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Supplementary Table 1. Levels of IP3R3 and signaling mediators by densitometric analysis of western blots. The optical density of each band was normalized to the same parameter derived from the corresponding tubulin control using ImageJ software and reported as relative protein level compared to the corresponding internal control. Three independent siRNA transfection of H1993 cells were analyzed, data were pooled and expressed as mean {plus minus} SE.

Purpose: Our aim was to test whether imaging with 18F-fluorothymidine (18F-FLT) PET/CT was able to detect the combined effects of EGFR and MET inhibitors in oncogene-driven non–small cell lung cancer (NSCLC) and to elucidate the mechanisms underlying the enhanced efficacy of drug combination.Experimental Design: NSCLC cells bearing MET amplification (H1993 and H820) were treated with EGFR and MET inhibitors either alone or in combination and then tested for cell viability and inhibition of signaling. Nude mice bearing H1993 tumors underwent 18F-FLT PET/CT scan before and after treatment with erlotinib and crizotinib alone or in combination (1:1 ratio) and posttreatment changes of 18F-FLT uptake in tumors were determined. The role of inositol trisphosphate receptor type 3 (IP3R3) in mediating the combined action of EGFR and MET inhibitors was tested by transfecting NSCLC cells with IP3R3-targeted siRNA.Results: Imaging studies showed a significant reduction of 18F-FLT uptake in response to combined treatment with EGFR and MET inhibitors that was higher than that obtained with single agents (ANOVA, F-ratio = 6.215, P = 0.001). Imaging findings were confirmed by analysis of surgically excised tumors. Levels of IP3R3 were significantly reduced in both cells and tumors after treatment with crizotinib, whereas EGFR inhibitors caused a reduction of IP3R3 interaction with K-Ras mainly through dephosphorylation of serine residues of K-Ras.Conclusions: Our findings indicate that 18F-FLT PET/CT is able to detect the enhanced efficacy of EGFR and MET inhibitors in oncogene-driven NSCLC and that such enhancement is mediated by IP3R3 through its interaction with K-Ras. Clin Cancer Res; 24(13); 3126–36. ©2018 AACR.

Supplementary Figure S1. Toxicity of combined treatment with erlotinib and crizotinib in H820 cells. Cell toxicity was determined by MTS assay in H820 cells treated with increasing doses of erlotinib and crizotinib used alone or in combination at 1:1 ratio for 72 h. Three independent experiments were performed and data are expressed as mean {plus minus} SE.

α1-Antitrypsin (AAT) deficiency is a common genetic disease presenting with lung and liver diseases. AAT deficiency results from pathogenic variants in the SERPINA1 gene encoding AAT and the common mutant Z allele of SERPINA1 encodes for Z α1-antitrypsin (ATZ), a protein forming hepatotoxic polymers retained in the endoplasmic reticulum of hepatocytes. PiZ mice express the human ATZ and are a valuable model to investigate the human liver disease of AAT deficiency. In this study, we investigated differential expression of microRNAs (miRNAs) between PiZ and control mice and found that miR-34b/c was up-regulated and its levels correlated with intrahepatic ATZ. Furthermore, in PiZ mouse livers, we found that Forkhead Box O3 (FOXO3) driving microRNA-34b/c (miR‐34b/c) expression was activated and miR-34b/c expression was dependent upon c-Jun N-terminal kinase (JNK) phosphorylation on Ser(574). Deletion of miR-34b/c in PiZ mice resulted in early development of liver fibrosis and increased signaling of platelet-derived growth factor (PDGF), a target of miR-34b/c. Activation of FOXO3 and increased miR-34c were confirmed in livers of humans with AAT deficiency. In addition, JNK-activated FOXO3 and miR-34b/c up-regulation were detected in several mouse models of liver fibrosis. This study reveals a pathway involved in liver fibrosis and potentially implicated in both genetic and acquired causes of hepatic fibrosis.

Background Invadopodia are actin-based cell-membrane protrusions associated with the extracellular matrix degradation accompanying cancer invasion. The elucidation of the molecular mechanisms leading to invadopodia formation and activity is central for the prevention of tumor spreading and growth. Protein tyrosine kinases such as Src are known to regulate invadopodia assembly, little is however known on the role of protein tyrosine phosphatases in this process. Among these enzymes, we have selected the tyrosine phosphatase Shp1 to investigate its potential role in invadopodia assembly, due to its involvement in cancer development. Methods Co-immunoprecipitation and immunofluorescence studies were employed to identify novel substrate/s of Shp1AQ controlling invadopodia activity. The phosphorylation level of cortactin, the Shp1 substrate identified in this study, was assessed by immunoprecipitation, in vitro phosphatase and western blot assays. Short interference RNA and a catalytically-dead mutant of Shp1 expressed in A375MM melanoma cells were used to evaluate the role of the specific Shp1-mediated dephosphorylation of cortactin. The anti-invasive proprieties of glycerophosphoinositol, that directly binds and regulates Shp1, were investigated by extracellular matrix degradation assays and in vivo mouse model of metastasis. Results The data show that Shp1 was recruited to invadopodia and promoted the dephosphorylation of cortactin at tyrosine 421, leading to an attenuated capacity of melanoma cancer cells to degrade the extracellular matrix. Controls included the use of short interference RNA and catalytically-dead mutant that prevented the dephosphorylation of cortactin and hence the decrease the extracellular matrix degradation by melanoma cells. In addition, the phosphoinositide metabolite glycerophosphoinositol facilitated the localization of Shp1 at invadopodia hence promoting cortactin dephosphorylation. This impaired invadopodia function and tumor dissemination both in vitro and in an in vivo model of melanomas. Conclusion The main finding here reported is that cortactin is a specific substrate of the tyrosine phosphatase Shp1 and that its phosphorylation/dephosphorylation affects invadopodia formation and, as a consequence, the ability of melanoma cells to invade the extracellular matrix. Shp1 can thus be considered as a regulator of melanoma cell invasiveness and a potential target for antimetastatic drugs.

Recurrent pregnancy loss (RPL) is an important reproductive health issue defined as two and more failed pregnancies...

Objective: To study the controversial association between human leukocyte antigen-G (HLA-G) 14 bp polymorphism and recurrent pregnancy loss (RPL). We performed a meta-analysis of studies in the literature that enrolled only women of European countries who experienced RPL spontaneously or after undergoing IVF.Design: Systematic meta-analysis of articles published before January 2019 pertaining the association of HLA-G genotype and RPL. The search was performed in electronic databases (PubMed, Web of Science, Scopus, EMBASE), without any language or publication year restriction.Setting: Academic hospitals and private clinics.Patient(s): Women who experienced RPL spontaneously or after undergoing IVF.Intervention(s): Genotyping of 14 bp polymorphism (insertion/insertion, insertion/deletion, deletion/deletion) in exon 8 of the HLA-G gene.Main Outcome Measure(s): Meta-analyses of the association between HLA-G 14 bp polymorphism in homozygosis (insertion/insertion) and heterozygosis (insertion/deletion) in women with RPL compared with pregnant controls with at least one live birth and no history of RPL.Result(s): Ten studies were analyzed comprising 1,091 women with RPL and 808 controls without RPL. Women with RPL showed significantly higher prevalence of HLA-G 14 bp insertion/insertion genotype compared with women without RPL (19.8% vs. 14.1%; odds ratio = 1.562; 95% confidence interval, 1.203-2.027), and this result was also confirmed when separately analyzing women with RPL during a spontaneous pregnancy (odds ratio, 1.562; 95% confidence interval, 1.203-2.027) and those undergoing IVF (odds ratio, 1.990; 95% confidence interval, 0.978-4.051).Conclusion(s): Women of European countries with the HLA-G 14 bp insertion/insertion genotype have a significantly higher prevalence of RPL. ((C) 2019 by American Society for Reproductive Medicine.)

Purpose: Our aim was to test whether imaging with 18F-fluorothymidine (18F-FLT) PET/CT was able to detect the combined effects of EGFR and MET inhibitors in oncogene-driven non–small cell lung cancer (NSCLC) and to elucidate the mechanisms underlying the enhanced efficacy of drug combination. Experimental Design: NSCLC cells bearing MET amplification (H1993 and H820) were treated with EGFR and MET inhibitors either alone or in combination and then tested for cell viability and inhibition of signaling. Nude mice bearing H1993 tumors underwent 18F-FLT PET/CT scan before and after treatment with erlotinib and crizotinib alone or in combination (1:1 ratio) and posttreatment changes of 18F-FLT uptake in tumors were determined. The role of inositol trisphosphate receptor type 3 (IP3R3) in mediating the combined action of EGFR and MET inhibitors was tested by transfecting NSCLC cells with IP3R3-targeted siRNA. Results: Imaging studies showed a significant reduction of 18F-FLT uptake in response to combined treatment with EGFR and MET inhibitors that was higher than that obtained with single agents (ANOVA, F-ratio = 6.215, P = 0.001). Imaging findings were confirmed by analysis of surgically excised tumors. Levels of IP3R3 were significantly reduced in both cells and tumors after treatment with crizotinib, whereas EGFR inhibitors caused a reduction of IP3R3 interaction with K-Ras mainly through dephosphorylation of serine residues of K-Ras. Conclusions: Our findings indicate that 18F-FLT PET/CT is able to detect the enhanced efficacy of EGFR and MET inhibitors in oncogene-driven NSCLC and that such enhancement is mediated by IP3R3 through its interaction with K-Ras. Clin Cancer Res; 24(13); 3126–36. ©2018 AACR.

We report that cells from human fetal dermis, termed here multipotent fetal dermal cells, can be isolated with high efficiency by using a nonenzymatic, cell outgrowth method. The resulting cell population was consistent with the definition of mesenchymal stromal cells by the International Society for Cellular Therapy. As multipotent fetal dermal cells proliferate extensively, with no loss of multilineage differentiation potential up to passage 25, they may be an ideal source for cell therapy to repair damaged tissues and organs. Multipotent fetal dermal cells were not recognized as targets by T lymphocytes in vitro, thus supporting their feasibility for allogenic transplantation. Moreover, the expansion protocol did not affect the normal phenotype and karyotype of cells. When compared with adult dermal cells, fetal cells displayed several advantages, including a greater cellular yield after isolation, the ability to proliferate longer, and the retention of differentiation potential. Interestingly, multipotent fetal dermal cells expressed the pluripotency marker SSEA4 (90.56 ± 3.15% fetal vs. 10.5 ± 8.5% adult) and coexpressed mesenchymal and epithelial markers (>80% CD90+/CK18+ cells), coexpression lacking in the adult counterparts isolated under the same conditions. Multipotent fetal dermal cells were able to form capillary structures, as well as differentiate into a simple epithelium in vitro, indicating skin regeneration capabilities.

Background: The two main mechanisms of resistance to EGFR tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) are the occurrence of T790M secondary mutation in the kinase domain of EGFR and METamplification. The aim of the present study was to test whether early changes of 18F fluorodeoxyglucose (18F-FDG) uptake in animal models bearing erlotinib-resistant NSCLC may have different imaging patterns of response to erlotinib depending on the molecular mechanisms underlying resistance. Animal tumor models were developed using NSCLC H1975 cells bearing the T790M mutation and H1993 cells with MET amplification. Nude mice bearing erlotinib-resistant H1975 and H1993 xenografts (four animals for each cell line and for each treatment) were subjected to 18F-FDG PET/CT scan before and immediately after treatment (50 mg/kg p.o. for 3 days) with erlotinib, WZ4002, crizotinib, or vehicle. A three-dimensional region of interest analysis was performed to determine the percent change of 18F-FDG uptake in response to treatment. At the end of the imaging studies, tumors were removed and analyzed for glycolytic and mitochondrial proteins as well as levels of cyclin D1. Results: Imaging studies with 18F-FDG PET/CT in H1975 tumor-bearing mice showed a reduction of 18F-FDG uptake of 25.87 % ± 8.93 % after treatment with WZ4002 whereas an increase of 18F-FDG uptake up to 23.51 % ± 9.72 % was observed after treatment with erlotinib or vehicle. Conversely, H1993 tumors showed a reduction of 18F-FDG uptake after treatment with both crizotinib (14.70 % ± 1.30 %) and erlotinib (18.40 % ± 9.19 %) and an increase of tracer uptake in vehicle-treated (56.65 % ± 5.65 %) animals. The in vivo reduction of 18F-FDG uptake was always associated with downregulation of HKII and p-PKM2 Tyr105 glycolytic proteins and upregulation of mitochondrial complexes (subunits I–IV) in excised tumors. Conclusions: 18F-FDG uptake is a reliable imaging biomarker of T790M-mediated resistance and its reversal in NSCLC whereas it may not be accurate in the detection of MET-mediated resistance.

Neutrophil extracellular traps (NETs), originally recognized as a host defense mechanism, were reported to promote thrombosis and metastatic dissemination of cancer cells. Here we tested the role of integrins α5β1 and ανβ3 in the adhesion of cancer cells to NETs. Neutrophil-like cells stimulated with calcium ionophore (A23187) were used as a stable source of cell-free NETs-enriched suspensions. Using NETs as an adhesion substrate, two human K562 cell lines, differentially expressing α5β1 and ανβ3 integrins, were subjected to adhesion assays in the presence or absence of DNAse 1, blocking antibodies against α5β1 or ανβ3, alone or in combination with DNAse 1, and Proteinase K. As expected DNAse 1 treatment strongly inhibited adhesion of both cell lines to NETs. An equivalent significant reduction of cell adhesion to NETs was obtained after treatment of cells with blocking antibodies against α5β1 or ανβ3 indicating that both integrins were able to mediate cell adhesion to NETs. Furthermore, the combination of DNAse 1 and anti-integrin antibody treatment almost completely blocked cell adhesion. Western blot analysis and immunoprecipitation experiments showed a dose-dependent increase of fibronectin levels in samples from stimulated neutrophil-like cells and a direct or indirect interaction of fibronectin with histone H3. Finally, co-immunolocalization studies with confocal microscopy showed that fibronectin and citrullinated histone H3 co-localize inside the web-structure of NETs. In conclusion, our study showed that α5β1 and ανβ3 integrins mediate cell adhesion to NETs by binding to their common substrate fibronectin. Therefore, in addition to mechanical trapping and aspecific adsorption of different cell types driven by DNA/histone complexes, NETs may provide specific binding sites for integrin-mediated cell adhesion of neutrophils, platelets, endothelial and cancer cells thus promoting intimate interactions among these cells.

Purpose: One of the hallmarks of cancer cells is the excessive conversion of glucose to lactate under normoxic conditions, also known as the Warburg effect. Here, we tested whether the targeted inhibition of EGFR may revert this effect and reactivate mitochondrial oxidative phosphorylation in non–small cell lung cancer (NSCLC). Experimental Design: Sensitive (HCC827) and resistant (H1975 and H1993) NSCLC cells were treated with a panel of EGFR or MET inhibitors, and then tested for changes of EGFR signaling, glycolytic cascade, and mitochondrial function. Silencing of key glycolytic enzymes was then performed with targeted siRNAs. Furthermore, tumor-bearing nude mice treated with EGFR inhibitors were evaluated with 18F-FDG PET/CT and tumors were analyzed for glycolytic and mitochondrial proteins. Results: Effective inhibition of EGFR signaling in NSCLC cells induced a dramatic reduction of hexokinase II (HKII) and phospho-pyruvate kinase M2 (p-PKM2, Tyr105) levels as well as an upregulation of mitochondrial complexes subunits (OXPHOS). Accordingly, a decreased lactate secretion and increased intracellular ATP levels were also observed in response to EGFR inhibitors. Downregulation of HKII and PKM2 by targeted siRNA transfection did not cause upregulation of OXPHOS but enhanced the effects of EGFR TKIs. Conversely, selective inhibition of AKT and ERK1/2 caused OXPHOS upregulation and glycolysis inhibition, respectively. Similar findings were obtained in tumors from animals treated with appropriate EGFR inhibitors. Conclusions: Our findings indicate that EGFR inhibitors may reactivate oxidative phosphorylation of cancer cells and provide a mechanistic clue for the rational combination of agents targeting EGFR-dependent proliferation and glucose metabolism in cancer therapy. Clin Cancer Res; 21(22); 5110–20. ©2015 AACR.

Dermatological therapy can be a combination of systemic and topical treatments although topical treatment is often the only therapy prescribed. As a consequence, dermatologists have to be familiar with the use of topical preparations, whether a drug or an active agent needs to be added or not. Tailor-made preparations have undoubtedly a great positive impact on patients, but their use is limited by two main factors: the poor confidence of practising dermatologists to prescribe magistral preparations and the difficulty in finding pharmacists able and willing to provide them. In a recent analysis of compound prescriptions, some mixtures prescribed by dermatologists were questionable either because of the number of active ingredients or the selected concentrations.

Peripheral subcutaneous panniculitis-like T-cell lymphoma (PSPTCL) is a rare form of cutaneous lymphoma recently proposed as a distinct clinicopathological entity. It usually presents with multiple indurated subcutaneous plaques or tumours, most commonly located on the extremities and trunk and clinically mimicking lobular panniculitis. Associated constitutional symptoms due to haemophagocytic syndrome may advance or, more often, complicate the clinical course in about 40-70% of cases. Finding of TIA-1+ and perforin + cytolytic granules in atypical pleomorphic lymphocytes suggests PSPTCL origin from granular cells of T-cell or natural killer cell phenotype. Cells have a CD3+ CD4+ CD8- or CD3+ CD4- CD8+ T-cell phenotype. Moreover, these lymphomas can express natural killer cell associated antigens, such as CD56, especially in gamma/delta variants. PSPTCL following an indolent clinical course with recurrent self-healing lesions have been described. The prognosis of most PSPTCL is poor even when treated with aggressive chemotherapy. This paper reports a case of PCTCL in a young woman with T-cytotoxic differentiation, with rapid progression unresponsive to several treatments.

Purpose: MET amplification is one of the mechanisms underlying acquired resistance to EGFR tyrosine kinase inhibitors (TKI) in non–small cell lung cancer (NSCLC). Here, we tested whether 3′-deoxy-3′-[18F]-fluorothymidine ([18F]FLT) positron emission tomography/computerized tomography (PET/CT) can detect MET-mediated resistance to EGFR TKIs and monitor the effects of MET inhibitors in NSCLC. Experimental Design: H1993 and H820 NSCLC cells with high and low levels of MET amplification, respectively, and HCC827-expressing MET, but without gene amplification, were tested for the effects of MET inhibitors on the EGFR pathway and proliferation both in vitro and in vivo. Nude mice bearing NSCLCs with and without MET amplification were subjected to [18F]FLT PET/CT before and after treatment with crizotinib or erlotinib (50 mg/kg and 100 mg/kg p.o. for 3 days). Results: H1993 cells showed high responsiveness to MET inhibitors and were resistant to erlotinib. Conversely, HCC827 cells showed high sensitivity to erlotinib and were resistant to MET inhibitors. Accordingly, H1993 tumors bearing MET amplification showed a mean reduction in [18F]FLT uptake of 28% and 41% after low- and high-dose treatment with crizotinib for 3 days, whereas no posttherapy changes of [18F]FLT uptake were observed in HCC827 tumors lacking MET amplification. Furthermore, a persistently high [18F]FLT uptake was observed in H1993 tumors after treatment with erlotinib, whereas HCC827 tumors showed up to 39% reduction of [18F]FLT uptake following erlotinib treatment. Imaging findings were confirmed by Ki67 immunostaining of tumor sections. Conclusions: [18F]FLT PET/CT can detect MET-mediated resistance to EGFR TKIs and its reversal by MET inhibitors in NSCLC. Clin Cancer Res; 20(18); 4806–15. ©2014 AACR.

Cutaneous adverse events commonly reported with tyrosine kinase inhibitors (TKIs) in the treatment of malignancies, represent an important clinical concern since they can limit the optimal use of these novel drugs. Although there are numerous reports in the literature of these events there are no practical guidelines on how they should be managed. The Sorafenib Working Group (SWG) was established with the objective of developing recommendations to allow the early detection, prevention and management of cutaneous adverse events in everyday clinical practice. The SWG was a multidisciplinary team made up of experts in the field who were closely involved in the sorafenib clinical development program. This review provides an overview of the nature and incidence of cutaneous adverse events which manifest with sorafenib treatment and provides recommendations for their early detection and effective management in clinical practice.

This paper investigates the ceramide composition of the psoriatic scale compared with that of normal human SC. A method was optimalized, based on TLC separation followed by densitometry, allowing the provision of good resolution and quantification of ceramide fractions from both normal and pathological specimens. Seven ceramide fractions were isolated and submitted to compositional analysis. The obtained results suggested a revisitation of previous ceramide designation. Therefore a simple classification is suggested, based on grouping ceramides carrying structural similarities under common codes. According to these rules, ceramides were grouped into five classes designated as: (1) Cer[EOS], which contains ester-linked fatty acids, omega-OH fatty acids and sphingosines; (2) Cer[NS], which contains non-OH fatty acids and sphingosines; (3) Cer[NP], which contains non-OH fatty acids and phytosphingosines; (4) Cer[AS], which contains alpha-OH fatty acids and sphingosines; (5) Cer[AP], which contains alpha-OH fatty acids and phytosphingosines. Analysis of ceramides from the psoriatic scale, compared to those from normal human SC, resulted in an impairment of the Cer[EOS] content as well as of the ceramides containing phytosphingosine, with concurrent increase in ceramides containing sphingosine, being the total amount maintained identical. Since one of the suggested pathways for phytosphingosine biosynthesis involves the water addition to the corresponding sphingosine double bond, we can speculate that the observed alteration is due to a deranged water bioavailability, associated with psoriasis.

Cholesterol embolization syndrome is due to dislodgment of cholesterol crystals from the atherosclerotic plaques lining the walls of major arteries resulting in an occlusion of small arteries. We describe a case of severe cutaneous cholesterol emboli syndrome following repeat coronary angiography showing by our observation that this syndrome is often unrecognized or misdiagnosed and that a better evaluation of risks factors in patients undergoing invasive procedures could prevent this severe complication.

An open clinical study was undertaken to evaluate the anti-photoageing efficacy of topical tretinoin. A length of cream of approximately 1 cm was applied to the face daily in the evening for 6 months: during month 1 of therapy 0.01% tretinoin cream was administered; 0.025% was given during month 2; and 0.05% was given in months 3-6. The clinical symptoms of photoageing (coarse wrinkling, fine wrinkling, skin thinning, mottled hyperpigmentation, laxity and xerosis) were evaluated before and after therapy. A total of 19.1% of patients withdrew from the study; only 5.6% were for treatment-related reasons. At the end of the treatment period all the clinical parameters, except xerosis, were improved. The amount of improvement varied, but only 4.2% of patients failed to show any improvement. Tolerability was excellent in 51.4% of patients, good in 44.4% and fair in 4.2%, and compliance was excellent in 47.0% of patients, good in 48.5% and fair in 4.5%. Tolerability and compliance were improved by applying the same amount of cream each day but increasing the concentration of tretinoin over the 6-month period. Silicone skin replicas of the same area of skin taken before and after treatment, analysed by scanning electron microscopy, profilometry and computer image analysis, showed a decrease in the width of wrinkles, and an improvement in skin texture and follicle density.

We describe a 38-year-old woman with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) who developed two symmetric paramalleolar ulcers with similar diameters, depths and borders, that were successfully treated with photodynamic therapy with 5-aminolevulinic acid (ALA-PDT).

Cutaneous toxicity is the most evident adverse effect of epidermal growth factor receptor (EGFR) inhibitors because of the specific role played by EGFR in skin biophysiology. Dermatological adverse reactions, mainly folliculocentric, have been widely reported in the literature. However, the mechanism of these reactions is not well defined and their management is still a matter of debate. In this paper keratinocyte differentiation, activation and pathways regulating gene expression are reviewed in order to improve the understanding of adverse skin reactions and obtain success in their management. The authors had the opportunity to treat skin reactions induced by cetuximab in a cohort of patients affected by metastatic colorectal carcinoma. The aims of this clinical approach were to control the signs and symptoms of skin toxicity in order to avoid delay in cancer therapy and to use nondrug agents for the treatment of drug-induced skin reactions.

Topical application of tretinoin induces skin changes characterized by erythema, dryness and desquamation. These changes are primarily due to a specific tretinoin action on keratinocyte differentiation and proliferation [1]. Tretinoin added to cultured keratinocytes is able to impair morphological structure and the expression of several markers of terminal differentiation such as filaggrin, loricrin and involucrin [2]. The reduction in the expression of these proteins is considered to be the cause of a reduced and less adhesive horny layer [3]. Interlamellar lipids also play an important role in lamellar cohesion and adhesion in the stratum corneum layers [4]. However, little information is available on the effects of retinoic acid (RA) on interlamellar lipid synthesis [5]. The synthesis of these specific lipids is a part of keratinocyte differentiation [6], so theoretically tretinoin may also affect keratinocyte lipid metabolism. The aim of this study was to verify in vivo whether the topical application of tretinoin induces any modification in lipid production. For this purpose lipids from cyanoacrylate strippings of the horny layer after 21 days tretinoin exposure were analysed and compared with those from a vehicle-treated area of the same subject. Stratum corneum samples were obtained from the volar forearm of ten healthy volunteers, all females (ages ranging from 28 to 35 years), treated with freshly prepared tretinoin (all-trans-RA) 0.05%, glycoalcoholic solution, once a day, Monday to Friday, for a period of 21 days (15 applications). The contralateral forearm was treated with vehicle (propylene glycol/ethanol, 1:1) as control. The clinical score was recorded 2 days after the final application, and transepidermal water loss (TEWL) was measured by means of an evaporimeter (EP1, Servomed, Stockholm, Sweden). Stratum corneum samples were obtained by a single

Iatrogenic Kaposi's sarcoma develops in patients undergoing immunosuppressive treatment and is considered to be induced by activation of latent HHV8. In most cases the first manifestation of Kaposi's sarcoma develops after 1 year from when the drug was first administered. In a recent study from Italy on HHV8 positivity in patients with Kaposi's sarcoma, it was found that 52% of the control group were positive (Masini C., et al. G Ital Dermatol Venereol 1999; 134: 315–320). For this reason we could expect a larger number of cases of iatrogenic Kaposi's sarcoma given the number of patients who undergo immunosuppressive treatment for one reason or another. Thus, we have to look to a contemporaneous presence of other factors that co-operate with the HHV8. We present a case of a 49-year-old woman, HHV8 and HCV positive, who develops a Kaposi's sarcoma after 9 months of steroid therapy (methylprednisolone 16 mg/die). The low dose of steroids prescribed to our patient and the fact that the first skin manifestation developed after a shorter period than average from the start of therapy do not explain the acute onset of an extensive Kaposi's sarcoma even taking into account the HHV8 positive status. Both HHV8 and HCV produce proteins, such as IL6 and IL8 which are able to control cell growth. It can be supposed that the contemporaneus presence of the two viruses created a sinergy for the onset of the Kaposi's sarcoma.

Summary In a previously reported study on the anti-photoageing effects of topical tretinoin, the following regimen produced good patient compliance: 0·01% for 1 month, 0·025% for 1 month; and 0·05% for 4 months. The majority of patients (60/89) enrolled in the initial study continued to apply the cream to the face, and a further 140 patients were enrolled for a long-term study (mean duration 2 years). The prolonged study showed that 91·4% of patients used tretinoin in an attempt to slow down skin ageing, and 8·6% sought subjective skin benefits. Of the 163 patients who completed the study, 58·8% sought an improvement of wrinkles, 30·1% skin trophism and 14·7% reduced pigmentation. The product was used throughout the year by 66·9% of patients, but 8·0% stopped using it during the summer. A daytime moisturizing cream was required by 77·9% of patients, and 82·8% used a sunscreen in the summer. Tretinoin was applied to other areas of the body by 63·8% of patients.

This report is concerned with gingival manifestations associated with a case of dermochondrocorneal dystrophy (DCCD) or Francois syndrome occurring in a 42-year-old woman. Our Department treated this patient for 15 years. Oral examination of (his case revealed a diffuse enlargement and severe inflammation of the attached gingiva. Systemic findings were similar to those reported in the literature for patients with DCCD. Firm papules 3 mm wide, localized on the face and on the dorsal surface of the hands, were associated with corneal involvement and progressive and severe articular disorder. Because they recurred after surgical ablation, the gingival lesions became an important problem in the management of the patient. After 10 years of unsuccessful treatment limited to scaling, oral hygiene control and mouth rinses with 0.2% chlorexidine solution, the patient was submitted to extraction of the remaining teeth, remodeling osteoplasty and cutaneous graft. An acrylic full denture was inserted. In a follow-up of 7 years, good results for the oral health of the patient were seen.

The efficacy of air bags has been proven in diminishing the rate of fatalities and severity of injury in motor vehicle crashes. Unfortunately, as with any developing technology, new problems have been encountered that are directly attributable to the deployment of the air bag itself. Most air bag-related injuries are minor and, surprisingly, more than 7% are burns typically involving the upper extremity or head or neck. Fortunately, these are superficial burns that usually require only expectant therapy, but a high degree of suspicion in these circumstances is needed to make the proper diagnosis.

Five patients who developed severe cutaneous complications after artificial hair implantation were subjected to a scalp biopsy. The pathology showed the presence of hyperplastic epidermal proliferations that produced infundibulum-like structures around the implanted fibers in the superficial dermis. A dense acute inflammatory infiltrate surrounded the artificial fibers in the superficial dermis. In the deep dermis a granulomatous infiltrate was present whereas in the hypodermis the inflammatory infiltrate was sparse and the fibers were embedded in fibroplasia. The pathology of a patient who did not present any skin inflammation after artificial hair implantation showed similar pathological features but the absence of acute inflammation suggesting that bacterial infections play a major role in the development of the cutaneous complications of hair implantation. Since definitive treatment of the infections is ineffective until the fibers are removed from the scalp, surgical treatment was required in 2 of our patients.

The case of a patient who developed multiple soft fibroma-like lesions on his lower extremities affected by lymphedema and Kaposi's sarcoma is reported. To the best of our knowledge, the coexistence of these three pathologic processes is unusual and has never been described before in the literature.

QUANTITATIVE ANALYSIS OF TUMOR INFILTRATING LYMPHOCYTES IN CUTANEOUS MELANOMA Stefania Motta, PhD, University of Milan-Istituto Clinico Humanitas, 20089 Rozzano Milano, Italy, Marcello Monti, MD, University of Milan-Istituto Clinico Humanitas, 20089 Rozzano Milano, Italy, Barbara Franceschini, PhD, University of Milan-Istituto Clinico Humanitas, 20089 Rozzano Milano, Italy, Carlo Russo, PhD, University of Milan-Istituto Clinico Humanitas, 20089 Rozzano Milano, Italy

Background and Design: In psoriasis the formation of the cornified layer is deranged and water flux is reportedly increased. We investigated three different forms of psoriasis: transepidermal water loss was measured on uninvolved skin and psoriatic plaques; lipids from plaques were extracted; and ceramide distribution in scale vs normal stratum corneum was compared. Moreover, the lipid biochemical results were compared with transepidermal water loss rates in the same lesions. To assess potential alteration in ceramide distribution, lipids from both psoriatic scale and normal stratum corneum were extracted by the Bligh-Dyer method, separated on high performance thin layer chromatography plates, and quantified by computerized densitometry. Water flux was measured as transepidermal water loss using an evaporimeter; results between uninvolved and involved psoriatic skin and age-matched control skin were statistically evaluated. Results: In comparison with normal stratum corneum, psoriatic plaque ceramides showed a different distribution; in particular, ceramide 1 was significantly decreased. The increased transepidermal water loss values of psoriatic plaques vs control skin and between Psoriatic involved vs uninvolved skin were significant. Conclusion: Our findings indicate that in psoriasis the altered ceramide distribution can be linked specifically to the defect in keratinocyte differentiation; the defect in skin barrier function may be attributed largely or in part to ceramide 1 reduction. (Arch Dermatol. 1994;130:452-456)

After an accident in a chemical plant in Seveso, Italy, on July 10, 1976, 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD) spread over a populated area. The event was exceptional because children were also affected and because the contamination took place not only through direct exposure but also through inhalation and the ingestion of contaminated foods, especially fruits and vegetables. This paper illustrates the early dermatologie lesions, the late acneic (chloracne) lesions, and their evolution during a 10-year period. Peculiar cutaneous findings, histologic data, and a comparison with previously reported similar accidents are also included.