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Changes in the autoimmune blistering response: a clinical and immunopathological shift from pemphigus foliaceus to bullous pemphigoid.

We describe a 64-year-old Brazilian man who developed bullous pemphigoid (BP) 12 years after pemphigus foliaceus (PF) was diagnosed. On his first presentation in 1992, histological examination revealed intraepidermal blistering and acantholysis at the granular layer, direct immunofluorescence (DIF) demonstrated intercellular deposits of C3 in the epidermis, and indirect immunofluorescence showed the presence of IgG antibodies against the intercellular spaces. In 2004, laboratory findings revealed a subepidermal blister with neutrophils and eosinophils (by histology), DIF demonstrated deposition of IgG and C3 along the basement membrane zone, salt-split skin showed IgG deposition in the epidermal side of the blister, and immunoblotting showed reactivity against BP180. The occurrence of two autoimmune blistering conditions in the same patient is a rare event, and may suggest an intermolecular epitope-spreading phenomenon.

Anti-Bacterial Agents↗

Autoimmune subepidermal blistering diseases in Uganda: correlation of autoantibody class with age of patients.

BACKGROUND: No data are available on the incidence and immunoreactivity of autoimmune subepidermal blistering skin diseases in East Africa. METHODS: All patients with frank blisters/erosions on the skin and/or mucous membranes that attended the Department of Dermatology at Mbarara University, Uganda, from May 2000 to June 2002, were investigated. The diagnosis was based on the clinical presentation and on the presence of circulating autoantibodies detected by indirect immunofluorescence microscopy on 1 m NaCl-split human skin and by Western blotting of recombinant and cell-derived forms of BP180, BP230, and type VII collagen. RESULTS: Twenty-two patients with autoimmune subepidermal blistering skin disorders were identified, including nine with bullous pemphigoid (41%), four with linear immunoglobulin A (IgA) disease (18%), three with mucous membrane pemphigoid (14%), two with linear IgG/IgA bullous dermatosis (9%), and one each with cicatricial pemphigoid and epidermolysis bullosa acquisita (5%). In addition, two patients with immunoreactivity to both the epidermal and dermal side of salt-split skin by indirect immunofluorescence microscopy, who were unreactive to type VII collagen, were provisionally diagnosed as "mixed pemphigoid" (9%). In patients with subepidermal blistering diseases, IgG reactivity correlated significantly with old age, whereas younger patients preferentially developed IgA autoantibodies (P = 0.024). CONCLUSIONS: The age of patients with autoimmune subepidermal blistering diseases appears to influence the immunoglobulin class of autoantibodies. The high frequency of IgA autoantibodies in Ugandan patients may be explained by the age distribution of the Ugandan population.

Adolescent↗

Passage of albumin from plasma to suction skin blisters.

To study the transvascular passage of albumin from the plasma to the skin interstitium, suction blisters were developed on the abdominal skin of six young, healthy male volunteers. Fifteen minutes after the blisters had been formed and the suction had been stopped, 125I-albumin was injected intravenously and the disappearance of the tracer protein from plasma was compared to its appearance in blister fluid as a function of time for up to 4 h after the injection. There was a linear increase of blister fluid activity with time after injection and the mean accumulation rate of the six subjects was 0.9 +/- 0.4 (SD)%/h. It is suggested that this figure is a representative measure of the leakage of albumin through the skin microvasculature. In previous studies, a ten-fold higher accumulation rate of albumin was found when the protein tracer was administered intravenously before the suction was induced. This difference was probably due to a suction-induced increase of the filtration gradient across the skin vessels. It is concluded that both methods might be used to obtain a comparative measure of the local rate at which albumin passes from plasma to the interstitial fluid of the skin.

Adult↗

Cefodizime in serum and skin blister fluid after single intravenous and intramuscular doses in healthy volunteers.

In gonorrhea therapy, cephalosporins are conventionally administered by intramuscular (i.m.) injection, which rather frequently leads to local side effects. To investigate whether the well-tolerated intravenous (i.v.) injection of cephalosporins may be of comparable gonocidal effect, levels of cefodizime, a new broad-spectrum cephalosporin, in serum and tissue fluid (suction blister and cantharides blister fluid) were determined in six healthy men. Single doses of 1 g of cefodizime were injected i.v. and i.m. according to a randomized crossover design. On i.m. injection the drug was completely bioavailable, and the peak concentration in serum was 75 +/- 8 micrograms/ml. The terminal half-life of serum levels was 2.4 h. Cefodizime concentrations in the blister fluids increased for 1.5 to 3 h after the i.v. dose and for at least 3 h on i.m. administration. The concentrations of non-protein-bound cefodizime in blister fluid already exceeded the MIC for 90% of Neisseria gonorrhoeae strains 10 min after i.v. injection and 20 to 30 min after the i.m. dose. At 6 h after each dose, active concentrations were still present in serum. The results suggest that cefodizime administered i.v. and i.m. has equivalent high cure rates in uncomplicated gonorrhea. This hypothesis should be tested further by a controlled clinical trial. If equivalent, i.v. administration excels because it is better tolerated locally.

Adult↗

Determination of protoporphyrin in plasma and suction-blister fluid from light-irradiated and non-irradiated skin in protoporphyria patients.

At various times of the year the protoporphyrin level in the plasma of 11 patients with erythropoietic protoporphyria was determined. In the winter as well as in the summer, when the patients received carotenoids, an area on one forearm of 10 patients was irradiated with artificial (xenon arc) light, after which suction blisters were made on both forearms. Protoporphyrin levels were determined in the plasma and in the blister fluid from both areas. Suction-blister fluid was considered representative of the interstitial fluid in the dermis. In the winter the protoporphyrin levels of the blister fluid from the irradiated area were significantly higher than in the fluid from the non-irradiated area. In the summer this difference was not significant. The possible implications of these findings are discussed.

Blister↗

92-kD gelatinase is produced by eosinophils at the site of blister formation in bullous pemphigoid and cleaves the extracellular domain of recombinant 180-kD bullous pemphigoid autoantigen.

Eosinophils are prominent in bullous pemphigoid (BP), and proteases secreted from these and other inflammatory cells may induce disruption of the basement membrane. We used in situ hybridization and immunohistochemistry to localize the sites of 92-kD gelatinase expression in BP lesions. In all samples (20/20), a strong signal for gelatinase mRNA was detected only in eosinophils and was most pronounced where these cells accumulated at the floor of forming blisters. No other cells were positive for enzyme mRNA. Both eosinophils and neutrophils, however, contained immunoreactive 92-kD gelatinase indicating that active expression occurred only in eosinophils. Degranulated eosinophils were also seen near blisters, and as demonstrated by gelatin zymography, immunoblotting, and ELISA, 92-kD gelatinase protein was prominent in BP blister fluid. No other gelatinolytic activity was specifically detected in BP fluid, and only small amounts of 92-kD gelatinase were present in suction blister fluids. As demonstrated in vitro, 92-kD gelatinase cleaved the extracellular, collagenous domain of recombinant 180-kD BP autoantigen (BP180, BPAG2, HD4, type XVII collagen), a transmembrane molecule of the epidermal hemidesmosome. Our results suggest that production and release 92-kD gelatinase by eosinophils contributes significantly to tissue damage in BP.

Autoantigens↗

Passive transfer of anti-laminin 5 antibodies induces subepidermal blisters in neonatal mice.

Patients with a recently identified subepithelial blistering disease have IgG anti-laminin 5 autoantibodies. To determine if such antibodies can be pathogenic in vivo, we developed and characterized rabbit anti-laminin 5 IgG, and passively transferred these antibodies to neonatal mice. Immune rabbit IgG specifically bound human and murine epidermal basement membranes, immunoblotted and immunoprecipitated all laminin 5 subunits from extracts of human and murine keratinocytes, and showed no reactivity to other keratinocyte proteins or epithelial basement membranes that do not contain laminin 5. Mice (n = 29) receiving purified anti-laminin 5 IgG developed, in a dose-related fashion, circulating anti-laminin 5 antibodies, deposits of rabbit IgG and murine C3 in epidermal basement membranes, and subepidermal blisters of skin and mucous membranes. No alterations developed in controls (n = 14) receiving identical amounts of normal rabbit IgG. Passive transfer of anti-laminin 5 (but not control) IgG to neonatal C5- (n = 3) or mast cell-deficient (n = 3) mice produced subepidermal blisters with the same clinical, histologic, and immunopathologic features as those documented in BALB/c mice. These studies establish an animal model of a human blistering disease that can be used to define disease mechanisms and treatment modalities.

Animals↗

Osmotic blistering in enamel bonded with one-step self-etch adhesives.

One-step self-etch adhesives behave as permeable membranes after polymerization, permitting water to move through the cured adhesives. We hypothesize that osmotic blistering occurs in bonded enamel when these adhesives are used without composite coupling. Tooth surfaces from extracted human premolars were bonded with 5 one-step self-etch adhesives. They were immersed in distilled water or 4.8 M CaCl(2), and examined by stereomicroscopy, field-emission/environmental SEM, and TEM. Water blisters were observed in bonded enamel but not in bonded dentin when specimens were immersed in water. They collapsed when water was subsequently replaced with CaCl(2). Blisters were absent from enamel in specimens that were immersed in CaCl(2) only. Water trees were identified from adhesive-enamel interfaces. Osmotic blistering in enamel is probably caused by the low water permeability of enamel. This creates an osmotic gradient between the bonded enamel and the external environment, causing water sorption into the interface.

Acid Etching, Dental↗

Treatment of a heel blister caused by pressure and friction.

The treatment of heel blisters caused by pressure, friction, and shear is a complex subject. Issues such as whether to aspirate the blister, how to protect it from further pressure, friction, and shear, and which dressing to choose, need to be considered. As nurses are increasingly aware of the emphasis that has been placed on the importance of ensuring that clinical practice is based on evidence, this article attempts to shed some light on these areas of debate. It is important to highlight that during the literature search very little research was found to exist on blister care and it became clear that further research into the nursing care of a blister caused by pressure would be useful to aid nursing care and patient comfort.

Aged↗

Skin breakdown and blisters from senna-containing laxatives in young children.

BACKGROUND: At the direction of the Food and Drug Administration, phenolphthalein was removed from all over-the-counter laxatives in 1999. Phenolphthalein was then replaced in most laxative products with the natural product senna from Cassia acutifolia Delile, which contains various anthraquinones. No data are available on the safety of senna use in children <6 years of age. OBJECTIVE: To describe the clinical outcomes of exposure to unintentional ingestion of senna-containing laxatives in young children. METHODS: All ingestion exposures of senna-containing laxatives in children <5 years of age from 6 poison centers over a 9-month period were evaluated. Inclusion criteria required 24-hour follow-up and the presence of diarrhea to confirm ingestion. Parents were told routinely that severe diaper rash was possible and to protect the perianal area with frequent cleansing and a barrier ointment if the child was wearing diapers. RESULTS: During the study period, 111 cases were reported: 19 children experienced no diarrhea, 4 were lost to follow-up, and 88 exposures were evaluated. Fifty-two children (59%) were </=2 years old. Fifty children remained in diapers, 28 children were fully toilet trained, and 10 wore diapers (pull-up pants) overnight. Twenty-nine children (33%) experienced severe diaper rash. The mean +/- SD time to recognition of the diaper rash was 15.6 +/- 8.6 hours. Ten children (11%) had blisters and skin sloughing. There was a significant increase in severe diaper rash (p < 0.05) and onset of blisters and skin breakdown (p < 0.05) in children wearing diapers versus those who were fully toilet trained. The mean time to onset of blisters was 14.5 +/- 6.8 hours. Skin burns and loss were seen primarily on the buttocks and perineum, loosely following the diaper area. CONCLUSIONS: Unintentional ingestion of senna-containing laxatives in young children may potentially cause severe diaper rash, blisters, and skin sloughing.

Blister↗

The effect of floor type on the development of breast blisters and feather follicle infections in broilers.

The effect of floor types on the incidence of feather follicle infection and breast blisters in caged broilers was investigated in two trials. Broilers were reared on corn-soybean meal diets with warm-room brooding. Floor types included litter (control), wire, wire covered with various types of plastic mats, and wire woven with surgical tubing. In trial 1, the effect of a coccidiostat (Monensin, 120 mg/kg of feed) was investigated for broilers not on litter, and in trial 2, the effect of supplemental zinc was investigated. Broilers were processed in the laboratory processing plant when they were 51 (trial 1) and 53 days old (trial 2). Carcasses were evaluated for the occurrence of fleshing downgrades, crooked keels, feather follicle infection, and breast blisters. Incidence of feather follicle infection was low for broilers on litter or wire floor and high for broiler reared on plastic mats. Breast blisters incidence was low for broilers on litter and plastic mats and was high for broilers on the wire floor. Overall carcass quality for litter (control) was superior to any of the cage treatments. Neither the coccidiostat nor supplemental zinc affected the incidence of breast blisters or feather follicle infection.

Animals↗

A prospective analysis of the treatment of friction blisters with 2-octylcyanoacrylate.

Early military investigations of blister treatment using tissue adhesives have shown promise, finding that these agents provide pain relief, prevent infection, and allow continuation of recruit training. A trial was conducted to compare a 2-octylcyanoacrylate treatment with military standard therapy. Patients were recruited during foot evaluation at the end of a 10-km military road march. Seventy-five patients were screened, and 51 were enrolled. Twenty-six patients received 2-octylcyanoacrylate and 25 received standard treatment. A total of 80 blisters were treated (40 in each group). The mean (SD) blister size in the 2-octylcyanoacrylate group was 12.7 (6.2) mm and in the standard group was 12.0 (5.7) mm. There were no statistically significant differences in any of the baseline variables. Baseline and repeated-measures visual analog scale scores demonstrated no statistically significant differences in initial or repeat pain scores. Both groups showed a similar change across time, with a nonsignificant trend toward improvement in the standard therapy group at 10 min (28.5 versus 24.9) and in the 2-octylcyanoacrylate group at 3 days (42.9 versus 50.1). Mean Likert scores were similar, indicating no difference in patient satisfaction. Time to resumption of normal activity was similar, with one patient in each group unable to return to activity at the time of follow-up. There was a trend toward an increased proportion of patients in the 2-octylcyanoacrylate group who were able to return to normal activity within 48 hours, but this did not reach statistical significance. 2-Octylcyanoacrylate was associated with a greater degree of procedural discomfort. No infected blisters were noted in either group.

Adult↗

Causes of enlarged sternal bursas (breast blisters).

Bursas were taken weekly as normal turkeys developed from 3 to 22 weeks of age, and no specific bacterial or viral infection was found. Bacterial cultures were also made of condemned bursas taken at processing. Of 144 collected, 91% were sterile; the others had staphylococci, streptococci, or coliform organisms. Experiments with turkeys measured the effect of strain, body conformation, completeness of breast-feather cover, protection of the keel with sheepskins, age when put on range, and type of range on the incidence of enlarged sternal bursas. The strain difference of 25.4% breast blisters against 11.5% was highly significant (99% confidence). In one experiment the effect of feather cover was significant at the 95% level of confidence (61.7% breast blisters with no feather cover, versus 40% with complete feather cover). The effect of protecting the breast with sheepskin was significant at the 95% level of confidence in reducing incidence of breast blisters. There was no significant difference in the incidence of breast blisters in relation to body conformation, age, or type of range.

Animal Feed↗

Ultrastructural analyses of the Caenorhabditis elegans DR 847 bli-1(n361) mutant which produces abnormal cuticle blisters.

The bli-1 gene of Caenorhabditis elegans has been previously described as a mutation which disrupts the structure of the adult-stage cuticle causing the formation of fluid-filled blisters. We investigated the blistering phenotype exhibiting n361 allele and observed a gradual blister formation in adult nematodes. In the course several fine changes occurred including a high electron density granulous material filling the intermediate layer, alterations on struts structure, and finally the total disappearance of the fibrous and basal layers. With the ethanolic phosphotungstic acid technique (E-PTA), which localizes basic proteins, reaction product was observed in the cortical layer of the wild strain, whereas in the mutant strain an irregular labelling pattern was observed in this region. The granulous material inside the intermediate layer of the mutant strain showed also a strong reaction. An imidazole-buffered osmium tetroxide solution was used to visualize lipids at ultrastructural level, however no dense product was detected in the cuticle of both strains of C. elegans. Based on these results we postulated that the blistering phenotype is due to an altered function of bli-1 gene, which is probably enzymatic.

Animals↗

Epidermis is the origin of high creatine kinase levels in skin blister fluid.

Creatine kinase (CK) isoenzyme, CK-BB, known as the brain fraction, is not normally present in serum but predominates in several normal and malignant tissues and body fluids. We recently reported increased CK-BB levels in suction blister fluid. In the present study the cellular origin of the enzyme in skin was studied from homogenates of blister top epidermis and blister base dermis as well as from homogenates of split skin dermatome shavings and isolated keratinocytes. The CK-BB in human skin was derived almost exclusively from the epidermis. Enzyme determinations from various spontaneous bullae suggest that all types of skin blisters initially contain high CK-BB levels.

Biopsy↗

Plasmin-like proteinase associated with high molecular weight complexes in blister fluid of bullous pemphigoid.

Blister fluid from tense bullae of 10 patients with bullous pemphigoid was investigated using a radial caseinolysis assay and zymography. Proteolytic activity, varying from 4.7 to 10 micrograms/ml, was found in 3 out of 10 patients, by using the caseinolysis assay. Zymography revealed that a major part of this caseinolytic activity co-migrated with plasmin standard. In addition, in the zymography, proteolytically active high molecular weight complexes were seen. This characteristic pattern was seen in the zymography of both positive and negative samples in the caseinolysis assay. These high molecular weight complexes were not seen in the zymography of the blister fluid of 2 patients with epidermolysis bullosa or in the suction blister fluid of 3 healthy control patients. These findings suggest that plasmin is generated at some phase of the blister formation, being possibly involved in the pathomechanism of bullous pemphigoid.

Blister↗

Penetration of cefonicid in skin suction blister fluid in healthy volunteers.

A new cephalosporin, cefonicid (1 g) was given intramuscularly to six healthy volunteers 12 h after induction of skin suction blister. Serum and blister fluid were collected during a 24 h period. Antibiotic assay was made by a microbiological method. The data were analysed by a one-compartment kinetic model. Despite the high protein binding, cefonicid penetrated in microbiologically effective concentration into blister fluid. Blister fluid concentrations were higher than the serum concentrations at 24 h.

Adult↗

Skin blister fluid--an access to the peripheral compartment. Implications of a study on the behaviour of bendroflumethiazide in rats.

Pharmacokinetics of bendroflumethiazide (bft) were investigated in blood and skin suction blister fluid (SBF) in rats dosed 10 mg/kg intravenously. Additionally tissue levels were determined. Mean volume of skin blisters amounted to 10 or 20 microliters, respectively. While blood levels were best described according to a three-compartment model (terminal half-life 152 min) bft concentrations in SBF raised to a maximum 60 min post administration and thereafter declined (half-life 143 min). There was no significant effect of blister volume on SBF level-time course. A close relation between bft concentrations in SBF and the calculated amount of drug in the deep compartment was observed (r = 0.987). Plasma protein binding of bft was 84% while binding to SBF amounted to 76%. The drug is evenly distributed between plasma and erythrocytes. Therefore concentrations of unbound drug in SBF exceed those in plasma during the terminal phase. The determination of tissue levels showed bft concentrations in liver, kidney, lung and heart to follow blood levels whereas the pharmacokinetic behaviour of skin and muscle is rather close to that of the deep compartment. The results suggest the determination of skin blister fluid levels to be valuable if a drug acts from the tissue compartment then SBF being closer to the biophase than blood.

Albumins↗