A comparison of bathwater and oral delivery of 8-methoxypsoralen in PUVA therapy for plaque psoriasis.

Bath-PUVA is an alternative to oral-PUVA for the treatment of psoriasis. This study compares the effectiveness of the two methods in two groups, each consisting of 17 patients with plaque psoriasis. Three patients who failed to improve with oral-PUVA were transferred to bath-PUVA and subsequently cleared. Another seven patients who returned with a further episode of psoriasis received the alternative treatment; this gave a group of 10 patients in whom a cross-over comparison was possible. In both comparisons bath-PUVA was as effective as, or more effective than, oral-PUVA and required, overall, less than 50% of the total UVA, although this saving was not as great as in previous reports. Bath-PUVA caused fewer immediate problems and was preferred by many patients. It is suitable for those taking other systemic medications and we recommend it as a valuable therapeutic option that should be available at all treatment centres.

The prevalence of epidermolysis bullosa in Scotland.

The prevalence of epidermolysis bullosa (EB) in Britain and most other countries is unknown. Patients suffering from the inherited forms of EB and living in Scotland have been traced. Two hundred and fifty-nine affected people from 76 families have been identified, of whom 211 were clinically assessed. One-third of these Scottish EB sufferers had never been seen by a dermatologist. In Lothian, where there appears to be a relatively high prevalence of EB, 75% of patients were unknown to their general practitioners. The point prevalence of all forms of EB at the outset of the study was 49.0 per million, comprising EB simplex 28.6 per million and dystrophic EB 20.4 per million. Extrapolation of accurate data available for the Lothians suggests that the point prevalence of all forms of EB in Scotland is in excess of these figures.

Urinary glycosaminoglycans excretion in Graves' disease.

Urinary excretion of glycosaminoglycans was measured in 10 patients with pretibial myxoedema, 7 of whom also had thyroid-associated ophthalmopathy, and in 3 additional patients with ophthalmopathy but no skin changes. Total uronic acid excretion was raised above control levels in only 2 patients, who had both eye and skin disease of recent onset. In these patients excretion was initially three times the control level but declined sharply in subsequent months. This decline was in the absence of effective treatment or spontaneous improvement and would appear to reflect the natural history of the disease. These data show that although glycosaminoglycans excretion may be disturbed in Graves' disease, it provides an unreliable reflection of clinical status and of the effectiveness of treatment.

Changes in skin pH after the use of baby wipes.

We recently made a double-blind safety-in-use comparison of four different brands of baby wipes using a panel of 302 infants over a period of 10 weeks. For the first two weeks, only soap and water was used to clean the babies' skin after each diaper change, and then for the eight-week test phase each baby was allocated one or other of the products for normal home use. The wipes differed in cleansing lotion formulation (emollients, preservative, pH) and fibrous composition. There were no clinically detectable differences in the effects of the wipes in terms of erythema, frequency of rashes, edema, and desquamation, but we recorded significant changes in the pH of pubic and buttock skin inside the diaper area. In particular, the brand of wipes with the lowest pH (2.8) in the lotion reduced the mean skin pH from 5.6 to 5.0 (p < 0.01), and those with a pH of 5.5 had no significant effect. Wipes of intermediate pH (3.7) gave a final skin pH of 5.4-but the downward trend was not statistically significant. These data indicate that skin pH can be depressed by such topical application, although the trial lasted only a fraction of the total time wipes might be used on each infant. Further research is necessary to evaluate the implications of these findings.

Skin fibroblast activity in pretibial myxoedema and the effect of octreotide (Sandostatin) in vitro.

The accumulation of glycosaminoglycans in the skin in pretibial myxoedema appears to be a response by local fibroblasts to a stimulating factor in the patient's serum, but the identity of the factor, its ability to stimulate skin fibroblasts as opposed to cultured thyroid cells, and the specificity of its effect to pretibial skin fibroblasts, are all controversial. We have studied fibroblasts cultured from the lesional skin of two women with pretibial myxoedema, and compared their proliferation and secretion of glycosaminoglycans with those of fibroblasts from the patients' forearms and from the forearm skin of two normal subjects. We found that in the presence of the patients' sera all six lines of fibroblasts secreted more glycosaminoglycans [205 +/- 21% (SD)] than with normal human sera (147 +/- 19%), or fetal calf serum (100%). Fibroblast proliferation showed the same pattern of differences: patients' sera 142 +/- 22%; normal human sera 116 +/- 9%, and fetal calf serum 100%. These experiments confirm the presence of a serum factor in pretibial myxoedema which is capable of stimulating the activity of skin fibroblasts in vitro, and show that its effects are not restricted to fibroblasts from pretibial skin or to those grown from the skin of the patients. Proliferation of normal fibroblasts cultured in medium supplemented with fetal calf serum was reduced by Sandostatin (octreotide), but it failed to inhibit their secretion of glycosaminoglycans. In contrast, secretion of glycosaminoglycans by a patient's pretibial skin fibroblasts was almost completely inhibited by 1 mM minoxidil. In the presence of patients' sera Sandostatin (0.1-10 micrograms/ml) reduced secretion of glycosaminoglycans by about 50%.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparison of morphoea and lichen sclerosus et atrophicus in vitro: the effects of para-aminobenzoate on skin fibroblasts.

To study the effects of para-aminobenzoate on the dermis, fibroblast cell lines derived from lesions of lichen sclerosus et atrophicus, from morphoea and from normal skin were incubated with Potaba in vitro. Monolayer cultures containing Potaba showed a dose-dependent inhibition of proliferation beginning at 1,000 micrograms/ml with total inhibition at 10,000 micrograms/ml. Mean ID50 values for the three groups were not significantly different. There was a similar dose dependent inhibition of glycosaminoglycan secretion in all 3 groups, except at 10,000 micrograms/ml where secretion by lichen sclerosus et atrophicus and morphoea fibroblasts was significantly more inhibited than normal lines. Inhibition of the glycosaminoglycan secretion at 10-1,000 micrograms/ml was a direct effect of the drug rather than an indirect effect of changes in cell density, and lichen sclerosus et atrophicus fibroblasts produced about 40% more GAG than the morphoea or normal lines growing at similar densities. Collagen synthesis was increased in both lichen sclerosus et atrophicus and morphoea cell lines, with increased non-collagenous protein in morphoea lines. These results confirm that there are differences between lichen sclerosus et atrophicus and morphoea, and suggest glycosaminoglycan secretion as a possible target for the therapeutic action of Potaba.

The metabolism of fibroblasts from normal and fibrotic skin is inhibited by minoxidil in vitro.

The effects of minoxidil in vitro were studied using fibroblasts grown from the lesional skin of patients with lichen sclerosus et atrophicus, morphoea and from the skin of normal individuals. The proliferation of all fibroblast lines over 3 days was inhibited in proportion to the concentration of minoxidil, being 20% or less of controls at 1 mM, where cell viability was only marginally reduced (84 +/- 2% vs. 88 +/- 2% (SEM) in controls). At 5 mM there was usually a net loss of cells and only 72% of those remaining were viable. In contrast, minoxidil at 0.1-1 mM stimulated the proliferation of foreskin keratinocytes by up to 130%. Contraction of collagen lattices containing the three types of fibroblasts was inhibited by 22-26% with 1 mM minoxidil after 5 days and by 50-94% with 5 mM. Secretion of glycosaminoglycans by normal fibroblasts showed concentration-dependent reduction, being 25 +/- 6% of that of untreated cultures with 1 mM minoxidil. These findings show that minoxidil has a range of inhibitory effects on both normal and abnormal skin fibroblasts in vitro, which contrast with its stimulation of skin epithelial cells, and support suggestions that it may provide a useful topical treatment for keloids and other fibroses.

Production of glycosaminoglycans by human skin fibroblasts growing in a collagen lattice.

Production of glycosaminoglycans (GAG) by human skin fibroblasts cultured in collagen lattices closely resembled that already described for the same cells grown as monolayers on plastic. There was no inhibition of GAG corresponding to that of DNA and protein synthesis and similar controls by cell density and anti-inflammatory drugs operated in 3-dimensional as in 2-dimensional culture. The bulk of the GAG synthesised in lattice culture was hyaluronic acid, as in monolayers.

Glycosaminoglycans production by cultured skin fibroblasts from the Pasini and Cockayne-Touraine forms of dominant dystrophic epidermolysis bullosa.

Qualitative and quantitative comparisons of glycosaminoglycans e (GAG) production were made on fibroblast lines cultured from the skin of six patients with the Pasini (albopapuloid) form of dominant dystrophic epidermolysis bullosa, six with the non-albopapuloid form (Cockayne-Touraine), eight lines from patients with simplex or recessive dystrophic epidermolysis bullosa and eight lines from normal individuals. A reasonable match of donor age and gender, site, and passage number was achieved. Contrary to an earlier report, the lines from the Pasini group were unexceptional in the amount of GAG they secreted and the proportions of sulfated and nonsulfated GAG showed no consistent difference from the Cockayne-Touraine or control lines. The Pasini lines secreted 77 +/- 18 (SEM) microgram GAG-uronic acid per 10(7) cells and the Cockayne-Touraine lines 81 +/- 12 micrograms at equivalent cells densities. Sulfated GAG represented averages of 19 +/- 4+ in Pasini lines, 17 +/- 5% in Cockayne-Touraine, and 14 +/- 3% in controls. These findings are consistent with current views of albopapuloid lesions as an unreliable clinical sign in epidermolysis bullosa and bring into question the validity of the Pasini entity.

Fibroblast-keratinocyte interactions in psoriasis: failure of psoriatic fibroblasts to stimulate keratinocyte proliferation in vitro.

We have tested in two ways the hypothesis that dermal fibroblasts direct the hyperproliferation of the overlying epidermis in psoriasis. First, culture medium from psoriatic and from normal skin fibroblasts was added to monolayer cultures of foreskin keratinocytes. Second, psoriatic and normal fibroblasts embedded in hydrated collagen lattices were co-cultured with monolayers of foreskin keratinocytes. There was no evidence in either study that psoriatic fibroblast products could stimulate the proliferation of the keratinocytes, or that normal fibroblast products inhibited their proliferation. A positive control for the fibroblasts was provided by leucocyte supernatants, which stimulated keratinocyte proliferation by up to 65%. Our data do not support a primary role for dermal fibroblasts in psoriasis.

Failure of a helium-neon laser to affect components of wound healing in vitro.

The red light of a helium-neon (He-Ne) laser has been reported to stimulate wound healing and cell growth. To investigate the nature of its influence on wound healing we have studied seven components of the healing process in vitro: human skin fibroblast, epithelial and endothelial cell proliferation, cellular migration from skin explants, collagen lattice contraction, collagen synthesis and glycosaminoglycans (GAG) secretion. We used a 5 mW He-Ne laser emitting a I mm diameter beam of wavelength 633 nm. Cellular proliferation was not affected by irradiation three times a day for 3 days. There was no effect on cellular migration or on the rate of collagen lattice contraction. The rate of collagen synthesis, measured as the incorporation of 3H-proline into collagenase-sensitive protein, was no greater than that of controls and GAG secretion did not increase in the irradiated group. We have not found any significant effects of He-Ne irradiation.

Increased urinary excretion of glycosaminoglycans in insulin-dependent diabetic patients with limited joint mobility.

Urinary excretion of glycosaminoglycans (GAG) was measured in 24-h specimens collected from 27 insulin-dependent male diabetic patients and 22 healthy males of similar ages. Excretion relative to creatinine was increased by 35% in the diabetic group as a whole (p less than 0.02). Twelve patients with limited joint mobility (cheiroarthropathy) formed a subgroup with high excretion (69% above controls), those without limited joint mobility being similar to controls. There was no correlation of retinopathy with GAG excretion.

Contraction of collagen lattices by skin fibroblasts: drug-induced changes.

A range of dermatologically useful drugs were added to human skin fibroblasts cultured in collagen lattices to assess possible effects on the rate of lattice contraction. Vitamin C, Vitamin E, phenytoin, sodium salicylate, D-penicillamine and dibutyryl c-AMP had no significant effect. Chlorhexidine acetate at 10 micrograms/ml arrested contraction after 24 h but this was related to its cytotoxicity. The antibiotics griseofulvin (2-16 micrograms/ml) and cycloheximide (5-30 micrograms/ml) caused dose-related inhibitions of contraction without affecting fibroblast viability. Four corticosteroids at 10 micrograms/ml inhibited contraction, clobetasol propionate having the greatest effect. On the other hand 4 retinoids at 10(-5) M enhanced contraction by up to 20%. As lattice contraction appears to model the contraction of skin wounds and there are broad parallels between the effects shown here of antiseptics, corticosteroids and the retinoids, and their reported influence on healing wounds, the lattice system may be a useful pharmacological screen for new compounds.

Urinary excretion of glycosaminoglycans in psoriasis.

Urine from patients with generalised plaque psoriasis contained substantially more precipitable glycosaminoglycans (GAG) and uronic acid than the urine of healthy controls. The difference was not related to sex, age, renal function, the hospital environment, or to the presence of arthritis. Successful topical treatment with tar or dithranol, or PUVA therapy, did not affect the rate of GAG excretion. Cellulose acetate electrophoresis of the GAG from patients and controls showed similar patterns dominated by chondroitin sulphate. There was no evidence to favour the skin lesions as the source of the additional glycosaminoglycans and the findings are consistent with the concept of psoriasis as a general disease.

Proliferation and glycosaminoglycans secretion in fibroblasts from psoriatic skin: differential responses to retinoids.

The effects of four retinoids, all-trans-retinoic acid (tretinoin), 13-cis-retinoic acid (isotretinoin), R0 10-1670 (etretin) and the arotinoid, R0 15-0778, on fibroblast proliferation and glycosaminoglycans (GAG) secretion in vitro were studied. Fibroblasts lines cultured from normal skin (HSF) were compared with those from lesional (PSA) and non-lesional (PSB) psoriatic skin. In general, the retinoids inhibited proliferation; the action was cytostatic, in rank order tretinoin greater than isotretinoin greater than etretin greater than arotinoid. The psoriatic cells tended to be more sensitive than the HSF lines, overall mean proliferation values (+/- SEM), as a percentage of untreated controls being: HSF 72 ++- 3, PSA 61 +/- 3 and PSB 54 +/- 3. Stimulation of GAG secretion at low concentrations (10(-7) M) of all four retinoids, declined as concentrations increased, and secretion was inhibited at 10(-4)M in PSB fibroblasts. Calculation of effects on GAG secretion due to changes in cell density confirmed the rank order for direct stimulation of secretion as arotinoid greater than etretin greater than isotretinoin greater than tretinoin. Electrophoresis of [3H]-labelled glycosaminoglycans secreted in the presence of 10(-7) M arotinoid showed that it was predominantly hyaluronic acid, as in untreated cells. These data confirm that different retinoids have contrasting levels of effects on mesenchymal cells and suggest a greater sensitivity to drugs in fibroblasts from psoriatic skin.

Toxicity of the anthraquinone glycoside P-1894B for human skin fibroblasts.

P-1894B inhibits prolyl hydroxylase in vitro and has been proposed as a topical treatment for dermal fibrosis. The drug had similar effects on two fibroblast lines from normal human skin and one line from a patient with lichen sclerosus et atrophicus. Exposure of logarithmically-growing cell monolayers for 72 h caused dose-dependent inhibition of proliferation at 0.05-0.5 microgram/ml but time-dependent cell death at 1-50 micrograms/ml. The epithelial cell line NCTC 2544 gave a similar result. Collagen lattices containing normal fibroblasts contracted more slowly in the presence of the drug at 0.1-0.5 microgram/ml, but this was clearly related to loss of viability. Collagen synthesis by monolayer cultures was unaffected at 0.05 and 0.1 microgram/ml P-1894B in one line of normal fibroblasts, but was reduced by 40% and 15%, respectively, in the other. The concentrations of P-1894B reported to be active against prolyl hydroxylase are therefore lethal to cultured skin cells. Although the effective use of dithranol as a topical anti-psoriatic agent, despite its cytotoxicity in vitro, is encouraging for P-1894B, further toxicological studies are imperative.

Contraction of collagen lattices by skin fibroblasts from dystrophic recessive epidermolysis bullosa and other dermatoses.

Skin fibroblasts incorporated into a lattice of hydrated collagen contract it to form a tissue-like structure. Fibroblasts from patients with skin diseases, including psoriasis, epidermolysis bullosa, and scleroderma, and from control subjects, all showed a similar ability to contract the lattice. The established skin epithelial cell line NCTC 2544, melanoma cells, and 3T6 mouse fibroblasts produced little contraction. Contraction required the presence of serum and was unaffected by the addition of fibronectin (10-20 micrograms/ml). Hyaluronic acid at 50-500 micrograms/ml had no effect, but contraction was inhibited at 1 mg/ml.

Werner's syndrome. Biochemical and cytogenetic studies.

Werner's syndrome is a rare condition of autosomal-recessive inheritance, showing some features of accelerated aging. We describe the clinical findings and laboratory studies in a 29-year-old man with this disorder, who presented because of a leg ulcer. Skin fibroblasts from our patient were difficult to culture and proliferated more slowly than those of controls. They produced less glycosaminoglycans than those of controls but synthesized more collagen, which was normal in type. The patient's urinary glycosaminoglycan level was slightly elevated, with hyaluronic acid as a major component. His peripheral blood lymphocytes showed no chromosomal instability and responded normally to mutagens.