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PubMed2002

Cytokine pattern in blister fluid and sera of patients with pemphigus.

BACKGROUND: Pemphigus is a chronic auto-immune blistering disease with four main variants, i.e. pemphigus vulgaris (PV), foliaceus (PF), erythematosus (PE) and vegetans. The common histological feature of this disease is acantholysis. OBJECTIVE: The aim of this study was to compare levels of some cytokines in blister fluid and sera of patients with pemphigus, using as control blister fluid of patients with bullous pemphigoid (BP) and bullous contact dermatitis (BCD). METHODS: Using an immuno-enzymatic assay (ELISA), we tested 16 sera and 6 blister fluids of patients with various forms of pemphigus (13 with PV, 1 with PF, 2 with PE), the sera of 16 healthy control subjects, 5 blister fluids of patients with BP and 5 blister fluids of patients with BCD, for the presence of some cytokines (IL-10, IL-8 and IFN-gamma). Intercellular antibodies were searched for and titred; desmoglein 1 and 3 antibody levels were independently evaluated to compare them with the severity of both cutaneous and oral involvement. RESULTS: The levels of IL-10 in the sera of patients with pemphigus were below the detection limits. IL-8 was significantly increased only in 4 samples of sera from pemphigus patients compared with controls, while IFN-gamma was detected at low levels in almost all patients compared with sera of controls. The cytokine levels in blister fluid of patients with pemphigus were significantly higher than in the sera. There was a difference between the expression of cytokines in blister fluid of control patients with BP and BCD compared with those of pemphigus patients. CONCLUSION: This report discusses the anti-inflammatory role played by IL-10 in the chronic form of pemphigus and the hypothesis of a possible role of IL-8 in neutrophil and lymphocyte-monocyte recruitment.

Adolescent↗
PubMed1989

The existence of a platelet-activating factor (Paf-acether)-like substance in blister fluid derived from patients with bullous pemphigoid as demonstrated by human platelet aggregation techniques.

We investigated the presence of a platelet-activating factor-like substance in blister fluid obtained from patients with bullous pemphigoid. Human platelet aggregation activity was present in blister fluids obtained from 4 out of 6 patients with bullous pemphigoid and in blister fluids obtained from 3 cases with contact dermatitis. Platelet aggregation activity of pemphigoid blister fluids was inhibited by pretreatment with lyso-PAF, which is a precursor metabolite of platelet activating factor. The activity of blister fluids obtained from patients suffering from contact dermatitis was not inhibited by precursor. Blister fluid obtained from burn lesions and blisters of normal skin induced in the suction revealed no platelet aggregation activity. These results suggest that platelet-activating factor or a similar substance is present in the blister fluid obtained from suffered bullous pemphigoid.

Adult↗
PubMed1989

[Leukotriene (LT) B4 and LTC4 in the blister fluid of bullous pemphigoid].

Leukotriene (LT) B4 and LTC4 content in the blister fluid of patients with bullous pemphigoid (BP) was determined by radioimmunoassay. In samples from BP patients, the former (3.80 +/- 1.99 ng/ml, n = 7) significantly exceeded the amount noted in second degree burn patients (0.93 +/- 0.82 ng/ml, n = 5) and that in the fluid of suction blisters produced on normal human skin (0.95 +/- 0.21 ng/ml, n = 2). The immunoreactive LTC4 (i-LTC4) content in the blister fluid of BP (3.63 +/- 1.13 ng/ml, n = 6) was not statistically significantly different from that observed in second degree burns (2.09 +/- 1.66, n = 5), but more than that in suction blisters on normal skin (0.86 +/- 0.20, n = 2). The chemotactic activity of BP blister fluid toward polymorphonuclear leukocyte (PMNL) of BP blister fluid was determined by injecting them into guinea pig skin and observing the time course of the increase in the number of infiltrated PMNL. Neutrophils and eosinophils had accumulated in the dermis at 3 hours and at 6 hours; the numbers of each had increased remarkably. Methanol-eluted samples of blister fluid used for the radio-immunoassay showed the same chemotactic activity. In addition, the dermis developed marked edematous changes. On the basis of the results presented above, LTB4 and i-LTC4 appear to be generated in skin lesions of BP, the former attracting PMNL to the dermis and the latter causing exudation of blister fluid. Both compounds directly enhance the exacerbation of BP skin lesions.

Animals↗
PubMed1998

Genetic interactions and cell behaviour in blistered mutants during proliferation and differentiation of the Drosophila wing.

In this work, we analyse the blistered function in wing vein development by studying genetic mosaics of mutant cells, genetic interactions with other genes affecting vein development and blistered expression in several mutant backgrounds. blistered encodes for a nuclear protein homologous to the mammalian Serum Response Factor and is expressed in presumptive intervein cells of third larval instar and pupal wing discs. Clones of blistered mutant cells proliferate normally but tend to grow along veins and always differentiate as vein tissue. These observations indicate that vein-determined wing cells show a particular behaviour that is responsible for their allocation to vein regions. We observe strong genetic interactions between blistered, veinlet and genes of the Ras signaling cascade. During disc proliferation, blistered expression is under the control of the Ras signal transduction pathway, but its expression is independent of veinlet. During the pupal period, blistered and veinlet expression become interdependent and mutually exclusive. These results link the activity of the Ras pathway to the process of early determination of intervein cells, by the transcriptional control of the blistered nuclear factor.

Animals↗
PubMed2002

Soluble interleukin 2 receptor and interleukin 1alpha in toxic epidermal necrolysis: a comparative analysis of serum and blister fluid samples.

BACKGROUND: Toxic epidermal necrolysis (TEN) is a rare but severe adverse drug disease, characterized by extensive skin and mucosal detachment with participation of different immunoinflammatory pathways, in particular with early participation of activated CD8+ T lymphocytes. OBJECTIVE: To further study the potential role of T lymphocytes in the early phase of keratinocyte necrosis. DESIGN: Prospective study. SETTING: University hospitals. PATIENTS: Thirteen patients with clinical and histopathologic criteria of TEN and 6 patients with second-degree burns. MAIN OUTCOME MEASURES: Measurement of soluble interleukin (IL) 2 receptor (sIL-2R) and IL-1alpha in serum samples and fluid of recent blisters. RESULTS: In the blister fluid of patients with TEN, we found significantly higher levels of sIL-2R than in patients with burns, whereas IL-1alpha levels were higher in the blister fluid of burned patients. No significant differences were found in serum samples of patients with TEN and burns, in either sIL-2R or IL-1alpha. In TEN we also found significantly higher levels of sIL-2R in the blister fluid compared with serum samples, pointing to a predominantly local production contrasting with the low concentration of sIL-2R in the blister fluid of burned patients. CONCLUSIONS: Our findings of elevated sIL-2R levels in blister fluid of patients with TEN are probably related to a local down-regulation of an immunologically mediated cytotoxic reaction and further support the involvement of activated T lymphocytes in the early blisters of TEN.

Adolescent↗
PubMed1991

Distribution kinetics of netilmicin in human blister fluid: effect of renal impairment.

The pharmacokinetics of netilmicin in plasma and blister fluid were compared in 10 healthy volunteers with normal renal function and 10 voluntary patients with varying degrees of renal impairment. Netilmicin kinetics in plasma were characterized by an open two-compartment kinetic model. For the study of the kinetics of the antibiotic in blister fluid a specific kinetic distribution model was employed. In the healthy volunteers the plasma kinetics of netilmicin showed a behaviour similar to that of other aminoglycoside antibiotics, although a high blister/plasma partition coefficient was obtained, with a mean value of 4.27 +/- 1.65. The transfer of netilmicin in blister fluid governed by the constants K1b/Vb and Kbl had mean values of 0.19 +/- 0.09 h-1 l-1 and 0.25 +/- 0.10 h-1, respectively. In the patients with renal impairment plasma and blister fluid antibiotic levels showed a progressive accumulation. In these patients the affinity of netilmicin for blister fluid was significantly altered. The blister/plasma partition coefficient in this group of patients had a mean value of 2.65 +/- 1.33; the decrease being statistically significant (p = 0.023) with respect to the value obtained in the healthy volunteers. Similarly, the exit constant of netilmicin from blister fluid (Kb1) showed a statistically significant increase (p = 0.035) in the patients with renal impairment. The findings point to a loss of affinity of netilmicin for blister fluid as a result of renal impairment. Linear and logarithmic relationships were established between some pharmacokinetic parameters and creatinine clearance. Dosage schedules are proposed for netilmicin in patients with renal impairment.

Adult↗
PubMed1985

Neurogenic inflammation in skin blisters.

The histology of blisters induced in rats using dry-ice or vacuum applied to the hind limb footpad has been examined by light microscopy and indirect fluorescence immunohistochemistry for substance P like immunoreactivity. The effects of sciatic nerve stimulation on plasma extravasation as determined by estimation of Evans Blue content of blister fluid has been examined. Plasma extravasation occurs in both blister types following nerve stimulation. Plasma extravasation may be induced by substance P released from nerve terminals in the blister wall. Vacuum induced blisters have less tissue and nerve damage and therefore less of an inflammatory response, but plasma extravasation is greater in thermal blisters. Blisters serve as a useful model for the study of neurogenic inflammation.

Animals↗
PubMed1995

Influence of aging, localization, glucocorticoids and isotretinoin on matrix metalloproteases 2 (MMP-2) and 9 (MMP-9) in suction blister fluids.

The expression of two matrix metalloproteases (MMPs), 72 kDa gelatinase (MMP-2) and 92 kDa gelatinase (MMP-9), was studied in suction blister fluids and serum using a zymographic method. Both of the enzymes were detectable in blister fluid and serum, but their expression varied. The MMP-2 levels in serum were only about half of the levels in blister fluid, while the level of MMP-9 was about the same in both serum and blister fluid. The overall level of MMP-2 in serum was much lower than the level of MMP-9 which was possibly derived from inflammatory cells. In subjects aged from 20 to 86 years, no marked changes were seen in the serum or blister fluid gelatinases. Interestingly, body site affected the level of MMP-9. The lowest level was recorded in fluid from blisters on the lower leg, in which the level was only about 19% of the level in fluid from blisters on the abdomen. The levels of MMP-2 and MMP-9 were not decreased by pharmacological doses of systemic prednisone or isotretinoin, which indicates that systemic glucocorticoid or retinoid treatment does not affect the basal levels of MMP-2 and MMP-9.

Adult↗
PubMed1992

Penetration of tinidazole into skin blister fluid following its oral administration.

Plasma and skin blister fluid concentrations of tinidazole following a single oral dose of 2 g drug, and after multiple doses of 0.25 g every 12 h, were determined. Skin blisters were produced by direct application of 0.25% cantharidin ointment to the skin. The maximum concentration in plasma of about 36 mg.l-1 was observed after about 2 h, whereas in skin blister fluid the peak occurred after about 6 h and was 30 mg.l-1. The half-life in plasma was slightly shorter than in blister fluid at 17 and 19 h, respectively, but the difference was not significant. The penetration of tinidazole into cantharidin-induced skin blister fluid, defined according to Wise as the ratio of the AUCs in blister fluid and plasma was 1.00. During routine treatment with tinidazole (0.25 g every 12 h), the concentrations in plasma and blister fluid collected before and 3 h after the morning dose exceeded the minimal inhibitory concentrations for susceptible pathogens. The results provide a pharmacokinetic basis for the proven efficacy of tinidazole in the treatment of protozoal and anaerobic infections.

Administration, Oral↗
PubMed1995

Effects of an antiperspirant with emollients on foot-sweat accumulation and blister formation while walking in the heat.

BACKGROUND: Friction blisters are a common injury in sports activities and military operations. Blisters can compromise performance, so it is important to devise preventive strategies to reduce these injuries. OBJECTIVE: This study investigated the influence of an antiperspirant with emollient additives on frequency and severity of friction blisters, hot spots, and irritant dermatitis. METHODS: Twenty-three healthy men walked on a treadmill (1.39 m/sec, 1% grade) in a warm environment (28 degrees C, 25% relative humidity) carrying a total mass of 21 +/- 1 kg. For 4 consecutive days before the walk, the subjects' feet were treated with either (1) an antiperspirant (20% aluminum zirconium tetrachlorohydrex glycine concentration plus water) with emollient additives, (2) emollient additives alone (placebo control), or (3) nothing (nontreated). In two separate trials (1 month apart) each participant received the antiperspirant treatment and both control treatments (emollient [placebo] and no treatment). RESULTS: No differences were seen among treatment conditions for sweat accumulation (p = 0.86), blister incidence (p = 0.36), hot spot incidence (p = 0.83), or blister severity (p = 0.31). Irritant dermatitis was not reported in any of the treatment conditions. CONCLUSION: The use of an antiperspirant with emollients reduces irritant dermatitis but does not reduce total foot-sweat accumulation, blister or hot spot incidence, or blister severity.

Adult↗
PubMed2004

Breast blisters in groups of slow-growing broilers in relation to strain and the availability and use of perches.

1. The relationship between perching behaviour, availability of perches and the incidence and severity of breast blisters in broilers was investigated together with a comparison between two slow-growing broiler strains. 2. Sixteen single strain groups (n = 60) of Labresse and i657 broilers were subjected throughout the experiment to one of three perch availabilities: 15 cm per bird (Labresse and i657), 7.5 cm per bird (i657 only), and 0 cm per bird (i657 only) with 4 replicates per treatment. The birds were housed indoors from one day old, and at 43 d of age 52 birds from each group were moved to outdoor housing facilities with access to grass-covered outdoor areas until slaughter at 84 d of age. 3. The use of perches was monitored via video recordings throughout the experimental period. The severity of breast blisters was recorded on a scale from 0 to 2 at slaughter. 4. Groups of i657 with 15 cm perch per bird used these more than groups with 7.5 cm perch per bird (19% vs 8% of birds perching at midnight). A positive association between access to perches and severe breast blisters (score 2) was found in the groups of i657, with odds ratios of 3.1 and 3.4 for 7.5 and 15.0 cm per bird, respectively, relative to the no-perch treatment. 5. Labresse were more likely to develop breast blisters than i657 (odds ratio 3.5), but used the perches less (0.1 vs 9.5 birds perching at midnight) and weighed less (2011 g vs 2246 g) than i657. 6. Males had a higher incidence of breast blisters than females (odds ratio 12.2), and this was most prominent in the Labresse strain (odds ratio 40.0). 7. In some broiler strains access to perches may be associated with an increase in the occurrence of severe breast blisters, but strain and sex of broiler chickens appear to have a much larger influence than access to perches on the incidence of breast blisters.

Animals↗
PubMed2004

Measurement of cytokine expression and Langerhans cell migration in human skin following suction blister formation.

Contact allergen-induced migration of epidermal Langerhans cells (LCs) to draining lymph nodes is dependent upon receipt by LCs of at least two cytokine signals provided by tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta. It has been reported previously that intradermal injection of healthy human volunteers with homologous TNF-alpha or IL-1beta each induces a significant reduction in LC frequency, as measured in epidermal sheets prepared from 6-mm punch biopsies. In the current experiments, we have compared the frequency of LCs in punch biopsies with those obtained concurrently in epidermal sheets from the roofs of suction blisters isolated from the sun-protected buttock skin of healthy adult volunteers. There was a significant, approximately 30%, reduction in CD1a(+) LC numbers in suction blister roofs compared with punch biopsies. Injection of homologous recombinant IL-1beta, a stimulus that provokes measurable epidermal LC mobilization in punch biopsy sites, failed to provoke further LC migration in suction blister sites. These data suggest that the mechanical trauma to the skin caused by the creation of suction blisters provokes the degree of cutaneous inflammation necessary for LC mobilization. The responsive cells (only a proportion of resident LCs, approximately 30%) have already migrated, thus addition of an exogenous cytokine signal (IL-1beta) is without further effect. It is not possible therefore to measure the regulation of LC mobilization by exogenous cytokines in suction blister roofs. However, this technique provides an opportunity to profile induced changes in the cutaneous cytokine environment, with cytokine expression measured by a multiple cytokine array system. Using this technique, intradermal injection of IL-1beta was found to cause a marked upregulation of proinflammatory cytokines including TNF-alpha, IL-6, IL-8, monocyte chemotactic protein-1 (MCP-1) and the anti-inflammatory cytokine IL-10 in fluid from suction blisters raised at the site of injection. In conclusion, the suction blister technique appears to be a powerful tool for measurement of induced changes in cutaneous cytokines.

Adult↗
PubMed1982

Basement membrane components and galactosylhydroxylysyl glucosyltransferase in suction blisters of human skin.

Basement membrane components and collagen biosynthesis were studied in suction blisters in human skin. The basement membrane components were characterized by immunofluorescence using specific antibodies to type IV collagen, laminin and fibronectin, and collagen biosynthesis was studied by assaying galactosylhydroxylysyl glucosylatransferase. In suction blisters, the separation of epidermis and dermis occurred above the lamina lucida, indicating that the basement membrane, composed of lamina lucida and lamina densa, forms a mechanically strong entity. During the regeneration phase of blisters, type Iv collagen and laminin were not observed in the old epidermal blister roof. This indicates that keratinocytes when separated from the underlying basement membrane or connective tissue do not synthesize laminin or type IV collagen. Galactosylhydroxylysyl glucosyltransferase activity could be demonstrated in blister fluid and was about the same as in serum when expressed on the basis of protein in fresh blisters. It increased by 2-3 fold during the repair of blisters, indicating that there was local production of this enzyme. Further studies revealed that pure epidermis contained galactosylyhdroxylysyl glucosyltransferase and hydroxyprolineand this suggests that epidermis may synthesize some collagen type which, according to these studies, is not type IV (basement memebrane) collagen.

Adult↗
PubMed1994

Increased serum and blister fluid levels of creatine kinase in patients with toxic epidermal necrolysis.

Keratinocytes have recently been reported to contain creatine kinase (CK) of brain-type isoenzyme. The aim of this study was to investigate whether necrosis of keratinocytes induced raised CK levels in toxic epidermal necrolysis (TEN). The serum and blister fluid levels of creatine kinase and its isoenzymes [muscular-type (MM), brain-type (BB), myocardial-type (MB)] were measured in 40 patients with TEN, 10 patients with other bullous dermatoses, and in suction blisters in five controls. The mean serum CK was significantly higher in TEN patients than in patients with other bullous dermatoses (mean +/- SD: 480 +/- 535 U/l vs. 107 +/- 44 U/l, P < 0.05). The MM-isoenzyme was predominant (94%). A positive correlation was found between the level of the serum CK and the percentage of body surface area (BSA) involved (r = 0.49, P < 0.001). The mean blister CK was significantly higher in TEN patients than in patients with other bullous dermatoses or controls (mean +/- SD: 728 +/- 437 U/l vs. 310 +/- 244 U/l and 268 +/- 194 U/l, respectively, P < 0.02). The isoenzyme distribution of blister CK in TEN patients was: 76.8% MM, 18.1% MB and 5% BB. Although a significant part of blister CK comigrating with CK-MB, after preincubation with protein A-Sepharose, appeared to be CK-BB/IgG complex, the CK-BB fraction constituted less than 25% of blister CK. Therefore, the CK present in increased amounts in serum and blister fluid in TEN was not directly produced by keratinocytes.

Adolescent↗
PubMed1995

Suction-induced blister fluid penetration of cefdinir in healthy volunteers following ascending oral doses.

The pharmacokinetics and suction-induced blister fluid penetration of cefdinir following single oral administrations of 200, 300, 400, and 600 mg were studied in 16 healthy young male volunteers according to a Latin square design. Plasma, blister, and urine samples were assayed by high-pressure liquid chromatography. We observed a nonlinear relationship (P = 0.02) between the dose and the maximum concentration in plasma as well as between the dose and the area under the concentration-time curve (AUC) in plasma (P < 0.001), which may be indicative of a limited absorption process. This resulted in a lower AUC value than expected as well as a smaller fraction of cefdinir excreted unchanged at a dose of 600 mg. Renal clearance decreased with increasing doses (P < 0.006; analysis of variance with the Latin square design and Games-Howell procedure). Maximal cefdinir concentrations in blister fluid were delayed compared with concentrations in plasma. Blister fluid penetration measured by the ratio of the AUC in blister fluid to the AUC in plasma was extensive (92.4 to 108.4%). Cefdinir concentrations in blister fluid remained equal to or higher than the concentrations in plasma from 6 to 12 h following cefdinir administration. On the basis of the concentrations in blister fluid and the in vitro MIC data, we estimated that cefdinir at 200 to 400 mg administered twice daily would be adequate to treat uncomplicated skin infections caused by Streptococcus pyogenes. Seven volunteers experienced episodes of light-to-moderate diarrhea. These adverse events occurred irrespective of dose.

Adult↗
PubMedDate not supplied

Protein concentration of subcutaneous interstitial fluid in the human leg. A comparison between the wick technique and the blister suction technique.

The wick technique and the blister suction technique are the most common methods for sampling of subcutaneous interstitial tissue fluid in man. The blister suction technique has the advantage of being less invasive than the wick technique, but the reliability of this method is still controversial. The aim of this study was to evaluate whether the simpler blister suction technique using large (8 mm) blisters could replace the wick technique in the investigation of patients with postreconstructive leg edema. Fifteen patients with ipsilateral leg edema following infrainguinal bypass surgery for lower limb atherosclerosis were investigated. The two different fluid sampling techniques were applied simultaneously on both legs. The concentration of total protein and albumin as well as colloid osmotic pressure of the subcutaneous interstitial tissue fluid in the leg were measured in all fluid samples. Agreement analysis was applied to compare the two methods, while the correspondence between the methods was estimated with linear regression analysis. The agreement index was found to be positive for all variables from the operated as well as from the contralateral control limb. Furthermore, all values were within the agreement limit. The best agreement between the two methods was found for colloid osmotic pressure on the operated side. According to the equation of linear regression, there was a slight overestimation of the wick values compared to the observed blister values. In conclusion, there was a good methodological agreement between the blister suction technique and the wick technique. The less invasive blister suction technique should be regarded as the method of choice for the investigation of subcutaneous interstitial tissue fluid in patients with postreconstructive leg edema.

Aged↗
PubMed1999

Reassessment of the suction blister model of wound healing: introduction of a new higher pressure device.

BACKGROUND: Negative pressure suction blisters have been used as a reproducible and minimally invasive in vivo model of wound healing in human subjects. Despite advantages over other available methods, this technology has not been employed widely in wound healing research. OBJECTIVE: To evaluate the efficacy of a higher pressure suction blister system as an in vivo human model for wound healing and to demonstrate the superiority of this method over previously used suction blister models of wound healing. METHODS: Four 5-mm-diameter suction blisters were induced on the medial upper arm of 18 healthy men and women using a negative pressure of 508 mmHg. Blister roofs were removed, exposing the underlying dermis. The time required for blister formation, the uniformity of wounds, and the degree of patient tolerance were assessed. The ability to monitor clinical re-epithelialization over 7 days was also evaluated by a team of investigators. RESULTS: Four partial-thickness wounds were produced on each subject in 25-45 min. Wound diameter and morphology were uniform in all subjects. Volunteers tolerated the procedure without complaints of pain or discomfort. Progressive, re-epithelialization was observed daily starting 24 h post-wounding. CONCLUSIONS: This higher pressure suction blister system is an effective model for wound healing. Compared to reported methods, it is a better tolerated, more reliable, and more efficient approach to studying in vivo wound repair in human subjects. It is especially well suited for screening the wound healing potential of new pharmacologic agents.

Adult↗
PubMed1989

Mediators of inflammation in blister fluids from patients with pemphigus vulgaris and bullous pemphigoid.

Blister fluids from 39 previously untreated patients with pemphigus vulgaris (PV) and 28 patients with bullous pemphigoid (BP) were tested for the presence of inflammatory mediators. Blister fluids from patients with PV and BP showed interleukin 1 (IL-1)- and interleukin 2 (IL-2)-like activity and contained prostaglandin (PG) E2, F1 alpha, and F2 alpha, thromboxane B2 (TB2), leukotriene (LT) B4 and C4, serine esterases, and proteolysis inhibitors. Pemphigus blister fluid was distinguished by high IL-1-like activity, proteolytic activity, and high concentrations of TB2 and LTB4, while pemphigoid blister fluid was characterized by higher IL-2-like activity, antiprotease activity, and high PGE2 concentration. We also determined that the content of mediators of inflammation varied depending on the duration of blister development. In both PV and BP, the initially high IL-1-like activity, amount of serine proteases, and concentrations of PGF2 alpha, TB2, LTB4, and LTC4 decreased by the fifth day of blister existence, whereas antiprotease activity as well as PGE2 and PGF1 alpha concentrations gradually increased as blisters developed. These findings suggest certain differences in the character of the bullous inflammatory process in pemphigus and pemphigoid.

Adult↗