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Poorly differentiated thyroid carcinoma (PDTC) and anaplastic thyroid carcinoma (ATC) are very rare tumors with extremely aggressive behavior. Their comprehensive genetic background is still unclear. Some of the main genetic changes of differentiated thyroid carcinomas, such as mutations in BRAF and RAS genes, as well as changes in CTNNB1, PIK3CA, TP53, AXIN1, PTEN or APC genes leading to the dedifferentiation of the tumors, are described.DNAs from fresh frozen thyroid tissues of 3 PDTCs and 5 ATCs were extracted. The next-generation sequencing (NGS) approach was used to target 94 genes involved in cancer. The samples were prepared using a TruSight Cancer panel and sequenced with a MiSeq sequencer. Analysis of variants was performed by the MiSeq Reporter and NextGENe software and stringent criteria for prioritization of the variants were used in the Illumina VariantStudio software.Using NGS, we identified 26 genetic changes in 18 genes, novel variants included.NGS is a useful tool for searching for new variants and genes involved in PDTC and ATC. It seems that each of these rare tumor types has its own specific genetic background. These data could be helpful for recognizing new genetic markers and targets for future personalized therapy.

Hirschsprung's disease (HD) can be associated with the development of neuroendocrine tumours such as medullary thyroid carcinoma (MTC). The RET proto-oncogene is the major gene responsible for both HD and MTC. Mutations in exon 10 (codons 609, 611, 618, 620) were found in patients with co-occurrence of HD and MTC. The aim of the study was to screen the MTC risk in patients with HD. The prospective and retrospective genetic analyses comprised 56 HD patients (41 males, 15 females, aged 0-47). The prospective subgroup of patients consisted of 34 patients (25 boys, 9 girls) operated on between June 2003 and December 2005. The retrospective subgroup comprised 22 patients (16 boys, 6 girls) of 194 patients who were operated on between December 1979 and May 2003, non-systematically chosen preferably for total colonic aganglionosis (TCA). DNAs were isolated from blood and resected segments of aganglionic bowel. The HD patients and nine available family members (2 HD) were tested for RET mutations in exons 10, 11, 13, 14, 15 and 16. Direct double-stranded fluorescent sequencing revealed typical germline heterozygous MTC risk RET mutations in 3/56 (5.4%) female HD patients: Cys609Tyr, Cys620Arg (both exon 10) and Tyr791Phe (exon 13). Two of these patients had TCA and one patient had classical type of HD. One TCA patient developed clinical stage of MTC and underwent total thyroidectomy (TTE). The other two RET positive HD patients (aged 7 and 25 years) are screened for calcitonin level and they are without TTE till now. Two family members (mothers of TCA patients) with detected RET mutation underwent prophylactic TTE with MTC finding. Results showed the benefit of systematic RET mutation screening in HD patients in order to identify the risk of MTC in preclinical stage of the disease in patients with HD and their family members. We recommend to investigate not only exon 10 but also exon 13.

UNLABELLED The peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor subfamily of transcription factors. PPARgamma2 plays a key role in regulation of adipocyte differentiation and energy homeostasis. Recent studies provide evidence that the Pro12Ala polymorphism is linked to obesity and type 2 diabetes mellitus, but the results are controversial and depend on ethnicity. The aim of this study was to determine allele frequencies and to study the influence of the polymorphism on biochemical and anthropometric parameters in a Czech healthy adult population, in type 2 diabetics, and in a group of obese women. RESULTS The frequency of the Pro12Ala PPARgamma2 gene polymorphism in Czech probands is similar to other central European populations. Frequency of the Pro12Ala substitution tends to be higher in obese women and diabetics compared with controls. The fasting insulin levels in the 12Ala carriers were significantly lower within the group of diabetics even after adjustment for age, BMI, and the length of diabetes duration. In obese women, higher WHR was found in subjects with the 12Ala allele. CONCLUSIONS This study indicates that the substitution Pro12Ala is not associated with a decreased obesity or diabetes risk in the Czech population. However, the present data show that fasting insulin concentrations are lower in diabetics with the 12Ala allele than in those without it. This finding provides evidence that the polymorphism may influence glucose homeostasis.

Background and Aim: Adiponectin is regarded as a possible link between adiposity and insulin resistance. Ghrelin and leptin are considered as signals of energy status. We evaluated the relationships between these peptides, androgens and insulin sensitivity in women affected by polycystic ovary syndrome. Methods: Thirty-six women with PCOS were examined with euglycemic hyperinsulinemic clamp (to determine M/I, index of insulin sensitivity). Leptin, ghrelin, adiponectin, androgens, and SHBG were determined. Statistics was done using correlation analysis and backward stepwise multiple regression. Results: The positive correlation of adiponectin with testosterone remains significant even after adjustment for BMI (p = 0.01), M/I (p = 0.009) and for both M/I and BMI (p = 0.02). In multiple regression with testosterone, M/I, leptin and ghrelin as independent variables, the model including testosterone (p = 0.03) and ghrelin (p = 0.002) explained 49% of the variability (p < 0.0012) of adiponectin. Conclusions: Both adiponectin and ghrelin can be involved in the pathophysiology of PCOS but their relation must be delineated further.

et al., [Purpose]: To investigate the prognostic value of BRAF V600E mutation for the recurrence of papillary thyroid cancer (PTC). [Patients and Methods]: This was a retrospective multicenter study of the relationship between BRAF V600E mutation and recurrence of PTC in 2,099 patients (1,615 women and 484 men), with a median age of 45 years (interquartile range [IQR], 34 to 58 years) and a median follow-up time of 36 months (IQR, 14 to 75 months). [Results]: The overall BRAF V600E mutation prevalence was 48.5% (1,017 of 2,099). PTC recurrence occurred in 20.9% (213 of 1,017) of BRAF V600E mutation–positive and 11.6% (125 of 1,082) of BRAF V600E mutation–negative patients. Recurrence rates were 47.71 (95% CI, 41.72 to 54.57) versus 26.03 (95% CI, 21.85 to 31.02) per 1,000 person-years in BRAF mutation–positive versus –negative patients (P < .001), with a hazard ratio (HR) of 1.82 (95% CI, 1.46 to 2.28), which remained significant in a multivariable model adjusting for patient sex and age at diagnosis, medical center, and various conventional pathologic factors. Significant association between BRAF mutation and PTC recurrence was also found in patients with conventionally low-risk disease stage I or II and micro-PTC and within various subtypes of PTC. For example, in BRAF mutation–positive versus –negative follicular-variant PTC, recurrence occurred in 21.3% (19 of 89) and 7.0% (24 of 342) of patients, respectively, with recurrence rates of 53.84 (95% CI, 34.34 to 84.40) versus 19.47 (95% CI, 13.05 to 29.04) per 1,000 person-years (P < .001) and an HR of 3.20 (95% CI, 1.46 to 7.02) after adjustment for clinicopathologic factors. BRAF mutation was associated with poorer recurrence-free probability in Kaplan-Meier survival analyses in various clinicopathologic categories. [Conclusion]: This large multicenter study demonstrates an independent prognostic value of BRAF V600E mutation for PTC recurrence in various clinicopathologic categories., Supported by National Institutes of Health (NIH) Grants No. R01CA134225 and RO1CA113507 (M.X.); by Grant No. UL1 RR 025005 from the National Center for Advancing Translational Sciences of NIH and NIH Roadmap for Medical Research (K.A.C.); and by the following funding to individual study centers: National Science Centre Poland Grants No. N N403 194340 (A.C.) and N N401 612440 (B.J.); grants from Griffith Health Institute (Australia; A.K.L.); Grants No. BFU2010-16025, RD06/0020/0060-RD12/0036/0030 FIS, ISCIII, and S2011/BMD-2328 TIRONET (Spain; P.S.); NIH Grant No. RO1-CA50706 and the Byrne Foundation (J.A.F.); Grant No. IG 9338 from the Fondazione Cassa di Risparmio di Perugia and Associazione Italiana per la Ricerca sul Cancro (Italy) and the Beadle Family Foundation (San Antonio, TX; E.P.); Grant No. IGA MH CR NT 13901- 4 (Czech Republic; B.B., V.S.); grants from the New South Wales Cancer Institute (C.J.O.) and Cancer Council of New South Wales (Australia; R.C.- B.); Grant No. MIUR 20074zw8la from the Ministero della Istruzione Universitaria e Ricerca Scientifica and the Associazione Italiana per la Ricerca sul Cancro (Italy; G.T.); NIH/National Institute on Aging Grant No. 5R03AG042334-02 (L.Y.); grants from the Ministero della Istruzione Universitaria e Ricerca Scientifica, the Associazione Italiana per la Ricerca sul Cancro, the Istituto Toscano Tumori, and the Ministero della Salute (Italy; D.V., R.E.); and Grant No. CB-2011-03-02 from the Korean Foundation for Cancer Research (South Korea; Y.K.S., T.Y.K.).

OBJECTIVE—To evaluate insulin sensitivity (IS) and β-cell function (βF) in lean and obese women with polycystic ovary syndrome (PCOS), either separately or by using a disposition index (DI). RESEARCH DESIGN AND METHODS—A total of 64 women with PCOS and 20 healthy women were examined by anthropometry, oral glucose tolerance tests (OGTTs), and insulin tolerance tests. Statistical analysis used one-way ANOVA, Kruskal-Wallis, and Mann-Whitney U tests, as appropriate. RESULTS—A significantly higher waist-to-hip ratio (P < 0.0001) was found in both lean and obese women with PCOS. Higher basal blood glucose (P < 0.004) and blood glucose values at 3 h of OGTT (P < 0.008) were found in lean and obese PCOS subjects in comparison with control subjects. Insulin resistance by homeostasis model assessment (P < 0.007) was significantly higher in obese PCOS than in control or lean PCOS subjects. Early-phase insulin secretion (insulinogenic index [ΔI/ΔG30–0, where I is insulin and G is glucose]; P < 0.0007) was significantly higher in both lean and obese PCOS subjects than in healthy women. All tested combinations of parameters of IS and βF (DIs) followed a physiological hyperbolic relationship. Significantly lower values of the fasting state–derived DIs were found (all P < 0.05) in obese PCOS subjects. Significantly higher values of all of these indexes derived from nonfasting values were found in lean PCOS as compared with control and obese PCOS subjects (all P < 10–3). CONCLUSIONS—Increased βF was found even in lean individuals with PCOS. Insulin hypersecretion is thus probably connected to the pathogenesis of PCOS.

The geneMTNR1Bencodes a receptor for melatonin. Melatonin receptors are expressed in humanβ-cells, which implies that genetic variants might affect glucose tolerance. Meta-analysis confirmed that the rs10830963 shows the most robust association. The aim of the study was to assess the rs10830963 in Czech GDM patients and controls and to study relations between the SNP and biochemical as well as anthropometric characteristics. Our cohort consisted of 880 women; 458 were diagnosed with GDM, and 422 were normoglycemic controls without history of GDM. Despite similar BMI, the GDM group showed higher WHR, waist circumference, abdominal circumference, and total body fat content. The risk allele G was more frequent in the GDM group (38.3 versus 29.4% in controls, OR 1.49 CI95% [1.22; 1.82];POR=0.0001). In spite of higher frequency, the G allele in the GDM group was not associated with any markers of glucose metabolism. In contrast, controls showed significant association of the allele G with FPG and with postchallenge glycemia during the oGTT. Frequency analysis indicates that rs10830963 is involved in gestational diabetes in Czech women. However, the association of the SNP with glucose metabolism, which is obvious in controls, is covert in women who have experienced GDM.

 Statistical evaluation NCSS 2004 programme (Statistical Solutions, Saugus, MA, USA) and Chi-square test Introduction: Thyroid carcinomas are the most often endocrine malignancy and their incidence is still growing. But the genetic predisposition to the development of thyroid cancer is still unknown. Beside mutations in the specific oncogenes, the genetic predisposition to the thyroid cancer could be influenced by risk variants in tumor suppressor genes encoding key proteins in regulation of cell cycle and cell surviving.

Hirschsprung disease (HSCR) is a congenital aganglionosis of myenteric and submucosal plexuses in variable length of the intestine. This study investigated the influence and a possible modifying function of RET proto-oncogene's single nucleotide polymorphisms (SNPs) and haplotypes in the development and phenotype of the disease in Czech patients. Genotyping of 14 SNPs was performed using TaqMan Genotyping Assays and direct sequencing. The frequencies of SNPs and generated haplotypes were statistically evaluated using chi-square test and the association with the risk of HSCR was estimated by odds ratio. SNP analysis revealed significant differences in frequencies of 11 polymorphic RET variants between 162 HSCR patients and 205 unaffected controls. Particularly variant alleles of rs1864410, rs2435357, rs2506004 (intron 1), rs1800858 (exon 2), rs1800861 (exon 13), and rs2565200 (intron 19) were strongly associated with increased risk of HSCR (p

The present paper informs about allele frequencies in three HLA locuses (HLA-DRB3, HLA-DQAl and HLA-DQH1). The Czech population results are com-pared with data of different european and South Ameri...

Laparoscopic greater curvature plication (LGCP) is an emerging bariatric procedure that reduces the gastric volume without implantable devices or gastrectomy. The aim of this study was to explore changes in glucose homeostasis, postprandial triglyceridemia, and meal-stimulated secretion of selected gut hormones [glucose-dependent insulinotropic polypeptide (GIP), glucagon-like peptide-1 (GLP-1), ghrelin, and obestatin] in patients with type 2 diabetes mellitus (T2DM) at 1 and 6 months after the procedure.Thirteen morbidly obese T2DM women (mean age, 53.2 ± 8.76 years; body mass index, 40.1 ± 4.59 kg/m2) were prospectively investigated before the LGCP and at 1- and 6-month follow-up. At these time points, all study patients underwent a standardized liquid mixed-meal test, and blood was sampled for assessment of plasma levels of glucose, insulin, C-peptide, triglycerides, GIP, GLP-1, ghrelin, and obestatin.All patients had significant weight loss both at 1 and 6 months after the LGCP (p ≤ 0.002), with mean percent excess weight loss (%EWL) reaching 29.7 ± 2.9% at the 6-month follow-up. Fasting hyperglycemia and hyperinsulinemia improved significantly at 6 months after the LGCP (p 0.05), with parallel improvement in insulin sensitivity and HbA1c levels (p 0.0001). Meal-induced glucose plasma levels were significantly lower at 6 months after the LGCP (p 0.0001), and postprandial triglyceridemia was also ameliorated at the 6-month follow-up (p 0.001). Postprandial GIP plasma levels were significantly increased both at 1 and 6 months after the LGCP (p 0.0001), whereas the overall meal-induced GLP-1 response was not significantly changed after the procedure (p 0.05). Postprandial ghrelin plasma levels decreased at 1 and 6 months after the LGCP (p 0.0001) with no significant changes in circulating obestatin levels.During the initial 6-month postoperative period, LGCP induces significant weight loss and improves the metabolic profile of morbidly obese T2DM patients, while it also decreases circulating postprandial ghrelin levels and increases the meal-induced GIP response.

Background: Sex hormone-binding globulin (SHBG) belongs to the factors contributing to the pathophysiology of type 2 diabetes mellitus (T2DM). We determined genotypic frequencies of the single nucleotide polymorphisms (SNPs) rs6259 and rs6257 in T2DM patients, offspring of T2DM patients, gestational diabetics, patients suffering from polycystic ovary syndrome (PCOS), and in healthy adult Czechs. 1687 volunteers entered the study. The aim was to compare genetic constellation between the groups and to study the possible association of the SNPs with biochemical and anthropometric markers of insulin sensitivity. Methods: TaqMan (LC480, Roche) was used for genotyping, statistical evaluation was carried out using Statgraphics Centurion version XVI and NCSS 2007. Results: The SNPs distribution was similar between the groups. We found lower SHBG concentrations in diabetics and PCOS patients. The rs6259 SNP was associated with SHBG levels: in the NN carriers, the concentration was significantly higher in comparison with DD and DN. Unexpected results were observed when association of the rs6259 SNP with insulin sensitivity was assessed. In spite of higher SHBG concentration, which is considered to be protective factor, the NN homozygotes exhibited systematically higher stimulated glucose levels during the 3-hour oral glucose tolerance test and lower Cederholm index of insulin sensitivity. Conclusions: Genetic analysis confirmed the association between rs6259 NN genotype and higher SHBG levels. Furthermore, the NN genotype showed higher stimulated glycemia and lower insulin sensitivity. This observation seems intriguing considering established protective effect of higher SHBG levels in relation to T2DM and should be verified on a larger group of probands.

PMID:23571588.-- et al., [Importance]: BRAF V600E is a prominent oncogene in papillary thyroid cancer (PTC), but its role in PTC-related patient mortality has not been established. [Objective]: To investigate the relationship between BRAF V600E mutation and PTC-related mortality. [Design, Setting, and Participants]: Retrospective study of 1849 patients (1411 women and 438 men) with amedian age of 46 years (interquartile range, 34-58 years) and an overall median follow-up time of 33 months (interquartile range, 13-67 months) after initial treatment at 13 centers in 7 countries between 1978 and 2011. [Main Outcomes and Measures]: Patient deaths specifically caused by PTC. [Results]: Overall, mortality was 5.3% (45/845; 95% CI, 3.9%-7.1%) vs 1.1% (11/1004; 95% CI, 0.5%-2.0%) ( P < .001) in BRAF V600E-positive vs mutationnegative patients. Deaths per 1000 person-years in the analysis of all PTC were 12.87 (95% CI, 9.61-17.24) vs 2.52 (95% CI, 1.40-4.55) in BRAF V600E-positive vs mutation-negative patients; the hazard ratio (HR) was 2.66 (95% CI, 1.30-5.43) after adjustment for age at diagnosis, sex, and medical center. Deaths per 1000 person-years in the analysis of the conventional variant of PTC were 11.80 (95% CI, 8.39-16.60) vs 2.25 (95% CI, 1.01-5.00) in BRAF V600E-positive vs mutation-negative patients; the adjusted HR was 3.53 (95% CI, 1.25-9.98). When lymph node metastasis, extrathyroidal invasion, and distant metastasis were also included in the model, the association of BRAF V600E with mortality for all PTC was no longer significant (HR, 1.21; 95% CI, 0.53-2.76). A higher BRAF V600E-associated patient mortality was also observed in several clinicopathological subcategories, but statistical significance was lost with adjustment for patient age, sex, and medical center. For example, in patients with lymph node metastasis, the deaths per 1000 person-years were 26.26 (95% CI, 19.18-35.94) vs 5.93 (95% CI, 2.96-11.86) in BRAF V600E-positive vs mutation-negative patients (unadjusted HR, 4.43 [95% CI, 2.06-9.51]; adjusted HR, 1.46 [95% CI, 0.62-3.47]). In patients with distant tumor metastasis, deaths per 1000 person-years were 87.72 (95% CI, 62.68-122.77) vs 32.28 (95% CI, 16.14-64.55) in BRAF V600E-positive vs mutation-negative patients (unadjusted HR, 2.63 [95% CI, 1.21-5.72]; adjusted HR, 0.84 [95% CI, 0.27-2.62]). Conclusions and Relevance: In this retrospective multicenter study, the presence of the BRAF V600E mutation was significantly associated with increased cancer-related mortality among patients with PTC. Because overall mortality in PTC is low and the association was not independent of tumor features, how to use BRAF V600E to manage mortality risk in patients with PTC is unclear. These findings support further investigation of the prognostic and therapeutic implications of BRAF V600E status in PTC. ©2013 American Medical Association. All rights reserved., Funding/Support: This project was supported by National Institutes of Health (NIH) grants R01CA134225 and R01CA113507 to Dr Xing. The statistical effort of Ms Carson for this project was supported by grant UL1RR025005 from the National Center for Advancing Translational Sciences and the NIH Roadmap for Medical Research. In addition, the studies at individual centers were supported as follows: the Ministry of Science and Higher Education Research grants N N403 194340 and N N401 612440 to Drs Czarniecka and Jarzab, respectively(Poland); grant NIDCR/NCI SPORE P50DE019032 to Dr Sindransky (United States); grants BFU2010–16025, RD06/0020/0060-RD12/0036/0030 FIS, ISCIII, and S2011/BMD-2328TIRONET to Dr Santisteban (Spain); grant NIH-R01-CA50706 and Byrne Foundation funding to Dr Fagin (United States); grants from Fondazione Cassa di Risparmio di Perugia and Associazione Italiana per la Ricerca sul Cancro (IG 9338) (Italy) and the Beadle Family Foundation (San Antonio, Texas) to Dr Puxeddu; grant IGA MHCRNT13901–4 to Drs Sykorova and Bendlova (Czech Republic); and grants from the New South Wales Cancer Institute to Dr O’Neill and from the Cancer Council of New South Wales to Dr Clifton-Bligh (Australia).

The goal of the study was to examine whether resting or post-exercise metabolic substrate levels are associated with differential exercise performance and long-term outcome in control subjects or heart failure (HF) patients with or without type 2 diabetes mellitus (DM). Methods and results Twenty five healthy controls matched with 97 patients with stable advanced HF were prospectively enrolled. Exercise capacity, age, gender, and HF aetiology were balanced between HFDM- and HFDM+ groups. Subjects underwent maximal bicycle spiroergometry with blood sampling to measure metabolites and neurohormones before and imme- diately after the exercise. HFDM+ patients had increased free fatty acids, glucose, and b-hydroxybutyrate compared with controls. HFDM+ patients had higher baseline copeptin (24+ 16 vs. 17+ 13 pmol/L, P , 0.05) but otherwise showed similar neurohumoral activation and exercise response to HFDM- patients. Peak oxygen consumption (VO2) was unrelated to post-exercise free fatty acids, glucose, lactate, or glycerol, but strongly correlated with post-exercise pyruvate (in all: r ¼ 0.62, P , 0.001). During the next 17+ 10 months, 36% of HF patients experienced an adverse event (death, urgent transplantation, or assist device insertion). From metabolic factors, only post-exercise glucose (hazard ratio (HR) 1.28, P ¼ 0.04), total body fat (HR 0.58, P , 0.001), and the presence of DM (HR 1.98, P ¼ 0.04) were predictive of the outcome. Conclusions With the exception of pyruvate, acute changes of metabolic substrates are not related to cardiac performance in HF, regardless of diabetic status. Inhibition of body fat depletion, attenuation of stress-related hyperglycaemia, or increas- ing dynamics of plasma pyruvate may represent therapeutic targets in advanced HF.

BACKGROUND Reliable and effective primary screening of mutation carriers is the key condition for common diagnostic use. The objective of this study is to validate the method high resolution melting (HRM) analysis for routine primary mutation screening and accomplish its optimization, evaluation and validation. Due to their heterozygous nature, germline point mutations of c-RET proto-oncogene, associated to multiple endocrine neoplasia type 2 (MEN2), are suitable for HRM analysis. Early identification of mutation carriers has a major impact on patients' survival due to early onset of medullary thyroid carcinoma (MTC) and resistance to conventional therapy. METHODS The authors performed a series of validation assays according to International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines for validation of analytical procedures, along with appropriate design and optimization experiments. After validated evaluation of HRM, the method was utilized for primary screening of 28 pathogenic c-RET mutations distributed among nine exons of c-RET gene. RESULTS Validation experiments confirm the repeatability, robustness, accuracy and reproducibility of HRM. All c-RET gene pathogenic variants were detected with no occurrence of false-positive/false-negative results. CONCLUSIONS The data provide basic information about design, establishment and validation of HRM for primary screening of genetic variants in order to distinguish heterozygous point mutation carriers among the wild-type sequence carriers. HRM analysis is a powerful and reliable tool for rapid and cost-effective primary screening, e.g., of c-RET gene germline and/or sporadic mutations and can be used as a first line potential diagnostic tool.

Background: To compare the prevalence of impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) in lean and overweight/obese women with polycystic ovary syndrome (PCOS), with the data from a normal population sample. Methods: PCOS-affected women fulfilling ESHRE diagnostic criteria underwent an oral glucose tolerance test. Prevalence of IGT and T2DM in control sample of white healthy females was extracted from the published data from NHANES II. Results: In 225 women with PCOS, IGT was present in 6/104 (5.8%) lean and in 15/121 (12.4%) overweight/obese women. T2DM was present in 1/104 (1.0%) lean and in 3/121 (2.5%) overweight/obese PCOS women. In a sample of 643 women from NHANES II, the crude rate of IGT was 5.9%. IGT was significantly more common only in the overweight/obese PCOS subgroup as compared to the NHANES II cohort (χ2 = 5.99, p < 0.01). Conclusions: IGT was found significantly more frequently only in overweight/obese PCOS subjects in comparison with healthy controls.

Objectives: Two cases of an extremely rare paraganglioma-like variant of medullary thyroid carcinoma (MTC) are reported. Methods: The patients were a 65-year-old male (case 1) and a 14-year-old female (case 2). Unilateral thyroid nodule and homolateral cervical lymphadenopathy was present in case 1; bilateral thyroid nodules were seen in case 2. Fine needle aspiration cytology (FNAC) was performed from thyroid nodules (in both cases) and from a cervical lymph node (in case 1). Results: The cytological smears contained predominantly ovoid to spindled epithelial cells arranged in cohesive three-dimensional clusters with sharp margins; isolated individual cells were seen only rarely. No colloid or other material was present in the background. The tumour cells showed significant nuclear atypia with occasional bizarre and/or binucleated cells. The nuclear chromatin was coarse and granular, sometimes with grooves and intranuclear inclusions. The cytoplasm was inconspicuous. Polygonal or triangular cells, amyloid and azurophillic cytoplasmic granules were absent in both cases. Calcitonin expression was demonstrated in case 2. Histological examination confirmed the paraganglioma variant of MTC in both cases. Mutation of RET proto-oncogene in exon 16 (Met918Thr) – germline in case 2 and somatic in case 1 was detected by sequencing of DNA in both cases. Conclusions: This is the first description of cytological findings in the paraganglioma-like variant of MTC. Despite its rarity, it can be reliably diagnosed by FNAC if material for immunocytochemistry is obtained.

ObjectivePolycystic ovary syndrome (PCOS) has been linked to a high risk of type 2 diabetes mellitus. Disturbances in the secretion of the incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) have been observed in states with impaired glucose regulation. This paper considers the secretion of GIP and GLP-1 after oral glucose load in a group of lean, glucose-tolerant PCOS women in comparison with age- and body mass index (BMI)-matched healthy women.DesignCase control.MethodsPCOS (n=21, 25.8±4.1 years, BMI 21.6±1.7 kg/m2) and control healthy women (CT, n=13, 28.5±7.2 years, BMI 20.3±2.5 kg/m2) underwent oral glucose tolerance test (OGTT) with blood sampling for glucose, insulin, C-peptide, total GIP, and active GLP-1. Insulin sensitivity was determined both at fasting and during the test.StatisticsRepeated measures ANOVA.ResultsGlucose levels and insulin sensitivity did not differ between PCOS and CT. PCOS had significantly higher levels of C-peptide (PPPPConclusionsIncreased total GIP and lower late phase active GLP-1 concentrations during OGTT characterize PCOS women with higher C-peptide secretion in comparison with healthy controls, and may be the early markers of a pre-diabetic state.

Mixed medullary-follicular carcinoma of the thyroid with a pleomorphic pattern is a rare malignant epithelial tumor characterized by clinical and immunohistochemical features of follicular and parafollicular thyroid cells. Different molecular mechanisms for mixed thyroid tumors have been suggested.We describe a 71-year-old man with a history of malignant melanoma with mixed medullary-follicular thyroid carcinoma. Cytology results of a fine needle aspiration biopsy were suspicious of a thyroid carcinoma. The patient underwent a total thyroidectomy for a solitary thyroid nodule in the right lobe. No lymph node metastases were present. Histopathology and immunohistochemistry revealed a mixed medullary and follicular cell carcinoma that showed characteristic patterns and calcitonin and thyroglobulin positivities in many of the tumor cells. The tumor was not associated with multiple endocrine neoplasia type 2. Detection of RET proto-oncogene point mutations in risk exons 10, 11, 13, 14, 15, and 16 was negative. Two polymorphisms, one in exon 11 G691S (GGT--AGT) and another in exon 15 S904S (TCC--TCG) were detected.A mixed differentiated thyroid tumor is a diagnostic challenge with fine needle aspiration. Definitive diagnosis remains the domain of histology because of the necessity of topographic information. The origins of this rare tumor entity are unclear. The possible association with the uncommon polymorphism G691S of the RET proto-oncogene is discussed.

BACKGROUND Diabetes mellitus type 2 (DM2) affects 10% of women with polycystic ovary syndrome (PCOS). We evaluated the sensitivity and specificity of clinical and fasting biochemical parameters in screening for impaired glucose tolerance (IGT) and DM2. METHODS Women with PCOS [n=244, age 27.4+/-7.5 years, body mass index (BMI) 27.5+/-6.9 kg/m(2)] and healthy women (n=57, age 26.8+/-5.8 years, BMI 21.3+/-2.1 kg/m(2)) underwent basal blood sampling and an oral glucose tolerance test (oGTT). RESULTS Insulin resistance was identified in 40.2% of PCOS women. Impaired fasting glucose (5.6-6.9 mmol/L) was found in 30 subjects (12.3%), but the oGTT revealed IGT in only six of these cases and DM2 in one subject. IGT was found in 23 (9.4%) and DM2 in four (1.6%) of the women with PCOS. The conventional upper limits for total cholesterol, triglycerides, systolic and diastolic blood pressure and fasting glucose revealed low sensitivity for the identification of impaired glucose metabolism. CONCLUSIONS No single parameter nor any combination of them showed an accuracy sufficient for screening of IGT or DM2 in PCOS patients. All PCOS patients should be screened using an oGTT to identify disturbances in glucose metabolism.

Medullary thyroid carcinoma (MTC) occurs mostly as a sporadic tumor or in connection with inherited cancer syndromes-multiple endocrine neoplasia (MEN) types 2A and 2B and familial MTC. Germline mutations in the RET proto-oncogene are found in most of the familial cases. Somatic mutations in the RET proto-oncogene are detected in 23%-69% of patients with sporadic MTC. The most frequent somatic mutation is Met918Thr in exon 16 and only a small percentage of mutations in other RET exons have been observed. In a very few cases double mutations were found. Genetic screening for somatic mutations in RET exons 10, 11, 13, 14, 15, and 16 in Czech patients with sporadic MTC was carried out by DNA sequencing. This study presents a new triplesomatic mutation Gly911Asp, Met918Thr, and Glu921Lys in exon 16 of the RET proto-oncogene detected in an 18-year-old Czech male patient. In the second case, a new double-somatic mutation Val591Ile in exon 10 with a concomitant somatic mutation Met918Thr in exon 16 was found in a 77-year-old Czech female patient. These both newly described somatic multiple mutations were revealed in a hemizygous status, the loss of heterozygosity in tumor tissues in comparison with germline DNA was confirmed.

We conducted pooled and meta-analyses of the association of the calpain-10 gene (CAPN10) polymorphisms SNP-43, Indel-19 and SNP63 individually and as haplotypes with type 2 diabetes (T2D) in 3237 patients and 2935 controls of European ancestry. In the pooled analyses, the common SNP-43 ¤ G allele was associated with modest but statistically signiWcant increased risk of T2D (odds ratio (OR) D 1.11 (95% conWdence interval (CI), 1.02–1.20), P D 0.01). Two haplotype combinations were associated with increased risk of T2D (1-2-1/1-2-1, OR D 1.20 (1.03–1.41), P D 0.02; and 1-1-2/1-2-1, OR D 1.26 (1.01–1.59), P D 0.04) and one with decreased risk (1-1-1/2-2-1, OR D 0.86 (0.75–0.99), P D 0.03). The meta-analysis also showed a signiWcant eVect of the 1-2-1/1-2-1 haplogenotype on risk (OR D 1.25 (1.05–1.50), P D 0.01). However, there was evidence for heterogeneity with respect to this eVect (P D 0.06). The heterogeneity appeared to be due to data sets in which the cases were selected from samples used in linkage studies of T2D. Using only the population-based case-control samples removed the heterogeneity (P D 0.89) and strengthened the evidence for association with T2D in both the pooled (SNP-43 ¤ G, OR D 1.19 (1.07–1.32), P D 0.001; 12-1/1-2-1 haplogenotype, OR D 1.46 (1.19–1.78), P D 0.0003; 1-1-2/1-2-1 haplogenotype, OR D 1.52 (1.12–2.06), P D 0.007; and 1-1-1/2-2-1 haplogenotype, OR D 0.83 (0.70–0.99), P D 0.03) and the meta-analysis (SNP-43 ¤ G, OR D 1.18 (1.05–1.32), P D 0.005; 1-2-1/1-2-1 haplogenotype, OR D 1.68 (1.33–2.11), P D 0.00001). The pooled and meta-analyses as well as the linkage disequilibrium and haplotype diversity studies suggest a role for genetic variation in CAPN10 aVecting risk of T2D in Europeans.

Variability in the number of tandem repeats of the insulin gene (INS VNTR) is known to influence several phenotypes, including polycystic ovary syndrome (PCOS), diabetes mellitus type 1, diabetes mellitus type 2, and birth weight. The presence of the class III allele of INS VNTR has been reported to be protective in diabetes mellitus type 1, but in contrary it increases the disease risk of PCOS and diabetes mellitus type 2. PCOS is a very common endocrinopathy in women of reproductive age. The etiology of PCOS is uncertain, but family history of this syndrome suggests a major genetic cause. The aim of this pilot study was to investigate the possible association of INS VNTR polymorphism with PCOS in Czech women. In PCOS, significantly higher WHR, BMI, G(0), G(180), I(30), Cp(0), Cp(30), Cp(60), AUC-I, AUC-Cp, and insulinogenic index and significantly lower AUC-G/AUC-I were found. No significant differences in INS VNTR genotype, phenotype, or allele frequencies were found between PCOS and controls. In spite of several differences in anthropometric and biochemical parameters (abdominal fat localization, increased beta-cell function, and lower insulin sensitivity in PCOS women), no effect of INS VNTR polymorphism was found on insulin secretion, insulin action, or any other screened parameter.