sodium-dodecyl-sulfate and Edema

Investigations of pruritogenic substances in humans have involved intradermal injections in normal skin; itching of inflamed skin has been little studied.. To develop an itch model with provocation of itch in experimentally inflamed skin as well as in normal skin, using subjects as self-controls.. In 32 non-atopic volunteers aged 21-30 years, the skin of five selected test sites on one volar forearm was pretreated for 24 h with large Finn chambers containing 1% sodium lauryl sulphate (SLS) used as a standard contact irritant to induce inflammation. Twenty microlitres of different pruritogenic substances [histamine, substance P, neurokinin A, neurokinin B, trypsin, platelet-activating factor (PAF) and serotonin] and saline as control were injected intradermally into the inflamed test sites and in corresponding non-treated sites on the opposite forearm. The test individuals scored itch intensity on a visual analogue scale for 20 min, and weal area was then measured. :. Histamine and substance P induced itch in both normal and inflamed skin compared with a saline reference. Neurokinin A, trypsin, PAF and serotonin only elicited itch in normal skin, and neurokinin B neither elicited itch in normal skin nor in inflamed skin. Itch was induced in normal and SLS-inflamed skin to a similar magnitude. However, weal area after histamine was significantly (P < 0.001) larger in inflamed skin when compared with normal skin.. Histamine and substance P elicited itch to the same degree in normal skin and inflamed skin pretreated with SLS despite a stronger weal response in inflamed skin. Mediators present in inflamed skin did not potentiate itch, a c-fibre-mediated neuronal response. The weal reaction is based on enhanced vascular permeability (protein extravasation). A greater skin perfusion in inflamed skin may therefore have increased the weal size. We propose an experimental model in humans for testing of itch involving both normal and inflamed skin. The model has the potential for use in evaluating new topical and systemic treatments of itch.

Topics: Adult; Dermatitis, Contact; Double-Blind Method; Edema; Erythema; Female; Histamine; Humans; Injections, Intradermal; Male; Neuropeptides; Pain; Pain Measurement; Pruritus; Severity of Illness Index; Sodium Dodecyl Sulfate

Variable types of skin irritation were induced in 8 human female volunteers, ranging from subclinical to visible erythema with slight oedema. Skin reactions were graded clinically and objectively using transepidermal water loss (TEWL), laser Doppler flowmetry (LDF) and improved reflectance spectroscopy. This last technique enables separation of in vivo erythemas into relatively deoxygenated (venous--deoxy hem) and oxygenated (arterial--oxy hem) haemoglobin components. Compared to uninvolved skin, an empty patch increased oxy hem by 197% +/- 121% (p < 0.05). Exposure to vehicles also changes skin biophysics. At sodium lauryl sulfate (SLS) and hydrochloric acid (HCl) exposed sites, a linear correlation between concentration and oxy hem, LDF and TEWL was found. These chemicals predominantly increased TEWL values. Nonanoic acid (NON) and imipramine (IMI) also raised oxy hem, LDF and TEWL values linearly at increasing concentrations. Although IMI 2.5% clinically was graded as a type ++ response, no significant increase in TEWL was found. The improved reflectance spectroscopic technique proved valid in skin irritation studies, with a higher sensitivity than laser Doppler flowmetry, and allowed irritant vascular reactions to be discriminated into arterial and venous components. Furthermore, our observations clearly demonstrate that clinically indistinguishable skin irritation reactions induce significantly different changes in barrier function (disruption) and vascular status.

Topics: Adult; Biomedical Engineering; Dermatitis, Irritant; Edema; Erythema; Fatty Acids; Female; Hemoglobins; Humans; Hydrochloric Acid; Imipramine; Irritants; Laser-Doppler Flowmetry; Middle Aged; Oxyhemoglobins; Skin; Sodium Dodecyl Sulfate; Spectrophotometry; Water Loss, Insensible

1. The limitations of the Draize rabbit skin irritation test for hazard evaluation for man are widely documented. Nevertheless it remains the prescribed method for determining acute skin irritations hazard. 2. While the use of human testing for risk assessment of irritants is well established, the use of predictive testing in man for hazard identification has not been explored widely, and this is the object of the research programme. 3. The experiment described in this report evaluates the sensitivity of four patch testing systems (Finn chamber, Hill Top patch, Van der Bend chamber, and Webril patch) using a total of six irritant substances. 4. Following preliminary range-finding experiments, test materials were applied to the upper outer arm for up to 4 h. Assessments were performed immediately after patch removal and at 1, 24, 48, and 72 h. 5. Webril and Hill Top patches generated the greatest levels of response, although responses with Finn and Van der Bend were observed. Hill Top patches are recommended for future development work. 6. The use of very small preliminary panels to predict the effects in larger panels using different volunteers was only of limited value as each volunteer was found to have different irritant thresholds.

Topics: Acetates; Acetic Acid; Adolescent; Adult; Aged; Cetrimonium Compounds; Cyclohexenes; Edema; Erythema; Ethanolamines; Female; Humans; Irritants; Limonene; Male; Middle Aged; Patch Tests; Risk Assessment; Sodium Dodecyl Sulfate; Sodium Hydroxide; Terpenes; Time Factors

Animal testing causes ethical problems and in view of EU regulations (e.g. EU-Guideline (76/768/EEC, February 2003)) or REACH the development of reliable in vitro assays has become even more important. Up to now, we use the modified local lymph node assay (IMDS) for toxicological hazard identification of sensitizing and irritant properties of chemicals in accordance with OECD Guideline 429. In this study, we investigated whether analyses of cell signaling pathways can provide a methodology for the detection of sensitizing compounds in vitro. Murine and human skin explants as well as reconstituted skin models (epidermal model EST-1000 and full-thickness model AST-2000) were exposed to sensitizing (oxazolone and DNFB) or irritant compounds (SDS and TritonX-100). Phosphorylation of MAP-kinases (p38, ERK1/2 and JNK1/2), STAT1 and PLCgamma were determined by cytometric bead array (CBA). In skin explants, all three MAP-kinases were exclusively activated after exposure to sensitizing compounds. For the reconstituted skin models phosphorylations of p38 and JNK1/2 were obtained after stimulation with allergens, whereas treatments with irritant compounds led to ERK1/2 activation. Activation of PLCgamma and STAT1 were never detected. In conclusion, MAP-kinase activation provides a promising in vitro tool for the discrimination between sensitizers and irritants.

Topics: Allergens; Animals; Dinitrofluorobenzene; Dose-Response Relationship, Drug; Edema; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Irritants; JNK Mitogen-Activated Protein Kinases; Lymph Nodes; Membranes, Artificial; Mice; Mice, Hairless; Mice, Inbred C3H; Octoxynol; Oxazolone; p38 Mitogen-Activated Protein Kinases; Phospholipase C gamma; Phosphorylation; Signal Transduction; Skin; Skin Irritancy Tests; Sodium Dodecyl Sulfate; STAT1 Transcription Factor; Tissue Culture Techniques

Easily applicable water-specific instruments measuring local oedema in skin are not available. The aim of this study is to demonstrate quantitative assessment of skin oedema with the dielectric technique by measuring increase of skin water content related to sodium lauryl sulphate (SLS)-induced irritant contact dermatitis.. Irritant skin reaction and resulting oedema were induced by an irritant patch test on volar forearms in 12 healthy volunteers with the application of 1% SLS for 6 h. After occlusion the volunteers were divided into two groups: the patch test site of group I (six volunteers) received no treatment other than a base cream for the skin reaction, while for group II (six volunteers) a strong corticosteroid (clobetasol propionate) was applied on the irritant skin. During a follow-up of 72 h, erythema was scored visually, and irritant-induced oedema was measured with a novel water-specific instrument MoistureMeter-D.. In the untreated irritant skin, a maximum increase of 45% in skin water content was found at 10 h postocclusion and water content was still elevated at 72 h. With these persons, the degree of oedema agreed well with the ultrasound-measured skin thickness (P=0.053). In the corticosteroid-treated skin, an increase of 8% in water content was measured during 72 h but there was no correlation between oedema and skin thickness. There was no correlation between erythema and oedema in untreated or corticosteroid-treated skin.. The new instrument can easily be applied for noninvasive quantitative evaluation of local oedema and fluid retention in irritant-exposed skin.

Topics: Adult; Anti-Inflammatory Agents; Body Water; Clobetasol; Dermatitis, Irritant; Edema; Electrochemistry; Erythema; Humans; Skin; Skin Diseases; Sodium Dodecyl Sulfate; Surface-Active Agents; Ultrasonography

In the evaluation of the severity of skin inflammations, visual scoring system is widely being used as a subjective method. However, it is well known that interobserver variations occur even between the interpretations of experienced dermatologists.. To develop a new objective and quantitative method for the evaluation of skin surface contours and for the assessment of severity in SLS-induced edema, a new investigatory tool based on the concept of "stereoimaging" was studied. Differences between binocular images were used by a stereoimage optical topometer (SOT) system capable of calculating skin surface topographic information, which was then used to produce a three-dimensional image.. We compared the results obtained by SOT with a visual scoring system after applying sodium lauryl sulfate (SLS) to skin. The degree of edema was assessed qualitatively by visual scoring (0: none, 1: mild edema, 2: moderate edema, 3: severe edema). To quantify the severity, five three-dimensional parameters (S(a), S(z), SL, SA and SV) were used in the SOT analysis.. The means of these five parameters increased significantly in visual grade 1+ compared to visual grade 0 and the means of the five parameters decreased significantly and proportionally as the visual grades increased from 1+ to 3+. A highly significant correlation was found to exist between the visual scoring results and the five SOT parameters.. SOT can be used to evaluate the severity of SLS-induced edema objectively and can be extensively applied to evaluate the degree of severity in other inflammatory skin conditions.

Topics: Adult; Edema; Humans; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Male; Middle Aged; Severity of Illness Index; Skin; Skin Diseases; Sodium Dodecyl Sulfate

These studies were conducted to investigate the potential use of a flow cytometric analysis method for the identification and differentiation of chemicals with the capacity to induce irritation, IgE- or T cell-mediated hypersensitivity responses. An initial study investigated the ability of equally sensitizing concentrations (determined by local lymph node assay) of IgE-mediated (Toluene Diisocyanate-TDI) and T cell-mediated (Dinitrofluorobenzene-DNFB) allergens to differentially modulate the IgE+B220+ population in the lymph nodes draining the dermal exposure site. Sodium lauryl sulfate (SLS) was also tested as a nonsensitizing irritant control. Female B6C3F1 mice were dermally exposed once daily for 4 consecutive days, with the optimum time point for analysis determined by examining the IgE+B220+ population 8, 10, and 12 days post-initial chemical exposure. At the peak time point, day 10, the IgE+B220+ population was significantly elevated in TDI (41%), while moderately elevated in DNFB (18%) exposed animals when compared to the vehicle (0.8%), and remained unchanged in SLS (2.2%) exposed animals when compared to the ethanol control (2.5%). Experiments in our laboratory and others have demonstrated that the draining lymph node B220+ population becomes significantly elevated following exposure to allergens (IgE- and T cell-mediated), not irritants, allowing for their differentiation. An existing mouse ear swelling assay was used to identify chemical irritants. Therefore, using the endpoints of percent ear swelling, percent B220+ cells, and percent IgE+B220+ cells, a combined irritancy/phenotypic analysis assay was developed and tested with tetradecane (irritant), toluene diisocyanate, trimellitic anhydride (IgE-mediated allergens), benzalkonium chloride, dinitrofluorobenzene, oxazolone, and dinitrochlorobenzene (T cell-mediated allergens) over a range of concentrations. Based upon the pattern of response observed, a paradigm was developed for continued evaluation: Irritant exposure will result in significant ear swelling without altering the B220+ or IgE+B220+ populations. Exposure to sensitizers (IgE-mediated or T cell-mediated) will increase the B220+ population and the percent ear swelling will remain unchanged or will significantly increase, depending on the irritancy capacity of the chemical. Both the IgE+B220+ and B220+ populations will become elevated at the same test concentration following exposure to IgE-mediated, hypersensitivity inducing alle

Topics: Administration, Cutaneous; Animals; Dermatitis, Contact; Dinitrofluorobenzene; Dose-Response Relationship, Drug; Edema; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin G; Irritants; Lymph Nodes; Mice; Phenotype; Sodium Dodecyl Sulfate; Specific Pathogen-Free Organisms; T-Lymphocytes; Toluene 2,4-Diisocyanate

Pit viper venoms contain multiple proteinases which cause considerable damage in tissues and systemic effects after envenomation. A proteinase, kallikrein-like enzyme, belonging to the serine group must play a very important role on systemic effects. The corresponding enzyme from Lachesis muta rhombeata venom was purified to homogeneity by a combination of isoelectrofocusing fractionation followed by one step of gel filtration HPLC. The enzyme focused with pI 5.0-6.5, it had a molecular mass of 32 kDa by gel filtration HPLC, had edematogenic activity, and induced a hypotensic effect in anesthetized rats. It exhibited strong N-alpha-tosyl-L-Arg methyl esterase (955.38 units/mg) and N-Bz-DL-Arg-pNA amidolytic (233.02 units/mg) activities, hydrolyzed tripeptide nitroanilide derivatives weakly or not at all, and cleaved selectively the A-alpha and B-beta chains of fibrinogen, apparently leaving the Y-chain unaffected. The 30 N-terminal amino acid sequence of the L. m. rhombeata protein showed greatest identity (74% in 26 amino acids) with Crotalus viridis kallikrein-like protein, but significant similarities in sequence were observed with enzymes from other snake venoms and pig pancreatic kallikrein.

Topics: Amino Acid Sequence; Animals; Crotalid Venoms; Dose-Response Relationship, Drug; Edema; Electrophoresis, Polyacrylamide Gel; Endopeptidases; Hypotension; Kallikreins; Male; Mice; Molecular Sequence Data; Molecular Weight; Peptide Fragments; Rats; Rats, Wistar; Sequence Analysis; Sequence Homology, Amino Acid; Snakes; Sodium Dodecyl Sulfate; Substrate Specificity

Topics: Animals; Body Water; Data Display; Dermatitis, Irritant; Edema; Erythema; Guinea Pigs; Laser-Doppler Flowmetry; Patch Tests; Regional Blood Flow; Severity of Illness Index; Skin; Sodium Dodecyl Sulfate; Tretinoin

Topics: Animals; Data Display; Dermatitis, Allergic Contact; Dermatitis, Atopic; Dermatitis, Irritant; Edema; Fatty Acids; Humans; Hydrochloric Acid; Image Processing, Computer-Assisted; Mice; Mice, Hairless; Nickel; Patch Tests; Skin; Sodium Dodecyl Sulfate; Sodium Hydroxide; Ultrasonography

Topics: Acetylcysteine; Antioxidants; Dermatitis, Irritant; Dimethyl Sulfoxide; Edema; Erythema; Humans; Ointments; Sodium Dodecyl Sulfate; Tumor Necrosis Factor-alpha

Vipera berus is widely distributed throughout the northern part of Europe and Asia. Characterization of several toxic effects of its venom in the mouse, as well as of in vitro enzymatic activities was performed. Vipera berus venom displayed in vitro proteolytic, fibrinolytic, anticoagulant, and phospholipase A2 activities. The i.p. LD50 of the venom for Swiss mice was 0.86 micrograms/g (95% confidence limits 0.71-1.01 microgram/g). Significant local tissue-damaging effects, including edema, hemorrhage and myonecrosis, were observed. The local edema was characterized by rapid onset, reaching a maximum after 0.5-1 hr, and with dose-dependent persistence. The hemorrhagic potency was measured by a skin test, giving a minimum hemorrhagic dose value of 3.2 micrograms. The venom also induced a moderate local myonecrosis, evidenced by histological evaluation of injected tissue (gastrocnemius), and by biochemical parameters (increase of plasma creatine kinase activity, and decrease of muscle residual MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide)-reducing activity). Characterization of the venom by SDS-polyacrylamide gel electrophoresis revealed 10 (reduced) or 11 (unreduced) main protein bands, which were further analyzed in relation to mol. wt and relative concentration by densitometry. A rabbit antiserum to V. berus venom recognized all main venom bands by immunoblotting. This antiserum cross-reacted to a variable extent with several crotaline venoms, as assessed by enzyme immunoassay.

Topics: Animals; Antibodies; Antigens; Cross Reactions; Crotalid Venoms; Disease Models, Animal; Dose-Response Relationship, Drug; Edema; Electrophoresis, Polyacrylamide Gel; Immunoblotting; Lethal Dose 50; Mice; Peptide Hydrolases; Rabbits; Sodium Dodecyl Sulfate; Viper Venoms

The intensity of the cutaneous response was assessed after application of a standard irritant to increasing areas of normal forearm skin. Twenty subjects were tested to determine the minimal irritant dose (MID) to dilutions of aqueous sodium dodecyl sulphate. Each subject was then treated under occlusion for a period of 24 h with different areas of filter paper (9, 25, 100, 225 and 400 mm2) soaked with the concentration required to give the individual's MID. At 25 and 48 h the degree of erythema was assessed using a 0-4 arbitrary scale, a 10-cm visual analogue scale (VAS) and an erythema meter. Cutaneous blood flow was measured with a laser-Doppler device and cutaneous oedema measured by pulsed A-scan ultrasound. The results at 25 and 48 h were almost identical. Both forms of visual assessment (arbitrary scale and VAS) showed an increase in perceived erythema with increasing area and this was confirmed by the erythema meter. Further area-related changes were noted with both cutaneous blood flow and ultrasound measurements.

Topics: Administration, Cutaneous; Adult; Dose-Response Relationship, Drug; Edema; Erythema; Female; Humans; Irritants; Male; Middle Aged; Patch Tests; Predictive Value of Tests; Skin; Sodium Dodecyl Sulfate; Ultrasonography

An inflammatory reaction was induced in vivo by injection of zymosan into the peritoneal cavity of the rabbit. The inflammatory exudate was found to contain oedema-inducing and neutrophil chemoattractant activity when assayed in rabbit skin in vivo, using 125I-albumin and 111In-neutrophils. This activity was additional to that of complement fragment C5a, which was removed by an affinity gel. Two chemoattractants were isolated by cation-exchange, gel-filtration and reversed-phase h.p.l.c. One of these, which ran as a single band of 6-8 kDa on SDS/PAGE, was subjected to N-terminal sequence analysis without reduction and alkylation of cysteine residues. Positive identification of 28 of the first 31 amino acids revealed a rabbit homologue of interleukin-8 (75% sequence identity with human interleukin-8). The demonstration of interleukin-8 as a major neutrophil chemoattractant in an inflammatory reaction in vivo provides the basis for further investigations into the role of this cytokine in the inflammatory process.

Topics: Amino Acid Sequence; Animals; Chemotactic Factors; Edema; Electrophoresis, Polyacrylamide Gel; Inflammation; Interleukin-8; Molecular Sequence Data; Neutrophils; Peritonitis; Rabbits; Sodium Dodecyl Sulfate; Zymosan

Inflammation was induced on the forearms of volunteers by twenty-four closed patch tests to either the irritant 10% sodium lauryl sulfate (SLS) or Toxicodendron extract. Each chemical was tested at eight sites on the ventral forearms of each volunteer in order to assess the variability of response among test sites in individual subjects. Inflammation was assessed about 10 minutes after patch tests were removed. The degree of inflammation elicited by both Toxicodendron and SLS was variable among subjects, but variation among individual test sites was much more marked in subjects tested with SLS (p less than 0.002). The marked variability of responses to irritation that occur in any single subject may explain why irritant patch test responses do not reliably identify the irritation-prone individual.

Topics: Adolescent; Adult; Allergens; Body Surface Area; Dermatitis, Contact; Dermatitis, Toxicodendron; Drug Evaluation, Preclinical; Edema; Erythema; Female; Forearm; Humans; Irritants; Male; Patch Tests; Plant Extracts; Plants, Toxic; Skin; Sodium Dodecyl Sulfate; Time Factors; Toxicodendron

Patch tests with aqueous solutions of the irritating detergent sodium lauryl sulfate (SLS) elicit varying degrees of inflammation from subject to subject and from site to site. For an investigation of the causes of this variability, two patch tests with 10% aqueous solutions of SLS were applied to adjacent areas of ventral forearm skin of eighteen volunteers. In one test the water vehicle was allowed to evaporate from the patch test unit before the patches were applied. After 22 hours the patch tests were removed, and 2 hours later the degree of inflammation was graded. Less inflammation was present at the site of the dry patch test in fifteen of eighteen subjects, and the score of inflammation between each pair was significantly less at the dry patch test site (p less than 0.001). These studies show that evaporation of water from aqueous solutions can influence the irritating potential of SLS on human skin.

Topics: Adult; Chemistry, Pharmaceutical; Dermatitis, Contact; Drug Evaluation, Preclinical; Edema; Erythema; Female; Forearm; Humans; Irritants; Male; Patch Tests; Pharmaceutical Vehicles; Skin; Sodium Dodecyl Sulfate; Time Factors; Volatilization; Water

A 5 h, semi-occlusive human patch test repeated daily for 5 days, was modified for use in the rabbit Using this method, the relative irritancy of three cosmetic samples of known human irritancy was determined in the rabbit. In addition, attention was paid to assessment methods and a detailed histological scoring system is described as well as two methods of visual assessment. In order to compare the effectiveness of these methods, the reaction to a series of concentrations of sodium lauryl sulphate was examined. All three assessment methods gave similar results and the merits of combining histological and visual assessment are discussed.

Topics: Acanthosis Nigricans; Animals; Cosmetics; Edema; Erythema; Female; Models, Biological; Parakeratosis; Patch Tests; Rabbits; Skin Tests; Sodium Dodecyl Sulfate; Ulcer

Topics: Animals; Basement Membrane; Blister; Cytoplasm; Edema; Female; Inflammation; Keratosis; Lipid Metabolism; Microscopy, Electron; Rats; Ribosomes; Skin; Sodium Dodecyl Sulfate