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Despite historical mischaracterization as a cosmetic condition, patients with the autoimmune disorder vitiligo experience substantial quality-of-life (QoL) burden. This systematic literature review of peer-reviewed observational and interventional studies describes comprehensive evidence for humanistic burden in patients with vitiligo. PubMed, EMBASE, Scopus and the Cochrane databases were searched through February 10, 2021, to qualitatively assess QoL in vitiligo. Two independent reviewers assessed articles for inclusion and extracted data for qualitative synthesis. A total of 130 included studies were published between 1996 and 2021. Geographical regions with the most studies were Europe (32.3%) and the Middle East (26.9%). Dermatology-specific instruments, including the Dermatology Life Quality Index (DLQI; 80 studies) and its variants for children (CDLQI; 10 studies) and families (FDLQI; 4 studies), as well as Skindex instruments (Skindex-29, 15 studies; Skindex-16, 4 studies), were most commonly used to measure humanistic burden. Vitiligo-specific instruments, including the Vitiligo-specific QoL (VitiQoL; 11 studies) instrument and 22-item Vitiligo Impact Scale (VIS-22; 4 studies), were administered in fewer studies. Among studies that reported total scores for the overall population, a majority revealed moderate or worse effects of vitiligo on patient QoL (DLQI, 35/54 studies; Skindex, 8/8 studies; VitiQoL, 6/6 studies; VIS-22, 3/3 studies). Vitiligo also had a significant impact on the QoL of families and caregivers; 4/4 studies reporting FDLQI scores indicated moderate or worse effects on QoL. In general, treatment significantly (P 0.05) improved QoL, but there were no trends for types or duration of treatment. Among studies that reported factors significantly (P ≤ 0.05) associated with reduced QoL, female sex and visible lesions and/or lesions in sensitive areas were most common. In summary, vitiligo has clinically meaningful effects on the QoL of patients, highlighting that greater attention should be dedicated to QoL decrement awareness and improvement in patients with vitiligo.

Introduction Surgical site infection (SSI) is one of the most common healthcare-associated infections (HAIs). However, there is a lack of data available about SSI in children worldwide, especially from low-income and middle-income countries. This study aimed to estimate the incidence of SSI in children and associations between SSI and morbidity across human development settings. Methods A multicentre, international, prospective, validated cohort study of children aged under 16 years undergoing clean-contaminated, contaminated or dirty gastrointestinal surgery. Any hospital in the world providing paediatric surgery was eligible to contribute data between January and July 2016. The primary outcome was the incidence of SSI by 30 days. Relationships between explanatory variables and SSI were examined using multilevel logistic regression. Countries were stratified into high development, middle development and low development groups using the United Nations Human Development Index (HDI). Results Of 1159 children across 181 hospitals in 51 countries, 523 (45·1%) children were from high HDI, 397 (34·2%) from middle HDI and 239 (20·6%) from low HDI countries. The 30-day SSI rate was 6.3% (33/523) in high HDI, 12·8% (51/397) in middle HDI and 24·7% (59/239) in low HDI countries. SSI was associated with higher incidence of 30-day mortality, intervention, organ-space infection and other HAIs, with the highest rates seen in low HDI countries. Median length of stay in patients who had an SSI was longer (7.0 days), compared with 3.0 days in patients who did not have an SSI. Use of laparoscopy was associated with significantly lower SSI rates, even after accounting for HDI. Conclusion The odds of SSI in children is nearly four times greater in low HDI compared with high HDI countries. Policies to reduce SSI should be prioritised as part of the wider global agenda. ©

Summary Aim To validate the Spanish version of the Emotional Labour Scale (ELS), an instrument widely used to understand how professionals working with people face emotional labor in their daily job. Method An observational, cross-sectional and multicenter survey was used. Nursing students and their clinical tutors (n = 211) completed the self-reported ELS when the clinical practice period was over. First order and second order Confirmatory Factor Analyses (CFA) were estimated in order to test the factor structure of the scale. Conclusions The results of the CFA confirm a factor structure of the scale with six first order factors (duration, frequency, intensity, variety, surface acting and deep acting) and two larger second order factors named Demands (duration, frequency, intensity and variety) and Acting (surface acting and deep acting) establishing the validity of the Spanish version of the ELS.

The aim of this review was to evaluate, by a thorough revision of the literature, the true efficacy of currently available topic and systemic cosmetic acne agents.The efficacy of currently available cosmetic acne agents has been retrospectively evaluated via thorough revision of the literature on matched electronic databases (PubMed). All retrieved studies, either randomized clinical trials or clinical trials, controlled or uncontrolled were considered.Scientific evidence suggests that most cosmetic products for acne may enhance the clinical outcome. Cleansers should be indicated to all acne patients; those containing benzoyl peroxide or azelaic/salicylic acid/triclosan show the best efficacy profile. Sebum-controlling agents containing nicotinamide or zinc acetate may minimize excessive sebum production. Cosmetics with antimicrobial and anti-inflammatory substances such as, respectively, ethyl lactate or phytosphingosine and nicotinamide or resveratrol, may speed acne recovery. Topical corneolytics, including retinaldehyde/glycolic acid or lactic acid, induce a comedolytic effect and may also facilitate skin absorption of topical drugs. Finally, the use of specific moisturizers should be strongly recommended in all acne patients.Cosmetics, if correctly prescribed, may improve the performance of the therapy, whereas wrong procedures and/or inadequate cosmetics may worsen acne. Cosmetological recommendations may allow clinicians to make informed decisions about the role of various cosmetics and to indentify the appropriate indications and precautions. The choice of the most effective product should take into consideration the ongoing pharmacological therapy and acne type/severity as well.

Although techniques such as biopanning rely heavily upon the screening of randomized gene libraries, there is surprisingly little information available on the construction of those libraries. In general, it is based on the cloning of 'randomized' synthetic oligonucleotides, in which given position(s) contain an equal mixture of all four bases. Yet, many supposedly 'randomized' libraries contain significant elements of bias and/or omission. Here, we report the development and validation of a new, PCR-based assay that enables rapid examination of library composition both prior to and after cloning. By using our assay to analyse model libraries, we demonstrate that the cloning of a given distribution of sequences does not necessarily result in a similarly composed library of clones. Thus, while bias in randomized synthetic oligonucleotide mixtures can be virtually eliminated by using unequal ratios of the four phosphoramidites, the use of such mixtures does not ensure retrieval of a truly randomized library. We propose that in the absence of a technique to control cloning frequencies, the ability to analyse the composition of libraries after cloning will enhance significantly the quality of information derived from those libraries. (C) 2000 Published by Elsevier Science B.V. All rights reserved.

He debido recurrir a una serie de nociones teóricas que permitieran desarrollar la tesis del presente trabajo: la institucionalización en el sistema literario español de una relación contractual entre autor y lector, relación a través de la cual se posibilita un cierto realismo en el programa escritural villeniano, que abarca la curva lírica 1971-1993. La literatura entendida como sistema permite ver la obra de Villena en una curva lírica sobreimpresa a la estética novísima y a la poesía de los '80, y con ello, las correlaciones de lo literario y de lo extraliterario El caso de Villena resulta interesante, porque su personaje social se imbrica con sus otros sujetos textuales, ya sea ante un gran público, ya sea desde la televisión o desde la columna periodística. Delimita así un campo mucho más amplio y complejo que el meramente textual, para la reconstrucción de una ideología.

The aetiopathogenic mechanisms of vitiligo are still poorly understood, and this has held back progress in diagnosis and treatment. Up until now, treatment guidelines have existed at national levels, but no common European viewpoint has emerged. This guideline for the treatment of segmental and nonsegmental vitiligo has been developed by the members of the Vitiligo European Task Force and other colleagues. It summarizes evidence-based and expert-based recommendations (S1 level).

Vitiligo skin shows different burning capacity in people with different phototype. In normal skin antioxidant status is correlated to skin phototype, but unexpectedly it appears that there is a gradual decrease in burning susceptibility of depigmented skin of individuals with increasing phototype (II→VI).To assess if the antioxidant response in the lesional vitiligo skin is involved in those protection mechanisms. Moreover, a possible correlation between cutaneous and systemic endogenous antioxidants in vitiligo patients has been investigated.We enrolled in the study 29 patients with active vitiligo, divided into five groups according to skin type (II to VI). We analysed reduced and oxidized glutathione (GSH and GSSG, respectively), ubiquinone (CoQ10), catalase (Cat), superoxide dismutases (Cu/Zn-SOD and Mn-SOD), GSH peroxidase (GSH-Px), as indexes of chemical and enzymatic antioxidants, in suction blister roofs as well as in peripheral blood mononuclear cells (PBMNCs).The vitiligo patients showed an imbalance of antioxidant network, both in depigmented skin and PBMNCs. Interestingly, in vitiligo skin a phototype-related increase of antioxidant enzyme activities (Cat, Mn-SOD and GPx) and GSH amount have been observed. Similarly in PMBNCs Cat and total SOD activities, as well as GSH content progressively increased from skin type II to skin type VI. Endogenous antioxidants in vitiligo skin are correlated to those in PBMNCs, suggesting that systemic and epidermal antioxidant network functionalities are connected.The correlation between antioxidant levels and clinical phototype confirmed the hypothesis that other factors than melanin determine largely the minimal erythema dose values in vitiligo lesional skin.

Recent studies suggest that paraoxonase-1 (PON1), complexed with high-density lipoproteins, is the major lactonase in the circulation. Using 5-hydroxy eicosatetraenoate δ-lactone (5-HETEL) as the substrate, we observed lactonase activity in serum from Pon1−/− mice. However, 6–12 carbon fatty acid γ- and δ-lactones were not hydrolyzed in serum from Pon1−/− mice. Serum from both wild-type and Pon1−/− mice contained a lactonase activity towards 5-HETEL and 3-oxo-dodecanoyl-homoserine lactone that was resistant to inactivation by EDTA. This lactonase activity was sensitive to the serine esterase inhibitor phenyl methyl sulfonyl fluoride and co-eluted with carboxylesterase activity by size-exclusion chromatography. Analysis of serum from the Es1e mouse strain, which has a deficiency in the carboxylesterase, ES-1, proved that this activity was due to ES-1. PON1 activity predominated at early time points (30 s), whereas both PON1 and ES-1 contributed equally at later time points (15 min). When both PON1 and ES-1 were inhibited, 5-HETEL was stable in mouse serum. Thus, while long-chain fatty acid lactones are substrates for PON1, they can be hydrolyzed by ES-1 at neutral pH. In contrast, medium-chain length fatty acid lactones are stable in mouse serum in the absence of PON1, suggesting that PON1 plays a specific role in the metabolism of these compounds.

Cutaneous pigmentation is regulated by a complex melanogenic network in which both keratinocytes and fibroblasts synthesize growth factors and cytokines. Solar lentigo (SL) is characterized by hyperpigmented lesions occurring on photodamaged skin areas. Despite the association of SL to ultraviolet (UV) exposure, the mechanisms underlying the development of these spots are not completely defined.To analyse the involvement of the fibroblast-derived growth factors, hepatocyte growth factor (HGF), keratinocyte growth factor (KGF) and stem cell factor (SCF) in SL hyperpigmentation; to evaluate whether the photoageing process occurring in fibroblasts could be responsible for the altered expression of these cytokines; and to investigate a new possible role of KGF in regulating pigmentation through the specific induction of melanogenic cytokines by keratinocytes.We performed immunohistochemical analysis of HGF, KGF and SCF on SL biopsies. We analysed the mRNA expression of these cytokines using an in vitro model of photoageing induced on fibroblasts. Finally, we evaluated the effects of KGF on the expression of melanogenic cytokines at the mRNA and protein levels on keratinocytes.We found positive staining for HGF, KGF and SCF in the upper dermis of SL lesions and a significant induction of the three cytokines in photoaged fibroblasts. We also demonstrated the contribution of KGF to pigmentation, showing its ability specifically to modulate the expression of SCF in keratinocytes.Fibroblasts may be persistently activated by UV exposure to release melanogenic growth factors; this inducible cytokine network acts both directly and indirectly through keratinocytes and may contribute to the hyperpigmentation of SL.

An imbalance of keratinocyte-derived cytok-ines and dysregulations of cytokine/receptor interactions both capable of affecting melano-cyte activity and survival has been demonstrated in vitiligo epidermis. Melanogenic growth factors such as stem-cell factor (SCF) and endothelin (ET)-1, as well as inflammatory cytokines with anti-melanogenic properties, such as IL-1, IL-6, and tumor necrosis factor (TNF)-α, are implicated. The over-expression of TNF-α may facilitate the apoptosis of keratinocytes, leading to a decrease in the production of ET-1 and SCF, thus enhancing melanocyte disappearance. At the junction of lesional vitiligo epidermis, melanocytes remain and express tyrosinase, S100α and ET-B receptor (ETBR), but not c-kit or MITF-M. At the center of lesional vitiligo epidermis, there is a complete loss of melanocytes expressing c-kit, S100α, ETBR, and/or tyrosinase. This deterioration in the expression of c-kit by melanocytes and its downstream effectors, including MITF-M, may be associated with the dysfunction and/or loss of melanocytes in vitiligo epidermis. The cytokine imbalance in vitiligo epidermis may be also related to an impaired keratinocyte senescence process.

Fetal skin fibroblasts produce a soluble “migration stimulating factor” (MSF) which is not made by their normal adult counterparts. MSF stimulates the migration of adult skin fibroblasts into 3D collagen gels, thus providing the basis of a convenient bioassay for its presence. We have previously reported that MSF stimulates hyaluronic acid (HA) synthesis by adult skin fibroblasts and that this effect on matrix deposition appears to be responsible for the observed increase in cell motility. In the present study, wound fluid samples were collected from 18 patients undergoing surgery for various nonmalignant conditions and these were then fractionated according to the protocol used to isolate MSF from fetal fibroblast-conditioned medium. Detectable levels of migration stimulating activity were present in 17 18 (94%) of these samples. Paired serum samples obtained both pre- and postoperatively from five patients with positive wound fluid samples were also analyzed for MSF activity; such activity was found in only 1 5 (20%) of the preoperative and 0 5 (0%) of the postoperative serum samples. These data suggest that the MSF present in wound fluid is not derived from a plasma transudate or from platelet degranulation, but may reflect the transient and localized reinitiation of MSF production by adult fibroblasts in response to wounding. Taken together with previous observations regarding the effect of MSF on fibroblast migration and HA synthesis, our data suggest a possible physiological function of MSF in the wound healing response. Previous studies have revealed that MSF is produced by a subpopulation of apparently persistent fetal-like skin fibroblasts obtained from breast cancer patients and is also found in the serum of these individuals. Wound fluid and serum samples were accordingly collected from patients undergoing surgery for various types of malignant conditions or with a previous history of cancer; detectable levels of MSF activity were found in 8 10 (80%) of these wound fluid samples, 2 3 (66.6%) of the preoperative, and 3 3 (100%) of the postoperative paired serum samples. These findings suggest that the presence of detectable serum levels of MSF is not restricted to breast cancer and may be a general feature of malignant disease.

The pathobiology of vitiligo has been hotly disputed for as long as one remembers, and has been a magnet for endless speculation. Evidently, the different schools of thought--ranging, e.g. from the concept that vitiligo essentially is a free-radical disorder to that of vitiligo being a primary autoimmune disease--imply very different consequences for the best therapeutic strategies that one should adopt. As a more effective therapy for this common, often disfiguring pigmentary disorder is direly needed, we must strive harder to settle the pathogenesis debate definitively--on the basis of sound experimental evidence, rather than by a war of dogmatic theories. Recognizing, however, that it is theories which tend to guide our experimental designs and choice of study parameters, the various pathogenesis theories on the market deserve to be critically, yet unemotionally re-evaluated. This Controversies feature invites you to do so, and to ask yourself: is there something important or worthwhile exploring in other pathogenesis scenarios than those already favoured by you that may help you improve your own study design, next time you have a fresh look at vitiligo? Vitiligo provides a superb model for the study of many fundamental problems in skin biology and pathology. Therefore, even if it later turns out that, as far as your own vitiligo pathogenesis concept is concerned, you have barked-up the wrong tree most of the time, chances are that you shall anyway have generated priceless new insights into skin function along the way.

Paraoxonase 1 (PON1) requires calcium for activity and is inactivated in the presence of EDTA. Because of this, studies to date have used serum or heparinized plasma for both activity and mass assays of PON1. Whole serum and EDTA plasma were analyzed by SDS-electrophoresis and Western blot using anti-PON1 monoclonal antibody 4C10. Because PON1 has one disulfide and one free cysteine residue, the samples were reduced with dithiothreitol before electrophoresis. Western blot identified a major PON1 band with a molecular mass of approximately 45 kDa and two minor bands of approximately 40 and 35 kDa in both serum and EDTA plasma. This established that PON1 is inactive, but structurally intact, in EDTA plasma and suggested that a mass assay could be developed based on SDS-electrophoresis and Western blot. Linearity was established for plasma and for a PON1 standard. Quantification was based on the major PON1 band at 45 kDa. The correlation between serum and plasma PON1 mass was 0.9553. The between-run variation was determined with a serum pool to be 7.8%. The mass of PON1 in serum was significantly correlated with arylesterase activity (r = 0.85). Thus, we have demonstrated the feasibility of measuring PON1 mass in either serum or EDTA plasma.

Aberrant activation of beta-catenin contributes to the onset of a variety of tumours. There are many tumours that display beta-catenin accumulation in the absence of mutations in its gene. Recently, abnormal accumulation of wild-type beta-catenin has been associated with mutational inactivation of the p53 tumour suppressor.To investigate the potential role of p53 and its homologue p63 in beta-catenin deregulation and to correlate this with disease outcome.We analysed a panel of 24 samples of mycosis fungoides (MF), the most frequent manifestation of cutaneous T-cell lymphoma (CTCL), for beta-catenin, p53 and p63 protein expression by immunohistochemistry. Based on the immunostaining results for beta-catenin protein, 11 positive cases were selected for laser microdissection, genomic DNA isolation and subsequent mutation analysis of beta-catenin exon 3 and p53 exons 4-8.Our findings revealed overexpression of beta-catenin, p53 and p63 in 46%, 38% and 17% of cases, respectively. The number of p53-positive cases of MF was significantly higher (P0.05) in the beta-catenin-positive group (73%). Sequence analysis demonstrated that wild-type beta-catenin accumulation in MF is not associated with mutational inactivation of the p53 gene and, more importantly, our data provide evidence that a common polymorphic form of p53 (Arg72Pro) is significantly associated with beta-catenin overexpression (P0.05). No significant differences in the three genotypes were observed between the CTCL cases and the control group, demonstrating that Arg72Pro polymorphism of the p53 gene is not associated with the risk of developing cutaneous lymphomas (P0.05).We found an association of beta-catenin and p53 overexpression without detection of structural alteration in the genes, suggesting that p53 mutation is not an important mechanism for beta-catenin activation in primary CTCL. Additionally, we speculate that the p53 codon 72 polymorphism may influence negative feedback control involving beta-catenin and p53.

We developed a simple and reliable analytical method for the quantification and the characterization of ceramides extracted from biological samples by high-performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry(ESI/MS/MS). The chromatographic separation of analytes was carried out in a RP8 column, eluting with a methanol-water mixture in gradient elution mode. The separated lipids were detected by total ion monitoring and characterised by MS/MS spectra; quantitative analysis was performed by integrating the extracted ion peaks obtained in the negative ion mode. Good repeatability was obtained for retention time (0.3–2%), peak area ratio (AS/AIS, 2–8%), as well as limit of detection (LOD, 5–26 pg) and quantification (LOQ, 13–53 pg). The method was validated for the analysis of N-palmitoyl-D-erythro-sphingosine (Cer16), N-stearoyl-D-erythro-sphingosine(Cer18), N-tetracosanoyl-D-erythro-sphingosine (N24:0, lignoceric ceramide,Cer24:0), and N-tetracos-159-enoyl-D-erythro-sphingosine (N24:1, nervonic ceramide, Cer24 : 1), giving good results. Lipid mixtures, extracted from skin and epidermal cells, were analysed for their content of the studied ceramides.

To assess the knowledge of the risks and benefits of oral contraceptives (OCs) in a heterogeneous group of women and to identify their sources of information.A self-administered questionnaire assessing demographics, contraception history, knowledge of risks and benefits of OCs, and information sources was given to literate English- and Spanish-speaking women waiting for appointments at 4 clinics serving distinct populations in Portland, Oregon.Approximately half of the 211 women studied were of the opinion that OCs decreased the risk of acne, dysmenorrhea, and menorrhagia and increased the risk of weight gain, headaches, and thrombosis. Less than 15% knew of the decreased risk of anemia, endometrial cancer, colon cancer and pelvic inflammatory disease, but 28% understood the decreased risk of ovarian cancer. Seven percent to 36% of women used their own experiences in assessing the effect of OCs on a variety of general and reproductive factors. Women relied primarily on printed information for knowledge of OCs' effects on cardiovascular health and cancer.Women in this heterogeneous population of women were unaware of several benefits of OCs. Women relied heavily on their own experiences in assessing the risks and benefits of OCs. Women cited printed information more frequently than medical personnel as major sources of information on cardiovascular and oncological risks and benefits of OCs. The Internet, however, played a minimal, if any role in educating women about OCs.

Peripheral sensory neuropathy is the main non-hematological side-effect related to cisplatin chemotherapy. The strong similarity between clinical and neuropathological aspects in peripheral neuropathy induced by cisplatin and neurologic syndromes due to vitamin E deficiency, prompted us to investigate the relationship between cisplatin neuropathy and plasmatic level of vitamin E (alpha-tocopherol). We measured vitamin E in the plasma of 5 patients (Group 1) which developed severe neurotoxicity after cisplatin treatment and in another group of 5 patients (Group 2) we analyzed the plasmatic level of vitamin E before and after 2 or 4 cycles of cisplatin treatment. The results showed that the patients of group 1 presented low plasmatic levels of vitamin E and that the patients of group 2 presented significantly lower levels of vitamin E after 2 or 4 cycles of cisplatin than before treatment. Our preliminary data suggest that an inadequate amount of the antioxidant vitamin E due to cisplatin treatment could be responsible of the peripheral nerve damage induced by free-radicals. Given the lack of toxicity of vitamin E, we need to systematically assess the possible neuroprotective role of vitamin E supplementation in patients treated with cisplatin chemotherapy.

Keratinocyte growth factor (KGF) is involved in the control of proliferation and differentiation of human keratinocytes. It binds to, and activates, the tyrosine kinase KGF receptor (KGFR), a splicing transcript variant of the fibroblast growth factor receptor 2. We have previously shown (C. Marchese et al., Cell Growth Differ., 8: 989-997, 1997) that differentiation of primary cultured keratinocytes triggered by high Ca2+ concentrations in the growing medium induced up-regulation of KGFR expression, which suggested that KGFR may play a crucial role in the control of the proliferative/differentiative program during transition from basal to suprabasal cells. Here we analyzed the process of modulation of the expression of KGFRs in the human keratinocyte cell line HaCaT, widely used as a model to study keratinocyte differentiation. Western blot and double immunofluorescence for KGFR and the K1 differentiation marker showed that cell differentiation and stratification induced by confluence and high cell density correlated with an increase in KGFR expression. KGFRs, present on suprabasal differentiated cells, appeared to be efficiently tyrosine-phosphorylated by KGF, which indicated that the receptors up-regulated by differentiation can be functionally activated by ligand binding. Bromodeoxyuridine incorporation assay revealed that a significant portion of suprabasal differentiated cells expressing KGFR seemed to be still able to synthesize DNA and to proliferate in response to KGF, which suggested that increased KGFR expression may be required for retention of the proliferative activity.

The reconstruction of the epidermal melanin unit ex vivo has been achieved during the last decade, using the combination of previous cell culture techniques. The system reviewed is basically a modification of the Pruniéras model, using the air-liquid interface to grow differentiated keratinocytes, with the addition of 5% melanocytes in the seeding suspension, as well as the use of a more adapted culture medium. Repeated UVB irradiation induces a stimulation of melanogenesis macroscopically, and increases melanin concentration and melanosome transfer in reconstructs. These results have been reproduced with skin of various phototypes. This model allows to study the physiology of the epidermal melanin unit as well as pathologic conditions, like vitiligo and nevi. Recent evidence of a complex interaction of keratinocytes and melanocytes in photoprotection was provided by the use of chimeric reconstructs and by comparing autologous reconstructs made with and without low phototype caucasoid melanocytes. Based on these findings, we suggest a novel interpretation of the concept of phototype.

IFSCC Magazine, 11 (2008) (4) 323–330 Presented as a podium presentation at the 25th IFSCC Congress 2008 in Barcelona, Spain. Several classes of pigments are responsible for coloration in birds. Melanin pigments most commonly appear in bird feathers and bare parts. They impart black, brown and chestnut hues. Carotenoids are a second group of coloring biochemicals in birds. These two types of pigment-based coloration are found in nearly every order of extant birds. In contrast, parrots harbor bright-colored pigments in their feathers, which have different structures. As many colored substances exhibit free-radical scavenging activity and parrots have quite a long lifespan, the aim of our study was to identify the structures of these substances, attempt their synthesis and study their performance and possible use in cosmetics while overcoming stability problems. Studies were also planned to compare their efficacy with that of known anti-aging molecules. We were able to identify a structural moiety that could be the basis for a new family of molecules with high functionality. This example could open up the field of research for new functional molecules derived from animal or vegetal metabolism, which so far have been neglected because of their high instability. Keywords: Anti-oxidant, octatrienol, photoaging, radical stability, retinol-like

To investigate the antioxidant status of cultured uveal melanocytes from patients with uveal melanoma and uveal melanoma cells to characterize some of the biochemical properties of these cells in respect to the normal cutaneous melanocytes.The fatty acid pattern of membrane phospholipids, intracellular vitamin E level, and superoxide dismutase (SOD) and catalase activities were studied in uveal melanocytes (n = 10) and uveal melanoma cell (n = 10) cultures, by gas chromatography mass spectrometry or by spectrophotometer.Among the uveal melanocyte cultures, two groups were differentiated, according to catalase activity: group A with catalase values comparable to those of cutaneous ones and higher SOD activity and group B with catalase values 2 SD lower (P0.001) and lower SOD activity. Vitamin E concentration was not significantly different between melanoma cells and melanocytes, whereas a significantly higher percentage of polyunsaturated fatty acids was found in melanoma cells and the B group of melanocytes (P = 0.022). In uveal melanoma cells SOD activity was significantly lower than that detected in uveal melanocytes (P0.005).These results show a different pattern of antioxidants in uveal melanocytes with respect to cutaneous ones, possibly related to the anatomic distribution. However, as in cutaneous melanocytes, two subgroups were identified on the basis of the antioxidant pattern that could be the expression of a constitutional increased susceptibility to oxidative stress in some subjects. Moreover, an imbalance of the antioxidants was observed in melanoma cells, possibly related to the disease status and progression.

To assess any synergistic stimulatory effect in vivo of Interleukin 3 (IL-3) and Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) upon white cell and platelet counts, toxicity and antitumour effect, we conducted this phase I study. IL-3 0.25, 0.5 or 5 micrograms/kg/day for 1, 4 or 7 days was given by continuous intravenous (i.v.) infusion to 35 patients with advanced malignancy. 21 of the 35 patients also received sequential or overlapping treatment with continuous i.v. infusion of GM-CSF 1 or 3 micrograms/kg/day for up to 10 days. Monotherapy with IL-3 producted significant dose related increases in platelets and white cell counts. Combinations of IL-3 and GM-CSF also produced increases in white cell counts, but these were no greater than would be expected following GM-CSF treatment alone. There was a trend for platelets to increase more in patients receiving IL-3 and GM-CSF than those receiving IL-3 alone, but this did not reach statistical significance. In general, IL-3 and combinations of IL-3 and GM-CSF were well tolerated and the most common side-effect was fever. A maximum tolerated dose was not reached and antitumour effects were not seen. Future studies using combinations of IL-3 5 micrograms/kg/day and GM-CSF 3 micrograms/kg/day may help to define the optimal therapeutic regimen.

Protein-protein interactions are of major importance in many cellular processes. When no enzymic activity is involved, assays for direct binding are required. One such example is the relatively weak interaction between oncogenic Pas and the GTPase-activating protein neurofibromin (NF1). The complex between the catalytic domain of NF1 and the GTP-form of oncogenic Pas protein dissociates rapidly; hence, equilibrium binding must be quantitated. Scintillation proximity assay (SPA) technology, a radioisotopic technique that requires no separation step, was used to characterize this interaction. Leu-61 Ras complexed with [3H]GTP was generated by nucleotide exchange in the presence of a GTP-regenerating system. A SPA signal was obtained when radiolabeled Pas was mixed with NF1 fused with glutathione S-transferase (GST), anti-GST, and protein A-coated SPA beads. This signal was abolished when any of the components were omitted and also by the addition of NaCl, which potently reduces the affinity of interaction between Pas and NF1. The neutralizing anti-Pas monoclonal antibody Y13-259 and the detergent n-dodecyl maltoside, a specific inhibitor of NF1 catalytic activity, both abolished the SPA signal from the NF1/Ras assay but neither affected a control SPA signal in which a [3H]GTP.GST-Ras fusion protein was bound to protein A-coated SPA beads. This technology could be readily extended to the measurement of other protein-protein interactions and could form the basis for high-throughput screens for the discovery of novel therapeutic agents.

We have previously reported that fetal and adult skin fibroblasts display distinctive migratory phenotypes on 3-D collagen substrata and that these behavioural characteristics may be quantified by a function defined as the cell density migration index (CDMI). Subsequent work indicated that this difference in migratory phenotype was due to the production by fetal fibroblasts of a migration stimulating factor (MSF) that is not produced by normal adult skin fibroblasts. We now present data indicating that: (a) unselected fibroblasts obtained from 14/14 (100%) of adult gingival explants expressed fetal-like CDMI values compared to only 1/10 (10%) of similarly explanted paired skin cells; (b) 12/12 (100%) of these gingival fibroblast lines also produced detectable quantities of MSF compared to 0/9 (0%) of the tested skin cells; (c) by microdissection studies, gingival fibroblasts obtained from different anatomical microdomains consisted of behaviourally distinct subpopulations, with cells derived from the papillary tips (PAP fibroblasts) displaying fetal-like CDMI values and persistent MSF production, whilst cells obtained from the deeper reticular tissue (RET fibroblasts) were adult-like with respect to these two criteria; (d) PAP fibroblasts were also smaller and achieved higher saturation cell densities compared to paired RET cells; (e) PAP fibroblasts passaged in vitro underwent a fetal-to-adult phenotypic transition characterized by the adoption of various RET cell characteristics, including the acquisition of CDMI values falling within the adult range and cessation in MSF production; and (f) early passage PAP fibroblasts incubated in the presence of an affinity-purified anti-MSF rabbit polyclonal antibody were induced to alter their migratory phenotype and exhibited CDMI values falling within the adult range. Statistical analysis indicated a highly significant correlation between the expression of a fetal-like CDMI and production of MSF (P < 0.00001, using the Fisher exact contingency test). Taken together, these observations suggest that the production of MSF by PAP fibroblasts is responsible for their characteristically fetal-like migratory behaviour. The existence of such inter- and intra-site phenotypic heterogeneity in populations of skin and gingival fibroblasts is discussed in the context of fibroblast lineage relationships and the possible contribution of persistently fetal-like fibroblast subpopulations to connective tissue function in wound healing.

Interactions between epithelial cells and fibroblasts play a central role in directing both the spatial and temporal pattern of differentiation during the course of embryonic development [1]. Such “epithelial-mesenchymal” interactions continue to modulate and integrate various diverse elements of cell behaviour in the adult [2]. The biochemical identity of the signal molecules mediating epithelial-mesenchymal interactions has been the subject of intense investigation. Grobstein [3] first speculated that insoluble components of the extracellular matrix (ECM) might be involved; this initial insight has been confirmed in subsequent studies and there is now a substantial literature indicating that ubiquitous matrix macromolecules (such as collagens and proteoglycans) influence a variety of fundamental cellular attributes. The mechanism by which the ECM exerts these different effects appears to depend upon the interaction of matrix macromolecules with specific cell surface receptors resulting in alterations in the organisation of the cytoskeleton and consequent changes in cell shape [4].

Understanding of the functional morphology and biology of the oral mucosa has advanced in recent year, particularly in such areas as epithelial-mesenchymal interaction, fibroblast heterogeneity, extracellular matrix, migration factors and growth factors, and the importance of these areas in the development and maintenance of the oral mucosa is increasingly appreciated. This article reviews current knowledge.

Ultraviolet (UV) irradiation is capable of producing a dose-dependent decomposition of skin surface lipids and particularly of squalene, with the concomitant generation of active lipoperoxides. The biological effects of UV-peroxidated squalene were tested, compared with those produced by synthetic lipoperoxides (cumene hydroperoxide), on some immunological parameters in vivo modified by UVB irradiation. Application of UV-peroxidated squalene as well as cumene hydroperoxide significantly inhibited the induction of contact hypersensitivity to dinitrofluorobenzene in mice, which was associated with a decrease in the number of ATPase positive cells. The effect was dose-dependent (over 40 micrograms for peroxidated squalene and over 20 micrograms for cumene) and relevant after 2 d of treatment. Down-regulation towards the applied hapten was demonstrated. The results indicate that UV-induced lipoperoxides of squalene are capable of inhibiting the induction of contact hypersensitivity in mice and suggest that, among the other photoproducts generated in humans, squalene peroxides may play a role as biochemical messengers of the biological effects of UV irradiation of the skin.

An assay to measure the ability to stimulate migration of fibroblasts into collagen gel was carried out on serum from treated and untreated breast cancer patients and from healthy controls. Migration stimulating activity was found in the serum of 10 (83%) of 12 untreated breast cancer patients immediately before surgical resection of the primary tumour and in 9 (75%) of them 4 days after resection; in 13 (93%) of 14 patients 1-13 years after tumour resection who had received adjuvant treatment; and in 2 (10%) of 20 healthy women matched for age. The migration stimulating activity in cancer patients' serum was indistinguishable from the migration stimulating factor produced in vitro by fetal and cancer patient skin fibroblasts in its behaviour in various biochemical fractionation procedures. The presence of this activity in the serum of treated breast cancer patients clearly distinguishes it from other oncofetal proteins, which all seem to be produced by tumours.

Fetal skin fibroblasts migrate into 3D collagen gels to a significantly greater extent than do adult cells. This enhanced motility of fetal fibroblasts appears to result from the production of a "migration stimulating factor" (MSF) which is not made by their normal adult counterparts. Adult skin fibroblasts retain responsiveness to MSF and cells exposed to this factor achieve the elevated levels of migration characteristic of fetal cells. MSF has been purified to homogeneity, has an apparent molecular mass of 70 kD and has been further characterized in terms of a number of biochemical parameters. Studies concerned with the mechanism of action of MSF indicate that it stimulates the production of a high molecular weight class of hyaluronic acid (HA). Concurrent exposure of cells to Streptomyces hyaluronidase blocks the stimulation of adult fibroblast migration by MSF. In a related series of experiments, we have shown that TGF-beta inhibits the effects of MSF on both cell migration and HA production. Taken together, these data suggest that the stimulation of fibroblast migration by MSF is dependent upon (and may directly result from) a primary induction of HA synthesis. We have previously reported that skin fibroblasts obtained from patients with sporadic and familial breast cancer, as well as the unaffected first-degree relatives of familial breast cancer patients, commonly display a fetal-like migratory phenotype. Subsequent work has indicated that (a) these fetal-like cells also produce MSF, and (b) detectable levels of MSF are present in the serum of sporadic breast cancer patients both prior to and following surgical resection of the primary tumor mass. On the basis of these and related observations, we have put forward an hypothesis suggesting that the disruption in normal epithelial-mesenchymal interactions caused by the persistent production of MSF by fibroblasts in the adult may contribute directly to the pathogenesis of an epithelial cancer. The demonstration of aberrant fibroblasts in sporadic cancer patients (both in our own and independent studies) is not consistent with the "germ-line genetic lesion" model commonly invoked to account for the presence of such cells in patients with hereditary cancer syndromes.(ABSTRACT TRUNCATED AT 400 WORDS)

Fetal skin fibroblasts migrate into 3D collagen gels to a significantly greater extent than do adult cells. This enhanced motility of fetal fibroblasts appears to result from the production of a “migration stimulating factor” (MSF) which is not made by their normal adult counterparts. Adult skin fibroblasts retain responsiveness to MSF and cells exposed to this factor achieve the elevated levels of migration characteristic of fetal cells. MSF has been purified to homogeneity, has an apparent molecular mass of 70 kD and has been further characterized in terms of a number of biochemical parameters. Studies concerned with the mechanism of action of MSF indicate that it stimulates the production of a high molecular weight class of hyaluronic acid (HA). Concurrent exposure of cells to Streptomyces hyaluronidase blocks the stimulation of adult fibroblast migration by MSF. In a related series of experiments, we have shown that TGF-beta inhibits the effects of MSF on both cell migration and HA production. Taken together, these data suggest that the stimulation of fibroblast migration by MSF is dependent upon (and may directly result from) a primary induction of HA synthesis.

Previous research in our laboratories has demonstrated that the human sebaceous gland is a target organ for several stress hormones including α-melanocyte-stimulating hormone (α-MSH) and corticotropin-releasing hormone (CRH). To further investigate the role of stress hormones in acne, we examined the expression of opioid receptors (ORs) and the biological actions of β-endorphin (β-ED), a natural high-affinity ligand for the μ-OR (MOR) and low-affinity ligand for the Δ-OR (DOR), in human sebocytes. RT-PCR, Western immunoblotting and immunofluorescence studies identified the MOR but not the DOR in the human sebocyte cell line SZ95. Expression of the MOR was also confirmed in the sebaceous gland in normal human adult skin and was detectable primarily in peripherally located sebocytes. SZ95 sebocytes did not express the β-ED precursor pro-opiomelanocortin (POMC) as shown by RT-PCR analysis and Western immunoblotting and stimulation of cells with prototypical POMC inducers such as tumour necrosis factor-α, α-MSH, dbcAMP, phorbol ester and CRH failed to induce POMC. On the functional level, β-ED significantly suppressed the growth of SZ95 sebocytes induced by epidermal growth factor in chemically defined calcium-rich medium. On the other hand, β-ED enhanced lipogenesis as shown by Nile red staining and gas chromatography. Accordingly, β-ED dramatically increased the amount of C16 : 0, C16 : 1, C18 : 0, C18 : 1 and C18 : 2 fatty acids in an extent similar to linoleic acid used as a positive stimulus. Our data demonstrate that human sebocytes express the MOR and respond to β-ED with reduced in vitro proliferation and increased lipogenesis. These data establish a further link between stress and acne, the latter in which excessive sebum production is an invariable feature.

Plasma levels of vitamin E (Vit E), polyunsaturated fatty acids of phospholipids (PUFA-PL), lipoperoxides as well as erythrocytes glutathione peroxidase activity (GSH-Px), were evaluated in 200 migrants coming from developing countries, some of which at high risk for serious infective diseases. HIV-1 and syphilis infections were also investigated. 114 subjects (57%) had blood levels of Vit E, PUFA-PL and GSH-Px significantly lower (p less than 0.001, p less than 0.01) than those of normal healthy individuals (n = 30), while lipoperoxides values were unchanged. 8 from this group were found to be HIV-1 positive, and 5 TPHA positive. In contrast, the remaining 86 migrants did not show any signs of infections and their blood parameters were normal enough. These results show that factors such as widespread poverty, inadequate housing, malnutrition, insufficient access to medical care, psychological stress are strictly correlated to the reduction of blood parameters which are critical for the normal cell function of mammalian cells. PUFA-PL deficiency may cause lesions likely due to faulty cellular membranes. The lack of Vit E and GSH-Px, which are considered major protective molecules against lipoperoxidation damage in vivo, has been involved in several human diseases. We suggest that low blood levels of Vit E, GSH-Px and particularly PUFA-PL may play a pathogenetic role in the onset and development of AIDS and other infections. In this connection, we have found that a deficiency of these blood parameters occurs in patients with AIDS (n = 50) and in 32% of HIV seronegative intravenous drug abusers (n = 100).

Plasma levels of vitamin E (vit E) and polyunsatured fatty acids of phospholipids (PUFA-PL) as well as erythrocyte glutathione peroxidase (GSH-Px) activity are significantly lower (p less than 0.001) in patients HIV sero-positive (AIDS and ARC cases) both affected and not affected with seborrheic dermatitis and in 32% of HIV sero-negative intravenous drug abusers (IVDA, A subgroup) than in controls. The deficiency of PUFA-PL (mainly C20:3 n-6, C20:4 n-6 and C22:6 n-3) which is associated with a significant increase (p less than 0.001) of saturated palmitic and stearic acids and monounsaturated oleic acid, cannot be correlated to an active lipoperoxidative process. In fact the levels of thiobarbituric acid-reactive materials (TBA-RM) are not increased in the plasma of HIV sero-positive patients and A subgroup of IVDA. It is likely that the reduction of PUFA-PL is due to an inhibition of hepatic microsomal desaturase enzymes (delta 6 desaturase, delta 5 desaturase, delta 4 desaturase) which are involved in both n-6 and n-3 pathways. Since IVDA represent, and not only in Italy, a major risk category for HIV infection, we suggest that reduced blood levels of vit E, GSH-Px and particularly PUFA-PL may be added to the list of risk factors favouring the onset and the development of AIDS.

The production of aflatoxins and the unsaturated/saturated ratios of different lipid fractions of lipid extracts of mitochondria and microsomes of Aspergillus parasiticus grown in cultures supplemented with carbon tetrachloride (CCl4) and linoleic acid hydroperoxide are described. The presence of a strong free radical scavenger (cysteamine) was capable of fully inhibiting the aflatoxin production induced by CCl4 and linoleic acid hydroperoxide. In the culture supplemented with CCl4 the unsaturated/saturated fatty acid ratio in phospholipids of microsomes and mitochondria falls drastically compared to the normal values of the control. The addition of cysteamine to CCl4 supplemented cultures reverts the ratios to normal values.