Molecular screening for point mutations and rearrangements is feasible in air-dried FNAs. Although the impact of detecting point mutations and rearrangements in FNAs has most likely been overestimated in previous studies, molecular FNA analyses improve presurgical diagnostics. The detection of BRAF mutations in FNA may improve the choice of surgery and postsurgical treatment. Further data are necessary to elucidate the true impact of detecting RAS and PAX8/PPARG mutations in FNAs. The inclusion of additional rare somatic mutations and miRNA markers might further improve the impact of molecular FNA diagnostics.

Testing for a panel of somatic mutations has led to an improvement of sensitivity/specificity for indeterminate/follicular proliferation FNAB samples. Further methodological improvements, standardizations, and further molecular markers should soon lead to a broader application of molecular FNAB cytology for the differential diagnosis of thyroid nodules and to a substantial reduction of diagnostic surgeries.

Background: The diagnostic limitations of fine needle aspiration (FNA), like the indeterminate category, can be partially overcome by molecular analysis. As PAX8/PPARG and RET/PTC rearrangements have been detected in follicular thyroid carcinomas (FTCs) and papillary thyroid carcinomas (PTCs), their detection in FNA smears could improve the FNA diagnosis. To date, these rearrangements have never been analyzed in routine air-dried FNA smears, but only in frozen tissue, formalin-fixed paraffin-embedded (FFPE) tissue, and in fresh FNA material. Fixed routine air-dried FNA samples have hitherto been judged as generally not suitable for testing these rearrangements in a clinical setting. Therefore, the objective of the present study was to investigate the feasibility of extracting RNA from routine air-dried FNA smears for the detection of these rearrangements with real-time polymerase chain reaction (RT-PCR). Methods: A new method for RNA extraction from routine air-dried FNA smears was established, which allowed analysis for the presence of four variants of PAX8/PPARG and RET/PTC 1 and RET/PTC 3, which were analyzed in 106 routine FNA smears and the corresponding surgically obtained FFPE tissues using real-time quantitative PCR (RT-qPCR). To assess RNA quality, an intron-spanning PAX8 cDNA was amplified. Results: Acceptable RNA quality was obtained from 95% of the FNA samples and 92% of the FFPE samples. PAX8/PPARG was detected in 4 of 96 FFPEs and in 6 of 96 FNAs. PAX8/PPARG was present in 4 of 10 FTCs and in 3 of 42 follicular adenomas (FAs). Similarly, RET/PTC was found in 3 of 96 FFPEs and in 4 of 96 FNAs. Two of 21 PTC samples and 3 of 42 FA samples carried this rearrangement. Conclusion: These data are the first to show the feasibility of extracting RNA from routine air-dried FNA smears for the detection of PAX8/PPARG and RET/PTC rearrangements with RT-qPCR. These promising methodological advances, if confirmed in larger series of FNA and FFPE samples, may lead to the introduction of molecular analysis of routine air-dried FNA smears in everyday practice.

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Sharks are very sensitive to stress and prone to a high mortality rate after capture. Since approximately 50 million of sharks are caught as bycatch every year, and current recommendations to reduce the impact of commercial fishing strongly support immediate release, it is imperative to better understand post-release mortality caused by the stress of capture and handling. Blood samples allow the assessment of stress levels which are valuable tools to reduce mortality in commercial, recreational and scientific fishing, being essential for the improvement in those conservation measures. Biochemical analyses are widely used for sharks as stress indicators, with secondary plasma parameters (lactate, glucose and ions) being the most often employed assays. However, it is virtually impossible to determine baseline plasma parameters in free-ranging sharks, since blood withdrawal involves animal capture and restrain, which are stressful procedures. This study aims at analyzing secondary parameters of five healthy tiger sharks captured with circular hooks and handlines in Fernando de Noronha (Northeastern Brazil) and comparing them with secondary parameters of three dead tiger sharks caught off Recife (also Northeastern Brazil). The results showed that the analysis of some plasma constituents in dead animals may be an efficient tool to assess stress and lethality. However, traditional parameters such as glucose and calcium, need to be used with caution. The results also demonstrated the extreme importance of urea and phosphorus for assessing stress response and mortality in tiger sharks, both parameters frequently neglected and of utmost importance for shark's homeostasis.

No abstract

miR-31 and miR-130b may have an effect on tumorigenesis of CBTNs by regulating proliferation and apoptosis and the cell cycle through cyclin D1.

Key words• ▶ thyroid cancer • ▶ diagnostics • ▶ mutations • ▶ fi ne-needle aspiration • ▶ thyroid cytology respectively [ 4 ] . In FTCs, PAX8 / PPARG rearrangements and RAS mutations have been detected in 25-63 % and 40-53 %, respectively [ 4 ] . Therefore, molecular testing for somatic mutations has emerged as the most promising approach for molecular FNA diagnosis [ 5 -7 ] . It allows improved discrimination of the "follicular proliferation/indeterminate" and "suspicious" FNA categories, leading to reduced numbers of diagnostic thyroidectomies and false negative FNAs [ 8 ] . Hitherto a variety of molecular techniques have been used to detect these mutations. These comprise LightCycler PCR using hybridization probes, quantitative (q)PCR and post-PCR melting curve analysis, allele specifi c PCR, direct sequencing, restriction fragment polymorphism analysis, colorimetric assays, and nested PCR [ 9 -15 ] . In archived FNA smears a similar sensitivity in the Introduction ▼ Thyroid cancer comprises 94.5 % of all endocrine malignancies, and ranks 13 th in frequency of occurrence among all cancers, accounting for 2.6 % of all cases [ 1 ] . The majority of thyroid cancers are well-diff erentiated malignancies originating from thyroid follicular cells. The most frequent histotypes are papillary thyroid carcinomas (PTCs), followed by follicular thyroid carcinomas (FTCs). Currently the best method to select suspicious nodules for surgery is ultrasound-guided fi ne needle aspiration (FNA) cytology [ 2 , 3 ] . However, despite high specifi city and sensitivity, FNA cytology has some inherent limitations, which can partly be overcome by molecular analysis, since RET / PTC rearrangements and BRAF point mutations have been detected in Impact of Diff erent Methodologies on the Detection of Point Mutations in Routine Air-dried Fine Needle Aspiration (FNA) Smears