Fluorescence characteristics and pharmacokinetic properties of a novel self-adhesive 5-ALA patch for photodynamic therapy of actinic keratoses.

Actinic keratosis (AK) can be treated by photodynamic therapy (PDT), which is becoming a well-established tool in dermatology. Normally a precursor of the photosensitiser is applied topically and converted into protoporphyrin IX (PPIX) in the cells. By activating PPIX with light, the dysplastic cells will be destroyed. We report the results of two clinical studies investigating the properties of a novel self-adhesive 5-ALA-patch (PD P 506 A) intended for PDT of mild to moderate AK on the face and head. The studies investigated the influence of patch application duration on PPIX-specific fluorescence and the pharmacokinetic properties of the 5-ALA patch. The PPIX fluorescence in AK lesions and normal skin after patch application (intraindividual comparison; application for 2, 3, 4, 5 h) was investigated in 13 patients using DYADERM Professional (Biocam). In the subsequent pharmacokinetic study 12 patients were treated with 8 patches each (4 h application). 5-ALA and PPIX were analysed in plasma (over 24 h) and urine (over 12 h). PPIX-specific fluorescence measured immediately after patch removal increased with increasing application duration to a maximum at 4-h application. The fluorescence in AK lesions was more intense than in normal skin. A small increase of 5-ALA plasma concentrations was observed in 10 of 12 patients after applying 8 patches for 4 h, which rapidly declined to normal values after patch removal. The maximum increase was 3.7-fold of the pre-dose 5-ALA plasma concentration. No PPIX-concentrations above the lower limit of quantification were observed. PPIX-specific fluorescence in AK lesions can be steered by application duration of this novel 5-ALA patch. Application is safe and well tolerable. The observed small rise in 5-ALA plasma concentrations is regarded clinically irrelevant. Clinical efficacy of the patch in PDT will be investigated in further clinical trials.

El óxido nítrico y la endotelina-1,2 en queratosisactínica y carcinoma de células basales:cambios en el cociente óxido nítrico/endotelina / Nitric oxide and endothelin-1,2 in actinic keratosis and basal cell carcinoma: changes in nitric oxide/endothelin ratio

Antecedentes: El óxido nítrico (ON) es un gas inorgánico con un radical libre que desempeña acciones citostáticas/citotóxicas sobre tejidos tumorales, como cánceres ginecológicos, de mama y de colon. El óxido nítrico actúa también como una molécula activa productora de señales multifuncionales en muchas células del cuerpo, como células endoteliales, macrófagos, monocitos, hepatocitos, mastocitos, y astrocitos. La endotelina-1 (ET-1) es un péptido compuesto de 21 aminoácidos que estimula la proliferación de las células del músculo liso vascular, fibroblastos y queratinocitos y desempeña un papel importante en la expresión de los protooncogenes (c-myc,c-fos) que preceden a la proliferación celular. Igual que el ON, la ET se secreta por diferentes por diferentes tipos de células, como macrófagos, monocitos, hepatocitos, células endoteliales, células del músculo liso vascular y varias células tumorales. Se observan valores elevados de ET-1 en cánceres pulmonares, de células hepáticas y de próstata. La queratosis actínica (QA) y el carcinoma de células basales (CCB) son tumores de piel frecuentes con hiperqueratinización, hiperpigmentación y proliferación de queratinocitos acentuadas. Objetivo: Investigar la concentración en plasma de ONx (nitritos/nitratos -productos finales del metabolismo de ON), ET, y el valor del cociente ONx/ET en pacientes con QA y GCB en comparación con controles sanos. Métodos: Se midieron ONx, ET y el cociente ONx/ET en 13 pacientes con QA, en 12 pacientes con CCB y en 16 controles sanos. Resultados: Los análisis de los datos indican un aumento significativo de las concentraciones en plasma de ONx, ET, y del cociente ONx/ET en pacientes con CCB respecto a los controles (p<0,001, p<0,005 y p<0,001, respectivamente). Las concentraciones plasmáticas de ET en la QA también aumentaron en comparación con los controles (p<0,001). Cuando se compararon los dos grupos del estudio (QA y CCB), se encontró un aumento significativo (p<0,001) en el cociente ONx/ET en CCB. Conclusiones: El aumento de las concentraciones en plasma de ET y ONx en QA y especialmente en CCB son probablemente el resultado y/o la razón de la hiperqueratinización, hiperpigmentación y proliferación de queratinocitos acentuadas. La mayor producción de ET y ON por los queratinocitos puede actuar como factor de crecimiento y citotóxico y potenciar los mitógenos, lo que puede acelerar aun más la proliferación de estos tumores de piel. Además, el aumento del cociente ONx/ET probablemente refleja la alteración del equilibrio entre estas dos sustancias, lo que origina daño celular y desarrollo y proliferación tumorales (AU)

Tyrosinemia type II: nine cases of ocular signs and symptoms.

PURPOSE: To report nine cases of tyrosinemia type II, with ocular signs and symptoms. METHODS: Participants included nine patients (18 eyes) who were followed for a mean follow-up period of 6.5 years (range, 2 to 8 years). Intervention included dietary restriction of tyrosine and phenylalanine, which led to resolution of ocular and cutaneous lesions, improved behavior in one patient, and may have prevented developmental delay in others. The main outcome measures were visual acuity and serum tyrosine levels to determine the response to dietary therapy. Intelligence testing and developmental screening were performed when appropriate. RESULTS: All patients presented with ocular signs and symptoms as the primary manifestation of disease and serve to illustrate the typical ocular findings of this syndrome. Three patients presented with cutaneous manifestations, and one patient had mild mental impairment at the time of diagnosis. CONCLUSIONS: Recalcitrant pseudodendritic keratitis may be the presenting sign in tyrosinemia type II. Awareness of the presenting signs and symptoms may accelerate the diagnosis and dietary intervention. Initiation of a tyrosine-restricted and phenylalanine-restricted diet in infancy is most effective in preventing cognitive impairment.

Plasma antioxidant defense in actinic keratosis and basal cell carcinoma.

BACKGROUND: Reactive oxygen species and lipid peroxides have been implicated in the pathogenesis of a variety of diseases, particularly cancer. There may be an inverse correlation between lipid peroxidation and antioxidant defense mechanisms. The aim of this study was to investigate whether certain plasma antioxidants (ascorbic acid, alpha-tocopherol, total thiol groups, ceruloplasmin, urate, albumin and erythrocyte glutathione) are altered in actinic keratosis (AK) and basal cell carcinoma (BCC). METHODS: Plasma samples and red blood cells (RBC) of 13 patients with AK, 12 with BCC and 16 healthy controls were investigated. RESULTS: Data analysis indicates significant decrease of ascorbic acid (P < 0.001), alpha-tocopherol (P < 0.05 and P < 0.001), total thiol groups (P < 0.001), ceruloplasmin (P < 0.001 and P < 0.05), and RBC glutathione (P < 0.05) values in both AK and BCC groups compared to controls. Comparison of AK and BCC groups evidenced a significant decrease of alpha-tocopherol and RBC glutathione (P < 0.05) in BCC patient. CONCLUSION: Plasma antioxidants are decreased in the AK and BCC, probably due to the long exposure to UV irradiation which is one of the most important factors in the etiology of AK and BCC and alpha-tocopherol and RBC glutathione (P < 0.05) are most altered in BCC.

Effects of topical treatment with fenretinide (4-HPR) and plasma vitamin A levels in patients with actinic keratoses.

Eighteen patients with facial actinic keratoses were treated with the retinoid fenretinide (4-HPR), applied topically twice-daily for 3 months. After 3 months of treatment, complete regression was observed in 56% and partial regression in 44% of cases. Eight patients relapsed within 3 months after drug discontinuation. Six months later, only two patients (11%) showed a treatment response (complete regression). Blood samples showed that 4-HPR was not absorbed and no local or distant adverse effects were observed. Baseline plasma retinol levels were lower than in healthy subjects, thus suggesting that reduced retinol levels might be involved in this pathology. These encouraging preliminary results suggest the need for further studies to evaluate the best dosage schedules and duration of 4-HPR topical application in actinic keratoses.

Skin, adipose tissue and plasma levels of acitretin with rare occurrence of esterified acitretin during long-term treatment.

In a previous study acitretin and its 13-cis-metabolite were monitored in the plasma and epidermis of healthy volunteers. They were given 50 mg of trans-acitretin daily. No drug accumulation was observed in the skin, nor in the plasma. The purpose of the present study was to extend the data from non-psoriatic to psoriatic (n = 11) subjects, treated for at least 1 month with 25 mg acitretin. Plasma, skin biopsies and subcutaneous fat samples were analysed using HPLC. Trough levels of acitretin in skin were below the quantification limit, increasing to 28 +/- 16 ng/g within 5 h after dosing. Fat tissue levels exceeded those of skin, with values of 98 +/- 71 ng/g within 5 h after drug intake. In 2 patients, additional samples were taken 3 days post-therapy. Here, concentrations were below the quantification limit in adipose tissue, confirming that acitretin is not stored in subcutaneous fat. Esterification of acitretin into etretinate was observed in 2 subjects. This observation illustrates the recently described new metabolic pathway for acitretin. On both occasions, the unexpected ethylester metabolite was extensively stored in fat tissue.

Micronutrient concentrations in paired skin and plasma of patients with actinic keratoses: effect of prolonged retinol supplementation.

Much of our knowledge about the relationship between micronutrients and cancer comes from studies in which plasma (serum) micronutrient levels have been correlated with cancer incidence; however, the relationship between the concentrations of micronutrients in the plasma and in the target tissues has not been established. Ninety-three subjects (62 males and 31 females ages 42-86, median age 69) with actinic keratoses were recruited for investigation of this relationship. The subjects were randomly assigned and received placebo or retinol (25,000 IU/day) intervention for 48 to 65 months as part of a skin cancer chemoprevention trial. Shortly before the end of the trial, three fasting plasma samples and one skin biopsy were obtained from each subject. The concentrations of lutein, zeaxanthin, beta-cryptoxanthin, lycopene, alpha-carotene, beta-carotene, cis-beta-carotene, retinol, retinyl palmitate, alpha-tocopherol and gamma-tocopherol in the plasma and skin were simultaneously measured using HPLC. The profiles of the eleven micronutrients in the plasma and skin were similar. Lycopene, beta-carotene and alpha-tocopherol were the predominant micronutrients in both plasma and skin, but the ratio of retinyl palmitate to retinol was much greater in the skin than plasma. The three fasting plasma concentrations from the same subject during a one-month period were very consistent; however, the between-person variations were very large. The retinol supplementation caused a significant increase in the plasma concentrations of retinol, retinyl palmitate, lutein and alpha-tocopherol, especially retinyl palmitate as well as the skin concentrations of retinol and retinyl palmitate.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum parathyroid hormone level is elevated in some patients with disorders of keratinization.

BACKGROUND AND DESIGN: After the chance of observation of an elevated parathyroid hormone (PTH) value in a patient with pityriasis rubra pilaris, the serum PTH level was measured in the next 14 patients seen with disorders of keratinization. Calcium metabolism in three affected patients was then studied in depth. RESULTS: Five of 15 patients had twofold or greater elevations in serum PTH values. The patients had four different disorders of keratinization: bullous congenital ichthyosiform erythroderma (two patients); lamellar ichthyosis (one patient); pityriasis rubra pilaris (one patient); and ichthyosis linearis circumflexa (one patient). At least one other patient with each diagnosis had normal PTH values. Two of three patients who were studied further had clear evidence of increased, biologically active PTH, consistent with secondary hyperparathyroidism. An elevated PTH level spontaneously became normal in one patient, and in a second patient it became normal with a high-calcium diet. CONCLUSIONS: These data provide the first indication that patients with various disorders of keratinization have an increased risk for secondary hyperparathyroidism. The exact prevalence, origin, and physiologic significance of this finding remain to be elucidated.

Phrynoderma: is it an EFA deficiency disease?

The clinical response to various therapeutic agents was evaluated in 31 patients with phrynoderma. A complete clinical response with vitamin B-complex was noted in an average period of 5.7 weeks. In patients treated with vitamin E, partial or total improvement was seen in an average period of 12.3 and 10.7 weeks respectively. Patients treated with safflower oil showed a partial improvement in an average period of 13.2 weeks. The essential fatty acid (EFA) nutriture of 30 patients was compared with 7 controls. Plasma phospholipid fatty acid composition was used as an indicator of EFA nutriture. The patients with phrynoderma fell into two groups. In the 23 children in one group (pattern A), the mean levels of linoleic (18:2 omega 6), arachidonic (20:4 omega 6) and eicosatrienoic (20:3 omega 9) acids were similar to the levels in the controls. The ratio of eicosatrienoic to arachidonic acids (20:3 omega 9/20:4 omega 6), which is considered an accurate measure of EFA nutritional status, was 0.12 and in the normal range, suggesting that the EFA nutriture is normal in phrynoderma. The ratio of linoleic to arachidonic acids (18:2 omega 6/20:4 omega 6) was also found to be normal, suggesting that the metabolism of linoleic to arachidonic acid is not affected in phrynoderma. In seven children in a second group (pattern B), the fatty acid profile was different from patients with pattern A. In these two groups no obvious differences were noted in clinical features and severity. In patients treated with safflower oil, the mean levels of vitamin E were elevated. On all the three treatment schedules, the levels of other fatty acids were not altered. The biochemical and clinical evidence obtained indicate that phrynoderma may not be directly associated with EFA deficiency but that vitamin B-complex may have an important role. The plasma phospholipid fatty acid profile seems to reflect neither the clinical situation nor the response to therapy.

Skin cancer in renal transplant recipients is associated with increased concentrations of 6-thioguanine nucleotide in red blood cells.

Of 108 renal transplant recipients (53 men and 55 women) treated with azathioprine (0.8-2.9 mg/kg/day) and prednisolone (10 mg daily), 10 men had actinic keratoses, and five of these had squamous cell carcinoma, on light-exposed areas of skin. The time from transplantation to diagnosis of these skin lesions varied from 1.2 to 9.0 (mean 5.1) years. The concentration of the active azathioprine metabolite 6-thioguanine nucleotide was 120-425 (mean 276) pmol per 8 X 10(8) red blood cells in the transplant patients who developed skin lesions and 54-203 (mean 130) pmol per 8 X 10(8) red blood cells in a matched control group of renal transplant recipients. This difference was statistically significant (P = 0.005). There was no statistically significant difference between patients and controls in azathioprine dosage, clinical features of immunosuppression, sunlight exposure or infection with human papilloma virus. The association of raised 6-thioguanine nucleotide concentrations in red blood cells with actinic keratoses and malignant skin tumours in these patients supports chemical carcinogenesis as a possible cause.

Cell-surface carbohydrates in proliferative epidermal lesions. Distribution of A, B, and H blood group antigens in benign and malignant lesions.

The distribution of A, B, and H blood group antigens was studied by means of peroxidase-antiperoxidase technique in normal skin and in lesions of carcinomas in situ (solar keratoses, Bowen's disease), squamous cell carcinoma, keratoacanthomas, and verrucae. In normal skin, the epidermis of persons of blood group O showed H antigens throughout the epidermis; of blood group A, H and A antigens; and of blood group B, H and B antigens. In lesions of solar keratoses, there were no antigens of blood groups in the irregular downward proliferations. In five of 11 cases of Bowen's disease, there were no antigens of blood groups in the epidermis. In eight out of 10 cases of squamous cell carcinoma, no antigens of blood groups were found in the islands of the neoplastic process, but in two cases they were present in a patchy distribution. In the benign lesions examined, the antigens of A, B, and H blood groups were always present, although in verrucae the staining was confined to the upper layers of the epidermis only.

Skin disorders and vitamin A metabolism disturbances in chronic dialysis patients: the role of zinc, retinol-binding protein, retinol and retinoic acid.

The authors have studied--in the plasma--the changes of zinc, retinol binding protein (RBP), retinol and retinoic acid with reference to the dermatological status of fifty chronically haemodialysed renal insufficiency patients divided into four subgroups (normal skin, dry skin, dry skin with keratosis, and only keratosis). The results of these groups were compared to those of thirty healthy subjects. The values of these variables do not show any significant difference in function of the dermatological subgroups; but, despite the considerable rise in the retinol binding protein and retinol levels in comparison with the controls (haemodialysis patients: RBP = 11.77 +/- 2.83 mumol X l(-1), retinol = 7 +/- 2.57 mumol X l(-1); controls; RBP = 2.76 +/- 0.62 mumol X l(-1), retinol = 2.16 +/- 0.53 mumol X l(-1] the electromicroscopic examination of skin biopsy samples from some of the patients did not reveal any sign of hypervitaminosis A in the lesions.

DNA repair in cells from patients with actinic keratosis.

UV-induced DNA repair was studied in peripheral blood lymphocytes from patients with multiple actinic keratoses (AK) requiring surgical therapy and from age-matched normal control individuals. The DNA repair activity in lymphocytes from AK patients at 4 h after UV-irradiation was 50% of that in control lymphocytes, but at 21 h the extent of DNA repair synthesis was similar to that in control cells.

Role of vitamin E in the aetiology of phrynoderma (follicular hyperkeratosis) and its interrelationship with B-complex vitamins.

1. A study was undertaken to investigate the role of vitamin E in the aetiology of phrynoderma (follicular hyperkeratosis). Fifty-six children with the disease and twenty-one normal children were investigated for this purpose. 2. Plasma vitamin E levels (mean +/- SE; mg/l) were found to be low in phrynoderma (3.7 +/- 0.19) in contrast with normal children (6.6 +/- 0.40) and therapy with a combination of vitamin E and B-complex brought about complete cure. 3. The increase in plasma vitamin E levels after the administration of vitamin E at a dose of 100 mg three times daily for 4 weeks was higher than that obtained when vitamin E at the same dose was administered together with vitamin B-complex for 4 weeks, suggesting an interaction between the two vitamins. Further studies are necessary to find out the exact nature of this interrelationship.

DNA repair deficiency in lymphocytes from patients with actinic keratosis.

DNA repair activity was measured in peripheral blood lymphocytes from 18 patients with Actinic Keratosis and 18 age-matched control subjects, by comparing the incorporation of 3H-thymidine into cells after irradiation with ultraviolet light with that into unirradiated cells. The incorporation was followed autoradiographically or by measuring acid insoluble radioactivity in cells labelled in the presence of hydroxyurea. The repair activity in lymphocytes from Actinic keratosis patients was only 47.1 percent (+/- 6.5%) of that in cells from the control subjects.

Dyskeratosis congenita: hematologic evaluation of a sibship and review of the literature.

Dyskeratosis congenita is a rare genodermatosis whose hematologic complications include pancytopenia of variable time of onset, a propensity for opportunistic infections, and neoplasia. A family in which the disorder segregated in 3 generations and involved 9 members is reported, and the hematologic data of the 46 previously reported cases are reviewed.