MicroRNA-370-5p inhibits pigmentation and cell proliferation by downregulating mitogen-activated protein kinase kinase kinase 8 expression in sheep melanocytes

In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of sheep with different hair colors;however,its function remains unclear.In this study,we investigated the roles of miR-370-5p in sheep melanocytes and found that the overexpression of miR-370-5p significantly inhibited cell proliferation(P<0.01),tyrosinase activity(P=0.001)and significantly reduced(P<0.001)melanin production.Functional prediction revealed that the 3′-untranslated region(UTR)of MAP3K8 has a putative miR-370-5p binding site,and the interaction between these two molecules was confirmed using luciferase reporter assays.In situ hybridization assays revealed that MAP3K8 is expressed in the cytoplasm of melanocytes.The results of quantitative RT-PCR and Western blotting analyses revealed that overexpression of miR-370-5p in melanocytes significantly inhibits(P<0.01)MAP3K8 expression via direct targeting of its 3′UTR.Inhibition of MAP3K8 expression by siRNA-MAP3K8 transfection induced a significant inhibition(P<0.01)of melanocyte proliferation and significant reduction(P<0.001)in melanin production,which is consistent with our observations for miR-370-5p.Target gene rescue experiments indicated that the expression of MAP3K8 in melanocytes co-transfected with miR-370-5p and MAP3K8-cDNA(containing sites for the targeted binding to miR-370-5p)was significantly rescued(P≤0.001),which subsequently promoted significant increases in cell proliferation(P<0.001)and melanin production(P<0.01).Collectively,these findings indicate that miR-370-5p plays a functional role in inhibiting sheep melanocyte proliferation and melanogenesis by downregulating the expression of MAP3K8.

Direct Transplant of Melanocytes from Normal Donor Area into Vitiligenous Recipient Area by Intralesional Injection of Melanocytes Using Spade Like Needle Technique

Background: Vitiligo is a common autoimmune inflammatory skin disease, where there are different surgical techniques for treatment of stable patches of vitiligo .Objective: To find non-costly, minimally invasive, simple technique by direct melanocytes transplant by spade needle technique in treatment of vitiligo. Patients and Methods: This interventional, therapeutic, comparative study was done in Department of Dermatology, Baghdad Teaching Hospital, Baghdad, Iraq from April 2014-March 2015. Twenty patients with localized, generalized and segmental vitiligo were included. Full history and examination for each patient was done with 4 (20%) males and 16 (80%) females and their ages ranged from 9 - 40 (23.15 ± 11.44) years. Forty one patches in 20 patients treated by spade grafting technique and the donor and recipient sites were demarcated and anesthesia done by xylocaine 2% with adrenalin 1:100,000. Transplantation was started by using disposable needle gauge 18 (the sharp end of needle was cut by a scissor to make it a spade like) with medical syringe 5 ml supplied with normal saline. The micro-pieces were taken from donor site and transplanted directly, easily and rapidly into dermis of recipient site and followed by pushing normal saline and the procedure was repeated to cover all recipient sites with 5 mm distance between injection points. The surface area of the lesions was calculated and the reduction rate was estimated every month till the end of the 4th month period of the treatment. Results: Including 41 patches in 20 patients with the surface area of the patches ranged from 1.5 - 90 cm2 (13.78 ± 17.57) cm2. The mean ±SD of surface area of lesions was decreased from 13.78 ± 17.57 cm2 at baseline visit to 13.61 ± 17.48 cm2 at the second visit (after 2 weeks ) which was statistically significant (p value ≤ 0.001). The mean surface area continued to be reduced till reaching 12.20 ± 15.68 cm2 at the third visit and 12.01 ± 15.55 cm2 at the fourth visit. All were statistically significant when compared to baseline visit. There was reduction in surface area 1.1% at two weeks, 9.93%, and 12.5% at the 2nd, 4th months respectively. Conclusions: Intradermal injection of melanocytes in patients with vitiligo by spade like needle was very quick and simple non-costly technique, and gave 12.5% reduction which could be repeated at different times until satisfactory re-pigmentation of vitiligenous skin is achieved.

Effect of Xiaobailing Decoction on Melanocytes in Vitro

To observe the effect of Xiaobailing Decoction （消白灵汤） on murine melanocytes in vitro and to explore the mechanism of Xiaobailing Decoction in the treatment of vitiligo. Methods： B-16F10 murine melanoma cells were cultured in 1640 medium and treated respectively with different concentrations （lmg/ml, 2mg/ml, 3mg/ml） of the Chinese drug Xiaobailing Decoction and its main components, the drugs for replenishing the lddney-yang, and the drugs for nourishing blood and activating blood circulation, etc. for 7 days. MMT assay was used to determine the proliferation of B-16F10 murine melanoma cells. NaOH cleavage assay was adopted to measure the melanogenesis of melanocytes. Results：Xiaobailing Decoction, the drugs for replenishing the kidney-yang and the drugs for nourishing blood and activating blood circulation at different concentrations significantly improved the proliferation of B-16F10 murine melanoma cells from the 3rd day to the 5th day （P〈0.05）, with Xiaobailing Decoction at the concentrations of lmg/ml having the most distinct action on promoting the proliferation of melanocytes on the 3rd day （P〈0.001）; And the drugs for replenishing the lodney-yang at the concentrations of 2mg/ml and 3mg/ml and the drugs for nourishing blood and activating blood circulation at 3mg/ml significantly increased melanogenesis of melanocytes （P〈0.05）. Conclusion： Xiaobailing Decoction can promote melanocytic proliferation and melanogenesis in vitro, and it is indicated that the drugs for replenishing the kidney-yang and the drugs for nourishing blood and activating blood circulation play an important role in treating vitiligo.

Depigmentory Effects of Keishibukuryogankayokuinin in Human Epidermal Melanocytes

Keishibukuryogankayokuinin (KBY) is a traditional Japanese herbal medicine widely used to treat skin pigmentation. The scientific basis for its use is, however, unclear, and studies evaluating its mechanism and effectiveness are sparse. In this study, we compared the tyrosinase inhibitory effects of KBY and Keishibukuryogan (KB, which has the same composition of KBY, except Coix Seed [CS]) and CS under exposure to UV radiation as well as under non-exposure conditions. Neonatal human epidermal melanocytes obtained from a darkly pigmented donor were used. These cells were cultured in a final concentration of 500 μg/ml or 1000 μg/ml, to which KBY, KB, and CS were added. After incubation for 72 h, cells were stained with Fontana-Masson stain and counted. Tyrosinase activity was measured by its dopa oxidase activity, and tyrosinase expression was estimated using real-time PCR. For UV radiation, cells were exposed to UVB radiation for 90 s per day for 3 days. Under non-exposure conditions, tyrosinase activity significantly increased with both KBY and KB but significantly decreased with CS, regardless of the concentration. In addition, tyrosinase expression significantly decreased but only with KBY at both concentrations. Under UV radiation exposure, tyrosinase activity significantly increased with KBY and KB at both concentrations while tyrosinase expression significantly decreased with KBY and KB;a significant increase was, however, observed with CS at both concentrations. These results suggest that taking KBY after sunburn is effective against skin pigmentation, and the combination of KB and CS is useful for skin depigmentation.

THE EFFECT OF Ge-132 ON ULTRASTRUCTURE OF CULTURED MELANOCYTES

Objective To elucidate the effect of Ge 132 on the growth of melanocytes.Methods Melanocyes from epidermis were cultured and purified;the second generation of the cell was used for study;the cells were divided into two groups randomly,to group A, Ge 132 was added to the media at 0.04mg/L;to group B,common culturing method was used without Ge 132.After 5d, the cells were seperated by digestion for study by transmission electronic microscope.Results Compared to group B, the vaculoes of the cells were increased,mitochondria distended, endoplasmic reticulum dilated and the number of melanosome declined in the group A.Conclusion Ge 132 can inhibit the melanocytes growth at a certain concentration and might be used for treating pigmented diseases.

Treatment of Gray Hair in Vitiligo Patients by Direct Melanocytes Transplant Using Needling Micrografting and Dermabrasion Techniques

Background: Melanocytes transplant for treatment of vitiligo is a common therapy using different surgical procedures. But there was no interest in repigmentation of grayness of hair in the treated vitiliginous area. Objective: To do melanocytes transplant from donor area into the recipient vitiliginous area with associated gray hair. Patient and Methods: This is a case interventional study was done in Department of Dermatology/Baghdad Teaching Hospital from February 2011-March 2012. Eleven patients were enrolled in this study, six males and five females with vitiligo in association of gray hair. Their ages ranged from 8 - 35 years with a mean ±SD of 20.90 ± 7.006. Melanocytes transplant in patients with vitiligo using needling micrografting technique for twelve patches and direct melanocytes transplant from normal donor area into vitiliginous recipient area by dermabrasion technique for eleven patches. Dressing was applied and patients were seen every two weeks for the first month and monthly for one year. Results: Repigmentation of the vitiliginous area was started after two weeks and was obvious at one month that progressed over time. The repigmentation of hair appeared usually after few months and was obvious after four months and the repigmentation of gray hair was quicker in patients with micrografting technique than those with dermabrasion technique. The mean rate of repigmentation was 18.3% at six months and 37.5% at twelve months in micrografting technique while the mean rate of repigmentation was 9.15% at six months and 18.55 at twelve months in dermabrasion technique. Conclusions: Direct transplant of melanocytes from normal donor area into recipient vitiliginous area with associated white hair is an effective procedure to induce repigmentation of gray hair.

Direct Melanocytes Transplant from Normal Donor Area into Vitiliginous Recipient Area by Dermabrasion Technique

Background: Vitiligo is an autoimmune pigmentory disorder, that affects all age group that is treated by many medical treatments but some of them might need surgical therapy. Objective: To evaluate the dermabrasion technique in the treatment of vitiligo by direct transfer of melanocytes from the dermabraded normal donor area to the vitiliginous recipient area. Patients and Methods: This is a case interventional study was done in Department of Dermatology/Baghdad Teaching Hospital from February 2011-March 2012. Nine Patients with vitiligo were enrolled in this study with different clinical types of vitiligo including 5 segmental, 2 generalized and 2 localized. The donor and recipient areas were anesthetized at the same time with xylocain alone. Dermabrasion of recipient area was done first by manual abrader and left for few minutes until the oozing was stopped. Then the donor area was similarly dermabraded and the dermabraded tissue including the epidermis and superficial epidermis was immediately transferred into the recipient area and dressing was applied. Removal of the dressing was done after 10 - 14 days from the operative time. Follow up was done every 2 weeks in the first month then monthly for six months to record the result of implantation and repigmentation. Results: The re-pigmentation started one month after the operation as small macules and this increased gradually over time: the mean rate of re-pigmentation was 13% at 2 months, 27.8% at 4 months and 36.78% at 6 months. In addition, sun light exposure was applied to enhance re-pigmentation. The pigmentation was diffuse and not follicular in shape. Conclusion: Direct transfer of melanocytes from normal donor area into vitiliginous recipient area by dermabrasion technique was easy, rapid and non-costly and gave 36.78% mean rate of pigmentation at 6 months follow up and without complications.

Ultrastructure of Amelanotic Melanocytes from Human Hair Follicles

To investigate the ultra structure of amelanotic melanocytes （AMMC）. Methods： The hair follicles obtained from normal human scalp by 0.50% collagenase type V treatment were washed with 0.1 mol/L phosphate buffer salt （PBS）. Hair-follicle cell suspensions were prepared by trypsin treatment and cultured in melanocyte medium. Remaining keratinocytes were removed by differential trypsinization. 100μg/ml geneticin was used to eliminate the contaminating fibroblasts. At third passage, the cells were trypsinized, and then washed in phosphate-buffered saline and processed for transmission electron microscopy. Results： Under transmission electron microscope, the cultured cells showed round or oval shape, with single large nuclear and the karyotheca were double deck. There were obvious euchromosome within the nucleus, and sparse heterochromosome. There were various organelles in the cytoplasm, including plentiful melanosomes with nearly similar size, mitochondria, rough endoplasmic reticule （RER） and ribosome. The electron density granules in most of the melanosomes disposed along concentric circularities. Golgi apparatus in the cells was inconspicuous. Conclusion： The ultra structure of AMMC from human hair follicles is different from that of epidermal melanocytes, and these characteristics determine the functional immature of AMMC.

Decoding the Molecular Mechanisms of BRAFV600E-Induced Nevi Formation

Melanocytes derived from neural crest cells harbor the BRAFV600E mutation,which is the predominant driver of nevus formation in humans.This mutation leads to malignant cell proliferation and subsequent cell cycle arrest,culminating in oncogene-induced senescence and nevus development.Nevertheless,emerging evidence has highlighted the heterogeneity of cellular senescence markers in BRAFV600E-induced senescent melanocytes.Moreover,the capacity of melanocytes within nevi to regain their proliferative ability raises questions about the molecular mechanisms by which BRAFV600E,via the mitogen-activated protein kinase signaling pathway,triggers nevus formation.This study provides an overview and discussion of the molecular mechanisms underpinning BRAFV600E-induced melanocyte nevus formation and the relevant animal models employed for their elucidation.It also highlights the significance of elucidating dynamic changes in cytoplasmic and nuclear substrates that interact with phosphorylated extracellular signal-regulated protein kinases 1 and 2 and underscores the value of using targeted BRAFV600E animal models created through gene editing technologies.

SFRP5 inhibits melanin synthesis of melanocytes in vitiligo by suppressing the Wnt/b-catenin signaling

Secreted frizzled-related protein 5(SFRP5)plays a pivotal role in regulating the development of many tissues and organs,however,as an inhibitor of Wnt signaling,the role of SFRP5 in vitiligo remains unknown.Hence,we speculated that SFRP5 might be associated with melanogenesis in melanocytes by regulating Wnt signaling in vitiligo.In this study,we found that SFRP5 was overexpressed in the skin lesions of patients with vitiligo.Compared with that in normal epidermal melanocytes(PIG1),the expression of SFRP5 was increased in vitiligo melanocytes(PIG3V).To investigate the effect of SFRP5 on melanin synthesis,PIG1 cells were infected with recombinant SFRP5 adenovirus(AdSFRP5),and PIG3V cells were infected with recombinant siSFRP5 adenovirus(AdsiSFRP5).The results showed that SFRP5 overexpression inhibited melanin synthesis in PIG1 cells through downregulation of microphthalmia-associated transcription factor(MITF)and its target proteins via suppression of the Wnt/b-catenin signaling pathway.Accordingly,SFRP5 silencing increased melanin synthesis and activated the Wnt signaling pathway in PIG3V cells.Moreover,SFRP5 overexpression also downregulated the transcriptional activity of T cell factor/lymphoid enhancer factor(TCF/LEF)in PIG1 cells.Furthermore,this inhibitory effect of SFRP5 on melanin synthesis was reversed by treatment with the b-catenin agonist,SKL2001.The inhibitory action of SFRP5 in pigmentation was further confirmed in vivo using a nude mouse model.Hence,our results indicate that SFRP5 can inhibit melanogenesis in melanocytes.Additionally,our findings showed that SFRP5 plays a vital role in the development of vitiligo,and thus may serve as a potential therapeutic target for vitiligo.

Prostaglandin induces the expression of matrix metalloproteinase-1 in ciliary melanocytes

Background Latanoprost, a prostaglandin F2α analog, has been shown to be an effective intraocular pressure lowering agent which acts by inducing ciliary muscle cells to synthesise matrix metalloproteinases. However, the response of ciliary melanocytes to latanoprost has never been reported. This research has investigated the ability of latanoprost to induce matrix metalloproteinase-1 expression in human ciliary melanocytes, and thereby advance the understanding of the mechanism of PGF2α in decreasing intraocular pressure. Methods In vitro human ciliary melanocytes were treated for 48 hours with five different concentrations of latanoprost （100, 150, 200, 500, and 1000 nmol/L）. Ciliary melanocytes treated with 0.01% ethanol （vehicle） were used as a control. The expression of matrix metalloproteinase-1 in ciliary melanocytes was determined by Western blotting and immunofluorescent staining. Results Western blotting showed that the expression of matrix metalloproteinase-1 in ciliary melanocytes was induced by latanoprost, and the level of expression was dependent on the concentration of latanoprost in the culture medium. Immunofluorescent staining showed that matrix metalloproteinase-1 was confined to the ciliary melanocyte cytoplasm. Conclusions Latanoprost induced the expression of matrix metalloproteinase-1 in human ciliary melanocytes in a dose-dependent manner. Ciliary melanocytes, as well as ciliary muscle cells, may also play an important role in uveoscleral outflow modulation.

Imiquimod:A potential option for inhibition of repigmentation of congenital melanocytic nevus after laser ablation

Congenital melanocytic nevi(CMN) are common skin tumors. Large and specially located nevi cannot be completely removed by surgery, posing the risks of both cosmetic deformities and potential malignancy.Nonsurgical treatments, such as laser therapy and physical dermabrasion, can overcome the limitations of surgery;however, the high rate of repigmentation remains an unresolved global challenge. We conducted a self-controlled observational study of a patient with a nevus on the chest. Two areas of the lesion were treated with an Er:YAG laser and 5% imiquimod cream was applied to one of these areas. After nearly 7-months of follow-up, we observed a significant difference in color between the two areas, suggesting that topical imiquimod may inhibit repigmentation and significantly enhance the effectiveness of laser treatment.

Paraneoplastic retinopathies: an update on pathogenesis, diagnosis and management

The paraneoplastic retinopathies are a heterogeneous group of disorders with significant visual consequences occuring in the setting of a systemic malignancy.These conditions may be characterized by the presence of antiretinal antibodies and may predate or follow the diagnosis of an underlying malignancy.Herein I review the clinical findings,pathophysiology,laboratory testing and management of the paraneoplastic retinopathies:cancer-associated retinopathy(CAR),melanoma-associated retinopathy(MAR),bilateral diffuse uveal melanocytic proliferation(BDUMP)and paraneoplastic vitelliform maculopathy(PVM).The pathophysiology of the paraneoplastic retinopathies varies from molecular mimicry resulting in anti-retinal antibody production(CAR,MAR)to relases of soluble factors either by the primary tumor(BDUMP)and/or immune system in response to the primary tumor(PVM)which result in retinal and/or retinal pigment epithelium dysfunction.For each condition,structural and functional multimodal retinal testing can be helpful in establishing the diagnosis.Treatment for the paraneoplastic retinopathies involves a combination of treating the underlying malignancy plus additional local and/or systemic immunosuppressive agents though no systemic therapeutic protocols exist.Despite these interventions,the retinopathy may be progressive or the retinopathy may be a harbinger of poor survival.Nevertheless,prompt diagnosis may help identify an underlying malignancy earlier and may thus improve cancer-related survival.

Effect of Q-switched alexandrite laser irradiation on dermal melanocytes of nevus of Ota

OBJECTIVE: To investigate the effect of Q-switched alexandrite laser irradiation on dermal melanocytes of nevus of Ota. METHODS: Multiple biopsies were carried out on 4 patients with nevus of Ota before and after laser irradiation. Altogether 11 samples were examined under light microscope and 14 under transmission electron microscope. RESULTS: Immediately after laser irradiation, the dermal melanocytes were destroyed, the melanosomes were degenerated with central vesicle formation within most of them, and intradermal round vacuoles appeared. The epidermis remained intact. Three months to 1 year after irradiation, the degenerated melanosomes and cell debris were scavenged mainly by macrophages. Dermal melanocytes gradually decreased. No fibrosis was found. CONCLUSION: Q-switched alexandrite laser can selectively destroy dermal melanocytes of nevus of Ota and treat the disease safely.

Effect of Q-switched Alexandrite laser irradiation on epidermal melanocytes in treatment of Nevus of Ota

To investigate injury to epidermal melanocyte by Q-switched Alexandrite laser Methods Multiple biopsies were performed on 5 patients with nevus of Ota from before irradiation to 1 year after irradiation Fourteen specimens were obtained for light microscopy, and 17 for transmission electron microscopy Results Melanosomes in epidermal melanocytes were both smaller in size and fewer in number than those in dermal melanocytes Immediately after irradiation, focal extracellular vacuoles of the basal layer could be observed under light microscopy Most epidermal melanocytes underwent mild or moderate injury in the form of vacuolated melanosomes, swollen mitochondria, dilation of endoplasmic reticulum, and expansion of extracellular space, retaining intact cell membranes Normal structures were restored 5 months to 1 year after irradiation, with no depigmentation or hyperpigmentation as seen by light microscopy Conclusion Injury of melanosomes in epidermal melanocytes is reversible

Mobilization of Melanocytes During NB-UVB Treatment of Vitiligo

Vitiligo,a common skin depigmentation disorder,is the result of complex interactions of genetic,immunological,environmental,and biochemical events.Treatments for vitiligo include drugs,phototherapy,surgical transplantation,and so on.Among them,the efficacy of narrow band-ultraviolet B has been confirmed.By inducing keratinocytederived factors and signalings,narrow band-ultraviolet B can trigger and/or promote the mobilization of melanocytes which migrate to lesional epidermis ultimately,leads to the repigmentation of white patches.The mobilization of melanocytes includes stages of activation,migration,proliferation,and differentiation.Elucidating processes that enable the specific mobilization of melanocytes and the signaling pathways and factors involved,will help the development of new drugs and methods for the treatment of vitiligo.

Not Follicular but Dermal Melanocyte Precursors Are Histopathologically Retained in Vitiligo

Although perifollicular repigmentation in the vitiligo lesions is owing to activation of follicular melanocyte stem cells and/or precursor cells followed by supplying matured melanocytes, the underlying mechanism of diffuse repigmentation on the whole vitiligo surface remains still unknown. In addition to the presence of remaining melanocytes, it is conceivable that dermal melanocyte precursor cells contribute to induce diffuse repigmentation after treatment. Therefore, we investigated here whether dermal and follicular melanocyte precursor cells were reduced or not in vitiligo lesions. We performed an immunostaining for Nestin and p75NGFR as dermal melanocyte precursor cells and MITF/Fzd4 as follicular melanocyte precursor cells and compared the positive cells number between lesions and non-lesions (n = 11). Although MITF+/Fzd4+ cells in the hair follicle were significantly decreased in number in the lesions, Nestin+ and p75NGFR+ cells were not. This result indicates that dermal precursor cells could be retained in the vitiligo lesions but be disturbed to differentiate into matured melanocytes.

Difficulties in diagnosing anorectal melanoma: A case report and review of the literature

BACKGROUND Anorectal melanoma is a tumour that is difficult to identify due to its rarity and variability of presentation.Insufficient data published in the literature do not allow for diagnostic and treatment guidelines to be established.Anorectal melanoma has the worst prognosis among mucosal melanomas and is frequently misdiagnosed by standard identification methods.CASE SUMMARY A 66-year-old woman presented with intermittent anal bleeding,pain,and tenesmus in the past month,with no associated weight loss.Colonoscopy revealed a cauliflower-like tumour with a diameter of 1.5 cm,with exulcerated areas and an adherent clot but without obstruction.Biopsy results identified an inflammatory rectal polyp with nonspecific chronic rectitis.Tumour markers CA 19-9 and CEA were within the normal range.After 6 mo,due to the persistence of symptoms,a pelvic magnetic resonance imaging scan was performed.A lesion measuring 2.8 cm×2.7 cm×2.1 cm was identified at the anorectal junction,along with two adjacent lymphadenopathies.No distant metastases were detected.Immunohistochemistry was performed on the second set of biopsies,and a diagnosis of anorectal melanoma was established.Surgical treatment by abdominoperineal resection was performed.Evolution was marked by the appearance of lung metastases at 1 mo postoperatively,detected on a positron emission tomography-computer tomography scan,and perineal recurrence after 5 mo.After molecular testing,the patient was included in an immunotherapy trial.CONCLUSION This case highlights the difficulty of establishing a definitive early diagnosis of anorectal melanoma,the importance of performing histological analysis on a wellrepresented biopsy specimen,and the poor prognosis,even with radical surgery.

In Vitro Evaluation of the Potential Antioxidant of Bidens segetum Mart. ex Colla (Asteraceae) in Melanocyte and Melanoma Cells

Bidens segetum Martius ex Colla known as the “pic&atilde;o do mato”, is an herbaceous plant that occurs in the Cerrado biome of some Brazilian states. Among the species of Bidens, we highlight B. pilosa known as “pic&atilde;o preto”, of which several activities are reported as antioxidant and antibacterial. Ethanolic extract from Bidens segetum (EEBs) showed an-tioxidant potential when analyzed by free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and antifungal activity against Cladosporium cladosporiodes and C. sphareospermum fungi. PFFR3.3 subfraction from EEBs has 81.5% of 5-O caffeoylquinic acid (5-CQA) and potential antioxidant (DPPH). However, PFFR3.3 did not decrease superoxide anion in metastatic melanoma cells by dihydroeth-idium assay (DHE). PP4 subfraction is a mixture of polyacetylenes that has antifungal (Cladosporium) and antioxidant activity, since reduced superoxide anion amount in melanoma cells after 5 min of treatment. However, no dose-response and time-response curve were observed, not even with the authentic standard (5-CQA). Complementary chemical studies will be performed to confirm the polyacetylenes and 5-CQA structures present in the EEBs from B. segetum and new methodologies should be performed to confirm the antioxidant activity of these com-pounds and the effects on melanocytes and melanomas.

Eruptive Lentigines after Adalimumab Therapy

Adalimumab, a TNF-alpha antagonist, is the first fully humanized recombinant immunoglobulin G1 (IgG1) monoclonal antibody. It is presently widely used in the systemic treatment of rheumatoid arthritis, inflammatory bowel disease, moderate and severe psoriasis and hidradenitis suppurativa. However, its administration is associated with a two-fold risk of severe and possibly fatal infections and in some rare cases with congestive heart failure, lymphoma, lupus-like syndrome, cytopenias, hepatotoxicity and development of demyelinating neurological disorders. Furthermore, the occurrence of various types of melanocytic skin lesions has been reported during treatment with adalimumab. In the present paper we report the case of a female psoriatic patient who developed eruptive lentigines following treatment with this compound.