Sjögren-Larsson syndrome is an inborn error of metabolism with its primary clinical manifestations being congenital ichthyosis, spastic hemiplegia or quadriplegia, and moderate to severe mental retardation. Definitive diagnosis can be made by measuring fatty aldehyde dehydrogenase activity in cultured skin fibroblasts. Treatment of dermatologic manifestations is best accomplished with topical emollients and frequent bathing or showering. Some evidence supports medium-chain triglyceride diets and oral retinoids. These modalities, however, remain controversial.

Peanuts. along with milk and egss. have been documented to account for approximately 80% of adverse reactions to foods in patients with atopic dermatitis. Over the past 3 years, we have evaluated 71 patients with atopic dermatitis, ranging from mild to severe in nature. These patients were initially evaluated by allergy prick skin testing and when appropriate had double-blind placebo-controlled food challenges done.Thirty-nine (55%) patients had a positive prick skin test to one of the foods tested. There were 80 food challenges performed with peanut, accountingfor 12 (32%) of the 38 positive challenges in 23 (31 %) patients. As in earlier studies, patients developed skin (97%), respiratory (55%), and gastrointestinal (32%) symptoms during the challenge. Of the five patients with histories of prior anaphylactic reactions four (80%) were to peanut. These studies indicate that children with all degrees of atopic dermatitis may benefit from evaluation for food hypersensitivity. They also show that peanut is a major food protein responsible for these reactions. P eanuts, milk, and eggs have been documented to account for approximately 80% of adverse reactions to foods in patients with atopic dermatitis. I Peanuts are a commonly eaten food and in many countries contribute significantly to the total dietary protein. 2 In the

An existing technique (Vindelov et al. Cytometry 3:323, 1983) has been modified for DNA flow cytometry of human epidermis obtained from 2 to 3 mm punch biopsies. By varying the length of time of digestion of the epidermal disc by trypsin from 5 to 70 Key terms: Trypsin digestion, biopsy, keratimin a controlled release of keratinocytes occurred beginning with the stratum basale and proceeding toward, but not including, the superficial layer of the epidermis, the stratum corneum. nocyte, isolated nucleiAn important prerequisite for performing flow cytometry (FCM) is the acquisition of a single cell or nuclear suspension. For epidermis, the production of single cell suspensions permits the study of a variety of cell types such as Langerhans cells, keratinocytes, keratinocyte subpopulations, and basal cells (2-11). Because of our interest in the possibility of using cell cycle kinetic data obtained from human epidermis as indicators of the best circadian time for maximal host tolerance to chemotherapy, i.e., chronochemotherapy (11, we modified a n existing technique (12) for use in DNA FCM of human epidermis.Dispersion of epidermal cells or their nuclei into suspension has been accomplished by several different methods including trypsin, dithiotheritol (DTT), EDTA, ultrasonication, or ultrasonication plus DTT (2). The trypsin detergent method described by Vindelov et al. (12) has the advantages of (a) frozen storage of the specimen, (b) production of clean nuclei after digestion of cytoplasmic proteins with trypsin, and (c) quantitative staining of DNA with propidium iodide.This report describes a modification of an existing technique (12) for the production of isolated nuclei of keratinocytes obtained from 2-3 mm punch biopsies of adult human epidermis. Based on the duration of the incubation time in trypsin, a controlled release of keratinocyte populations can be achieved. Trypsin digestion dissolved epidermal cell cohesion beginning in the basal cell layer with progression toward the superficial-most layer after longer incubation. MATERIALS AND METHODS BiopsiesThe skin biopsies used in this study were obtained from the scalp during hair transplantation surgery. Epidermal-Dermal Separation and Freezing of Isolated EpidermisTo facilitate the removal of epidermis from dermis, skin biopsies were first individually placed in 0.5% cold acetic acid for 48 h. With the aid of a dissecting microscope, the epidermis was separated from the dermis using watchmaker's forceps. The disc of epidermis was then placed in fresh DMSO solution and frozen rapidly to -70" C. Intact biopsies as well as the peeled dermis and epidermis were stained with hematoxylin and eosin (H & E) and examined by routine light microscopy. Trypsinization and StainingThe trypsin-detergent method of Vindelov (12) was used with the modification that digestion times in trypsin were varied from 5 to 70 min. In addition to varying the digestion time, other variables which were studied included mechanical vortexing at 1 min, 3 min and 5 min intervals during th...

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