Search

MEN2A is a hereditary cancer-predisposing syndrome that affects patients with germline RET mutations. The effects of this oncogenic tyrosine kinase in the context of primitive stem cells are not known. In order to study these events, we generated a MEN2A induced Pluripotent Stem Cell (iPSC) line from a patient with RET mutation and an isogenic counterpart by CRISPR-Cas9 correction of the mutation. Whole exome sequencing of iPSC before and after CRISPR-Cas9 genome edition revealed no major exonic off target effect of the CRISPR correction. However, an integrative differential gene expression analysis of iPSC with oncogenic RET C634Y and its gene-corrected iPSC with RET Y634C as well as RET wt iPSCs revealed activation of the Early Growth Response 1 (EGR1) transcriptional program in RET-mutated iPSC, a pathway shown to be involved in RET-induced oncogenesis. These data constitute the first proof of concept of the feasibility of the use of an iPSC and its genome-corrected counterpart to unravel the molecular mechanisms underlying the development of the hereditary MEN2A cancer predisposing syndrome., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study we analyzed the expression levels of markers of epithelial-to-mesenchymal transition (EMT) in several papillary thyroid carcinomas (PTCs) and the relation with tumor genotypes and clinicopathological characteristics. The role of fibronectin-1 (FN1) was investigated by analyzing the effects of FN1 silencing in two human thyroid cancer cell lines. Most of EMT markers were significantly over-expressed in a group of 36 PTCs. In particular, FN1 mRNA levels were higher in tumor vs non-tumor tissue (117.3, p < 0.001) and also in aggressive and BRAF(V600E) samples. Similar results were observed (and confirmed at the protein level) when FN1 expression was analyzed in a validation group of 50 PTCs and six lymph node (LN) metastases. Silencing of FN1 in TPC-1 and BCPAP thyroid cancer cells significantly reduced proliferation, adhesion, migration, and invasion in both cell lines. Collectively, our data indicate that FN1 overexpression is an important determinant of thyroid cancer aggressiveness., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

An increased incidence of obesity is registered worldwide, and its association with insulin resistance and type 2 diabetes is closely related with increased morbidity and mortality for cardiovascular diseases. A major clinical problem in the management of obesity is the non-adherence or low adherence of patients to a hypocaloric dietetic restriction. In this study, we evaluated in obese mice the effects of shifting from high-calorie foods to normal diet on insulin sensitivity. Male C57BL/6JOlaHsd mice (n = 20) were fed with high fat diet (HFD) for a 24-week period. Afterward, body weight, energy, and food intake were measured in all animals, together with parameters of insulin sensitivity by homeostatic model assessment of insulin resistance and plasma glucose levels in response to insulin administration. Moreover, in half of these mice, Glut4 mRNA levels were measured in muscle at the end of the high fat treatment, whereas the rest of the animals (n = 10) were shifted to normocaloric diet (NCD) for 10 weeks, after which the same analyses were carried out. A significant reduction of body weight was found after the transition from high to normal fat diet, and this decrease correlated well with an improvement in insulin sensitivity. In fact, we found a reduction in serum insulin levels and the recovery of insulin responsiveness in terms of glucose disposal measured by insulin tolerance test and Glut4 mRNA and protein expression. These results indicate that obesity-related insulin resistance may be rescued by shifting from HFD to NCD.

Novel therapeutic approaches are required for the less differentiated thyroid cancers which are non-responsive to the current treatment. In this study we tested an innovative formulation of nanoliposomes containing sildenafil citrate or tadalafil, phosphodiesterase-5 inhibitors, on two human thyroid cancer cell lines (TPC-1 and BCPAP). Nanoliposomes were prepared by the thin layer evaporation and extrusion methods, solubilizing the hydrophilic compound sildenafil citrate in the aqueous phase during the hydration step and dissolving the lipophilic tadalafil in the organic phase. Nanoliposomes, made up of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine monohydrate (DPPC), cholesterol, and N -(carbonyl-methoxypolyethylene glycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-mPEG2000) (6:3:1 molar ratio), were characterized by a mean diameter of ~100 nm, a very low polydispersity index (~0.1) and a negative surface charge. The drugs did not influence the physico-chemical properties of the systems and were efficiently retained in the colloidal structure. By using cell count and MTT assay, we found a significant reduction of the viability in both cell lines following 24 h treatment with both nanoliposomal-encapsulated drugs, notably greater than the effect of the free drugs. Our findings demonstrate that nanoliposomes increase the antiproliferative activity of phosphodiesterase-5 inhibitors, providing a useful novel formulation for the treatment of thyroid carcinoma.

Objective: We report the case of a female patient with multiple endocrine neoplasia type 2A (MEN2A) who was found to have a double mutation in the RET (rearranged during transfection) proto-oncogene., Methods: RET mutational analysis was performed by Sanger DNA sequencing., Results: The proband was a compound heterozygote for the RET germline mutations Val648Ile and Val804Leu on exons 11 and 14, respectively. Genetic analysis of family members showed the presence of the Val648Ile mutation in all except 1 daughter who carried the Val804Leu mutation. However, none of them showed any clinical, biochemical, or histologic signs of neoplastic disease either in the thyroid or adrenal gland. Furthermore, a daughter and the proband's sister who underwent a prophylactic thyroidectomy did not show pathologic evidence of C-cell disease., Conclusions: We hypothesize that the combined effect of the 2 mutations may have induced the development of pheochromocytoma (PHEO) in our patient. Thus, in the presence of single RET-induced mild medullary thyroid cancer (MTC) phenotype, the search for additional genetic anomalies may lead to the discovery of rare but potentially more aggressive double mutation genotypes.

Recent studies have revealed in normal thyroid tissue the presence of the transcript of several phosphodiesterases (PDEs), enzymes responsible for the hydrolysis of cyclic nucleotides. In this work, we analyzed the expression of PDE5 in a series of human papillary thyroid carcinomas (PTCs) presenting or not BRAF V600E mutation and classified according to ATA risk criteria. Furthermore, we tested the effects of two PDE5 inhibitors (sildenafil, tadalafil) against human thyroid cancer cells. PDE5 gene and protein expression were analyzed in two different cohorts of PTCs by real-time PCR using a TaqMan micro-fluid card system and Western blot. MTT and migration assay were used to evaluate the effects of PDE5 inhibitors on proliferation and migration of TPC-1, BCPAP, and 8505C cells. In a first series of 36 PTCs, we found higher expression levels of PDE5A in tumors versus non-tumor (normal) tissues. PTCs with BRAF mutation showed higher levels of mRNA compared with those without mutation. No significant differences were detected between subgroups with low and intermediate ATA risk. Upregulation of PDE5 was also detected in tumor tissue proteins. Similar results were obtained analyzing the second cohort of 50 PTCs. Moreover, all tumor tissues with high PDE5 levels showed reduction of Thyroglobulin, TSH receptor, Thyroperoxidase, and NIS transcripts. In thyroid cancer cells in vitro, sildenafil and tadalafil determined a reduction of proliferation and cellular migration. Our findings demonstrate for the first time an overexpression of PDE5 in PTCs, and the ability of PDE5 inhibitors to block the proliferation of thyroid cancer cells in culture, therefore, suggesting that specific inhibition of PDE5 may be proposed for the treatment of these tumors.

Effects of flavonoids extracted from Citrus reticulata (mandarin) juice on proliferation and migration of 3 human anaplastic thyroid carcinoma (ATC) cell lines were evaluated. Flavonoid components of Mandarin juice extract (MJe) were analyzed by uHPLC. Proliferation of CAL-62, C-643, and 8505C cells, measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, was significantly reduced by MJe in a concentration- and time-dependent way, with maximal effect elicited at 0.5 mg/ml concentration after 48 h. Cytofluorimetric analysis showed a block in the G2/M phase of the cell cycle, accompanied by low cell mortality owed to autophagic death. The extract caused also a reduction of cell migration, associated with decreased activity of the metalloproteinase MMP-2. These findings demonstrate that the flavonoid fraction of mandarin juice exerts in vitro antiproliferative effects on ATC cells, associated with a reduction of migration, suggesting for such a functional food a potential use as adjuvant in the treatment of thyroid cancer.

Background: Narrowband ultraviolet B phototherapy (nbUVB) is an established treatment modality for patients with severe atopic dermatitis (AD) and is known to increase serum vitamin D levels (SVD). The relationship between SVD and AD remains unclear., Objectives: To assess SVD and disease severity in patients with AD before and after nbUVB and establish whether a change in SVD correlates to response to nbUVB. Methods: A single-centre, open observational study recruited 21 male and 17 female patients with AD between November and April. Eczema severity was measured using the SCORAD tool, and serum 25-hydroxyvitamin D3 levels were determined before and after nbUVB, which was administered thrice weekly., Results: Nine patients had severe AD, 23 moderate and six mild, as indicated by SCORAD measurements. Seventeen patients completed the study. Median SVD increased from 45 nmol/l pre-treatment to 169 nmol/l post-treatment (95% CI 2.9–5.0 times baseline) (P < 0.0001). Mean SCORAD reduction following nbUVB was significant at 21.9 units (95% CI, 14.9–28.9) (P < 0.0001). No evidence of a relationship between change in SCORAD and change in SVD was found., Limitations: A significant number of patients failed to complete the study., Conclusions: Patients with AD are at risk of vitamin D deficiency. Correlation between this and disease severity has been postulated, but this study does not provide confirmatory evidence.

Background: Cutaneous squamous cell carcinoma (cSCC) is increasing in incidence but mortality rates are low. Identifying high-risk tumours is important when rationalizing clinical review for patients with cSCC., Objectives: To assess the accuracy of death certification in cases of reported fatal cSCC and to identify risk factors for fatal cSCC., Methods: A retrospective, observational study of cases of fatal cSCC over 11 years (1993-2004) in Leeds, identified in cancer registry and death certification data., Results: Fifty-eight patients were recorded by the registry as having fatal cSCC in this period. Review of case notes and pathology specimens, where available (34 cases), confirmed that 21/34 patients had died of cSCC. Five were on the ear and none on the lip. Four patients had been treated for leukaemia or lymphoma and one was a renal transplant recipient. On pathology review five patients proved to have had malignant adnexal tumours rather than cSCC, and one a melanoma. In addition, three patients had disease of the ear canal or vulva., Conclusions: A proportion of deaths were falsely attributed to cSCC as a result of inaccurate histological diagnosis. Some fatalities were related to tumours in sites known to be at higher risk, and a significant proportion was postulated to be related to immunosuppression. In those cases attributed to cSCC in which this could be assessed, the majority were American Joint Committee on Cancer stage 2 and only 24% were in high-risk sites., (© 2013 British Association of Dermatologists.)

Acquired port-wine stains (PWSs) are rare but well recognized, and have often been reported in association with localized trauma. Various drugs have also been implicated as aetiological factors. We report the case of a 71-year-old man with an extensive congenital PWS affecting the left side of the face, upper chest and upper back. At the age of 69 years, he began to develop similar areas on his right hand and forearm, left hand and left knee. This corresponded temporally to being started on simvastatin and metformin, both of which may promote angiogenesis., (© The Author(s). CED © 2011 British Association of Dermatologists.)

The ingredients previously reported to cause protective glove allergy are presented and evaluated for strength of evidence. Allergens that have caused both delayed hypersensitivity and contact urticaria are considered for rubber, plastic, leather, and textile gloves. The current guidelines regarding glove manufacture are described. A list of materials confirmed by the industry to be used in glove production is presented together with a suggested series for investigating patients with delayed type hypersensitivity and contact urticaria secondary to glove use.

Background: Narrowband ultraviolet (UV) B phototherapy (nbUVB) is a widely used modality in the treatment of psoriasis and is generally accepted to be safe in pregnancy. Previous studies have described photodegradation of serum folate after exposure to UVA radiation but the effect of UVB is not known. Insufficient folate levels predispose to neural tube defects in pregnancy. In addition, folate deficiency may increase the risk of colorectal carcinoma and influence serum homocysteine levels, which are thought to be a risk factor for cardiovascular disease., Objective: We sought to determine whether exposure to nbUVB reduces folate levels., Methods: This single-center, open observational study was conducted in a teaching hospital setting. Serum and red cell folate levels were measured in patients with psoriasis attending for nbUVB at baseline and after a minimum of 18 exposures., Results: A total of 35 patients completed the study. Mean serum and red cell folate levels at baseline were 6.34 and 410 ng/ml, respectively. After nbUVB, mean serum and red cell folate levels were 6.35 and 423 ng/ml, respectively. This was not significantly different from baseline measurements (P = .99 serum folate and P = .57 red cell folate)., Limitations: This study examined the effect of a standard course of 18 exposures of nbUVB on folate levels, rather than prolonged treatment. Adult patients with psoriasis only were investigated., Conclusion: A standard course of nbUVB does not have an effect on serum and red cell folate levels in patients with psoriasis.

Previous studies report the benefit of UVA1 phototherapy in treating acrosclerosis in patients with systemic sclerosis. We carried out a retrospective study to examine the effectiveness of UVA1 phototherapy in scleroderma affecting acral and proximal sites in patients with this disease. Patients with systemic sclerosis (diffuse type, n=5; limited type, n=3) underwent low-dose UVA1 radiation (30-40 J/cm(2)) thrice weekly. In all patients skin lesions improved, demonstrated by a fall in the modified Rodnan skin score ranging from 8 to 18 points in diffuse systemic sclerosis and 6 to 10 points in limited disease. This study would suggest that UVA1 phototherapy is effective for scleroderma affecting proximal and acral sites in patients with systemic sclerosis.

Chronic actinic dermatitis (CAD) is an uncommon photodermatosis in which patients typically exhibit photosensitivity together with contact allergy. Improvement of the photosensitivity over time is well recognized in a proportion of patients but the concurrent resolution of contact allergy is rare. We report on a 46-year-old female with longstanding CAD who demonstrated resolution of both of these features.

Milia en plaque is a term used to describe an aggregation of milia occurring on an erythematous base usually localized in the retroauricular area. In most reported cases, no aetiological factors have been identified. We report the first case of milia en plaque associated with discoid lupus erythematosus occurring in the retroauricular site.

Cutaneous carcinosarcomas are rare biphasic malignant tumors with a malignant epithelial component together with malignant stroma. Five cases treated in the Yorkshire region of England between 2003 and 2006 are presented. The patients were male with an age range from 58 to 84 years. Four lesions occurred on the face and one on the trunk. Each tumor had an epithelial component resembling a typical nodular basal cell carcinoma. The stromal components demonstrated atypical spindle cells and tumor giant cells, undifferentiated stroma, or osteoid formation. All underwent surgical excision and none showed evidence of recurrence (follow-up 3-17 months, one death from unrelated causes). The recent literature concerning the pathogenesis and prognosis of these unusual tumors is reviewed.

Topical photodynamic therapy (PDT) is well established for the treatment of malignant and pre-malignant skin lesions and now it is increasingly used to treat benign dermatological conditions. The studies reporting the use of PDT in hidradenitis suppurativa are reviewed. Though small in number, there is considerable variation in the application of topical photosensitisers, light sources used and treatment regimes. In addition there is often limited information about patient selection in terms of disease severity and measuring precise patient outcome. These issues need to be addressed in future studies in order to accurately determine the role of PDT in this disease.

Renal transplant recipients are at increased risk for development of nonmelanoma skin carcinoma, owing to a number of causes, including ultraviolet exposure. It has been shown that, despite education, there is poor compliance by these patients with the advice given for protecting their skin from the sun. This repeat study was conducted to determine whether there had been an improvement in compliance over the last 6 years. Two hundred twenty-seven patients were invited to complete the questionnaire used in the previous study. This questionnaire was designed to establish whether patients understood the need for extra care, whether they recalled any education about protective measures, and what actual measures were taken. There was a significant increase in the proportion of patients taking appropriate precautions. Hence there has been a significant improvement in the compliance of renal transplant recipients in Yorkshire with skin protection measures since this was originally audited in 1998.

Solar urticaria is a well defined although uncommon photosensitivity disorder, and is said to be the underlying cause of chronic urticaria in approximately 0.5% cases. In contrast, solar angioedema is seldom reported. We describe two patients with postulated solar angioedema, associated with clinical and/or phototest features of solar urticaria. Recognition of solar provocation of angioedema has important consequences for patient management.

Cyclosporin and mycophenolate mofetil (MMF) are immunosuppressant agents now used frequently in the field of organ transplantation. More recently cyclosporin has been used for the treatment of a number of dermatological conditions, including severe psoriasis and eczema. Extensive diffuse morphoea is very difficult to treat. PUVA, UVA and a number of immunomodulating drugs have been used to attempt improvement but are most beneficial only in early disease. Combination treatments are often used in psoriasis, for example, but are not reported in morphoea. We present the case of a patient treated initially with cyclosporin and PUVA and subsequently with MMF and PUVA, with considerable improvement in his condition.

The data base management structure described in this paper combines the features of on-line accessibility and multiple-record management of INQUIRE, a software product of the Infodata Corporation, with the off-line efficiency of SAS, the Statistical Analysis System vended by the SAS Institute. The design process advocated herein involves the consolidation of information throughout the clinic setting into a set of single-entry source documents whereby each datum is collected only once, except for key identifiers, and a system allocation to real-time processing only for those data elements required for immediate access by users. Other data are processed in batch and stored off line but are catalogued in an on-line INDEX as part of the INQUIRE structure. The resultant software product is referred to as a "data engine," which has the capability of low-cost efficiency, being run entirely in batch, but allows growth to full on-line services, and is designed to function as the control node of a network.