Clinical utility of the imunohistochemical co-expression of p53 and MDM2 in thyroid follicular lesions.

In order to investigate the possible correlation between p53 and MDM2 co-expression with clinicopathological features of differentiated thyroid cancer (DTC) and its use as diagnostic and/or prognostic markers, we used immunohistochemistry to evaluate 317 thyroid samples including 208 DTC and 94 benign nodules, in addition to 15 normal tissues. MDM2 and p53 expression were highly associated (r = 0.7161; p < 0.0001). The co-expression of p53-MDM2 was observed more frequently in malignant lesions (p < 0.0001) and helped characterize follicular patterned lesions distinguishing FVPTC from FA (p < 0.0001) and FVPTC from FTC (p < 0.0001). In addition, p53-MDM2 co-expression was associated with characteristics of less aggressiveness. It was more frequent in patients ≤45 years old (p = 0.0035), with unique tumors (p = 0.0095), tumors <2 cm (p < 0.0001), tumors without extrathyroid invasion (p = 0.0425), without metastasis at evolution (p = 0.0179), and in patients evolving free of disease after treatment (p = 0.0485). We suggest that p53-MDM2 co-expression profile analysis might help establishing diagnostic and determining prognostic of DTC patients.

Impact of image analysis and artificial intelligence in thyroid pathology, with particular reference to cytological aspects.

OBJECTIVE: Thyroid pathology has great potential for automated/artificial intelligence algorithm application as the incidence of thyroid nodules is increasing and the indeterminate interpretation rate of fine-needle aspiration remains relatively high. The aim of the study is to review the published literature on automated image analysis and artificial intelligence applications to thyroid pathology with whole-slide imaging. METHODS: Systematic search was carried out in electronic databases. Studies dealing with thyroid pathology and use of automated algorithms applied to whole-slide imaging were included. Quality of studies was assessed with a modified QUADAS-2 tool. RESULTS: Of 919 retrieved articles, 19 were included. The main themes addressed were the comparison of automated assessment of immunohistochemical staining with manual pathologist's assessment, quantification of differences in cellular and nuclear parameters among tumour entities, and discrimination between benign and malignant nodules. Correlation coefficients with manual assessment were higher than 0.76 and diagnostic performance of automated models was comparable with an expert pathologist diagnosis. Computational difficulties were related to the large size of whole-slide images. CONCLUSIONS: Overall, the results are promising and it is likely that, with the resolution of technical issues, the application of automated algorithms in thyroid pathology will increase and be adopted following suitable validation studies.

Prevalencia y malignidad asociada de las citologías de categoría Bethesda III de nódulos tiroideos según el grupo «atipia citológica» o «atipia arquitectónica» / Prevalence and associated malignancy of Bethesda category III cytologies of thyroid nodules assigned to the "cytological atypia" or "architectural atypia" groups

Objetivo: Conocer la prevalencia de citologías de categoría Bethesda III, su tasa de malignidad y analizar si existen diferencias en la segunda citología, tasa de malignidad, tipo de carcinoma y estadio TNM entre los grupos atipia citológica (AC) y atipia arquitectónica (AA). Pacientes y método: Estudio retrospectivo de 973 citologías. Las de categoría Bethesda III se agruparon en AC cuando se apreció atipia nuclear pero no diagnóstica o sospechosa de malignidad, y en AA cuando se trató de extensiones con poca celularidad, pero con un patrón predominantemente microfolicular y con coloide mínimo o ausente. Se correlacionaron los resultados citológicos y anatomopatológicos. Resultados: Hallamos 87 (8,9%) citologías de categoría Bethesda III (34 AC, 53 AA). Se realizó una segunda citología en 23 casos (16 AC, 7 AA) con resultado benigno en el 68,7% de los AC y 71,4% de los AA. Fueron intervenidos quirúrgicamente 64 casos (23 AC, 41 AA) y 15 (23,4%) resultaron malignos: 39,1% AC vs 14,6% AA (p=0,029). Hubo un falso negativo en la segunda citología. El carcinoma papilar variante folicular fue el más frecuente (60%). No hubo diferencias entre AC y AA respecto al tipo de neoplasia ni estadio TNM. Conclusiones: Nuestra prevalencia de citologías de categoría Bethesda III está dentro de lo recomendado. La tasa de malignidad es significativamente superior en el grupo AC, pero sin diferencias en el resultado de la segunda citología, tipo de carcinoma hallado ni estadio TNM. La división de la categoría Bethesda III es útil para proporcionar una mejor estratificación del riesgo de malignidad

Objective: To ascertain the prevalence of Bethesda category III cytologies and their malignancy rate, and to analyze differences in the second cytology, malignancy rate, type of carcinoma, and TNM stage between the cytological atypia (CA) and architectural atypia (AA) groups. Patients and method: A retrospective study of 973 biopsies. Bethesda category III cytologies were classified as CA when nuclear atypia was seen but they were not diagnostic or suspicious of malignancy, and as AA when smears had few cells but had a predominantly microfollicular pattern and minimal or absent colloid. The cytological and pathological results were correlated. Results: There were 87 (8.9%) Bethesda category III cytologies (34 CC, 53 AA). Second cytologies were performed in 23 patients (16 with CA, 7 with AA), and a benign result was found in 68.7% of CA and 71.4% of the AA group. Sixty-four patients (23 CA, 41 AA) underwent surgery and 15 of these (23.4%) had a malignant disease: 39.1% CA vs 14.6% AA (P=.029). There was a false negative result in the CA group. The follicular variant of papillary thyroid carcinoma was the most common malignancy (60%). There were no differences in type of carcinoma or TNM stage between CA and AA patients. Conclusions: The reported prevalence of Bethesda category III cytologies was as expected. The malignancy rate was significantly higher in the CA group, but there were no differences in the result of the second cytology, type of carcinoma found, or TNM stage. The division of Bethesda category III cytologies is useful to provide a better stratification of the risk of malignancy

Immunoreactions for P53 isoforms are associated with ultrastructural proliferative profiles in benign thyroid nodules.

BACKGROUND: P53 isoforms originate from the alternative initiation of P53 gene translation through usage of an internal promoter located in intron 4. All P53 isoforms are spliced in intron 9 and may modulate cell proliferation and cell fate outcome in response to DNA damage. AIM: To examine immunoexpression of P53 isoforms in benign proliferative lesions occurring in multinodular thyroids and to assess the ultrastructural phenotype of P53 distribution in the thyrocytes of those lesions by electron microscopy. MATERIALS AND METHODS: By immunohistochemistry and transmission electron microscopy (TEM), we evaluated 38 multinodular thyroids containing a total of 102 benign lesions: 38 nodular goiters (NG; colloid=20, parenchymatous=18), 52 follicular adenomas (FA) and 12 Hashimoto's thyroditis (HT). FA were classified into 10 normo-follicular, 9 macro-follicular, 28 micro-follicular and 5 solid variants. RESULTS: Immunoreaction for P53 isoforms was observed in approximately 50% of all lesions, except macrofollicular variant FA (33%). At TEM analysis, immunoreactive NG, FA and TH lesions showed signs of proliferation by simultaneous appearance of dispersed chromatin, increased amounts of cytoplasmic organelles and dilation of the rough endoplasmic reticulum. TEM signs of apoptosis and proliferation were also detected in FA, but with different rates compared to NG. CONCLUSION: The immunohistochemical expression of P53 isoforms in NG, FA and HT suggests their role in the development of these lesions. Ultrastructural findings support the hypothesis that P53 immunoexpression correlates with reactive proliferative changes in thyrocytes.

[Parameters of thyroid glands functional units in diffuse toxic and nodular euthyroid goiter in population of border areas of Azerbaijan and Georgia].

The purpose of the study--estimate of thyroid tissue quantitative alteration features in different types of goiter on the operative material with a data of morphometry (stereometry) data, histology and electron microscopy in the border (neighboring) regions of Georgia and the Azerbaijan. Cohort study group made up thyroid specimens from 187 patients with nodular euthyroid and diffuse toxic goiter operated in the clinics of Azerbaijan Medical University (Baku) and the National Institute of Endocrinology in Tbilisi for the period 2010 - 2012. Results of the follicules and capillaries morphometric study indicate common clinico-morphologic manifestations of thyroid disorders in the studied regions of Georgia and Azerbaijan, as well as to: 1) determinate capacity to the hyperplastic processes in the thyroid parenchyma under the nodular euthyroid goiter, 2) the disbalance between the clinical manifestation of the disease and its growth potential, which is important to define the "biological behavior" of thyroid nodules, including relapse.

Ultrasound-guided fine-needle aspiration biopsy of the thyroid: methods to decrease the rate of unsatisfactory biopsies in the absence of an on-site pathologist.

PURPOSE: The rate of unsatisfactory samples from ultrasound-guided fine-needle aspirations of thyroid nodules varies widely in the literature. We aimed to evaluate our thyroid ultrasound-guided fine-needle aspiration biopsy technique in the absence of on-site microscopic examination by a pathologist; determine factors that affect the adequacy rate, such as the number of needle passes and needle size; compare our results with the literature; and establish an optimal technique. MATERIALS AND METHODS: We performed a retrospective review of cytopathology reports from 252 consecutive thyroid ultrasound-guided fine-needle aspiration biopsies performed by a radiologist between 2005 and 2010 in our hospital's radiology department. Sample adequacy, the number of needle passes, and needle size were determined. There was an on-site cytologist who prepared slides immediately after fine-needle aspiration but no on-site microscopic assessment of sample adequacy to guide the number of needle passes that should be performed. Cytopathology biopsy reports were classified as either unsatisfactory or satisfactory samples for diagnosis; the latter consisted of benign, malignant, and undetermined diagnoses. RESULTS: Seventy-seven biopsies were performed with 1 needle pass, 124 with 2 needle passes, and 51 with 3 needle passes. The rates of unsatisfactory biopsies were 33.8%, 23.4% (odds ratio [OR] 0.599 [95% confidence interval {CI}, 0.319-1.123]; P = .110), and 13.7% (OR 0.312 [95% CI, 0.124-0.788]; P = .014), respectively. CONCLUSION: In a hospital in which there is no on-site pathologist, a 3-pass method increases the specimen satisfactory rate by 20% compared with 1 pass, achieves similar rates to the literature, and provides a basis for further improvement of our practice.

Oncocytes, oxyphils, Hürthle, and Askanazy cells: morphological and molecular features of oncocytic thyroid nodules.

Our understanding of oncocytic change in thyroid nodules is evaluated in light of the recent progress in understanding the mitochondrial DNA, its mutations, and somatic mutations that affect mitochondrial function. These changes are largely unrelated to the genetic events that result in proliferation and neoplastic transformation of thyroid follicular epithelial cells. The criteria for diagnosing lesions that are composed predominantly of oncocytic cells are the same as those applied to follicular lesions that do not contain oncocytic cells, including follicular variant papillary carcinomas, based on nuclear morphology, immunohistochemical profiles, and molecular markers.

Scanning electron microscopy of thyroid cells under fully hydrated conditions--a novel technique for a seasoned procedure: a brief observation.

Technical information for handling fine-needle aspiration samples from thyroid lesions for WETSEM electron microscopy is presented. The use of wet SEM technology maintains cytological features of the thyroid cells, in the atmospheric electronic microscope chamber without the need for solidification. Images are presented from normal and pathological thyroid specimens showing subcellular elements unavailable to the cytopathologist by light microscopy. Of 24 samples, 18 were adequate for clinical evaluation. In 16 of these 18 specimens, we could find features compatible with the final histological or cytological diagnosis (post-hoc). In two cases, the cell features were too unique to be interpretable. Because this procedure is relatively simple, there is potential for the use of this technology as an adjunct to light microscopy in clinical and research settings.

Morphological changes of follicular cell basal borders and basement membranes in benign and malignant nodular lesions of the thyroid gland: an ultrastructural study.

Microfollicular nodular lesions of the thyroid gland may represent a differential diagnosis problem. Firstly, nodular areas of follicular hyperplasia have to be distinguished from follicular adenomas. On the other hand, nodular microfollicular areas exhibiting large pale nuclei, occasionally found in hyperplastic nodules and follicular adenomas, must be discriminated from latent papillary carcinomas with predominant follicular architecture. The diagnosis of follicular carcinoma still requires the detection of vascular and/or capsular microinvasion. A more refined study was planned to search for additional descriptors useful for diagnosis The authors report the results of an ultrastructural investigation carried out on 220 thyroid nodular lesions and 50 specimens of macroscopically nonnodular glands. An infolding arrangements of the thyreocyte basal border (TBB) and follicular basement membrane (FBM) was demonstrated in 50/50 nonnodular thyroid tissue specimens and 53/67 (79.1%) hyperplastic nodular lesions (p<.005). A linear arrangement of the TBB and FBM was found in 85/121 (70.2%) follicular adenomas and in 32/32 differentiated carcinomas (p<.001). In the last group, 12/32 (37.5%) cases showed focal discontinuities of FBM. In conclusion, the benign thyroid nodules show a prevalently infolding arrangements of TBBs, whereas the majority of proliferative lesions display a linear morphology. In absence of an infiltrating pattern there is no morphological evidence of discriminating potentially malignant vs. benign lesions. The linear distribution of TBBs and FBMs places the case in a group of borderline lesions that involve a more careful postsurgery investigation.

Extramedullary hemopoiesis in a thyroid nodule with extensive bone metaplasia and mature bone formation.

Thyroid nodules may undergo a wide range of degenerative change such as infarction, hemorrhage, or fibrosis, which may be localized or extensive and associated with calcification or even ossification. However, the detection of true bone formation in a thyroid nodule is a very rare occurrence. Extramedullary hemopoiesis (EMH) has been described in almost every organ of the body, mainly in tissues active in hemopoiesis in embryonic life. It is extremely rare for EMH to occur in the thyroid gland especially in patients without known chronic anemia. We describe a case of a cold thyroid nodule with histologically proven extensive bone metaplasia and formation of mature bone with foci of hemopoietic tissue in a young woman without chronic anemia, which, to the best of our knowledge, is the first to be reported in the English language literature.

Cold thyroid nodules show a marked increase in proliferation markers.

Thyroid follicular adenomas and adenomatous thyroid nodules are a frequent finding in geographical areas with iodine deficiency. They occur as hypofunctioning (scintigraphically cold) or hyperfunctioning (scintigraphically hot) nodules. Their predominant clonal origin suggests that they result from clonal expansion of a single cell, which is very likely the result of a prolonged increase in proliferation compared with non-affected surrounding cells. To test whether increased cell proliferation is detectable in cold thyroid nodules, we studied paraffin-embedded tissue from 40 cold thyroid nodules and their surrounding normal thyroid tissue for the occurrence of the proliferating cell nuclear antigen (PCNA) and Ki-67 (MIB-1 antibody) epitopes as markers for cell proliferation. All 40 thyroid nodules were histologically well characterized and have been studied for molecular characteristics before. The labeling index (number of labeled cells versus total cell number) for nodular and surrounding tissue was calculated. In 33 cold thyroid nodules a significant (p < or = 0.05) increase in the labeling index for PCNA was detectable. In 19 cold thyroid nodules a significant (p < or = 0.05) increase in the labeling index for Ki-67 was detectable. Moreover, surrounding tissues with lymphocyte infiltration showed a significantly higher labeling index for both PCNA and Ki-67 compared with normal surrounding tissue. These findings are first evidence that an increased thyroid epithelial cell proliferation is a uniform feature common to most cold nodules. However, the increase of proliferation markers shows a heterogeneity that is not correlated with histopathologic, molecular, or clinical characteristics.

Resolution of hyperthyroidism in a pregnant woman with toxic thyroid nodule by percutaneous ethanol injection.

Overt hyperthyroidism was found in a 35-year-old pregnant woman at the 13th week of gestation who was referred to us for tachycardia, tremors, and weight loss. Clinical signs, symptoms, and laboratory findings led to the diagnosis of toxic thyroid nodule. She was treated with ultrasound guided percutaneous ethanol injection (PEI) and, after 2 weeks of treatment, the woman was completely euthyroid. These findings suggest that during pregnancy PEI appears to be a rapid and safe therapy for toxic nodular goiter and an effective alternative to the administration of antithyroid drugs.

Serum thyroglobulin level and thyrotropin receptors in plasma membranes from thyroid nodular relapse.

The parameters of 125I-bTSH binding to plasma membrane as well as serum thyroglobulin (Tg) level in 13 euthyroid females, suffering from scintigraphically functioning thyroid nodule, were investigated. Thyroid tissue samples of five euthyroid patients were obtained by surgical reintervention, which was done because of scintigraphically functioning nodular goitre relapse (Group 1). Data were compared with two control groups. One of them (Group 2) consisted of 5 euthyroid females in whom partial thyroidectomy was done because of solid functioning thyroid nodule. Long-term follow-up during postoperative period confirmed normal morphofunctional status in them. The second control (Group 3) is consisted of 3 patients suffering from goitre relapse now. The percent of specific 125I-bTSH binding and the affinity of its binding to plasma membranes from nodular goitre relapse were lower (2.05% +/- 0.82; Kd1: 0.56 +/- 0.34 10(-9) M; mean and S.D., resp.) than to the membranes obtained from surrounding perinodular tissue (0.32 +/- 0.89; Kd1: 0.20 +/- 0.77 10(-9) M, resp.). However, the level of high affinity TSH receptors in plasma membranes prepared from nodular goitre relapse was higher than that in perinodular, assumed to be normal tissue (11.21 +/- 0.87 fmol/mg prot. versus 6.29 +/- 2.24 fmol/mg prot., mean and S.D.). Plasma membranes obtained from goitre relapse were characterised by high total number of receptor and binding sites (N1 + N2) for TSH. In Group 1 and 3 elevated serum Tg levels were confirmed during follow-up period with no correlation with TSH, which characterises a normal physiological status of regulation.

Influence of frequency of previous pregnancies on the prevalence of thyroid nodules in women without clinical evidence of thyroid disease.

In a region with insufficient alimentary iodine supply (Kiel, Northern Germany) the prevalence of thyroid nodules was studied by means of ultrasonography in 212 healthy women (36-50 years old) in four groups of 53 women each with 0, 1, 2, and 3-5 previous pregnancies. Goiters were found in 28.3% (15 of 53) of the women without children and in 28.9% (46 of 159) of the women with children. There was no significant increase of goiter prevalence according to the number of pregnancies. We detected thyroid nodules in 21.2% (45 of 212). Only 9.4% (5 of 53) of the women without previous pregnancies had thyroid nodules, but 25.1% (40 of 159) of the women with pregnancies in the past had nodules, the difference being statistically significant (p < 0.05). We observed nodules in 11 of 53 women with 1 child (20.7%), in 11 of 53 women with 2 children (20.7%), and in 18 of 53 women with 3-5 previous pregnancies (33.9%). We propose that, in regions with borderline or insufficient alimentary iodine supply, accentuated iodine deficiency during pregnancies due to increase of iodine requirement is a probable cause for the higher prevalence of thyroid nodules in women with previous pregnancies.

Benign follicular thyroid nodule composed of signet-ring-like cells with PAS-negative thyroglobulin accumulation in dilatated rough endoplasmic reticulums.

A benign follicular thyroid nodule composed of signet-ring-like cells in a 46-year-old Japanese euthyroid woman is reported. Thyroglobulin was positive, and periodic acid-Schiff (PAS) and other mucin stainings were negative in the cytoplasm of signet-ring-like cells. Ultrastructurally their cytoplasm contained many dilatated rough endoplasmic reticulums (rERs), the largest of which compressed the nucleus toward the cellular apical portion. Functional disturbance of rER or Golgi apparatus might be the cause of this characteristic cellular change.

[Functional microscopic imaging of the thyroid gland with the help of analytical ion microscopy]. / Imagerie microscopique fonctionnelle de la glande thyroïde A l'aide de la microscopie ionique analytique.

Secondary ion mass spectrometry (SIMS) microscopy provides a direct mapping of 127I in thyroid follicles. Thyroid tissue should be fixed and embedded in methacrylate resin, then cut in 3 microns-thick sections which are placed on a gold coverslip. The histological structure of the tissue is determined by the phosphorus ion (31P) which is present in large amounts in nuclei and phosphorylated molecules in the cytoplasm. An image processing system is used to superimpose images of 127I and 31P: this system allows measurement of local concentration of 127I in the follicular cell and follicular lumen compartments. A study in 8 subjects with normal thyroid glands showed that the level of 127I within follicular cells (430 +/- 250 micrograms/g; m +/- SE) was 6 to 7 times lower than the level in the follicular lumen (2.780 +/- 230 micrograms/g). In simple goiter (9 patients with macrofollicular adenomas), follicular lumen (346 +/- 17 micrograms/g) and cellular (68 +/- 6 micrograms/g) concentrations of 127I were decreased fivefold but the ratio of concentrations remained similar to that seen in normal tissue. In hyperfunctioning nodules (2 microfollicular and 3 macrofollicular adenomas), follicular cell and follicular lumen 127I levels varied widely and showed considerable overlap (2 to 16,000 micrograms/g and 1-21,000 micrograms/g, respectively). In benign cold nodules (1 microfollicular adenoma and 2 macrofollicular adenomas), follicular cell and follicular lumen 127I levels were barely detectable (3 +/- 0.6 micrograms/g and 6 +/- 0.7 micrograms/g respectively). In the 9 malignant nodules studied, no difference in 127I level profile was found with benign nodules.(ABSTRACT TRUNCATED AT 250 WORDS)