were undetectable. Other examples of noninfectious granulomatous disease have been reported in patients with primary hypogammaglobulinemia, but this is the first case of caseating cutaneous granulomatous disease to be reported in a patient with X-linked infantile hypogammaglobulinemia.

Seasonal variation in the proliferation fraction of Australian common nevi.

Image cytometry was used to assess seasonal variation in the proliferation fraction of Australian common nevi. Twenty pairs of nevi were evaluated. One member of each pair had been excised during the Australian summer, and the other member of the pair had been excised during the winter. The nevi were matched for age, sex, and body site. From each nevus, 6-microns sections were Feulgen-stained and evaluated with a CAS 200 image analyzer, which was used to obtain DNA histograms. Proliferation fractions were calculated from the histograms. The proliferation fraction of the nevi removed during the winter was 1.65 +/- 0.32%, whereas the proliferation fraction of the nevi removed during the summer was 1.95 +/- 0.42% (p = 0.41). For nevi from sun-exposed sites only, the proliferation rate of nevi excised during the winter was 1.81 +/- 0.39%, and 2.58 +/- 0.39% for nevi excised during the summer (p = 0.11). For nevi from sun-protected sites, there was no difference between winter and summer. When nevi excised during the summer were compared by site, sun-exposed nevi had a proliferation rate of 2.60 +/- 0.48% (p = 0.04). For nevi excised during the winter, there was a much smaller difference between sun-exposed and sun-protected nevi.

Image cytometry in early graft-versus-host disease.

Image cytometry was used to measure the size, shape, and texture of lymphocyte nuclei in 24 skin biopsy specimens retrospectively selected from patients with early, nonspecific, graft-versus-host disease (GVHD)-like eruptions. Half of these patients had progressed to overt GVHD, whereas half had never developed more than a mild nonspecific eruption. Significant differences were detected in the nuclear texture of the eruptions of the two groups, but the differences were not consistent enough to suggest that image cytometry might play a significant role in recognition of early GVHD.