Fine needle aspiration (FNA) is the primary diagnostic tool for evaluating thyroid nodules. However, 10%-21% of FNAs yield non-diagnostic (ND) results (Bethesda category I). Repeated ND findings occur in 25%-67.5% of cases, creating diagnostic uncertainty and increased healthcare costs. Our aim was to evaluate whether cumulative analysis of repeated ND cytology samples enables reclassification into a diagnostic Bethesda category.

Molecular testing has emerged as a pivotal tool for the preoperative assessment of cytologically indeterminate thyroid nodules. In this cross-sectional study, we evaluated the diagnostic utility of a targeted next-generation sequencing (NGS) 4-gene panel, including BRAFV600E, TERT promoter mutations, RET fusions, and NTRK3 fusion, for enhancing the cytological diagnosis of thyroid nodules prior to surgical intervention. A total of 827 thyroid nodules subjected to fine-needle aspiration and subsequent histopathological confirmation were analyzed, among which 773 (93.5%) were classified as malignant or noninvasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP). The observed prevalence of molecular alterations was: BRAFV600E, 68.3% (526/770); TERT promoter mutations, 10.3% (79/770); RET fusions, 10.3% (79/770); and NTRK3 fusion, 3.9% (30/770). Notably, the 4-gene NGS panel demonstrated brilliant diagnostic performance for indeterminate cytological nodules (Bethesda categories III-V), achieving a sensitivity of 87.9%, specificity of 96.3%, positive predictive value (PPV) of 99.7%, negative predictive value (NPV) of 35.9%, and overall accuracy of 88.2%. These findings indicate that the targeted NGS 4-gene panel provides high diagnostic precision in distinguishing benign from malignant nodules. Its implementation offers a cost-effective, efficient molecular diagnostic strategy that may reduce unnecessary diagnostic procedures and facilitate optimized clinical management.

The global rise in thyroid cancer incidence has intensified the demand for reliable, rapid and minimally invasive diagnostic tools as current clinical practice often result in unnecessary surgical procedures for benign thyroid nodules. In this context, this work proposes a highly sensitive label-free biosensor, based on a heterojunction gate-all-around ferroelectric p-n-i-n tunnel field effect transistor (HJ-GAA-Fe p-n-i-n TFET) for early thyroid cancer detection. The sensing mechanism leverages variations and the dielectric properties and charge states of thyroid cells, which modulates electrostatic environment of the device. A ferroelectric gate stack introduces a negative capacitance effect that amplifies biomolecule induced electrical perturbations, while a nanocavity beneath the gate metal provides a dedicated region for thyroid cell immobilization. TCAD based simulations are used to evaluate clinically relevant sensitivity metrics including drain current, ON-OFF current ratio and threshold voltage. Furthermore, the proposed biosensor's reliability is assessed through response time, limit of detection (LOD), temperature stability, fill factor variation, early-stage detection capability, steric hindrance effect and noise immunity. The influence of both charged and neutral thyroid cancer cells is systematically analyzed. The proposed biosensor achieved a drain current sensitivity of 5.2 x 10[8], an ON-OFF ratio sensitivity of 3.23 x 10[6], a threshold voltage shift of 0.25 V, and selectivity of 12.41 of cancerous thyroid cells, indicating strong potential for accurate and early thyroid cancer diagnostic purpose.

Indeterminate thyroid nodules carry a wide range of malignancy prevalence, necessitating incorporation of molecular testing to guide management. Molecular test results help guide prognosis to a certain extent, but additional prognostic factors need to be identified. Limited data have linked PLEKHS1 promoter (PLEKHS1p) mutations with higher aggressiveness of thyroid cancer. Hence, we aimed to evaluate the diagnostic and prognostic value of PLEKHS1p mutations in preoperative thyroid nodules undergoing molecular testing. We assessed PLEKHS1p mutations in 9279 patient samples from April 2023 to June 2024 among indeterminate thyroid nodules ((B)ethesda III/IV) with Afirma GSC-(S)uspicious results as well as among B V/VI cytology thyroid nodules. We also analyzed a subset of 20 consecutive cases positive for PLEKHS1p mutations with surgical resection for histology and for co-occurring molecular alterations from the Afirma testing. PLEKHS1p mutations were positive in 60/9279 (0.6%) of patient samples with three times higher frequency in B VI compared to B III cytology nodules (1.36% vs 0.47%, p < .01). Among the 60 samples harboring PLEKHS1p mutations, 28 had the C593T hotspot mutation, 38 the G590A mutation, and six samples had both; four samples had concomitant TERTp mutations and 18 samples had concomitant BRAFp.V600E alterations. Our study demonstrated an overall low frequency of PLEKHS1p mutations, but this frequency was highest among malignant (B VI) cytology thyroid nodules. The frequency of PLEKHS1p mutations did not strongly correlate with the severity of thyroid cancer but the surgical sample size was limited. Further research is needed to clarify the role of PLEKHS1p mutations in thyroid nodules and cancer.

A relatively uncommon occurrence of multiple unusual distant metastasis in a patient with thyroglobulin-elevated negative iodine scintigraphy (TENIS) (subcutis, skeletal muscle, kidney, and pericardium) is presented. A 65-year-old man with papillary thyroid carcinoma post two cycles of radioactive iodine therapy on subsequent follow-up showed elevated serum thyroglobulin (>300 ng/mL) and negative iodine scintigraphy (TENIS). [18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) for disease restaging revealed extensive metastatic disease involving cervical and mediastinal nodes, bones, liver, subcutis, skeletal muscle, kidney, and pericardium. [68Ga]-fibroblast activation protein inhibitor (FAPI)-04 PET/CT was done to evaluate for fibroblast activation protein (FAP) expression, and the feasibility of targeted radionuclide therapy showed nil to very low FAP expression at the metastatic sites, except for the few cervical and mediastinal nodes, subcutaneous nodule in the left arm, and L3 vertebral lesion. In addition to illustrate the rare presentation in TENIS, this case demonstrates the superiority of [18F]-FDG PET/CT over FAPI PET/CT in this patient. However, FAPI-PET/CT may be used as a theranostic tool for assessing radionuclide therapy prospects in selected cases.

Thyroid cancer (TC) is the most common endocrine malignancy, and challenges persist in preoperative diagnosis of indeterminate nodules and postoperative monitoring when thyroglobulin (Tg) assays are compromised by interfering anti-Tg antibodies (Tg-Ab). Extracellular vesicles (EVs) carry molecular cargo reflective of cells of origin and are increasingly explored as biomarker sources. In this study, we investigated whether thyroid-derived EVs retain the expression of thyroid-specific thyrotropin-receptor (TSHR), a suitable target in immunoaffinity-based EV isolation, and explored the presence of Tg in EV cargo as potential surrogate for serum Tg. EVs from thyroid cell lines (Nthy-Ori 3-1, TPC-1, OCUT2) and plasma of patients with benign, malignant tumors and recurrent TC were isolated by differential ultracentrifugation and characterized via nanoparticle tracking and Dot and Western blot analyses. EVs derived from Nthy-Ori 3-1 and TPC-1 cell lines were positive for surface TSHR and vesicular Tg, but not OCUT2. All plasma-derived EVs were positive for TSHR and Tg, while their electrophoretic profiles from vesicles differed compared to tissue lysate. Tg was detectable in EVs isolated from recurrent TC samples, even in Tg-Ab positive cases. Together, these results support the use of TSHR for targeted EV isolation and point to vesicular Tg as a potential recurrence marker.

Nuclear atypia has been proposed as a useful criterion for subclassifying thyroid nodules with atypia of undetermined significance (AUS), with potential implications for malignancy risk. In our analysis of nodules initially classified as AUS and subsequently confirmed as malignant, clinical outcomes were generally favourable and did not differ according to the presence of nuclear atypia. These findings suggest that while nuclear atypia may have diagnostic relevance, its prognostic impact may be limited.

Introduction Thyroid cancer is the most common endocrine malignancy, and its diagnosis relies on thyroid ultrasound and fine needle aspiration (FNA). To ensure uniform and complete communication between radiologist and clinician, a standardized thyroid ultrasound reporting protocol was implemented in December 2019 across a regional thyroid network in the Netherlands. This study aims to quantitatively assess the adherence to the protocol and qualitatively assess the reflection of the stakeholders on this protocol in its current form. Methods A sample was collected of ultrasound reports (n = 100) from three hospitals three years before and after implementation, and analyzed focusing on the description of key points in the standardized report. Additionally, semi-structured interviews were conducted. The interview structure is based on the grounded-theory approach using the Capability, Opportunity, Motivation - Behavior (COM-B) model for instituting change. Results The quantitative analysis showed that Thyroid Imaging Reporting and Data System (TI-RADS) classification pre-implementation was described in 0.0% of reports, and only in 37.5% of reports post-implementation. Lymph nodes were described pre-implementation in 80.0% of reports and 95.0% of reports post-implementation. Qualitative analysis revealed that the primary issue with using the standardized ultrasound report revolves around time constraints. Collaboration among endocrinologists, surgeons, and radiologists was mentioned to be essential for developing a comprehensive protocol with widespread acceptance. Encouraging the involvement of new radiologists is crucial. To maintain protocol quality and utilization, it is recommended that there is a culture of open communication for feedback within the thyroid network. Conclusion The findings underscore the importance of continuous improvement efforts and clear communication channels to enhance reporting practices and ensure quality assurance for thyroid ultrasounds between, and within health care institutions. Clinical Relevance: Effective implementation of a standardized thyroid ultrasound reporting protocol within a regional network in the Netherlands enhances diagnostic accuracy and treatment planning for thyroid nodules and cancer, optimizing patient care across multiple healthcare institutions and maximizing healthcare resources.

Thyroglossal duct cyst carcinoma is an uncommon entity, and the optimal management strategy continues to be debated, particularly regarding the indication for total thyroidectomy in addition to the Sistrunk procedure. We report the case of a 38-year-old woman who presented with a painless midline neck mass. Imaging studies revealed a cystic suprahyoid lesion with a vascularized mural nodule containing calcifications, raising suspicion for malignancy. The patient underwent a Sistrunk procedure, and histopathological examination demonstrated a 1.5 cm intracystic papillary thyroid carcinoma with capsular invasion and negative surgical margins. Given these intermediate-risk features and abnormal scintigraphic findings, completion total thyroidectomy was performed; final pathology revealed no evidence of synchronous thyroid malignancy. The patient was referred for radioactive iodine therapy and remains under close follow-up without evidence of recurrence. This case highlights the importance of maintaining a high index of suspicion when mural nodules are detected within a thyroglossal duct cyst and underscores the need for individualized management based on histopathological risk factors and thyroid imaging findings.