Patterns of cyclin A and B1 immunostaining in papillary thyroid carcinoma.

INTRODUCTION AND OBJECTIVES: Cyclin A, encoded by CCNA (cyclin A) gene with locus in chromosome 4q27, and cyclin B1, encoded by CCNB1 (cyclin B1) gene with locus in chromosome 5q12, are proteins that play a key role in the passage through the restriction point in G2 phase of the cell cycle. The aim of the study was to analyse immunohistochemically the expression of cyclins A and B1 in different variants of papillary thyroid carcinoma (PTC). MATERIAL AND METHODS: The immunostaining patterns of the proteins in question in the tissue of 40 resected PTC (20 cases of classic variant of PTC, 9 cases of PTC follicular variant and 11 cases of other non-classic variants of PTC) were investigated. RESULTS: On analyzing cyclin A and B1 expression, positive staining in 90% cases of PTC were observed. The study revealed a significant difference in expression of cyclins A and B1 between classic and non-classic variants of PTC. The expression of both examined cyclins was weaker in the classic variant of PTC. In the group of follicular variant of PTC, the expression of cyclins was of medium intensity and in the group of other non-classic variants of PTC, the expression was clearly higher. CONCLUSIONS: The results of the presented study suggest that cyclins A and B1 expression may have a characteristic pattern of immunostaining for particular variants of PTC. If the obtained results are confirmed in a larger group of patients, the diagnostic panel constructed of the antibodies against these proteins may increase the diagnostic accuracy in PTC cases.

Lateral spread of heat during thyroidectomy using different haemostatic devices.

INTRODUCTION: The presented study is an attempt to comprehensively analyze the lateral spread of heat during thyroidectomy. Obtained results may be valuable in other surgical disciplines in which thermal analysis is difficult or impossible. OBJECTIVE: The aim of the study was to evaluate the temperature distribution in the operating field during thyroidectomy performed with the use of modern haemostatic instruments, and to define the safety margin for the investigated devices. MATERIALS AND METHODS: Ninety-three patients were thyroidectomised due to thyroid neoplasm. During all the operations the thermovisual measurements were carried out along with continuous intraoperative neuromonitoring of the recurrent laryngeal nerve (CIONM). Investigated patients were divided into 5 groups, named according to the applied haemostatic technique: LigaSure (N=17); ThermoStapler (N=20); Focus (N=19); SonoSurg (N=17) and Monopolar (N=20). RESULTS: At maximal performance settings, the highest working temperature was observed for the ThermoStapler, while the lowest temperature was recorded for the Monopolar. Safety margin and working time were increased in Focus and SonoSurg, compared to LigaSure and ThermoStapler. The differences in the necrosis thickness were negligible. The largest distance of the midline of the active blade from isotherm of 42ºC observed in the study was 5.51 mm; none of investigated devices used at a bigger distance had influence on the morphology of the electric signal of CIONM. CONCLUSION: The thermo-visual camera allows non-invasive, safe, and real-time monitoring and analysis of temperature distribution in the operation area during thyroidectomy. Proposed minimal safety margin for the analysed devices is 5.51 mm.

A rare case of Hodkin's lymphoma of the mediastinum imitating retrosternal goiter--retrospective analysis of the diagnostic process.

Primary extranodal sites of development of lymphoid neoplasms are rare and concern about 5% of patients with Hodgkin's lymphoma. Extranodal development is more common in non Hodgkin's lymphoma and may reach 33%. A 27-year-old woman was diagnosed by a cardiologist for a short breath. On the physical examination no other abnormalities were observed. Echocardiography, performed by cardiologist, revealed a large tumor, overlaying the right ventricle and compressing the pulmonary trunk. Chest X-ray, ultrasound and CT-scan confirmed diagnosis. In fine needle aspiration clear, lucid fluid was obtained. Scintygraphy of the neck and thorax showed accumulation of the marker in the properly placed but enlarged thyroid gland. Patient was qualified for surgical treatment - cervicotomy and sternothomy were performed. The histopatological exam of the tumor revealed Hodgkin's lymphoma of the mediastinum (classical subtype NS-1). Following the surgery, adjuvant therapy was instituted. After the treatment PET-CT-scan did not show any kind of non-physiological radiomarker's accumulation in the monitored regions of the body and in in three-years follow-up the patient shows no signs of recurrence.

Comparison of lateral thermal spread using monopolar and bipolar diathermy, and the bipolar vessel sealing system ThermoStapler™ during thyroidectomy.

UNLABELLED: Electric devices enabling the maintenance of haemostasis during surgery have found application in modern thyroidectomy procedures. The haemostatic effect is associated with generation of heat, which apart from the intended result may bring about thermal tissue injury. The aim of the study was to determine the thermal spread around the active tip of electric devices in the operating field during total thyroidectomy, and the safe temperature range during the operation of studied devices. MATERIALS AND METHODS: Over 14 months from December 2009 until January 2011, 76 total thyroidectomy procedures were analysed. The surgeries employed mono- and bipolar diathermy as well as the ThermoStapler™ bipolar vessel sealing system. During the procedures, the thermal spread around the active tips of used electric devices was recorded with the use of high-definition camera. Comparable 5-second periods of electric device use at two power ranges (30 W and 50 W) were selected from the recorded material. The highest temperature of the active tip of electric devices was determined, and the 42°C isotherm was found with the use of computer image analysis, thus determining the safe distance of important anatomic structures from the active tip of the electric device. RESULTS: The temperature spread around the active tips of electric devices was recorded and the 42°C isotherm was determined. The diameter of this isotherm at the end of operation differed statistically significantly depending on the type of electric devices and power settings. The highest temperature, at both power ranges, was recorded for the bipolar vessel sealing system, while the lowest - for bipolar diathermy; at the same time a significantly lower 42°C isotherm diameter was found for ThermoStapler™ as compared with other devices. In all studied cases, the largest heat spread was found for monopolar diathermy. CONCLUSIONS: The mean safe distance of the active tip of an electric device from important anatomic structures is 5 mm and depends on the device type and its power settings. Monopolar diathermy causes the strongest heating of surrounding tissues, and the ThermoStapler™ bipolar vessel sealing system, despite producing the highest temperature during operation, causes relatively small thermal injury to the surrounding tissues.

Aberrant methylation as a main mechanism of TSGs silencing in PTC.

In the present study the role of tumour suppressor genes (TSGs) hypermethylation and genetic instability of LOH/MSI type in thyroid tumorigenesis was assessed. Expression, methylation status and presence of LOH/MSI were analyzed for 8 TSGs selected from imprinted (IR) and non-imprinted (NIR) chromosomal regions in papillary thyroid carcinomas (PTCs) and nodular goitres (NGs). The results show that methylation-induced gene silencing occurs at an early step of thyroid carcinogenesis and involves multiple genes. Genetic changes of LOH/MSI type are less frequent. In PTC samples, the lack of significant differences in the frequency of LOH in IR and NIR suggests that it is not a key mechanism changing the pattern of gene expression. Co-methylation observed both in NG and PTC raises a possibility that, in thyroid tissue, methylation-induced silencing may occur not only in malignant transformation but also in functional context. We did not recognize any of the studied TSGs - in regard to aberrant methylation status or LOH/MSI frequency - as a selective molecular marker in thyroid tumorigenesis.

Metastases of breast cancer to the thyroid gland in two patients - a case report.

INTRODUCTION: Metastatic cancer is rarely found in the thyroid (only 2-3% of malignant tumours found in that gland); primary sources usually including breast, kidney, and lung tumours. CASES REPORTS: Two cases of advanced breast cancer with thyroid metastases in female patients are presented. The similarities between these two cases included: 1) postmenopausal age; 2) diagnosis based on result of FNAB (numerous groups of cells with epithelial phenotype strongly implying metastatic breast cancer); 3) thyroid function - overt hyperthyroidism in the first woman and subclinical hyperthyroidism in the second one; 4) the presence of nodular goitre in clinical examination, the occurrence of many nodular solid normoechogenic lesions with calcifications in both thyroid lobes in US; and 5) negative antithyroid antibodies. The main difference was the time of establishing diagnosis; in the first woman - before mammectomy, parallel to diagnostics of breast tumour, and in the second woman four years after mammectomy, during cancer dissemination (with right pleural effusion and lung metastasis). In the first case, mammectomy was followed two weeks later by thyroidectomy. The second patient was disqualified from thyroid surgery due to systemic metastatic disease. CONCLUSIONS: 1. Fine needle aspiration biopsy of the thyroid gland should obligatorily be performed in patients with breast cancer and nodular goitre, even without any clinical data of metastatic disease. 2. The clinical context of cytological findings is of critical value. 3. In patients with breast cancer accompanied by multinodular goitre, we recommend that more punctures be performed during FNAB than is routinely done. (

Real-time, high-resolution ultrasonography of the vocal folds--a prospective pilot study in patients before and after thyroidectomy.

PURPOSE: The aim of the study was to evaluate the functionality of vocal folds (VF) by real-time, high-resolution ultrasonography (US) and to correlate the imaged features to results of laryngological examination (LE). METHODS: The study group comprised 50 patients (41 females and nine males), qualified to thyroidectomy. All the patients had LE and US examination before and 2 days, 2 months, and 3 months after the surgery. We used high-resolution US imaging to identify VFs and, subsequently, a pulsed Doppler and Doppler gate to quantify the tissue displacement velocity in the vibrating VF section. RESULTS: LE revealed unilateral VF paralysis in two patients. VF dysfunction was diagnosed in other four subjects. In simultaneously performed US examination, changes in VF displacement velocity (VFDV) were observed in ten patients. In two subjects, VFDV was below 30 cm/s- patients with VF paralysis, diagnosed in LE. In a further eight cases, we observed VFDV decrease by 50%, comparing to preoperative values. Both US-imaging and LE, performed after the 3-month follow-up, confirmed the transitional character of the above-mentioned pathologies. CONCLUSIONS: US imaging of the VFs correlated with LE results, while being a minimally invasive, easily reproducible, and inexpensive method of examining VF functionality. Thanks to many recording options, it may soon become a perfect tool for an early identification of postoperative VF dysfunction with its later monitoring. To our knowledge, it is the first application of US and Doppler gate modes for VFDF quantification; however, an analysis on a larger group of patients is necessary to standardize the technique.

Evaluation of selective embolization of thyroid arteries (SETA) as a preresective treatment in selected cases of toxic goitre.

BACKGROUND: in recent years, an increasing interest in the application of selective embolization of thyroid arteries (SETA) in the treatment of thyroid diseases is observed. In the present report, we analyse the value, safety and possible indications for preresective SETA in cases of large toxic goitres. PATIENTS AND METHOD: the study group comprised 10 patients with large toxic goitre (thyroid volume 254 +/- 50 mL), including one patient with cervicomediastinal goitre and one patient with anti-thyroid drug intolerance in state of overt thyrotoxicosis. All the patients underwent SETA of the superior and/or inferior thyroid arteries, followed by thyroidectomy, performed up to thirty-six hours after SETA (23.1 +/- 11 h). After embolization, selective angiographies of thyroid arteries were performed to ensure that the targeted arteries had been completely occluded. RESULTS AND CONCLUSION: in all the patients, SETA decreased blood flow through the thyroid. Preresective SETA reduced blood loss during and after thyroidectomy and decreased the operating time, but the differences were too small to justify routine applications of preresective SETA as an adjunct to surgical treatment of toxic goitre. On the other hand, SETA is a safe and minimally-invasive technique, which may become an attractive option for quick preparation to surgery in selected patients with toxic goitre, who present anti-thyroid drug intolerance or refuse radioactive iodine treatment.

Selective embolization of thyroid arteries as a preresective and palliative treatment of thyroid cancer.

Although many tumours of head and neck have been successfully embolized, the number of publications on the application of selective embolization of thyroid arteries (SETA) is limited. The aim of the present study is to evaluate the safety, efficacy and possible indications and contraindications for preresective or palliative SETA in thyroid cancer. The study group comprised 20 patients with thyroid tumours: 7 cases of advanced inoperable anaplastic thyroid cancer (ATC) and 13 cases of differentiated thyroid carcinoma (DTC). All the patients underwent SETA of the superior and/or inferior thyroid arteries. After SETA, selective angiographies of thyroid arteries were performed to ensure that the targeted arteries had been completely occluded. In all the cases, SETA decreased the blood flow through the thyroid. Preresective SETA limited bleeding during surgery and decreased operating time. We observed a massive increase of thyroglobulin (Tg) concentrations in cases of DTC that started 36-48 h after SETA and did not occur in cases of ATC. Although SETA had no influence on the mortality of ATC patients, they reported improvements in swallowing, breathing and decrease of the pain. Concluding, SETA is minimally invasive and safe method limiting blood flow through thyroid tumours. In DTC patients, SETA causes ischaemic necrosis of the gland which results in important increases in serum concentrations of Tg. Therefore, thyroidectomy should be performed during the first 36 h after preresective embolization. Moreover, SETA may become an attractive option of palliative treatment for ATC patients with intractable bleeding, pain or signs of tracheal and oesophageal compression.

Influence of L-thyroxine administration on poor-platelet plasma VEGF concentrations in patients with induced short-term hypothyroidism, monitored for thyroid carcinoma.

Angiogenesis is a process of new blood vessel development from pre-existing vasculature. It is a crucial process in normal physiology, as well as in several pathological conditions. The vascular endothelial growth factor (VEGF) represents a family of specific endothelial cell mitogens, involved in normal angiogenesis and in tumour development. The aim of the present study was to estimate the influence of L-thyroxine (L-T4) administration on poor-platelet plasma (P-PP) VEGF concentrations in patients with induced short-term hypothyroidism, monitored for differentiated thyroid carcinoma. In the present study, P-PP concentrations of VEGF, thyroglobulin, thyrotropin and free thyroid hormones were investigated in a population of 24 hypothyroid patients, who were withdrawn from L-T4 treatment for 5 weeks and studied before and after 2 months of L-T4 therapy. Only healthy female patients with no evidence of metastasis in whole body scintigraphy were included in the study. They were then compared with 20 healthy control subjects, matched for age, sex and body mass index (BMI). The patients had significantly lower plasma VEGF concentrations before treatment with L-T4 than after administration of that hormone. There was no significant difference in plasma VEGF levels, either between the patients treated with L-T4, and the controls, or between the patients untreated with L-T4, and the controls. Even short-time changes in thyrometabolic profile exert an important influence on P-PP VEGF concentrations, even if there is no thyroid tissue.

Selective embolization of thyroid arteries (SETA) as a palliative treatment of inoperable anaplastic thyroid carcinoma (ATC).

OBJECTIVES: Anaplastic thyroid carcinoma (ATC) is one of the most aggressive solid tumors in humans. Despite intense application of multimodality of treatment with surgery and/or external beam radiotherapy and chemotherapy, the survival rates remain low--generally the mean survival is about six (6) months after diagnosis. Rapid development--particularly over the last decade--of interventional radiology, provides methodology that allows examining thyroid arterial embolization as an alternative approach to ablating thyroid tissue. The aim of the present study was to evaluate selective embolization of the thyroid arteries (SETA) as a possible alternative for the palliative treatment of advanced, inoperable ATC. PATIENTS AND METHODS: The study group comprised five (5) patients with advanced stage of inoperable ATC. All the patients underwent SETA of the superior and/or inferior thyroid arteries. SETA was performed using polyvinyl alcohol particles, ranging from 500-710 microm in diameter. After SETA, selective angiography of thyroid arteries was performed to ensure that the targeted arteries were completely occluded. CONCLUSIONS: The results of the present study suggest that SETA is minimally invasive and save method of palliative treatment of ATC and, as such, may be recommended in cases of intractable hemorrhage and pain caused by ATC progression.

Patterns of cyclin E, retinoblastoma protein, and p21Cip1/WAF1 immunostaining in the oncogenesis of papillary thyroid carcinoma.

PURPOSE: Uncontrolled cell proliferation, a hallmark of cancer, may result from an increased expression of cell cycle up-regulators, and/or from a reduced expression of cell cycle down-regulators. In the present study, we analyzed, by immunohistochemistry, the expression of a panel of three proteins: cyclin E and two cell cycle inhibitors, p21(Cip1/WAF1) and retinoblastoma protein (pRb) product, in different stages of papillary thyroid carcinomas (PTC). EXPERIMENTAL DESIGN: We investigated immunostaining patterns of the proteins in question in 51 resected PTC in pathologic stages, ranging from pT(1a) to pT(4), taking into consideration their relation to clinicohistopathologic factors. RESULTS: We observed a significant, progressive loss of expression of p21(Cip1/WAF1) with advancing tumor grade. The differences reached values of significance between pT(1a) [papillary thyroid microcarcinomas (PMC)] and pT(2) and between PMC and pT(4) stages of PTC. pRb presented a similar immunostaining pattern to that of p21(Cip1/WAF1) and the differences reached values of significance between pT(1a) and pT(2), and between PMC and pT(4) stages of PTC. The results of cyclin E immunostaining corresponded to our recently published result, and a negative correlation was observed between the immunostaining index of cyclin E and pRb. CONCLUSIONS: The results of the present study suggest that cyclin E expression and suppression of pRb and p21(Cip1/WAF1) may be characteristic patterns of immunostaining for PTC with a tendency to early metastasizing. If our results are confirmed in a larger study, the diagnostic panel, constructed of the antibodies against these proteins, may become a valuable tool in predicting the metastatic potential in PTC.

Increased thymidine kinase activity in human thyroid toxic adenomas: effects of exposure to epidermal growth factor in vitro.

UNLABELLED: The activity of thymidine kinase (TK-EC 2.7.1.21)--an enzyme functioning as a part of the pyrimidine salvage pathway of DNA synthesis--is closely related to growth processes. The aim of the study was to measure TK activity in homogenates of human thyroid tissue of the following types: non-toxic nodular goiter (NTNG)--macroscopically unchanged tissue, non-toxic adenoma (NTA), and toxic adenoma (TA) (obtained from patients, who--before the surgery--had been treated with thyrostatic drugs for thyrotoxicosis). Thyroid tissue was obtained from female patients subjected to subtotal thyroidectomy at the Department of Endocrine Surgery, Medical University of Lódz. Thyroid homogenates were incubated in the presence of epidermal growth factor (EGF), used in five different concentrations (0.1 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1000 ng/ml). TK activity was estimated by chromatographic measurements of the amount of the main reaction product--deoxythymidine monophosphate. RESULTS: 1) We did not observe any significant difference between TK activity in the homogenates of the thyroid tissue collected from NTNG and NTA; TK activity was clearly higher in the homogenates of adenomatous tissue, collected from the patients with TA; 2) EGF increased TK activity in the homogenates of the macroscopically unchanged tissue, collected--during surgery--from the patients with NTNG, as well as in homogenates of thyroid tissue from NTA; 3) In case of hyperactive thyroid tissue, obtained from TA, EGF tended to increase TK activity, however, without any statistical differences. Our results confirm TK increased activity in hyperactive thyroid tissue. At the same time, the obtained data suggest a certain role of EGF in goiter formation in humans.

Expression of insulin-like growth factor I (IGF-I) gene and of genes for IGF-binding proteins 1, 2, 3, 4 (IGFBP-1-IGFBP-4) in non-neoplastic human thyroid cells and in certain human thyroid cancers. Effect of exogenous IGF-I on this expression.

AIM: The aim of this study was to examine the expression of the IGF-I gene and of genes for IGFBP-1, -2, -3, and -4 in cells from nodular goiters (NG), and from different human thyroid carcinomas (papillary--PTC, anaplastic--ATC, and medullary--MTC), cultured in monolayers. The influence, exerted by exogenous IGF-I on the expression of these genes, was also investigated. METHODS: Thyroid tissue specimens were obtained from 65 patients during subtotal or total thyroidectomies. After approximately 2-3 weeks of culture, thyroid cells were incubated for 24 hours with IGF-I in concentrations of: 0, 1, 10 and 100 ng/ml. The total mRNA was isolated according to the method described by Chomczynski and Sacchi with our own modifications. Afterwards, mRNA encoding IGF-I, IGFBP-1-IGFBP-4 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were amplified, using the reverse transcription-polymerase chain reaction (RT-PCR); GAPDH gene served as a control gene. PCR products were electrophoresed and then submitted to densitometric analysis. RESULTS AND CONCLUSIONS: Our study has shown that in carcinoma cells (ATC, PTC, MTC), IGF-I reveals a stimulatory influence on the expression of its own gene, that effect being most distinctive in ATC cells. These facts indicate an important role of IGF-I in the pathogenesis and invasiveness of the analyzed malignant neoplasms.

Cyclin E expression in papillary thyroid carcinoma: relation to staging.

Cyclin E plays a pivotal role in the regulation of G1-S transition and relates to malignant transformation of the cells. However, the clinical significance of cyclin E expression in patients with papillary thyroid carcinoma (PTC) remains unknown. We examined by immunohistochemistry the expression of cyclin E in 41 resected PTCs in pathologic stages from pT1a to pT4 and analyzed its relation to clinicohistopathologic factors. The positive staining was divided into 3 grades: no expression if less than 10%, expression if 11-50% and overexpression if more than 50% of the nuclei of tumor cells were stained positively. Cylin E expressions were observed in 75.6% of analyzed PTCs but only 60% of papillary microcarcinomas (PMCs) were immunopositive for cyclin E expression. However, cyclin E staining was observed in 90.4% of PTCs in a group with TNM higher than pT1a. The staining index was significantly different between the PMCs and the rest of the cancers investigated (14.91% +/- 14.4% vs. 34.03% +/- 23.44%, respectively; p < 0.005) and we observed positive relation between the staining index and factor T of staging of PTCs. All the lymph node metastases coexisted with cyclin E expression and most, but not all, of them coexisted with cyclin E overexpression. These findings indicate that cyclin E may play a key role for the oncogenesis and biologic behavior of PTC. If our results are confirmed in a larger study, a high level of cyclin E expression may become a new prognostic marker for PTCs.