Skin biopsies

Hematology and clinical chemistry should be considered in more severe exanthema, as systemic involvement has been described [1]. A skin biopsy may sometimes be needed for differential diagnosis. 

Kadotanl, T. Sato, H. Ohama, K. and Takahara, H. "A Technical Note on the Antenatal Chromosome Analysis by Trans-abdomlnal Fetal Skin Biopsy". Jap. Jour. Htiman Genet., (1971), 16, 42-47. 

Epidermal nerve fiber analysis by immunocytochemical techniques using the panaxonal marker protein gene product 9.5 (PGP 9.5) allows the study of epidermal innervation by small fiber C and A5 nerve fibers (McCarthy et al. 1995 Holland et al. 1997). Studies of skin biopsies of HIV infected patients with DSP or ATN showed reduction in the number of epidermal fibers in distal areas of the lower extremities with an inverse correlation between neuropathic pain intensity and epidermal nerve fiber density (Polydefkis et al. 2002) (Fig. 4.3). There were also fewer epidermal fibers in HIV seropositive patients without clinical evidence of neuropathy, suggesting that HIV infection may be associated with the loss of cutaneous innervation even before the onset of sensory symptomatology (McCarthy et al. 1995). 

McCarthy BG, Hsieh ST et al (1995) Cutaneous innervation in sensory neuropathies evaluation by skin biopsy. Neurology 45(10) 1848-1855... 

Eluorescence of both collagen (Monnier etal., 1986) and IgG (Jones etal., 1988) are associated with diabetic micrOangiopathy. Monnier etal. (1986) found that measured collagen-linked fluorescence in skin biopsies from both patients with type 1 diabetes ( = 41, age range 29-52 years) and controls ( =25, age range 28-41 years) was significantly correlated with the severity of retinopathy as well as arterial and joint stifihess. Jones et al. (1988) found increased fluorescence of serum IgG in diabetic patients with... 

Lampkin SR, Allred DC. Preparation of paraffin blocks and sections containing multiple tissue samples using a skin biopsy punch. I. Histotechnol. 1990 13 121-123. 

Skin biopsy of lesional skin is useful in confirming the diagnosis. 

Annex 12 Skin biopsy on fatal cases for diagnosis of Ebola... 

Th-17 cells appear to be involved in protection against bacterial pathogens. In addition, Th-17 cells may also be crucial in the pathogenesis of various chronic inflammatory diseases that were formerly categorized as Th-1-mediated disorders. Whereas IL-17 may play an important role in the pathogenesis of psoriasis and contact hypersensitivity, its role in atopic dermatitis is still unclear [36]. In skin biopsy specimens recovered from acute and chronic skin lesions from patients with atopic dermatitis, IL-17 was preferentially associated with acute lesions [37]. 

In addition to the pivotal studies, several publications (Table 6.2) used other methods to test the response of individuals with celiac disease who were introduced to oats. These studies did not fulfill the selection criteria of pivotal studies namely an in vivo oats challenge with an intestinal/skin biopsy to assess the biological response to the introduction of oats into an otherwise gluten-free diet. Instead, they used various in vitro techniques to assess the immune response to avenin, or serology without an intestinal mucosal biopsy. Most of the methods used duodenal mucosal cultures prepared from biopsies or intestinal T cell lines obtained from individuals with celiac disease. Other studies measured the immunogenic reaction in peripheral lymphocytes or measured the presence of various antibodies in individuals with verified celiac disease who included oats in their diet, in comparison with a reference group (Table 6.2). Some of these studies used patients that were previously included in pivotal studies. These studies are identified with an asterisk ( ) in Table 6.2. 

A 9-year-old boy is referred for evaluation of his hearing. A note from his school principal explains that he is inattentive in class. Initial physical examination indicates that he is at the 10th percentile for height, has coarse facial features, and is somewhat macrocephalic however, the remainder of the examination is within normal limits. Audiometry results confirm partial bilateral deafness, which is sensorineural in etiology. An IQ examination shows that he is in the 60th percentile for intelligence. Family history of mucopolysaccharidoses prompts specialty testing, which indicates elevated levels of dermatan sulfate and heparan sulfate in both a skin biopsy and urine sample. 

Biochemical analysis of a skin biopsy from this patient would most likely indicate a deficiency of which of the following enzymes ... 

Topically administered local anesthetics are useful in dermatology for preparation of the skin prior to minor surgical procedures, such as skin biopsies, laser treatment of vascular malformations, and curettage of mol-luscum contagiosum lesions, particularly in young children and needle-phobic adults. The topical anesthetic may be used alone or may be applied prior to intrader-mal injection of a local anesthetic to reduce the pain caused by the needle. Two recently approved drugs in... 

The assessment of DNA adducts may provide a sensitive indicatCH of previous exposure. The enzyme-linked immunosorbent assay (ELISA) has a lower limit of detection of about 0.08 femtomol per microgram of DNA (Perera et oL, 1982). This assay requires (1) the development of an antibody specific for a certain chemical metabolite bound covalently to DNA and (2) the isolation of DNA from some tissue sample, ag., skin biopsy, or lymphocytes of an exposed individual. It is anticipated that further refinement of such immunologic techniques may lower the threshold of sensitivity by one or two orders of magnitude. One such refined test is the ifitrasensitive ymatic radioimmunoassay (USE-RIA), purported to be about five times more sensitive than ELISA (Hsu et oL, 1981 Shamsuddin et oL, 1985 Harris et oL, 1985). Quantification by the development of monoclonal antibodies to aflatoxin Bj metabolites bound to DNA (Groopman et oL, 1982 Sizaret et oL, 1982) has now been reported. 

Fibroblasts are typically grown from a small skin biopsy sample collected during an outpatient visit or as part of a planned surgical procedure following routine culturing techniques. Cell cultures may also be derived from umbilical cord or, for prenatal diagnostic purposes, from amniocytes obtained by amniocentesis. In our laboratory the analysis for each patient is performed in triplicate. 

The lysosomal disorder SASD is characterized by accumulation of the free acid monosaccharide sialic acid in the lysosomal compartment of the cell. Diagnosis is based on the demonstration of abnormal excretion of free, not OGS-bound sialic acid in urine, coupled with accumulation of free sialic acid in cultured fibroblasts, and on microscopic evidence of vacuoles (increased and swollen lysosomes filled with light electron-lucent material in skin biopsy and peripheral blood lymphocytes). The inheritance is autosomal recessive. There are different clinical forms of this disorder an adult form, called Salla disease (SD) or Finnish sialuria (OMIM 604369) infantile SASD (ISSD OMIM 269920) and an intermediate form, severe Salla disease [3,16]. 

Primary fibroblasts from skin biopsy samples are grown at 37°C in the presence of 5% C02 in Dulbecco s modified Eagle s medium (DMEM) supplemented with 10% foetal calf serum (FCS) and 1% penicillin/streptomycin. For passaging, confluent cells are washed with phosphate-buffered saline (PBS) and incubated for 5-10 min at 37°C in the presence of trypsin. 

Fibroblasts cultivated from skin biopsy samples. 

Human skin fibroblasts are cultured from skin biopsy samples. The dermis is cut into small pieces (0.5 mm on each side) and placed in a dish containing DMEM, 10% (v/v) FCS, and 1% (v/v) antibiotic-antimycotic solution. When these primary cultures are confluent they are split into novel dishes. Cells between passages three and six are used for experiments. 

Human skin fibroblasts are cultured from skin biopsy samples. The dermis is cut into small pieces (0.5 mm on each side) and placed into a dish in DMEM containing 10% (v/v) FCS and 1% (v/v) antibiotic-antimycotic solution. When these primary cultures are confluent they are split and cells between passage three and six are used for experiments. For the cholesterol efflux assay, cells are grown in 24-well plates to 60-80% confluence and are labeled with [1,2-3H]-cholesterol (1 pCi/well) for 24 h. Cells are then washed with DMEM and incubated for 4 h at 37°C with DMEM containing BSA (0.2%, v/v) and either 0 (negative control) or 5-30 pg/ml apoA-I. The efflux medium is collected and centrifuged to remove cell debris. Cells are solubilized in 0.1 mol/1 NaOH and the radioactivity in the efflux media and in the cell lysates is determined by scintillation counting [11, 30, 75]. 

First described in 1919 by Hurler, mucopolysaccharidosis I (MPS I, the Hurler syndrome) leads to accumulation of partially degraded dermatan and heparan sulfates (Fig. 4-11).317,352 353 A standard procedure in the study of diseases of this type is to culture fibroblasts from a skin biopsy. Such cells cultured from patients with the Hurler syndrome accumulate the polysaccharide, but when fibroblasts from a normal person are cultured in the same vessel the defect is "corrected." It was shown that a protein secreted by the normal fibroblasts is taken up by the defective fibroblasts, permitting them to complete the degradation of the stored polysaccharide. 

A 27-year-old woman, a pharmacist, had dermatitis on three separate occasions a few hours after she started to take oral deflazacort 6 mg for vesicular hand eczema (185). On each occasion, her symptoms included a widespread macular rash mainly on the inner aspects of her arms and legs and buttocks. She also had severe scaling, fever, nausea, vomiting, malaise, and hypotension. A skin biopsy was consistent with erythema multiforme, and direct immunofluorescence showed granular deposits at the dermoepidermal junction. Patch tests to the commercial formulation of deflazacort 6 mg (1% aqueous solution) and to pure deflazacort (1% aqueous solution) were positive, but there were no cross-reactions to other glucocorticoids. 

The authors of the last report commented that generalized delayed type hypersensitivity to systemic administration of a glucocorticoid is rare. Despite the potent immunosuppressive effect of glucocorticoids on immunocompetent cells, the clinical features, the skin biopsy specimen, and the positive delayed skin test reactions strongly suggested an immunological mechanism T cells were clearly involved and the high concentrations of interleukins 5, 6, and 10 were consistent with a T helper type 2 reaction. The raised concentrations of interleukin-5 were probably responsible for the blood and tissue eosinophilia. 

An 84-year-old woman with polymyalgia rheumatica and a 79-year-old woman with undifferentiated connective tissue disease and leukocytoclastic vasculitis were given prednisolone 20 mg/day with subsequent dosage reductions. The first patient developed a raised purpuric rash and lymphedema of the left leg within 5 months and the second developed large purple nodules on the soles of her feet and the backs of her hands accompanied by periorbital and peripheral edema. Skin biopsies showed Kaposi s sarcoma, and both patients had raised IgG antibody titers to human herpesvirus-8. 

A 54-year-old woman fitted with a Mirena system soon developed fever and progressive skin lesions. A skin biopsy confirmed Sweet s syndrome. She was treatment with topical and oral glucocorticoids, but the condition relapsed on reduction of the dose. Her symptoms finally resolved on removal of the intrauterine system and she remained symptom free after 9 months. 

A skin biopsy from a 58-year-old man showed necrosis of keratinocytes with lymphocytic and eosinophilic infiltration. Liver enzymes were normal. After withdrawal the rash disappeared. After 3 weeks, rechallenge with acarbose 50 mg caused the skin changes to reappear. 

A 38-year-old woman developed erythematous dermatitis, in a skin biopsy consistent with necrolytic migratory erythema, and angular cheilitis, which disappeared after withdrawal of glucagon infusion after 2 months. 

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