High‐frequency p16INK4A promoter methylation is associated with histone methyltransferase SETDB1 expression in sporadic cutaneous melanoma

Epigenetic mechanisms participate in melanoma development and progression. The effect of histone modifications and their catalysing enzymes over euchromatic promoter DNA methylation in melanoma remains unclear. This study investigated the potential association of p16^INK4A promoter methylation with histone methyltransferase SETDB1 expression in Greek patients with sporadic melanoma and their correlation with clinicopathological characteristics. Promoter methylation was detected by methylation‐specific PCR in 100 peripheral blood samples and 58 melanoma tissues from the same patients. Cell proliferation (Ki‐67 index), p16^INK4A and SETDB1 expression were evaluated by immunohistochemistry. High‐frequency promoter methylation (25.86%) was observed in tissue samples and correlated with increased cell proliferation (P = 0.0514). p16^INK4A promoter methylation was higher in vertical growth‐phase (60%) melanomas than in radial (40%, P = 0.063) and those displaying epidermal involvement (P = 0.046). Importantly, p16^INK4A methylation correlated with increased melanoma thickness according to Breslow index (P = 0.0495) and marginally with increased Clark level (I/II vs III/IV/V, P = 0.070). Low (1–30%) p16^INK4A expression was detected at the majority (19 of 54) of melanoma cases (35.19%), being marginally correlated with tumor lymphocytic infiltration (P = 0.078). SETDB1 nuclear immunoreactivity was observed in 47 of 57 (82.46%) cases, whereas 27 of 57 (47.37%) showed cytoplasmic immunoexpression. Cytoplasmic SETDB1 expression correlated with higher frequency of p16^INK4A methylation and p16^INK4A expression (P = 0.033, P = 0.011, respectively). Increased nuclear SETDB1 levels were associated with higher mitotic count (0–5/mm² vs >5/mm², P = 0.0869), advanced Clark level (III‐V, P = 0.0380), epidermal involvement (P = 0.0331) and the non‐chronic sun exposure‐associated melanoma type (P = 0.0664). Our data demonstrate for the first time the association of histone methyltransferase SETDB1 with frequent methylation of the euchromatic p16^INK4A promoter and several prognostic parameters in melanomas.     

Incidence,aetiology, treatment outcome and complications of maxillofacial fractures. A retrospective study from Northern Greece

AimTo analyse the pattern of maxillofacial injuries and treatment outcomes in Northern Greece.MethodsA tertiary referral single centre hospital; retrospective chart review. Demographics, aetiology, fixation technique (Rigid Internal Fixation: RIF; Maxillomandibular Fixation: MMF) post-surgical infections, aesthetics and occlusion were recorded.ResultsOne thousand and ten males and 229 females were operated between 1998 and 2008. Mean age was 29.6 ± 13. Mean number of plates per patient was 3.96 ± 2.28. For those with midfacial fractures (n = 379) mean was 4.02 ± 2.05. For those with mandibular fractures (n = 333), mean was 2.74 ± 0.94 while those with combined mandible and midface fractures (n = 216) were treated using 5.74 ± 2.87 plates per patient. Among those treated with plates, an unadjusted 22% increased risk for post-surgical infection per plate used (OR = 1.22, 95%CI: 1.13–1.32) was found. Patients with mandibular fractures were more satisfied with their post-surgical facial appearance in contrast to those with midfacial or combined midfacial and mandibular fractures. Female patients were less satisfied with their post-surgical facial appearance than males.DiscussionThis study verified a young males predominance, a shift towards more assault related fractures –especially in females– and similar post-surgical results for MMF and RIF modalities in mandibular fractures. In those patients treated with RIF, placement of fewest plates possible to obtain stability better serves aesthetics at the same time reducing risk for post-surgical infections and malocclusion.     

Decomposition of Ag-based soldering alloys used in space maintainers after intra-oral exposure. A retrieval analysis study

Objective. The aim of this study was to evaluate the elemental alterations of Ag soldering alloys used in space maintainers after intra-oral exposure. Materials and methods. Twenty devices were fabricated by using two different soldering alloys; US (Dentaurum Universal Silver Solder, n = 10) and OS (Leone Orthodontic Solder, n = 10). All devices were manufactured by the same technician. Surface morphology and elemental quantitative analysis of the soldering alloys before and after intra-oral placement in patients was determined by scanning electron microscopy and energy-dispersive X-ray microanalysis (SEM/EDX). Statistical analysis was performed by t-test, Mann Whitney tests and Pearson's correlation. For all tests a 95% confidence level was used (α = 0.05). Results. Both soldering alloys demonstrated substantially increase in surface roughness after intra-oral aging. Statistical analysis illustrated a significant decrease in the Cu and Zn content after treatment. OS demonstrated higher Cu release than US (p < 0.05). The remaining relative concentrations of Cu and Zn after the treatment did not show any correlation (p > 0.05) with intra-oral exposure time, apart from Zn in OS (r = 0.840, p = 0.04). Conclusions. Both soldering alloys demonstrated a significant Cu and Zn reduction after intra-oral exposure that may raise biocompatibility concerns.     

Sulfated Progesterone Metabolites That Enhance Insulin Secretion via TRPM3 Are Reduced in Serum From Women With Gestational Diabetes Mellitus

Serum progesterone sulfates were evaluated in the etiology of gestational diabetes mellitus (GDM). Serum progesterone sulfates were measured using ultra-performance liquid chromatography–tandem mass spectrometry in four patient cohorts: 1) the Hyperglycemia and Adverse Pregnancy Outcomes study; 2) London-based women of mixed ancestry and 3) U.K.-based women of European ancestry with or without GDM; and 4) 11–13 weeks pregnant women with BMI ≤25 or BMI ≥35 kg/m² with subsequent uncomplicated pregnancies or GDM. Glucose-stimulated insulin secretion (GSIS) was evaluated in response to progesterone sulfates in mouse islets and human islets. Calcium fluorescence was measured in HEK293 cells expressing transient receptor potential cation channel subfamily M member 3 (TRPM3). Computer modeling using Molecular Operating Environment generated three-dimensional structures of TRPM3. Epiallopregnanolone sulfate (PM5S) concentrations were reduced in GDM (P < 0.05), in women with higher fasting plasma glucose (P < 0.010), and in early pregnancy samples from women who subsequently developed GDM with BMI ≥35 kg/m² (P < 0.05). In islets, 50 µmol/L PM5S increased GSIS by at least twofold (P < 0.001); isosakuranetin (TRPM3 inhibitor) abolished this effect. PM5S increased calcium influx in TRPM3-expressing HEK293 cells. Computer modeling and docking showed identical positioning of PM5S to the natural ligand in TRPM3. PM5S increases GSIS and is reduced in GDM serum. The activation of GSIS by PM5S is mediated by TRPM3 in both mouse and human islets.     

A somatic mutation in the thyrotropin receptor gene in a patient with an autonomous nodule within a multinodular goiter

Thyrotropin (TSH) is the prime regulator of thyroid cell growth and function and acts through the thyrotropin receptor (TSHR) located on the surface membrane of thyrocytes. Somatic heterozygous mutations that cause TSHR activation in the absence of TSH have been found in toxic adenomas and in hot nodules of multinodular goiters. Clinically and histologically heterogeneous nodules can share common gain-of-function mutations. Mutation prevalence varies greatly and is inversely related to iodine intake of the population. We report a Greek patient presenting with subclinical hyperthyroidism due to a fast-growing autonomous hyperplastic nodule in a long-standing multinodular goiter. Direct DNA sequencing showed that the hot nodule harbored a somatic heterozygous activating TSHR mutation: substitution of glutamine for leucine in the third transmembrane helix. This mutation (L512Q) was recently described in two solitary toxic adenomas. This report expands the spectrum of mutations shared by dissimilar hot nodules, supporting a common mechanism for nonautoimmune thyroid autonomy. The identification of the L512Q substitution demonstrates that gain-of-function TSHR mutations are encountered in Greece, although iodine deficiency has been significantly corrected over the last three decades.     

The analgesic effect of gabapentin and mexiletine after breast surgery for cancer

We investigated the analgesic efficacy of mexiletine and gabapentin on acute and chronic pain associated with cancer breast surgery in 75 patients. They were randomized to receive, in a double-blinded manner, mexiletine 600 mg/d, gabapentin 1200 mg/d, or placebo for 10 days. Anesthesia was standardized, and all patients had access to routine postoperative analgesics on demand. The visual analog scale score assessed pain at rest and after movement. Three months later, all patients were interviewed to identify intensity of chronic pain and analgesic requirements. Mexiletine and gabapentin reduced codeine consumed from the second to tenth day by 50% (P = 0.029; P = 0.018 and P = 0.035 for mexiletine versus control and gabapentin versus control comparisons, respectively). Total paracetamol consumption was also reduced during the same time (P = 0.0085; P = 0.007 and P = 0.011 for the mexiletine and gabapentin groups when compared with the control, respectively). Pain at rest and after movement was reduced by both drugs on the third postoperative day. Pain after movement also was reduced by gabapentin between the second and fifth postoperative day. Three months later, the incidence of chronic pain, its intensity, and need for analgesics were not affected by either treatment. However, burning pain was more frequent in the control group (P = 0.033). IMPLICATIONS: Patients undergoing breast surgery for cancer may develop chronic pain. We evaluated the effect of mexiletine and gabapentin on the acute and chronic pain after breast surgery for cancer. Both drugs reduced the postoperative analgesic requirements, and particularly, gabapentin reduced pain after movement. The overall incidence of chronic pain was unaffected except for burning pain.     

Successful conservative treatment of neutropenic enterocolitis complicating taxane-based chemotherapy: a report of five cases

Five cases of acute neutropenic enterocolitis complicating taxane-based chemotherapy are described. During a 34-month period, our department administered 4,600 courses of taxane-based (paclitaxel and docetaxel) chemotherapy to 800 cancer patients. Seven to 10 days postchemotherapy in five patients (0.1% of the given courses), neutropenic fever, abdominal pain, rebound tenderness, and grade II-IV diarrhea (bloody in two cases) developed. Two patients had oral candidiasis, and in two others septic shock developed. Computed tomography scans of the abdomen revealed in all patients thickening of the colon wall and pericolic edema, and a pericolic abscess was revealed in three of them. Both clinical and radiologic findings supported the diagnosis of acute neutropenic enterocolitis. All patients were successfully treated with broad-spectrum antibiotics and recombinant human granulocyte colony-stimulating factor. In conclusion, acute neutropenic enterocolitis is a severe complication of taxane-based chemotherapy. Early diagnosis and appropriate conservative treatment leads to complete recovery. Although rare, this infection is less often associated with other chemotherapeutic regimens.     

Early to bed: a study of adaptation among sexually active urban adolescent girls younger than age sixteen

OBJECTIVE: To examine the association between sexual activity among urban adolescent girls and four global measures of psychosocial adaptation (academic motivation, school achievement, depressive symptoms, and expectations about the future). METHOD: Data derived from the Social and Health Assessment, a self-report survey administered in 1998 to students in the public school system in New Haven, CT (149 classes at 17 middle and high schools). RESULTS: Of 1,413 respondents (57% black, 28% Hispanic; mean age 13.4 +/- 1.7 years), 414 (29%) acknowledged prior sexual intercourse; the proportions of sexually active girls in 6th, 8th, and 10th grades were 14%, 30%, and 50%, respectively. In multivariate analyses of covariance, sexual activity was significantly associated with all four measures of psychosocial adaptation (p < .001). Other correlates of at least one measure of maladaptation included socioeconomic status, sensation seeking, and lower school grade (p < .001 for each), peer pressure (p < .01), and black ethnicity, and the interaction of sexual activity by lower school grade (p < .05 for each). CONCLUSIONS: Compared with their sexually naive peers, sexually active adolescent girls had lower scores on global measures of psychosocial adaptation. These findings have clinical, policy, and research relevance to a vulnerable population at high risk of teenage pregnancy and sexually transmitted diseases.