Extracto de Polypodium leucotomos en dermatitis atópica: Ensayo multicéntrico, aleatorizado, doble ciego y controlado con placebo / Polypodium leucotomos Extract in Atopic Dermatitis: A Randomized, Double-Blind, Placebo-Controlled, Multicenter Trial

Background: Several studies have reported an association between tumor necrosis factor Introducción: Para el control de la lesión inflamatoria de la dermatitis atópica, y secundariamente del prurito, se utilizan corticosteroides tópicos, pero su empleo está limitado por las reacciones adversas. Objetivos: El objetivo primario del estudio fue evaluar si el tratamiento diario con extracto de Polypodium leucotomos permitiría reducir el uso de corticosteroides tópicos en niños y adolescentes con dermatitis atópica. Secundariamente se valoró el consumo de antihistamínicos orales, así como la evolución de la enfermedad. Pacientes y métodos: Se ha realizado un ensayo clínico en fase IV , multicéntrico, aleatorizado y doble ciego de extracto de Polypodium leucotomos, controlado con placebo, en 105 pacientes de 2 a 17 años de edad, con dermatitis atópica de intensidad moderada e indicación de corticosteroides tópicos. Los pacientes recibieron durante 6 meses extracto de Polypodium leucotomos o placebo por vía oral añadidos al protocolo terapéutico habitual, y se calculó el porcentaje de días en que se utilizaban corticosteroides tópicos u otros tratamientos para la dermatitis atópica. Resultados: El extracto de Polypodium leucotomos redujo de modo no significativo el uso de corticosteroides tópicos (11±12% de días), comparado con placebo (12±11%). El porcentaje de días en los que los pacientes requirieron antihistamínicos orales fue significativamente menor con extracto de Polypodium leucotomos (mediana de 4,5% días) que con placebo (13,6%) (p=0,038). También se redujo el porcentaje de pacientes que tomaron antihistamínicos orales. Conclusiones: El tratamiento prolongado con extracto de Polypodium leucotomos aporta beneficios relevantes para los pacientes en edad pediátrica con dermatitis atópica que precisan tratamiento farmacológico para controlar la lesión inflamatoria y reducir el prurito (AU)

Introduction: Topical corticosteroids are used to treat inflammation and relieve itching in atopic dermatitis, but their use is limited by adverse reactions. Objectives: The main aim of this study was to investigate whether daily treatment with Polypodium leucotomos extract would reduce the use of topical corticosteroids in children and adolescents with atopic dermatitis. We also analyzed oral antihistamine use and changes in disease severity. Patients and methods: We performed a phase IV randomized, double-blind, placebo-controlled, multicenter trial involving 105 patients aged between 2 and 17 years who were receiving topical corticosteroids to treat moderate atopic dermatitis. The patients were randomized to receive, in addition to their standard treatment, Polypodium leucotomos extract or placebo (both in capsule form) for 6 months. The percentage of days on which topical corticosteroids and other atopic dermatitis treatments were used was calculated. Results: Use of Polypodium leucotomos extract did not significantly reduce the mean (SD) percentage of days on which topical corticosteroids were used (11% [12%] vs 12% [11%] for placebo). A significant reduction was, however, observed for oral histamine use (median percentage of days, 4.5% in the Polypodium leucotomos group and 13.6% in the placebo group [P= 0.038]). The percentage of patients who used oral antihistamines was also lower in the Polypodium leucotomos group. Conclusion: Long-term treatment with Polypodium leucotomos extract has benefits for children and adolescents with atopic dermatitis who require pharmacologic treatment to reduce inflammation and relieve itching (AU)

Polypodium leucotomos extract in atopic dermatitis: a randomized, double-blind, placebo-controlled, multicenter trial.

INTRODUCTION: Topical corticosteroids are used to treat inflammation and relieve itching in atopic dermatitis, but their use is limited by adverse reactions. OBJECTIVES: The main aim of this study was to investigate whether daily treatment with Polypodium leucotomos extract would reduce the use of topical corticosteroids in children and adolescents with atopic dermatitis. We also analyzed oral antihistamine use and changes in disease severity. PATIENTS AND METHODS: We performed a phase IV randomized, double-blind, placebo-controlled, multicenter trial involving 105 patients aged between 2 and 17 years who were receiving topical corticosteroids to treat moderate atopic dermatitis. The patients were randomized to receive, in addition to their standard treatment, Polypodium leucotomos extract or placebo (both in capsule form) for 6 months. The percentage of days on which topical corticosteroids and other atopic dermatitis treatments were used was calculated. RESULTS: Use of Polypodium leucotomos extract did not significantly reduce the mean (SD) percentage of days on which topical corticosteroids were used (11% [12%] vs 12% [11%] for placebo). A significant reduction was, however, observed for oral histamine use (median percentage of days, 4.5% in the Polypodium leucotomos group and 13.6% in the placebo group [P= .038]). The percentage of patients who used oral antihistamines was also lower in the Polypodium leucotomos group. CONCLUSION: Long-term treatment with Polypodium leucotomos extract has benefits for children and adolescents with atopic dermatitis who require pharmacologic treatment to reduce inflammation and relieve itching.

Estrés oxidativo y suplementación antioxidante de la dieta en el envejecimiento, la aterosclerosis y la disfunción inmunitaria / Oxidative stress and antioxidant diet supplementation in ageing, atherosclerotic and immune dysfunction processes

El aumento de la longevidad en los países industrializados requiere costosos servicios sanitarios y asistenciales, pues muchas personas ancianas sufren un proceso de envejecimiento patológico ligado a enfermedades degenerativas crónicas y déficits funcionales de larga duración. Por otra parte, numerosos estudios sugieren que más personas podrían evitar el envejecimiento patológico si consumieran dietas ricas en antioxidantes. Así sería posible proteger más eficazmente al organismo contra el estrés oxidativo, que contribuye al envejecimiento normal y tiene un papel aún más importante en la aterosclerosis, la inmunodepresión y otros procesos degenerativos que a menudo forman parte del envejecimiento patológico. De acuerdo con lo anterior, y según los datos revisados, la suplementación de la dieta de los sujetos de edad madura o avanzada con antioxidantes (como la tioprolina, la N-acetilcisteina y los antioxidantes fenólicos de la cúrcuma) podría aumentar las probabilidades de prevenir o frenar los mencionados procesos degenerativos, ayudando así a conseguir una mayor longevidad con adecuada preservación funcional (AU)

Anapsos (Polypodium leucotomos) modulates lymphoid cells and the expression of adhesion molecules.

Anapsos is a medical prescription registered in the Health Ministry of Spain, that is obtained from the rhizomes of the fern Polypodium leucotomos. An immunomodulating effect of Anapsos on certain lymphocyte subsets and cytokines has already been described in the literature. The current study extends and supports part of the aforementioned results of the product on the immune system, showing the ability of Anapsos to stimulate proliferation and activation of T and natural killer lymphocytes, as well as an important down-regulating effect on CD11, CD18 and CD62-L adhesion molecules, both on peripheral blood mononuclear cells and on U-937 and HL-60 cell lines.

An hydroalcoholic extract of curcuma longa lowers the apo B/apo A ratio. Implications for atherogenesis prevention.

It is generally accepted that free-radical induced blood lipid peroxidation and especially peroxidized LDL play a central role in the pathogenesis of atherosclerosis and related cardiovascular disease. Moreover, recent research highlights the key contribution of apolipoprotein B (apo B) to atherogenesis as the main inductor of one of its earlier steps, i.e. macrophage proliferation. This has led us to investigate the apo B response to a very effective phenolic lipid-antioxidant, namely an hydroalcoholic extract of Curcuma longa, which according to our previous work does not show any toxic effects and decreases the levels of blood lipid peroxides, oxidized lipoproteins and fibrinogen. The present study shows that a daily oral administration of the extract decreases significantly the LDL and apo B and increases the HDL and apo A of healthy subjects. This and recent data on the increased anti-atherogenic action of the physiological antioxidant tocopherol in the presence of phenolic co-antioxidants (which eliminate the tocopheroxyl radical), justifies planned clinical research to test the usefulness of the curcuma extract as a co-antioxidant complement to standard treatments to prevent or retard atherosclerosis.

Melanocytes respond to mechanical stretch by activation of mitogen-activated protein kinases (MAPK).

Cells of human epidermis are permanently targeted by mechanical stimuli. Besides mechanical forces from external sources the body itself generates mechanical forces via muscle contractions and growth processes. Recently, it was demonstrated that mechanical stretch is connected to enhanced proliferation in epidermal cells. The underlying biochemical events are still a matter of debate. Here we show that mechanical stretch leads to activation of both ERK1/2 and SAPK/JNK in human melanocytes and keratinocytes. In response to a 5 min single stretch ERK1/2 becomes moderately induced in melanocytes and peaked 30 min after the stimulus. In keratinocytes strong activation of ERK1/2 is present directly after the stimulus. SAPK/JNK shows the same activation pattern in both cell species--a slow but steady activation. The different kinetics of both MAPK suggest that different signalling cascades were activated. Future studies should evaluate the relevance of stretch-dependent MAPK activation in triggering the cell proliferation.

Efectos farmacológicos y nutricionalesde los extractos de Curcuma longa L.y de los cucuminoides / Pharmacological and nutritional effects of Curcuma longa L. extracts and curcuminoids

La Curcuma longa L., es una planta de origen asiático muy usada comúnmente como una especia en la cultura asiática. El principal componente es la curcumina, uno de los ingredientes activos responsables de su actividad biológica. Se sabe que esta sustancia es estable en el estómago y en el intestino delgado; su elevada lipofilia le permite una rápida absorción gastrointestinal por difusión pasiva. Tras su administración, es metabolizada y excretada principalmente por bilis y heces, y también por orina. Sus principales metabolitos también son bioactivos. Desde antiguo, se han descrito muchas propiedades para los extractos de Curcuma longa y para la curcumina. Se conoce su actividad antibacteriana, antifúngica y antiparasitaria, y recientemente se ha demostrado su capacidad para inhibir la integrasa del HIV-1. También se han demostrado efectos específicos en otros tejidos y órganos, como la piel, el sistema gastrointestinal y respiratorio y en el hígado. Todas estas propiedades son debidas a distintos mecanismos de acción. Se ha demostrado que la cúrcuma posee efectos antiinflamatorios, a través de la modulación del metabolismo de los eicosanoides, tiene capacidad inmunomoduladora, principalmente alterando el perfil de las citoquinas Thl de los linfocitos T helper, y actividad hipolipidémica, disminuyendo el colesterol, los triglicéridos y los fosfolípidos plasmáticos así como en las LDL. Hay muchos estudios que demuestran la capacidad de la cúrcuma para estabilizar membranas y para prevenir la peroxidación lipídica, un proceso fundamental en el establecimiento, la progresión y las complicaciones de muchas patologías como las enfermedades hepáticas, renales, cardiovasculares, neurodegenerativas, en la diabetes y en las cataratas. Las últimas investigaciones sobre los efectos biológicos de los extractos de cúrcuma y de los curcuminoides están encaminados a estudiar su actividad anticancerosa, principalmente frente al cáncer de piel, colon y duodeno (AU)

Phototoxic effects of Hypericum extract in cultures of human keratinocytes compared with those of psoralen.

Extracts of Hypericum perforatum (St. John's wort) are used in the treatment of depression. They contain the plant pigment hypericin and hypericin derivates. These compounds have light-dependent activities. In order to estimate the potential risk of phototoxic skin damage during antidepressive therapy, we investigated the phototoxic activity of hypericin extract using cultures of human keratinocytes and compared it with the effect of the well-known phototoxic agent psoralen. The absorbance spectrum of our Hypericum extract revealed maxima in the whole UV range and in parts of the visible range. We cultivated human keratinocytes in the presence of different Hypericum concentrations and irradiated the cells with 150 mJ/cm2 UVB, 1 J/cm2 UVA or 3 h with a white light of photon flux density 2.6 mumol m-2 s-1. The determination of the bromodeoxyuridine incorporation rate showed a concentration- and light-dependent decrease in DNA synthesis with high hypericin concentrations (> or = 50 micrograms/mL) combined with UVA or visible light radiation. In the case of UVB irradiation a clear phototoxic cell reaction was not detected. We found phototoxic effects even with 10 ng/mL psoralen using UVA with the same study design as in the case of the Hypericum extract. These results confirm the phototoxic activity of Hypericum extract on human keratinocytes. However, the blood levels that are to be expected during antidepressive therapy are presumably too low to induce phototoxic skin reactions.

The mechanism of melanocyte dendrite formation: the impact of differentiating keratinocytes.

In human epidermis one dendritic melanocyte interacts with about 36 keratinocytes and supplies them with melanin. In contrast to the vivo situation melanocytes in culture are far less dendritic. In the present study different culture systems were tested in order to observe the mechanism of melanocyte dendrite formation. In particular, we focused on the role of keratinocytes in this process. Time lapse studies revealed that only differentiated keratinocytes enhance melanocyte dendricity. Differentiated keratinocytes form connected cell sheets, which attach to part of the melanocyte plasma membrane. By contraction and retraction of keratinocyte units, new dendrites were drawn out from the melanocytes. Melanocytes remain passive during this process, which is indicated by the observation that sometimes extended dendrites could not withstand the tension and shear.

Increase with age of serum lipid peroxides: implications for the prevention of atherosclerosis.

There is considerable support for the concept that oxygen free radicals and related lipid peroxides play a key role in the pathogenesis of normal senescence and of age-related chronic degenerative diseases, including atherosclerosis. This has led to a great deal of interest regarding peroxidized LDL, which seems to be more atherogenic than LDL. In contrast, the relationship of total serum or plasma lipid peroxides (which also have a marked atherogenic action) with both aging and atherogenesis are not well understood. In view of the above, we have determined the level of serum lipid peroxide (expressed as thiobarbituric acid reactive substances) in a sample of 100 healthy men and women ranging in age from 20 to 70 years. Our data show that there is an age related increase in the concentration of lipid peroxide, with men showing higher or about equal values than women until about 60 years, after which age women show the higher values. Our data also suggest that in certain men and women, aging is linked to a decline in the competence of the oxyradical-detoxifying mechanisms, which results in increased serum lipid peroxidation. Further research is needed to find out if lowering the serum peroxide levels of aging subjects by diet supplementation with antioxidants will decrease that risk. An adequate intake of antioxidants seems especially indicated in post-menopausal women because of their apparent greater sensitivity to age related oxygen stress.

Melanocytes in vitro: how do they undergo mitosis?

Human melanocytes of the adult skin are slow-cycling cells with a highly dendritic morphology. Nevertheless in vitro proliferation can be achieved using adequate stimulators. Time lapse studies revealed the morphologic changes during melanocyte mitosis: dendrites are drawn back into the cell body, the cell becomes spherical and detaches from the support. Cell division takes place while the cell is suspended. Consecutively the two cells reattach to the support and form new dendrites. About 1% cells per culture are detached from the support and ca. 70% of these cells are viable and putative within mitosis. By every medium change mitotic cells become withdrawn supporting selection of G0-cells, Therefore we recommend centrifugation of exhausted medium in order to add mitotic cells back to the culture.

Candida albicans suppresses transcription of melanogenesis enzymes in cultured melanocytes.

Human skin can be colonized by different yeasts that may have an impact on skin pigmentation. In order to study this effect normal human melanocytes were cultured with different yeasts. Reverse transcription polymerase chain reaction (RT-PCR) analysis gives evidence that Candida albicans suppresses the transcription of melanogenesis enzymes.

Effects of the antioxidant turmeric on lipoprotein peroxides: Implications for the prevention of atherosclerosis.

Extracts from the rhyzome of Curcuma longa are widely used as food additives in India and other Asiatic and Central American countries. It has been shown that these extracts ("turmeric"), as well as "curcumin" and related phenolic compounds isolated from Curcuma, have a powerful antioxidant action when tested in in vitro systems. Moreover, previous research from our laboratories has shown significant decreases in the levels of lipid peroxides in the blood of both mice and human subjects administered "turmeric." Our present research complements the previous data, showing that a daily intake of turmeric equivalent to 20 mg of the phenolic antioxidant curcumin for 60 days decreases the high levels of peroxidation of both the HDL and the LDL, in vivo, in 30 healthy volunteers ranging in age from 40 to 90 years. The effect was quite striking in the persons with high baseline values of peroxidized compounds in these lipoproteins, while no apparent change took place in the persons having low baseline values. In view of current concepts on the atherogenic role played by peroxidized HDL, and especially by peroxidized LDL, as inducers of foam and smooth cell proliferation in the arterial wall, this preliminary experiment suggests that the Curcuma phenolic antioxidants, because of their high antioxidant activity and lack of toxicity, might be a useful complement to standard hypo-lipidemic drugs in the prevention and treatment of atherosclerosis.

Measurement of anti-inflammatory effects of glucocorticoids on human keratinocytes in vitro. Comparison of normal human keratinocytes with the keratinocyte cell line HaCaT.

There are only few objective in vitro methods available for the testing of anti-inflammatory pharmaceutical products. One possibility is in the stimulation of cytokine production in cultivated human keratinocytes by UV light and the subsequent testing of suppressing activities. From the dermatological aspect the interleukins 6 and 8 are especially interesting because they are elevated in psoriatic skin. In the present work three glucocorticoids were tested in cultures of normal human keratinocytes and in the permanent keratinocyte cell line HaCaT. Both cell species produced IL-6 and IL-8 spontaneously, albeit in very small amounts. After UV irradiation the interleukin production increased in a dose dependent manner. The IL-6 and IL-8 induction could be suppressed by each of the glucocorticoids tested. The thymidine incorporation rate of the cells was not affected by the glucocorticoids indicating that the observed suppression of cytokine induction was not the result of a generalised cell damage. The response of both HaCaT keratinocytes and primary human keratinocytes to UV irradiation and glucocorticoid application was similar indicating the possible use of the generally available HaCaT cells for the pharmacological testing of anti-inflammatory activities in vitro.

Transcription of melanogenesis enzymes in melanocytes: dependence upon culture conditions and co-cultivation with keratinocytes.

Eumelanogenesis of human skin melanocytes requires at least three enzymes: tyrosinase, TRP 1, and TRP 2. The regulation of these enzymes on transcriptional level was detected in a semiquantitative attempt. The total RNA of melanocytes was reverse-transcripted and followed by a PCR with degenerated primers for all three enzymes. The amplification products were related to each other densitometrically. We examined five different culture conditions: 1) melanocytes in a popular phorbolester containing F-10-medium, 2) melanocytes in a co-culture medium with EGF, 3) melanocytes in a co-culture medium with high calcium, 4) melanocytes co-cultured with keratinocytes in EGF containing co-culture medium, and 5) melanocytes co-cultured with keratinocytes in co-culture medium with high calcium. Melanocytes cultured in phorbolester containing F-10-medium featured transcripts of tyrosinase, TRP 1, and TRP 2 in the ratio 45:45:10. The same results were obtained for melanocytes co-cultured with keratinocytes under the two different culture conditions. In melanocytes cultured alone in co-culture media only TRP 1-transcripts were present. It is likely that under co-culture conditions a keratinocyte-derived factor supports the transcription of all three enzymes. For melanocytes in the phorbolester-containing melanocyte medium a proteinkinase C dependent regulation of transcription seems possible.

In vitro studies on the immunomodulating effects of polypodium leucotomos extract on human leukocyte fractions.

The in vitro effect of Anapsos, a water based extract of the naturally occurring fern Polypodium leucotomos (calagualine), on human leukocyte fractions was investigated. Calagualine inhibited interleukin-2 secretion and concanavalin A (Con A) stimulated proliferation of T-lymphocytes in a concentration dependent manner. In contrast, a greatly enhanced secretion of IL-1 alpha, IL-1 beta and tumor necrosis factor alpha was induced suggesting a stimulation of monocytes and dendritic cells also present in this system. Endotoxin induced stimulation was excluded. Also in the absence of Con A, calagualine stimulated cytokine production. The presented data show for the first time that calagualine exerts an immunomodulating effect on leukocyte fractions, paving the way for further detailed studies related to possible clinical efficacy.

Levels of dopachrome tautomerase in human melanocytes cultured in vitro.

Several reports have been published about the level of activity and possible functions of dopachrome tautomerase (DCT) in mouse melanoma cells. Data about the levels of this activity in human melanocytes in culture are still scarce, and, as far as we know, a comparison between mouse and human melanocytes, or between normal and malignant melanocytes, has never been published. We have measured the tyrosinase and DCT activities, as well as the melanin content, in mouse Cloudman melanoma cells, two lines of human melanoma, and three lines of normal human melanocytes obtained from fetal skin. Although more cell lines should be tested to draw a general conclusion, our results suggest that normal melanocytes contained much higher tyrosinase activity and melanin content but lower DCT activity than malignant melanocytes. The two lines of human melanoma cells tested had lower levels of DCT activity than Cloudman melanoma cells. Finally, the low level of DCT activity found in normal human melanocytes cultured in vitro cannot be explained by any of the necessary stimulatory factors added to the cell culture media.

[Cell and tissue culture models in dermatology. The Frankfurt Center of Dermatology establishes models]. / Zell- und Gewebekulturmodelle in der Dermatologie. Die im Frankfurter Zentrum der Dermatologie etablierten Modelle.

In the course of the Third Frankfurt Talks, for the first time a congress in dermatology was dedicated exclusively to cell and tissue culture models. The complexity of a whole organ, in this case the skin, could be reduced to single aspects without losing the holistic context. Ways of managing this were discussed on an interdisciplinary level by dermatologists, physiologists, pharmacologists and biologists. The results are also expected to be useful to the clinician. Focus points of in vitro investigations for dermatology are wound closure models and the use of in vitro skin for transplantation in the therapy of non-healing ulcers and vitiligo. As an alternative to animal experiments, cultures of human cells are gaining increasing influence in drug testing. The effect of glucocorticosteroids on normal skin fibroblasts, keratinocytes and permanent cell lines is discussed as an example, and in vitro models of diseases such as psoriasis are established. Additionally, basic events such as differentiation and ageing have been modelled in cell cultures of melanocytes and keratinocytes. Mechanical stress, UV radiation and nicotine are discussed as inductors.

Effect of minoxidil on the mobility and differentiation of cultivated human keratinocytes.

The mode of topical action of minoxidil (U-10,858, CAS 38304-91-5) ist not entirely clear. The influence of minoxidil on the ultimate differentiation of keratinocytes was investigated. Human keratinocytes (HaCaT-cells) were incubated with minoxidil containing culture medium (10-250 micrograms/ml). Minoxidil inhibited in a concentration dependent manner cell mobility whereas the adhesion area and the cell circumference were increased. The minoxidil treated cultures had a higher desmosome density compared to parallel control cultures and they expressed the suprabasal keratins 1 and 10 (indicating progress in differentiation) earlier and to a larger extent than the controls. Keratins were revealed immunohistochemically and by two-dimensional polyacrylamide gel electrophoresis. The results suggest that minoxidil supports the differentiation of human keratinocytes.