Putative mechanism of the itch-scratch circle: repeated scratching decreases the cutaneous level of prostaglandin D2, a mediator that inhibits itching

In atopic dermatitis, scratching of the skin as a reaction to itching causes injury to the skin, which, in turn, further increases the itching resulting in the establishment of the so-called itch-scratch circle. We have shown that prostaglandin (PG) D2 plays an inhibitory role against pruritus in mice with atopic-like dermatitis; therefore, we examined the relationship between scratching and the cutaneous PGD2 level using an artificial scratching model with a wire brush. Mechanical scratching induced a temporary increase of the skin PGs levels (PGE2, PGD2, 6-ketoPGF1alpha, PGF2alpha). The skin PGD2 level and the ability of PGD2 production decreased at 48 h after repeated scratch, compared to that of normal skin, not so after single scratch. Immunohistochemical analysis and Western blotting revealed a decrease in the levels of cyclooxygenase-1 (COX-1) and hematopoietic PGD synthase in mechanically scratched skin. The reduced ability of the skin for PGD2 production following mechanical scratching could be caused by this decrease in the expression levels of COX-1 and PGD2 synthase. The results suggest that repeated scratching in mice decreases the ability of the skin to produce PGD2, which is an endogenous mediator that inhibits pruritus, resulting in the establishment of the itch-scratch circle.     

Differences in c-AMP levels in epithelial cells from pelvic and pectoral regions of the toad skin

Arginine vasopressin (AVP) stimulated active Na+ transport (JNa+) and osmotic water flow (JH2O) across the pelvic skin but only JNa+ across the pectoral skin of the toad, Bufo woodhouseii. Isolated epithelial cells from the pelvic skin had a maximal c-AMP level of 11.16 pmoles/mg protein after 5 min of AVP treatment while that of pectoral skin was 3.64 pmoles/mg protein. The c-AMP level of both skin areas fell to unstimulated values after 20 min of AVP treatment; however, JH2O (pelvic skin) and JNa+ (pelvic and pectoral skin) remained elevated during 3 hr of treatment. Dibutyryl c-AMP and theophylline stimulated JH2O across the pelvic but not the pectoral skin. Maintaining toads in water for 12-24 hr resulted in a substantial lowering of JH2O across the pectoral skin which was not reversible by treatment with c-AMP and theophylline.     

Temporal relationship of hormonal peaks to ovulation and sex skin deturgescence in the baboon

The purpose of this study was to determine the temporal relationship of peak levels of oestradiol (E₂), LH and progesterone to ovulation and sex skin deturgescence in the baboon. A total of 55 baboons were used in these studies. Hormonal levels were measured in 47 cycles and ovulation was documented by laparoscopic examination in 26 of these cycles. A temporal relationship of ovulation to sex skin deturgescence was established in 57 cycles. The mean interval from E₂ peak to ovulation was 41.4±2.3 hr, the interval from E₂ peak to LH peak was 17.3±2.0 hr and that from LH peak to ovulation was 18.4±2.0 hr. Eleven baboons showed an LH peak on the day of the E₂ peak. The number of days to the first sign of sex skin deturgescence after ovulation was 2.07±0.14 days (range 0–5 days). Nineteen cycles (33.3%) showed sex skin deturgescence 1 day after ovulation, another 19 cycles (33.3%) showed sex skin deturgescence 2 days after ovulation, and only 13 cycles (22.8%) showed sex skin deturgescence 3 days after ovulation. Sex skin deturgescence was observed on day 0, 4 or 5 postovulation in only two baboons.     

Proliferative processes in the epithelium of the autotransplant and surrounding skin of mice]

Proliferative processes were studied in epithelial cells of skin autografts and the surrounding skin in mice of CBA line. In the animals subjected to the operation but without cortisone injection (Fig. 1, a and Fig. 2, delta), a high proliferative activity in the skin epithelial cells surrounding the autograft was observed during the whole course of the experiment. In the cells of the autograft, the mitotic activity, after inhibition, restored quickly up to the original level, and in 10 days it sharply increased and did not differ from that in the epithelium of the surrounding skin. The amount of DNA-synthesising cells in the epithlium of the autograft at first remained at the original level, and then, after a short sharp rise, it decreased up to its level in the surrounding skin and then was changing nearly in the similar fashion with it. After repeated injection of cortisone both indices of proliferative activity (Fig 1, delta and Fig. 2, delta) in the epithlium of the autograft and of the surrounding skin gradually increased although slower than in the controls.     

The effect of the size of the irradiation field on the severity and outcome of the radiation injury of swine skin

Swine skin areas of 12.56 cm2, 50 cm2 and 100 cm2 were exposed to beta-particles from 90Sr + 90Y (100 Gy). The increase in size of the exposed site caused a considerable increase in the degree of the affection and a change in the regeneration rate. Epithelialization of 12.56 cm2 skin field was completed by the 14th-16th week, and it was absent after 4.5 years in the fields of 50 and 100 cm2.     

Wound healing of human skin transplanted onto the nude mouse: I. An immunohistological study of the reepithelialization process

Two months after transplantation of human skin onto the nude mouse, excisional wounds were made through the entire thickness of the skin, at the center of the graft, using a 2-mm punch. At various time intervals thereafter, ranging from 2 days to 9 weeks, the graft sites were harvested and processed for an immunohistological study. With a monoclonal antibody directed against HLA-ABC antigens, it was demonstrated that the healing epidermis is of human origin. Moreover, with three different monoclonal antibodies directed against human keratins, named respectively AE₁, AE₃, and KL₁ and with an anti-involucrin antiserum, it is reported that the keratinization and involucrin distribution patterns observed in normal human epidermis (1) are reconstituted, 2 months after transplantation, in the major part of the grafted epidermis, (2) undergo changes during the reepithelialization process, and (3) are restored in the healed epidermis 9 weeks after injury. This study indicates that the nude mouse/human skin model could be a valuable tool to study a major aspect of regeneration such as the reepidermization of human skin without recourse to human volunteers.     

Opposite actions of different doses of arginine-vasotocin and 1-deamino-arginine-vasotocin on sodium ion transport in skin of the frog Rana temporaria

Active transport of sodium ions across the isolated abdominal skin of the frog Rana temporaria after application of arginine-vasotocin (AVT) and 1-deamino-arginine-vasotocin (1dAVT) was studied by measurement of the short-circuit current (SCC). The maximal increase in the SCC values (26 and 19 microA/cm2) was observed after addition of 10 nM AVT or 100 nM 1dAVT, respectively, to the frog skin basal surface. An increase of concentration of AVT to 100 nM and of IdAVT to 1 microM terminated the sodium transport in the frog skin. A preliminary addition of an antagonist of arginine-vasopressin V1a-receptors to the Ringer's solution at the frog skin basal surface led to a rise in the SCC values in response to administration of ineffective doses of AVT or 1dAVT. V2-receptor antagonists did not affect the frog skin reaction to administration of these doses of AVT or IdAVT.     

Use of the in vivo skin comet assay to evaluate the DNA-damaging potential of chemicals applied to the skin

The aim of the present study was to evaluate both sensitivity and specificity of an in vivo skin comet assay using chemically treated, hairless mouse dorsal skin as a model. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, 0.0125-0.2%), 4-nitroquinoline-1-oxide (4NQO, 0.01-0.25%), mitomycin C (MMC, 0.0125-0.05%), benzo[a]pyrene (B[a]P, 0.25-2%), and 7,12-dimethylbenz[a]anthracene (DMBA, 0.25-1%) were each applied once to the dorsal skin of hairless male mice; after 3h, epidermal skin cells were isolated, and the alkaline comet assay was performed. The assay was performed after 24h for only the B[a]P and DMBA. Furthermore, B[a]P and DMBA were evaluated by alkaline comet assay using liver cells after both 3 and 24h. The mean percent of DNA (%DNA) in tail in the 0.05-0.2% MNNG and 0.1-0.25% 4NQO treatment groups was markedly higher than in the control group at 3h post-application. Although the mean %DNA values in the tail in the B[a]P and DMBA groups were the same as the controls at 3h post-application, the 2% B[a]P and 1% DMBA groups showed significantly higher values versus controls 24h after application. No significant increases in the mean %DNA in the tail were observed in the MMC group. No clear increases in %DNA in the tail were observed in the B[a]P and DMBA groups at 3 or 24h after application in the liver. These results suggest that the in vivo skin comet assay is able to accurately identify DNA-damaging potential with a skin-specific response and is a useful method to detect the DNA-damaging potential of genotoxic chemicals on the skin.     

Vaccination with irradiated cercariae of Schistosoma mansoni preferentially induced the accumulation of interferon-gamma producing T cells in the skin and skin draining lymph nodes of mice

Cytokine response to schistosomula of Schistosoma mansoni was evaluated in the skin of mice during the initial 72 h following infection. These studies showed a significant increase in the levels of IL-4 and IL-10 message in the skin in areas of cercarial penetration. The IL-4 message was detectable in the skin as early as 8 h after infection and the message for IL-10 appeared from 16 h after infection. However, mRNA for IFN-gamma was undetectable in the skin samples for up to 72 h after infection with normal cercariae. In sharp contrast, vaccination with irradiated cercariae induced IFN-gamma and IL-2 responses in the skin within 24 h. Analysis of the cytokine profile of cells isolated from the skin during these early time points showed that T cells are probably not a source of IL-4 or IL-10 in the skin of mice infected with normal cercariae. However, in vaccinated animals, the majority of the IFN-gamma is derived from skin-residing T cells. In vaccinated animals, responses in the skin were mirrored in the skin-draining lymph nodes as well. Analysis of the CD4/CD8 ratio showed a significant decrease in the skin following vaccination suggesting an increase in CD8+ cells. Interestingly however, when vaccinated animals were challenged with normal cercariae, there was a significant reduction in IFN-gamma response in the skin and its draining lymph nodes. These results show that vaccination with irradiated cercariae of S. mansoni, preferentially induce the accumulation of IFN-gamma producing T cells in the skin and skin-draining lymph nodes of mice.     

Expression of angiotensin II AT2 receptors in the rat skin during experimental wound healing.

We localized and characterized angiotensin II AT1 and AT2 receptors in the skin of 2-week-old rats during experimental wound healing. Both AT1 and AT2 were present in the skin. Three days after wounding, the expression of angiotensin II receptors was significantly enhanced in the dermis as well as in a localized band within the superficial dermis of the skin surrounding the wound. The major proportion of this increase was due to angiotensin II AT2 receptors. Our results suggest a physiological role for AT2 receptors in the process of tissue repair.     

Sequential mechanisms of cyclophosphamide-induced skin allograft tolerance including the intrathymic clonal deletion followed by late breakdown of the clonal deletion

The cellular basis of the transplantation tolerance in a model system of BALB/c (Mls-1b) mice rendered cyclophosphamide (CP)-induced tolerant to DBA/2 (Mls-1a) skin allograft was investigated by assessing V beta 6+ T cells. From our results, three major mechanisms that are essential to the CP-induced skin allograft tolerance were sequentially elucidated. The first mechanism was destruction of donor-Ag-stimulated T cells in the periphery by CP treatment. The second mechanism was intrathymic clonal deletion of donor-reactive T cells, such as V beta 6+ T cells, correlating strongly with intrathymic mixed chimerism. The clonal deletion, however, was not always essential for the maintenance of the skin allografts, because DBA/2 skin survived even after the clonal deletion terminated and V beta 6+ T cells reappeared in the periphery of the recipient BALB/c mice. The third mechanism was generation of tolerogen-specific suppressor T cells, especially in the late stage of the tolerance. In contrast, the clonal anergy that is evidenced by the specific suppression of mixed lymphocyte reaction in the recipient BALB/c mice after injecting with DBA/2 spleen cells alone was not considered as a significant mechanism in prolonging skin allograft survival because such anergic mice showed accelerated rejection of the skin allografts. These results may suggest practical hierarchy of the mechanisms of CP-induced allograft tolerance.     

Effect of chloroquine wash-out period of photosensitizers in the skin and selected organs in rats

In the last decade, photodynamic therapy has become an alternative method for the diagnosis and therapy of tumors. In human medicine hematoporphyrin derivatives, sulfonated hydrophilic meso-tetraphenylporphyrin (TPPS4) and an oligomer of hematoporphyrin (Photosan 3), are widely used. Chloroquine is used for the treatment of porphyria cutanea tarda for its power to release porphyrins from the liver tissue. The kinetics of two porphyrin photosensitizers TPPS4 and Photosan 3 in the skin and some organs as well as the effect of chloroquine on the porphyrin excretion and their accumulation in skin and organs of Wistar rats were studied. TPPS4 exhibited maximum fluorescence in skin 48 h after application with decreasing to basal level from the 8th to the 14th day. Maximum fluorescence was reached at 72 hours after Photosan 3 application and it decreased to basal level during 96 hours after application. TPPS4 caused significantly higher fluorescence compared to Photosan 3. Chloroquine after oral administration did not change the fluorescence of skin, but it significantly decreased the TPPS4 concentration in rat organs if chloroquine treatment started 3 days or 2 weeks after TPPS4 application. Chloroquine significantly decreased the serum TPPS4 concentration during the period of 28 days. Fluorescence of skin was significantly higher and lasted longer after application of TPPS4 compared to Photosan 3. Chloroquine after oral administration did not influence the fluorescence of the skin, but it significantly decreased the TPPS4 concentration in rat organs. This effect could be useful in photodynamic therapy for mobilizing exogenous porphyrins from tissues after parenteral photodynamic therapy.     

Efficacy of two ethanol-based skin antiseptics on the forehead at shorter application times

Background  

Recent research suggests that alcohol-based skin antiseptics exhibit their efficacy on the resident skin flora of the forehead in less than 10 minutes. That is why we have looked at the efficacy of two ethanol-based skin antiseptics applied for 10, 2.5 and 2 minutes on skin with a high density of sebaceous glands. Each experiment was performed in a reference-controlled cross-over design with at least 20 participants. Application of isopropanol (70%, v/v) for 10 minutes to the forehead served as the reference treatment. The clear (skin antiseptic A) and coloured preparations (skin antiseptic B) contain 85% ethanol (w/w). Pre-values and post-values (immediately after the application and after 30 min) were obtained by swabbing a marked area of 5 cm² for about 10 s. Swabs were vortexed in tryptic soy broth containing valid neutralizing agents. After serial dilution aliquots were spread on tryptic soy agar. Colonies were counted after incubation of plates at 36°C for 48 h. The mean log₁₀ reduction of bacteria was calculated. The Wilcoxon matched-pairs signed-ranks test was used for a comparison of treatments.     

Effect of helium-neon laser radiation on the morphology of experimental allergic contact dermatitis

Clinical and morphological studies of guinea-pig skin in the areas of dermatitis induced by application of dinitrochlorobenzene (DNCB) have shown the decreased skin response to DNCB after treatment with helium-neon laser (wave length 632.8 nm, power density 8-10 mvt /cm2). The ultrastructural signs of cell metabolism activation in the epidermis and derma as well as activation of capillary transport have been discovered in allergic contact dermatitis areas after treatment with helium-neon laser.     

Augmentation of blood flow in delayed random skin flaps in the pig: effect of length of delay period and angiogenesis

Skin capillary blood flow and angiogenesis were studied by radioactive microsphere and morphometry technique, respectively, in delayed random skin flaps in the pig. Skin flaps were delayed for 2, 3, 4, 6, or 14 days. Blood flow was measured 6 hours after complete raising of acute and delayed random skin flaps on the opposite flanks of the same pig. It was observed that the capillary blood flow increased significantly (p less than 0.05) within 2 days of delay compared to the acute skin flaps. This capillary blood flow further increased by about 100 percent between days 2 and 3, started to plateau after day 3, and remained unchanged between days 4 and 14 of delay. This increase in capillary blood flow was mainly in the distal portion of the delayed skin flaps. There was no indication of an increase in the density of arteries in all delay periods studied. Our observations did not support the hypotheses that the delay phenomenon involves angiogenesis or long-term adaptation to ischemia, as have been hypothesized previously. The possible mechanism of delay is discussed.     

Electron microscopical studies of Strongyloides ratti infective larvae: loss of the surface coat during skin penetration

Previous indications using radiolabelled larvae that Strongyloides ratti free-living infective larvae lose a surface coat during penetration of the skin were further investigated by transmission electron microscopy of the cuticle of S. ratti infective larvae in the free-living stage, after penetration of mouse skin, and after migration to the lungs. These studies demonstrated the presence of a faint electron-dense surface coat external to the epicuticle on free-living worms which was absent from larvae recovered from the skin and lungs. When free-living infective larvae were incubated in 10% CO2 at 37 C and then examined with phase-contrast microscopy, worms were observed in the process of losing this coat. These observations confirm the hypothesis that S. ratti infective larvae lose a surface coat during penetration of the skin.     

豚鼠皮肤光过敏试验方法的建立

目的探索并建立豚鼠光过敏试验的方法。方法采用TBS作为阳性对照物,同时设立阴性对照组。分别于第1天、第4天、第7天对豚鼠皮肤进行UV-A紫外光诱导,诱导剂量紫外光强度为30 J/cm2;于第一次诱导后28 d对豚鼠皮肤采用UV-A紫外光激发,激发强度为9 J/cm2。结果阴性对照组24 h、48 h、及72 h豚鼠光过敏试验阳性率均为0,阳性对照组为24 h、48 h、及72 h豚鼠光过敏试验阳性率均为100%。结论本研究成功建立了豚鼠皮肤光过敏试验模型,TBS可作为阳性对照物。     

Depression of prolylendopeptidase activity in the delayed hypersensitive guinea pig skin lesion induced by bovine gamma-globulin

Prolylendopeptidase activity was increasingly depressed with time from 6 to 24 hr after the start of sensitization in the delayed hypersensitive guinea pig skin lesion induced by bovine gamma-globulin as an antigen. The remarkably depressed activity of the enzyme in the violently inflamed skin began to be restored slowly 48 hr after sensitization, and its activity was ultimately recovered to the original level by 504 hr after a single sensitization in vivo. Depression of the enzymatic activity is caused by a novel prolyendopeptidase inhibitor, whose amino acid composition is 7 Glu, 1 Ser, 2 Gly, 1 Ala, 2 Pro, and 1 Val, generated by inflammation.     

Nondébridement of laser char after two carbon dioxide laser passes results in faster reepithelialization

Skin repair following laser injury can be accelerated by using techniques that promote rapid reepithelialization. In this article, the benefit of intraoperative nondébridement of laser debris after two laser passes is discussed. After carbon dioxide laser resurfacing of the face, skin specimens were examined using indirect immunofluorescence with antibodies to specific epidermal and basement membrane proteins. Biopsy specimens obtained immediately after resurfacing showed a greater injury to epidermal and basement membrane proteins when skin was wiped with saline-soaked gauze after laser passes than when there was no débridement after two passes. Later examination of skin specimens obtained from nine patients 2 days after carbon dioxide resurfacing showed that nondébrided, occluded skin had faster reepithelialization than the other treatments. Nondébridement of the skin at the time of resurfacing along with the use of postoperative occlusive dressings led to the rapid reestablishment of a multilayered epidermis only 2 days after resurfacing. Nondébridement along with occlusive dressings results in rapid reepithelialization of the skin after two carbon dioxide laser passes for skin rejuvenation.