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The effect of kinin agonists and antagonists on the pain response of the human blister base.

1. The effect of bradykinin (BK) and some analogues of BK on the human blister base was studied. 2. BK produced reproducible dose-related increases in pain responses. A characteristic delay, which was not dose-related occurred between application of BK and the resultant response. 3. The rank order of potency of several kinin analogues on the pain response was BK much much greater than sigma-cyclo-(Lys1-Gly6)-BK = sigma-cyclo-kallidin greater than des-Arg9-BK. 4. No increase in pain response was seen with repeated application of the selective B1 receptor agonist des-Arg9-BK to the same blister base at 4 h intervals. The B1 receptor antagonist des-Arg9-Leu8-BK was without effect against BK-induced responses. 5. The B2 receptor antagonists, D-Arg-Arg-Pro-Hyp-Gly-Thi-Ser-D-Phe-Thi-Arg-TEA and D-Pro-Phe-Arg-heptylamide produced significant antagonism of the bradykinin-induced pain responses at doses which had no effect against 5-hydroxytryptamine or potassium chloride. 6. It is concluded that the kinin receptor mediating pain on the human blister base is of the B2 type.

Blister↗

Absence of arylsulphatases A and B in suction blister fluid interstitial fluid and serum.

Suction blisters were raised on human skin. The blister fluid was collected for enzymic assay and the epidermis over the bullae was removed before further suction of interstitial fluid. Preliminary work indicated the presence of arylsulphatases A and B in epidermis, and the same assay procedure with 4-methylumbelliferyl sulphate as substrate was used. There appeared to be no enzymatic activity in blister fluid, interstitial fluid and serum.

Arylsulfatases↗

[Coagulation factors in human born blisters (author's transl)].

Following clotting factor assays were performed on the fluid of burn blisters of 11 patients with severe burns: Fibrinogen levels, Factor II, V, X and XIII, thrombin time, fibrin split products, plasminogen, antithrombin III, IgA, IgM, IgG, ethanol gelation test and total proteins. The results showed, that a quantity of Factor II, X and XIII, antithrombin III, plasminogen and IgG had left the circulation. On the contrary we found only small concentrations of Factor V, fibrinogen, IgM and IgA in the fluid of burn blisters. This distribution suggested that the losses of plasma proteins into the burn blisters were correlated to their concentration and their molecular weight. The decrease of plasma coagulation proteins during the first days after severe burns was probably partly due to losses through the walls of the vessels, because of the increased capillary permeability.

Antithrombins↗

Influenced of food on the kinetics of 8-methoxypsoralen in serum and suction blister fluid in psoriatic patients.

The influence of food on the kinetics of 8-methoxypsoralen (8-MOP) in serum and suction blister fluid was evaluated in a cross-over study in 19 psoriatic patients under PUVA treatment. The peak serum concentration of 8-MOP was reached 1.5 h after ingestion on an empty stomach, and in suction blister fluid the maximum concentration was already present in the first sample taken after 2 h, the time when UVA radiation was given. The postprandial kinetics of 8-MOP in serum and suction blister fluid differed, the highest levels being reached, respectively, at 2.4 and 3 h after intake, i.e. in both body fluids after irradiation had started. The side effects of 8-MOP, such as nausea and dizziness, in the two groups were similar. The present results indicate that to optimize the therapeutic effect of PUVA in individual patients, 8-MOP should be given on an empty stomach.

Blister↗

Correlations between theophylline concentrations in plasma, erythrocytes and cantharides-induced blister fluid and peak expiratory flow in asthma patients.

Theophylline levels in plasma, erythrocytes and cantharides-induced blister fluid (BF) have been correlated with peak expiratory flow (PEF) in asthmatic patients. Nine asthmatic men received an IV infusion of 240 mg theophylline at a constant rate for 30 min. The theophylline concentrations in erythrocytes and blister fluid were 37% and 78% of those in plasma. There was not a strong correlation between erythrocyte or blister fluid penetration and the clinical effect of theophylline.

Asthma↗

Alteration of lipoproteins of suction blister fluid by UV radiation.

Suction blister fluid is a good representative of the interstitial fluid feeding the epidermal cells. Lipoproteins contained in the suction blister fluid of healthy volunteers are readily photo-oxidized by UV radiation in the wavelength range 290-385 nm. Absorbed light doses equivalent to one minimal erythemal dose absorbed by skin exposed to UVB (e.g. 290-320 nm) are sufficient to induce lipid peroxidation and modification of apolipoproteins A-I, A-II and B. Albumin, which is known to protect serum fully from oxidative stresses, is not so effective against photo-oxidation. Although tryptophan (Trp) residues of proteins contained in the suction blister fluid are photo-oxidized, apolipoprotein A-II, which does not contain any chromophoric Trp residue, is also altered by the UV radiation. With regard to results obtained with a model reconstituted fluid, it is suggested that the radical chain reaction of the lipid peroxidation can transfer the initial photodamage at sites not directly susceptible to photo-oxidation. The biological implications of these results are discussed.

Apolipoprotein A-I↗

Subepidermal blistering disease with autoantibodies to both the p200 autoantigen and the alpha3 chain of laminin 5.

Anti-p200 pemphigoid and anti-laminin 5 mucous membrane pemphigoid are two distinct autoimmune blistering skin diseases. Patients with anti-p200 pemphigoid demonstrate circulating autoantibodies to an unknown 200-kd acidic noncollagenous glycoprotein of the lower lamina lucida, whereas anti-laminin 5 mucous membrane pemphigoid is characterized by an autoimmune response against the major basement membrane adhesion molecule laminin 5. In this report, we describe a patient who developed a recurrent nonscarring blistering eruption, affecting both skin and mucous membranes. Immunoblotting of dermal extracts and extracellular matrix of cultured keratinocytes revealed circulating autoantibodies directed to the p200 antigen and the alpha3 chain of laminin 5, respectively. The unusual clinical and immunologic profile of this case suggests that molecular specificity of circulating autoantibodies affects the clinical presentation of autoimmune subepidermal blistering skin diseases.

Antibody Specificity↗

Blistering distal dactylitis: a case series in children under nine months of age.

Blistering distal dactylitis (BDD) is a distinct clinical entity that is infrequently reported in the literature. Characteristically, blistering distal dactylitis is described as a localized infection involving the volar fat pad of the distal phalanx of the digits, and it usually presents as a fluid-filled blister. The usual causative organism is group A beta-hemolytic Streptococcus, but less commonly, Staphyloccous aureus and Staphyloccoccus epidermis are present. The normal age range is reported to be 2 to 16 years old, but there are case reports of this infection in adults. Only one case has been reported in the literature in a child younger than 24 months of age. In this report we describe three cases in children younger than 9 months old. These three cases indicate that BDD can and does occur in children younger than 2 years of age.

Anti-Bacterial Agents↗

Cantharidin blisters: a technique for investigating leukocyte trafficking and cytokine production at sites of inflammation in humans.

A skin blister technique is described which allows the investigation of acute inflammation in humans in vivo. Filter paper discs are placed on the skin, impregnated with cantharidin and covered with impermeable film held by adhesive tape. The assembly is easily applied, unobtrusive, stable and may be worn during normal activities. The blister formed at 24 h contains approximately 5x10(5)-5x10(6) cells, predominantly neutrophils and macrophages. Inflammatory cytokines and chemotactic factors are detectable in the blister fluid. The technique is useful for characterizing the acute inflammatory response in health and disease.

Blister↗

Identification and pharmacological characterization of platelet-activating factor and related 1-palmitoyl species in human inflammatory blistering diseases.

Through its pro-inflammatory effects on leukocytes, endothelial cells, and keratinocytes, the lipid mediator platelet-activating factor (PAF) has been implicated in cutaneous inflammation. Although the 1-alkyl PAF species has been considered historically the most abundant and important ligand for the PAF receptor (PAF-R), other putative ligands for this receptor have been described including 1-acyl analogs of sn-2 acetyl glycerophosphocholines. Previous bioassays have demonstrated a PAF-like activity in lesions of the autoimmune blistering disease bullous pemphigoid. To assess the actual sn-2 acetyl glycerophosphocholine species that result in this PAF agonistic activity, we measured PAF and related sn-2 acetyl GPCs in fresh blister fluid samples from bullous pemphigoid and noninflammatory (suction-induced) bullae by mass spectrometry. We report the presence of 1-hexadecyl as well as the 1-acyl PAF analog 1-palmitoyl-2-acetyl glycerophosphocholine (PAPC) in inflammatory blister fluid samples. Because PAPC is the most abundant sn-2 acetyl glycerophosphocholine species found in all samples examined, the pharmacological effects of this species with respect to the PAF-R were determined using a model system created by transduction of a PAF-R-negative epidermoid cell line with the PAF-R. Radioligand binding and intracellular calcium mobilization studies indicated that PAPC is approximately 100x less potent than PAF. Though a weak agonist, PAPC could induce PAF biosynthesis and PAF-R desensitization. Finally, intradermal injections of PAF and PAPC into the ventral ears of rats demonstrated that PAPC was 100x less potent in vivo. These studies suggest possible involvement of PAF and related species in inflammatory bullous diseases.

Animals↗

Hereditary callosities with blisters. Report of a family and review.

A family with calluses of the soles associated with blistering is described. Electron microscopic study of a bulla showed an intraepidermal blister with cytolysis of keratinocytes and clumping of tonofilaments. Review of the literature and our own experience with keratoderma palmaris et plantaris revealed no similar patients with this combination of findings. The appearance of the soles is similar to pachyonychia congenita, but the lack of nail and mucous membrane changes is not consistent with that disorder. Treatment with isotretinoin caused reduction in the size of the calluses but exacerbated the blistering.

Adult↗

Blood blisters of the oral mucosa (angina bullosa haemorrhagica).

Angina bullosa haemorrhagica is a benign phenomenon that is characterized by the sudden appearance of a blood blister on the oral mucosa in the absence of an identifiable cause or systemic disorder; local trauma has been suggested to be the most likely contributory factor. No treatment is required. Angina bullosa haemorrhagica affects mainly middle-aged and elderly people. There is no strong predilection for either men or women. In this article nine patients with such blisters are described. Blood blisters apparently are more common than has been suggested in the literature.

Adult↗

Eosinophil-colony stimulating activity in blister fluid of bullous pemphigoid.

We examined eosinophil-colony stimulating activity in blister fluid or serum obtained from patients with bullous pemphigoid using the methylcellulose semi-solid hematopoietic stem cell culture system, and demonstrated that the blister fluid of patients with bullous pemphigoid generated strong eosinophil-colony stimulating activity. This evidence suggests that eosinophil-colony stimulating activity in blister fluid probably influences the induction of eosinophilia in bullous pemphigoid.

Aged↗

[Swollen leg with blisters].

We report the case of a 84-year-old woman suffering from strong pain in her right leg initially resembling thrombosis of deep veins. Eight hours after admission a superficial blister developed at the calf with following hemorrhagic aspect and spontanous eruption of clear yellowish fluid. Later on a new blister appeared at the thigh. The patient died 33 hours after admission of streptococcal toxic shock syndrome. The latter was based on a necrotizing fasciitis. Streptoccus pyogenes (group A) could be cultivated from the blood and fluid of the blister. We discuss the clinical presentation of necrotizing fasciitis with "pain out of proportion" as characteristic complaint and the appropriate management.

Aged, 80 and over↗

High level of thymus and activation-regulated chemokine in blister fluid and sera of patients with bullous pemphigoid.

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by eosinophilia and high serum IgE levels. The accumulated evidence suggests that various cytokines are involved in the lesional skin of patients with BP. Recently, thymus and activation-regulated chemokine (TARC/CCL17), a CC chemokine, was identified as a selective chemoattractant for CC chemokine receptor 4 (CCR4)-expressing cells. OBJECTIVE: In this study, we examined the involvement of TARC in patients with BP. METHODS: We determined the fluid and serum TARC levels in patients with BP by enzyme-linked immunosorbent assay and compared the serum TARC levels with the eosinophil numbers in peripheral blood. We also compared the serum TARC levels in five patients with BP before and after they were treated. Moreover, we examined TARC, CCR4 and CXC chemokine receptor 3 (CXCR3) expression in the lesional skin of patients with BP by immunohistochemical procedures. Furthermore, we measured CCR4 positivity in CD4+ CD45RO+ cells of peripheral blood mononuclear cells (PBMCs) in patients with BP and healthy control subjects. RESULTS: The fluid TARC levels in patients with BP were significantly higher than those in blisters from burn patients or suction blisters of healthy control subjects. The serum TARC levels in patients with BP were also significantly higher than those in pemphigus vulgaris (PV) patients and healthy control subjects, and decreased after the treatment. The serum TARC levels in patients with BP significantly correlated with the eosinophil numbers in peripheral blood (r = 0.72, P < 0.002). Immunohistochemistry showed a strong reactivity of TARC in the epidermal keratinocytes (KCs) of BP. Moreover, both CCR4 and CXCR3 were expressed on the dermal infiltrating CD4+ T cells mainly beneath the bullae of patients with BP. Fluorescence-activated cell sorting analysis showed a higher percentage of CCR4 positivity in CD4+ CD45RO+ cells of PBMCs in patients with BP than that in healthy control subjects, while there was no significant difference of CXCR3 positivity in CD4+ CD45RO+ cells of PBMCs between patients with BP and healthy control subjects. CONCLUSIONS: These findings strongly suggest that TARC may be one of the important chemokines that are involved in the pathogenesis of BP.

Adult↗

Blister fluid T lymphocytes during toxic epidermal necrolysis are functional cytotoxic cells which express human natural killer (NK) inhibitory receptors.

Toxic epidermal necrolysis (TEN) is a rare life-threatening adverse drug reaction characterized by a massive destruction of the epidermis. Immunohistological studies of skin biopsies of TEN showed infiltrates of predominantly CD8+ T lymphocytes even though other authors reported a prominent involvement of cells of the monocyte-macrophage lineage. The aim of this study was to characterize phenotypically and functionally the cells present in the cutaneous blister fluid of four patients with TEN. We first determined that lymphocytes were predominant in blister fluid obtained early, while monocytes/macrophages later became the most important population. We then showed that this lymphocyte population, mainly CD3+CD8+, corresponded to a peculiar cell subset as they expressed cutaneous leucocyte antigen, killer inhibitory receptors KIR/KAR and failed to express CD28 molecule. Functionally, we determined that blister T lymphocytes had a cytotoxic T lymphocyte (CTL)- and NK-like cytotoxicity. The role of this cytotoxic lymphocyte population present at the site of lesions during TEN remains to be understood.

Adult↗

Modified technique of suction blistering for epidermal grafting in vitiligo.

BACKGROUND: Suction blister epidermal grafting is a useful modality of treatment of resistant and stable vitiligo; however, it requires expensive and heavy suction apparatus. This study is an attempt to develop a cheap and small apparatus which can be assembled in the physician's own office. PATIENTS AND METHODS: The method was tried in 22 vitiligo/leukoderma patients. The apparatus consisted of a cylindrical funnel connected with a three-way tap, and suction was given by a 50-mL syringe. The pressure inside the suction cup was retained by changing the position of lock of the three-way tap. The pressure was measured by connecting the three-way tap to a vacuum gauge. The apparatus remained adhered to the donor area because of negative pressure. The blister was formed in about 1.5 h. The roof of the blister was grafted onto the dermabraded recipient site. RESULTS: The pigmentation was complete in 20 out of 22 patients. There were no complications. CONCLUSIONS: The technique is inexpensive and easy and obviates the need of cumbersome and heavy equipment.

Adolescent↗

Animal models for skin blistering conditions: absence of laminin 5 causes hereditary junctional mechanobullous disease in the Belgian horse.

Recent achievements in the genetic correction of keratinocytes isolated from patients with junctional epidermolysis bullosa have paved the way to a gene therapy approach for the disease. Because gene therapy protocols require preclinical validation in animals, we have characterized spontaneous animal models of junctional epidermolysis bullosa. In this study we have elucidated the genetic basis of the hereditary junctional mechanobullous disease in the Belgian horse, a condition characterized by blistering of the skin and mouth epithelia, and exungulation (loss of the hoof). Immunofluorescence analysis associated the condition to the absent expression of the gamma2 chain of laminin 5 and designated Lamc2 as the candidate gene. Comparative analysis of the nucleotide sequence of the full-length gamma2 cDNA isolated by reverse transcription polymerase chain reaction amplification of total RNA purified from the epithelium of a junctional epidermolysis bullosa foal and a healthy control disclosed a homozygous basepair insertion (1368insC) in the affected animal. Mutation 1368insC results in a downstream premature termination codon and is predicted to cause absent expression of the laminin gamma2 polypeptide. Our results also show that: (i) the horse junctional epidermolysis bullosa genetically corresponds to the severe Herlitz form of junctional epidermolysis bullosa in man; (ii) the amino acid sequence and structure of the horse laminin gamma2 chain are virtually identical to the human counterpart; (iii) the moderate eruption of skin blisters in the affected animals with respect to the human Herlitz junctional epidermolysis bullosa patients correlates with the protection provided by hair. Our observations suggest that the affected foals are a convenient source of epithelial cells from tissues that cannot be obtained from human junctional epidermolysis bullosa patients, and imply that hairless strains of animals with recessive skin disorders would be the best models for in vivo gene therapy approaches to skin blistering diseases.

Animals↗