The depigmenting effect of α-tocopheryl ferulate on human melanoma cells

Oral vitamin E (alpha-tocopherol, alpha-T) supplementation has been reported to improve facial hyperpigmentation. alpha-Tocopheryl ferulate (alpha-TF) is a compound of alpha-T and ferulic acid connected by an ester bond; ferulic acid is also an antioxidant, and could scavenge free radicals induced by ultraviolet (UV) radiation, and thus maintain the long-lasting antioxidative effect of alpha-T. Our aim was to see whether alpha-TF might be useful as a whitening agent and an antioxidant to improve and prevent facial hyperpigmentation following UV exposure. In this study, the inhibitory effect of alpha-TF on melanogenesis was examined biochemically using human melanoma cells in culture. The results show that alpha-TF, solubilized in ethanol or in 0.5% lecithin, inhibited melanization significantly, as did alpha-T at a concentration of 100 microg/mL, without inhibiting cell growth. This phenotypic change was associated with inhibition of tyrosinase and 5, 6-dihydroxyindole-2-carboxylic acid polymerase activities, and the degree of inhibition was dose dependent. No significant effect on DOPAchrome tautomerase activity was observed. alpha-TF did not directly inhibit tyrosinase activity of the large granule fraction extracted from human melanoma cells, and Western blotting revealed that there were no changes in protein content or in molecular size of tyrosinase, tyrosinase-related protein (TRP)-1 or TRP-2. Therefore, the inhibition of tyrosinase activity by alpha-TF might be due to effects at the post-translational level, and possibly by a secondary molecule activated by alpha-TF. These results suggest that alpha-TF is a candidate for an efficient whitening agent which suppresses melanogenesis and inhibits biological reactions induced by reactive oxygen species.     

α-MSH-stimulated tolerogenic dendritic cells induce functional regulatory T cells and ameliorate ongoing skin inflammation

The neuropeptide α-melanocyte-stimulating hormone (α-MSH) is a well-known mediator of skin pigmentation. More recently, it has been shown that α-MSH also exerts a strong anti-inflammatory and immunosuppressive activity. To elucidate the mechanisms underlying α-MSH-induced immunosuppression, we investigated whether α-MSH affects dendritic cell/T cell communication, as especially this interaction has an important role in the regulation of immune responses. Here, we show that α-MSH, by binding to MC-1R, induced tolerogenic dendritic cells, which were capable of expanding CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) in vitro, as well as in vivo. Notably, those α-MSH-induced Tregs were functional as they efficiently inhibited cutaneous contact allergy and ongoing psoriasis-like skin inflammation in mice. Furthermore, α-MSH induced tolerogenic dendritic cells capable of generating functional Tregs in human blood. Interestingly, human Tregs expanded via α-MSH-stimulated dendritic cells suppressed the proliferation and cytokine secretion of pathogenic T-helper-17 (Th17) cells from individuals with psoriasis. Taken together, these data indicate that α-MSH induced immunosuppressive Tregs in vitro and in vivo, which inhibited disease progression in a mouse model of psoriasis-like skin inflammation and suppressed the activation and proliferation of effector T cells from subjects with psoriasis.     

High doses and low doses of α2-interferon plus Zidovudine in the management of Kaposi's sarcoma associated with human immunodeficiency virus infection

Background α-2-lnterferon (α₂-IFN) is recommended as an effective agent for the treatment of epidemic Kaposi's sarcoma (EKS). Objective Trial of efficacy of low dose α₂,-IFN plus Zidovudine (AZT) for the treatment of EKS. Study design Non-randomised study. Subjects Ten homosexual or bi-sexual male patients with cutaneous lesions of Kaposi's Sarcoma, divided into two equal groups. Intervention High doseα₂-IFN (10-30 million units/three times per week) alone, or combination of low doseα₂-IFN (5 million units three times per week) with AZT (500 mg/day). Results High dose, IFN: 3 stable disease and 2 disease progression. Low dose, IFN plus AZT: 1 complete remission, 2 partial remissions and 2 stable disease. Outcome Preliminary results suggest the usefulness of low dose α₂-IFN plus AZT in the treatment of cutaneous lesions of EKS without visceral involvement.     

α-Melanozyten-stimulierendes Hormon

α-Melanocyte-stimulating hormone (α-MSH) is a tridecapeptide that is produced by the skin itself from the precursor proopiomelanocortin. It crucially mediates ultraviolet light-induced tanning after binding to melanocortin-1 receptors (MC-1R) expressed on the surface of epidermal melanocytes. The potent pigment-inducing and also cytoprotective actions of α-MSH are the rationale for the performance of first phase II clinical trials with Nle₄-D-Phe₇-α-MSH (NDP-α-MSH), a subcutaneously administered synthetic and superpotent α-MSH analogue, in patients with photodermatoses such as erythropoietic protoporphyria. Since α-MSH has shown promising anti-inflammatory and antifibrotic properties in numerous preclinical studies, it will be most interesting to evaluate these effects in further clinical pilot studies with NDP-α-MSH. In addition to α-MSH analogues, truncated tripeptides such as KDPT which do not bind to MC-1R but have sustained anti-inflammatory properties are currently emerging as another novel therapeutic strategy in dermatology.     

Disseminated histoplasmosis, invasive pulmonary aspergillosis, and other opportunistic infections in a homosexual patient with acquired immune deficiency syndrome

A homosexual man with the acquired immune deficiency syndrome had an unusually wide array of opportunistic infections. Despite antibiotic treatment over a period of two and a half years, the patient died. Perianal herpetic ulcers, oral candidiasis, cytomegalovirus infection, and disseminated infections with both Histoplasma capsulatum and Mycobacterium avium-intracellulare were diagnosed during illness. An autopsy revealed invasive pulmonary aspergillosis and a cerebellar lesion caused by cytomegalovirus. The latter was probably responsible for the patient's gait disturbance.     

Addison's disease, diffuse skin, and mucosal hyperpigmenation with subtle "flu-like" symptoms--a report of two cases

Abstract:   Addison's disease, or chronic adrenocortical insufficiency, is the overproduction of adrenocorticotropic hormone by the pituitary gland as a compensatory mechanism for decreased cortisol production by the adrenal glands. Classically, patients affected with Addison's disease develop weakness, anorexia, electrolyte imbalances: decreased sodium and chloride with increased serum potassium resulting in hypotension, and hyperpigmentation of the skin and mucous membranes. Herein this case report, we focus on the subtle findings of diffuse hyperpigmentation and intermittent but repetitive "flu-like" symptoms in two patients to correctly identify the diagnosis of Addison's disease effectively and efficiently.     

Extra‐facial melasma: clinical,histopathological, and immunohistochemical case‐control study

Background Extra‐facial melasma is a prevalent dermatosis in some populations with special characteristics in relation to its clinical aspects and probable etiopathogenic factors. Few studies have attempted to address this alteration of pigmentation, which has become a challenge in clinical Dermatology. Objective To assess the clinical histopathological and immunohistochemical characteristics of extra‐facial melasma, comparing affected, and unaffected sites. Methods Case‐control study with 45 patients in each group (melasma and disease‐free volunteers), assessing their clinical characteristics. In 36 patients, biopsies were performed on the lesion and the normal perilesional skin. Specimens were stained with HE and Fontana‐Masson, and melanocytes analysed by immunohistochemistry. Objective measurements were accomplished by a specifically designed image analysis software. Results The melasma group had a mean age ± SD of 56.67 ± 8 years, the majority of them were women (86.7%) and 82.1% of the female cases had reached menopause. There were no significant differences between groups in terms of presence of comorbidities, use of medications or hormone therapies. For extra‐facial melasma patients, family history of this dermatose and of previous facial melasma was significantly higher than in the control group (P < 0.05). The HE staining showed increased rectification and basal hyperpigmentation, solar elastosis, and collagen degeneration in the pigmented area (P < 0.05). There was a significant increase in melanin density in melasma biopsies, but the immunohistochemical tests did not detect a difference between the groups in terms of number of melanocytes. Conclusion Extra‐facial melasma appears to be related to menopause, family history, and personal history of facial melasma, in the studied population. Histopathology revealed a pattern similar to what has been described for facial melasma, with signs of solar degeneration, and a similar number of melanocytes, when comparing patients, and controls, suggesting that the hyperpigmentation is most likely the result of abnormal melanin production or distribution.     

Analysis of mast cell subpopulations (MCT, MCTC) in cutaneous inflammation using novel enzyme-histochemical staining techniques

Abstract In order to gain insights into the dynamics of mast cell subpopulations in normal and diseased skin, a novel enzyme-histochemical double and triple staining method was employed that allowed the detection of metachromasia (toluidine blue) and the mast cell proteases tryp-tase and chymase within the same cell. Cryostat sections were used of skin biopsies from the following specimens: normal skin (N=4), psoriasis (N=13), atopic eczema (N=7), lichen planus (N=6), interferon α2a injection sites (N=l) of a leukemic infiltrate and corresponding normal skin of the same patient before and after treatment. (i) Equal numbers of tryptase-and chymase-positive mast cells (MCTC) were obtained in all normal and diseased specimens in papillary and reticular dermis, with threefold increases around appendages, (ii) Tryptase-positive mast cells (MCT) were absent in normal skin, but were markedly increased in a disease-specific pattern within the papillary dermis, the inflammatory infiltrate and around appendages, (iii) Marked increases of MCT were also noted at interferon injection sites within the leukemic infiltrate, but not in the normal skin of the same patient. These data suggest that disease-dependent mast cell dynamics involve only MCT in cutaneous inflammation and that MCT numbers are controlled by distinct, disease-specific local tissue factors.     

热处理对体外人黑素细胞及HaCaT细胞分泌α黑素细胞刺激素的影响

【摘要】 目的 探讨热处理后体外培养的人表皮黑素细胞和人角质形成细胞（HaCaT细胞）分泌α黑素细胞刺激素（α-MSH）的变化。方法 将体外培养的黑素细胞及HaCaT细胞分别给予热处理（42 ℃,每天60 min,连续3 d）,实时荧光定量PCR法（RT-PCR）及酶联免疫吸附测定法（ELISA）检测两种细胞分泌的α-MSH的变化。结果 黑素细胞热处理组（1.5862 ± 0.3262）与对照组（1.0000 ± 0.2715）相比,α-MSH mRNA表达均明显增加（P < 0.05）;HaCaT细胞热处理组（1.8013 ± 0.1216）与对照组（1.0000 ± 0.2532）相比,mRNA水平也有明显增加（P < 0.05）。ELISA结果：黑素细胞热处理组（0.3142 ± 0.0469）蛋白表达水平明显高于对照组（0.2557 ± 0.0330,P < 0.05）;HaCaT细胞热处理组（0.4632 ± 0.0345）蛋白水平也明显高于对照组（0.4389 ± 0.0399,P < 0.05）。结论 热处理可促进人黑素细胞及HaCaT细胞α-MSH分泌。 【关键词】 热; 黑素细胞; HaCaT细胞; α促黑素     

Attachment, spreading and migration of melanoma cells on vitronectin

Abstract Recent in situ studies suggest the α_vβ₃ integrin is a tumour progression marker in melanoma. We analyzed 5 human melanoma cell lines for their expression of the vitronectin binding α_vβ₃ and α_vβ₅, integrins using flow cytometry. The role of these receptors in cell attachment, spreading and migration was investigated using attachment assays, video time lapse spreading and migration assays and with function blocking monoclonal antibodies. Cell lines derived from later stages of tumor progression exhibited high levels of α_vβ₃ expression, whereas no similar correlation with α_vβ₅ expression was identified. Cell attachment, spreading and migration response on vitronectin correlated well with the expression level of the α_vβ₃ but not the α_vβ₅ vitronectin receptor. Blocking of the α_vβ₃ integrin resulted in a significant decrease in cell attachment, spreading and motility whereas the function blocking antibody against the α_vβ₅ integrin only inhibited cell attachment in cell lines with the highest level of expression of this integrin. Taken together, our study indicates that the level of expression of the α_vβ₃ and α_vβ₅ integrins is heterogeneous in melanoma cell lines and that the α_vβ₅ integrin, if present, may function only during the initial cell attachment whereas the α_vβ₃ plays an important rôle in cell spreading and cell migration as well.     

应用流式细胞仪研究深圳地区健康成人及HIV/AIDS患者的免疫状况

目的：研究深圳地区健康人、HIV无症状感染者及AIDS患者的免疫状态，以了解艾滋病早期T细胞亚群的变化过程。方法：应用流式细胞仪三色荧光单克隆抗体（CD4－FITC／CD8－PE／CD3－PC5）检测34例正常健康成人，12例HIV感染无症状者，23例AIDS患者的外周血CD4^ 、CD8 ^ T淋巴细胞计数及CD4^ /CD8^ 比值。结果：HIV感染无症状者及AIDS患者的CD4^ 淋巴细胞及CD4^ /CD8比值均明显低于健康成人（P＜0．01）；HIV感染无症状者CD8^ T淋巴细胞明显高于正常健康人和AIDS患者（P＜0．01）；CD4^ T淋巴细胞随病程进展不断下降，AIDS患者与HIV感染无症状者之间存在着显著差异。结论：T淋巴细胞免疫状况可作为评价AIDS病程进展程度的重要指标。     

A case of mosquito hypersensitivity terminating as malignant histiocytosis

A patient with a 5 year history of hypersensitivity to mosquitoes developed a skin nodule on the chest, high fever, and hepatosplenomegaly. The diagnosis of malignant histiocytosis was made, based upon histological examination of the skin and cytological study of the pleural effusion. Immunological studies revealed the impaired activity of both T and B cells. After four weeks of no response to chemotherapy, the patient died. Autopsy revealed malignant histiocytes invading the bone marrow, liver, spleen, lung, adrenal glands, lymph nodes and skin. The possible relationship of immunodeficiency to malignant histiocytosis is discussed.     

Streaky pigmentation in a patient with acquired immune deficiency syndrome (AIDS)

We report a case of hyperpigmentation due to bleomycin treatment in a patient with acquired immune deficiency syndrome (AIDS). Although this type of hyperpigmentation has been previously seen in patients with cancer who are receiving bleomycin, this is, to our knowledge, the first reported case of bleomycin-induced hyperpigmentation in an AIDS patient and should be added to the growing list of cutaneous eruptions seen in these patients.     

Tazarotene versus tazarotene plus hydroquinone in the treatment of photodamaged facial skin: A multicenter,double‐blind,randomized study

Objectives: To compare the efficacy and tolerability of tazarotene plus hydroquinone versus tazarotene alone in the treatment of facial photodamage. Methods: Patients with facial mottled hyperpigmentation of at least moderate severity and an overall integrated assessment of photodamage score of at least moderate applied tazarotene 0.1% cream each evening and either hydroquinone 4% cream or placebo cream each morning for up to 24 weeks. Results: Among 131 patients enrolled, 114/124 (92%) with exit data completed. Both regimens were highly effective in reducing photodamage, with tazarotene plus hydroquinone showing superiority over tazarotene alone for some efficacy measures. The incidence of ?1‐grade improvement from baseline (on a scale of none, minimal, mild, moderate, or severe) was significantly greater with tazarotene plus hydroquinone than with tazarotene alone for lentigines (weeks 12–24, p?0.01) and mottled hyperpigmentation (week 16, p?0.05). The incidence of ?50% global improvement was also significantly superior with the combination regimen as early as week 8 (p?0.01). Both regimens were associated with good tolerability and high patient satisfaction (no significant between‐group differences). Conclusions: The adjunctive use of hydroquinone can enhance the efficacy of tazarotene in reducing dyspigmentation associated with photodamage.     

Expression of galanin in melanocytic tumors

IntroductionGalanin is a neuropeptide with wide-ranging effects, especially within the endocrine and nervous systems. Galanin and its receptors are present in human skin. Galanin is expressed in different neural, endocrine and neuroendocrine tumors and, on the other hand, several neuropeptides, particularly α-MSH, seem to play a role in the pathogenesis of melanoma.ObjectiveTo investigate the expression of galanin in cutaneous melanomas and melanocytic nevi and correlate it with α-MSH expression and several prognostic factors for melanoma.Material and methodsWe performed an observational and retrospective study of the immunohistochemical expression of galanin and α-MSH in samples of cutaneous melanomas diagnosed in the last 5 years in the San Jorge Hospital, Huesca (Spain). Different types of melanocytic nevi were also analyzed.ResultsA total of 130 pigmented lesions were studied: 38 primary cutaneous melanomas, 6 cutaneous melanoma metastases and 86 melanocytic nevi. Immunostaining with galanin and α-MSH was significantly higher in melanomas than in melanocytic nevi (p < 0.001), although spindle cell and blue nevi showed significant expression of α-MSH. More than 50 % of nodular melanomas and 90 % of superficial spreading melanomas were positive for galanin and α-MSH, and the latter also showed the highest percentage of positive cells for galanin (mean 35.09 ± 28.16) as well as for α-MSH (mean 67.64% ± 35.38). A positive correlation of 71 % was found for immunostaining of both neuropeptides in melanomas. No significant correlation was observed between galanin expression and age, gender, location of the lesions, Breslow index, Clark level and mitotic index.ConclusionOur study shows the expression of galanin in cutaneous melanoma and its significant correlation with α-MSH immunostaining.     

γ-melanocyte stimulating hormone (γ-MSH)-like immunoreactivity is present in certain normal human keratinocytes

Abstract Using ihe indirect immunohistochemical approach, the occurrence of γ-melanocyte stimulating hormone (γ-MSH)-like immunoreactivily in human normal keratinocytes is described. The positive cells were observed in each layer of the epidermis (except stratum corneum) and often, at the level of the stratum spinosum, also around the orifices of cutaneous accessory organs, such as sweat glands and sebaceous glands/ hair follicles. Combining these data with our previous investigations, the results support the possibility that locally produced γ-MSH could be involved in cutaneous immune response, pigmentation and epithelial pro-liferation, as well as neuromodulation.     

Basal cell carcinomas display extensive abnormalities in the hemidesmosome anchoring fibril complex

Abstract We have employed a panel of antibodies directed against several newly-defined and well-characterized components of the epidermal basement membrane (BM) to investigate the biology of basal cell carcinoma (BCC) by indirect immunofluorescence and to determine whether alterations in BM components may play a significant role in BCC tumor invasion. We found that the 230 KD bullons pemphigoid antigen (BPA) was either not detected (13/16) or significantly diminished (3/16) in BCC tumor BM. While the 180 KD BPA revealed less intense staining of the normal overlying epidermal BM than did the 230 KD BPA, the 180 KD BPA was uniformly undetectable in BCC tumor BM (16/16). Epiligrin was either not detected (9/15) or minimally detected (6/15) in BCC tumor BM. α₆ integrin was not detected (15/16) or minimally detected (1/16) in BCC tumor BM, whereas β₄ integrin was uniformly undetectable in BCC tumor BM (16/16). Type VII collagen was also not detected (9/16) or was significantly diminished (4/16) in BCC tumor BM. Laminin and type IV collagen were both at least as strong in BCC tumor BM as in adjacent normal BM. All of these components were present both in the epidermis of normal skin as well as in the normal epidermal BM overlying BCC tumor nests. Our findings reveal extensive alterations in numerous components of the hemidesmosome anchoring fibril complex of BCC's. As this complex is thought to play an important part in epidermal cell adhesion to the BM, our findings suggest that these extensive BM abnormalities may facilitate or contribute to BCC tumor invasion.     

Decreased expression of α2β1 integrin in scleroderma fibroblasts

Abstract Systemic scleroderma (SSc) is a complex connective tissue disorder of unknown etiology. In early stages of the disease, libroblasts are activated to produce large amounts of collagen with subsequent fibrosis. Collagen metabolism of fibroblasts is modulated by their contact with the extracellular matrix (ECM), which involves distinct receptors on the cell surface, mainly belonging to the integrins. We investigated the expression of collagen receptor α₂β₁, in SSc and normal fibroblasts, since this receptor has been shown to be utilized by fibroblasts for adhesion to and reorganization of collagen I. 9 strains of scleroderma fibroblasts grown as monolayer cultures were first analyzed with respect to their collagen I expression. 6 of these strains were similar to controls (“low” producers) and 3 strains showed up to 2–3 × higher levels of collagen I mRNA expression (“high” producers). Northern hybridization using a cDNA probe specific for the α₂ integrin subunit revealed a decrease of the corresponding mRNA in SSc fibroblasts as compared to controls (75% versus 100%). “High” collagen producing cell strains displayed the lowest values for α₂ integrin mRNA. The decrease of α₂ integrin subunit expression at the mRNA level in selected fibroblasts was further substantiated by radioimmunoprecipitation using specific mAbs directed against α₂ integrin subunit. No significant changes in β₁ integrin expression could be observed-neither at mRNA nor at the protein level. Our data indicate a correlation between excessive synthesis of collagen and low levels of α₂ integrin subunit expression in SSc fibroblasts. Further experiments should clarify whether this observation is a phenomenon specific for scleroderma or whether it reflects an “activated” state of fibroblasts.