PTEN protein expression in postmenopausal steroid receptor positive early breast cancer patients treated with adjuvant tamoxifen.

PURPOSE: Since one of possible causes of resistance to antiestrogen therapy in steroid receptor positive (SR+) breast cancer (BC) patients is an alteration of PTEN (phosphatase and tensin homolog deleted on chromosome 10) signaling pathways, the aim of this study was to determine the PTEN protein expression in postmenopausal patients with steroid SR+ BC treated with adjuvant tamoxifen, to investigate the association of PTEN protein expression with tumor histology, size and grade, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) statuses and disease outcome. METHODS: This was a retrospective analysis of 78 postmenopausal stage I/II SR(+)BC patients treated with adjuvant tamoxifen. PTEN protein expression and ER, PR and HER2 status were determined using immunohistochemistry. RESULTS: The distribution of PTEN protein expression according to tumor histology was as follows: PTEN+ status in 27/43 (62.8%) patients with ductal and in 26/35 (74.3%) patients with lobular carcinomas; and PTEN(-) status in 16/43 (37.2%) patients with ductal and in 9/35 (25.7%) patients with lobular carcinomas. Disease relapse was observed in 38/78 patients: 14/53 (26.4%) of PTEN(+) BC subgroup and 24/25 (96%) of PTEN(-) subgroup (x(2), p=0.018). There were no significant associations between PTEN protein expression and tumor histology, size and grade, and ER, PR and HER2 expression. Patients with PTEN(-) had significantly shorter disease-free interval (DFI) and overall survival (OS) (for both, log rank test, p <0.01) compared to PTEN(+) BC patients. CONCLUSION: Our results suggest that PTEN protein expression might be of prognostic significance in postmenopausal SR(+) BC patients treated with adjuvant tamoxifen.

Evaluation of galectin-8 expression in thyroid tumors.

The expression of galectin-8 (gal-8) has been shown to be altered during neoplastic transformation of certain cell types. This is the first study aimed to analyze the expression of this protein in normal and pathological human thyroid tissue. A total of 41 archival thyroid tissue samples (5 follicular adenomas, 31 papillary carcinomas, 5 follicular carcinomas) together with 36 adjacent hyperplastic or normal thyroid tissues were analyzed by immunohistochemistry. Galectin-8 was expressed in the majority of papillary carcinomas (27/31; 87%). Positive but weaker staining was also found in some of the follicular thyroid carcinomas (2/5; 40%) and adenomas (2/5; 40%). This protein was not detectable in five normal thyroid tissue samples, whereas hyperplastic areas adjacent to tumor were weakly positive in 9 out of 31 cases (29%). High gal-8 immunostaining in papillary thyroid carcinoma indicates that gal-8 may potentially serve as a marker of papillary thyroid carcinoma. However, it does not seem to be helpful in the differential diagnostics of follicular carcinoma and adenoma. Further studies are required to determine biological functions and molecular mechanisms underlying the increased expression of gal-8 protein in thyroid lesions, particularly, in papillary thyroid carcinoma.

Changes in the balance between proliferation and apoptosis during the progression of malignancy in thyroid tumours.

The aim of this study was to gain better insight into molecular changes which reflect disturbances in the balance between proliferation and apoptosis during progression of thyroid malignancy from papillary microcarcinoma (PMC) via clinically manifest papillary carcinoma (PTC) to anaplastic carcinoma (ATC). The apoptosis related molecules (Bcl-2, Bax) and proliferation related marker (PCNA) were analysed immunohistochemically in 120 archival cases comprising PMC (n=34), PTC (n=52) and ATC (n=34). In addition, in situ apoptotic cell death was analysed by the TUNEL method. The average Bcl-2 staining score did not differ between PMC and PTC (p>0.05), but was significantly lower in ATC (p<0.05).The Bax score was higher in PTCs and ATCs than in PMCs (p<0.05). Due to these changes, the Bcl-2/Bax ratio showed a marked decrease from PMC to ATC (p<0.05), while proliferation activity increased significantly from PTC to ATC (p<0.05). Despite high Bax expression, the rate of apoptotic cell death was low in the investigated carcinomas, especially in ATC, i.e. the increase in proliferative activity was not counterbalanced with appropriate cell death. Differences were found in the expression of apoptotic molecules (Bcl-2 and Bax), their ratio (Bcl-2 /Bax) and in the rate of apoptotic cell death and proliferative activity between PMC, PTC and ATC, indicating that disturbances in the balance between apoptosis and proliferation, in favour of the latter, occur gradually during the progression of malignancy in thyroid tumours.

The incidence of familial nonmedullary thyroid cancer in a large case series.

PURPOSES OF THE STUDY: In contrast to familial medullary carcinoma, familial nonmedullary thyroid carcinoma (FNMTC) is less frequent and has been less investigated. The aim of this study was to determine the frequency of FNMTC and analyse the main demographic and clinical characteristics of the patients. MATERIAL AND METHODS: Data on 1411 patients surgically treated for nonmedullary thyroid carcinoma, in the Center for Endocrine Surgery in Belgrade, from 1995 to 2006 were analysed. The possible presence of malignant tumours of the thyroid gland was investigated in their closest relatives in order to identify cases of FNMTC. Only data on first-degree relatives (parents and children) and second-degree relatives (grandparents, grandchildren and siblings) were taken into account in the analysis. RESULTS: Thirteen patients (11 females and 2 males) (0.92% of those with nonmedullary carcinoma of the thyroid gland) had a familial form of the disease. In five families two members had a tumour, and in one family three members. In five out of six families it was a papillary carcinoma and in one family a follicular carcinoma. Patient age varied from 20 to 79 years, with a mean age of 40 years. The tumour size ranged from 5 to 60 mm (mean 25 mm). In two of the thirteen cases the tumour penetrated the capsule of the thyroid gland. In four cases the tumour was multicentric and bilateral, and in a further two metastases were present in regional lymph nodes. During the follow-up period, which lasted from 2 to 12 years (mean 8.5 years), two relapses were detected. CONCLUSION: Familial nonmedullary carcinoma of the thyroid gland occurs very rarely.

Surgery for thyroid Hürthle cell tumours--a single institution experience.

AIMS: The objective of the study was to report a series of patients with Hürthle cell tumours. METHODS: We reviewed medical records of single institution from January 1982 to December 2002, including follow-up information. RESULTS: We identified 199 patients with Hürthle cell tumours (HCT), 88 patients with Hürthle cell carcinoma (HCC) and 111 patients with Hürthle cell adenoma (HCA). The HCC group had significantly longer duration of the disease and larger tumours (4.8 vs 3.8 cm) compared with HCA group. Gender appeared to play significant role in patients with HCT (women outnumbered man by 7:1; p < 0.01). Surgical management for 80% of patients with HCA consisted of hemithyroidectomy and total thyroidectomy in 87% patients in the HCC group. Temporary laryngeal nerve palsy and temporary hypoparathyroidismus were not seen in HCA group, in HCC group were confirmed in 2.27 and 3.41%, respectively. Four patients with HCC relapsed and two died of HCC. CONCLUSIONS: HCC has outlook for favorable outcome when treated radically with total thyroidectomy.

Immunohistochemical localization of galectin-3 in malignant and benign human thyroid tissue.

BACKGROUND: Galectin-3 is an endogenous beta-galactoside binding lectin with putative roles in development, immunomodulation, transformation and metastasis. The purpose of this study was to analyze galectin-3 expression in a series of human thyroid neoplastic lesions. METHODS: A total of 76 cases, including 47 specimens of thyroid malignancies, 14 follicular adenomas and 15 specimens of normal thyroid tissue, were analyzed immunohistochemically using a monoclonal antibody to galectin-3 and avidin-biotin-peroxidase complex (ABC) method. RESULTS: The immunohistochemical staining results showed galectin-3 expression in neoplastic cells of all 20 cases of papillary carcinoma, 11 out of 15 follicular carcinomas, both oxyphilic carcinomas, all 10 anaplastic carcinomas and 5 out of 14 follicular adenomas. Galectin-3 localization was mostly cytoplasmic, but also membraneous or nuclear in some cells. Follicular cells in normal thyroid tissue were negative. CONCLUSIONS: The results of this study indicate that galectin-3 gene is expressed at the protein level in most thyroid carcinomas and some adenomas. Galectin-3 expression was not clearly correlated with histopathological aggressiveness, dedifferentiation state or determination of malignancy of the follicular tumour. The role of galectin-3 in thyroid tumour biology remains to be elucidated.

Unusual variants of the tributaries of the main pancreatic duct revealed by postmortem and endoscopic pancreatography.

Analysis of 97 human postmortem pancreatograms and 103 endoscopic pancreatograms revealed a total of 3 cases (1.5%) with isolated variants in the branches of the main pancreatic duct. In the first one, tortuous branches were present in the upper head-body region. The second case involved joint tributaries bridging over an unstenosed segment of the main pancreatic duct. In the third case, we found three branches from the uncinate process running down to the main duct. In all of these cases no pathological substrate was found, and they should be considered as anatomical variants observed during interpretation of a radiogram.

Differential expression of KCNQ1 K+ channel in tubular cells of frog kidney.

The aim of this study was to evaluate KCNQ1 K+ channel expression in the frog kidney of Rana esculenta. KCNQ1 K+ channel, also known as KvLQT1, is the pore forming a-subunit of the IKs K+ channel, a delayed rectifier voltage-gated K+ channel, which has an important role in water and salt transport in the kidney and gastrointestinal tract. The expression of KCNQ1 K+ channel along tubular epithelium differs from species to species. In the present study the expression of KCNQ1 K+ channel in the frog kidney has been demonstrated by immunohistochemistry. The presence of KCNQ1 K+ channel was demonstrated in the epithelial cells of distal convoluted tubule and collecting duct. However, the pattern of expression of KCNQ1 K+ channel differs between distal convoluted tubules and collecting duct. All epithelial cells of distal convoluted tubules revealed basolateral expression of KCNQ1 K+ channel. On the contrary, only the single cells of collecting duct, probably intercalated cells, showed diffuse cell surface staining with antibodies against KCNQ1 K+ channel. These findings suggest that KCNQ1 K+ channel has cell-specific roles in renal potassium ion transport.

Changes in the balance between proliferation and apoptosis during the progression of malignancy in thyroid tumours.

The aim of this study was to gain better insight into molecular changes which reflect disturbances in the balance between proliferation and apoptosis during progression of thyroid malignancy from papillary microcarcinoma (PMC) via clinically manifest papillary carcinoma (PTC) to anaplastic carcinoma (ATC). The apoptosis related molecules (Bcl-2, Bax) and proliferation related marker (PCNA) were analysed immunohistochemically in 120 archival cases comprising PMC (n=34), PTC (n=52) and ATC (n=34). In addition, in situ apoptotic cell death was analysed by the TUNEL method. The average Bcl-2 staining score did not differ between PMC and PTC (p>0.05), but was significantly lower in ATC (p<0.05).The Bax score was higher in PTCs and ATCs than in PMCs (p<0.05). Due to these changes, the Bcl-2/Bax ratio showed a marked decrease from PMC to ATC (p<0.05), while proliferation activity increased significantly from PTC to ATC (p<0.05). Despite high Bax expression, the rate of apoptotic cell death was low in the investigated carcinomas, especially in ATC, i.e. the increase in proliferative activity was not counterbalanced with appropriate cell death. Differences were found in the expression of apoptotic molecules (Bcl-2 and Bax), their ratio (Bcl-2 /Bax) and in the rate of apoptotic cell death and proliferative activity between PMC, PTC and ATC, indicating that disturbances in the balance between apoptosis and proliferation, in favour of the latter, occur gradually during the progression of malignancy in thyroid tumours.

[The incidence rate of thyroid microcarcinoma during surgery benign disease].

Thyroid microcarcinoma are well-differentiated tumors less than 1 cm in diameter. A retrospective analysis was performed on patients operated of benign thyroid disease at the Center for endocrine surgery, Institute of endocrinology, Clical Center of Serbia in Belgrade, from January 1st to December 31st 2004, in order to establish the incidence of microcarcinoma. Indications for surgery were euthyroid multinodular goiter in 201 patients, thyroiditis in 31, thyroid adenoma in 178, Graves disease in 89 and Plummers disease in 79 patients. The results of this study, demonstrate that in 13.4% of the patients operated for goiter, 6.4% operated for thyroiditis, 5.6% for thyroid adenomas, 9.0% for Graves disease and 7.0% of the patients operated for Plumers disease, the presence of a microcarcinoma was noticed in the definitive histopathologic examination. The results obtained are in line with the current knowledge of high incidence of thyroid microcarcinoma.

The efficacy of the thyroid peroxidase marker for distinguishing follicular thyroid carcinoma from follicular adenoma.

AIM: Expression of thyroid peroxidase (TPO) in the thyroid gland tissue is well known as a sensitive marker of the thyroid malignancy. We have evaluated immunohistochemical assay of TPO for distinguishing follicular thyroid carcinoma from follicular adenoma. MATERIALS AND METHODS: Sections of formalin-fixed tissues obtained from 92 patients with thyroid tumors (52 follicular carcinomas and 40 follicular adenomas including the Hurthle cell type) were analyzed using a monoclonal antibody (TPO mAb 47) and the avidin-biotin peroxidase complex immunohistochemical technique. Lesions with staining of more than 80% of the follicular cells/specimen were considered benign, while less than 80% were considered malignant. RESULTS: TPO immunostaining correlated with the histopathological diagnosis in 24/40 cases of follicular adenomas and 41/52 cases of follicular carcinomas, giving a specificity of 60% and a sensitivity of 79%. CONCLUSION: These results suggest that immunohistochemical assay of TPO expression has limited value for the differential diagnosis of follicular thyroid carcinoma from thyroid follicular adenoma.

Galectin-3 expression in papillary microcarcinoma of the thyroid.

AIMS: Galectin-3 is a beta-galactoside binding protein, recently recognized as a promising molecular marker of thyroid malignancy. As reported in several studies, galectin-3 is highly expressed in papillary thyroid carcinoma, but its expression has not been investigated in papillary microcarcinoma, which is a variant of papillary thyroid carcinoma. METHODS AND RESULTS: Using a monoclonal antibody to galectin-3 and the avidin-biotin-peroxidase complex (ABC) immunohistochemical technique, we analysed galectin-3 expression in 63 cases of papillary microcarcinoma. The results showed immunohistochemical reactivity for galectin-3 in 51 (80.9%) cases. Intensity of staining varied from strong or moderate to weak. Galectin-3 localization was mostly cytoplasmic, but also membranous or nuclear in some cells. Immunohistochemical expression of galectin-3 was not found in 12 (19.1%) cases. Most galectin-3 negative microcarcinomas (10/12) were of the non-classical type, i.e. without papillary architecture. Neither the frequency nor the intensity of a positive reaction was related to tumour size. CONCLUSIONS: Galectin-3 gene is expressed at the protein level in most papillary microcarcinomas, although with slightly lower frequency than that reported for clinically evident papillary thyroid carcinoma. The presence of galectin-3 in clinically silent microcarcinomas may indicate that galectin-3 is not related to growth or aggressiveness of papillary thyroid microcarcinomas but rather plays some other role in thyroid tumour biology.

Galectin-3 and proliferating cell nuclear antigen (PCNA) expression in papillary thyroid carcinoma.

BACKGROUND AND AIM: To examine the relationship between galectin-3 and cell proliferation in thyroid tumor tissue. Galectin-3, a beta-galactoside binding protein, has recently been recognized as a promising molecular marker of thyroid malignancy, due to its high expression in thyroid carcinomas and absence from normal or benign thyroid tissue. However, its exact role in thyroid tumor biology is still unknown. PATIENTS AND METHODS: We examined the relationship between galectin-3 and cell proliferation by comparative immunostaining for galectin-3 and proliferating cell nuclear antigen (PCNA) in paraffin-embedded tissues from 126 cases of papillary thyroid carcinoma. RESULTS: Positive cytoplasmic immunostaining for galectin-3 was found in 115 (91.3%) cases. Nuclear staining for PCNA was detectable in 93 (74.4%) cases. A low level of PCNA staining (less than 10% positive cells) was found in 36 (28.6%) cases, moderate staining for PCNA (more than 10% but less than 30% positive cells) in 35 cases (27.8%), while highly increased PCNA expression (more than 30% positive cells) was found in 32 (25.4%) cases. Moderate or strong galectin-3 expression, found in 99 cases, was associated with highly increased PCNA staining in 28.3% of them but with no detectable PCNA expression in 24.3% of them. CONCLUSION: These results suggest that overexpression of galectin-3 is not clearly related to proliferative activity of papillary thyroid carcinoma cells as assessed by PCNA immunostaining.

[Endoscopic ultrasonography (EUS) in preoperative localization of neuroendocrine tumors (NET) of the pancreas].

BACKGROUND: Preoperative localization of pancreatic neuroendocrine tumours (NET) is usually very difficult. Noninvasive, imaging tests, such as abdominal ultrasound, CT or MRI are not sensitive enough as well as selective angiography. The aim of the study was to clarify the usefulness of the EUS in preoperative localization of the pancreatic NET. METHODS: From September 1998 March 2005, EUS was performed in 1600 patients. Among them, in 10 (0.7%), this examination was carried out due to previous biochemical tests, which diagnosed the pancreatic NET. We studied the location, the size and echo-pattern of the neoplasm. The results were compared with operation and histology or EUS- FNA guided pancreatic biopsy in 9/10 patients. All EUS examinations were performed using Olympus GIF-130 videoecho-endoscope with 7,5 /12MHz switchable radial probe. RESULTS: EUS correctly localized the pancreatic NET in 7/8 cases, (sensitivity:87.5%). In 2 patients, EUS accurately exclouded pancreatic NET. There were no false positive findings (specificity 100%). Six tumours were benign (75%), and two were malign (25%). We localized 6 insulinomas and single pancreatic carcinoid tumour. The median tumour size detected by EUS was 21mm. CONCLUSION: EUS is highly accurate in preoperative localization of the pancreatic NET-s and We confirmed it in our study. EUS presents the method of choice for preoperative localization of the pancreatic NET.

Tolerance to heat of rats treated by beta-blocker. Role of submaxillary salivary glands.

Effect of beta-adrenoceptor blockade on the changes in rectal temperature (RT), heat production (HP) and noradrenaline (NA) concentration in the submaxillary salivary glands (SMSG) of rats exposed to heat (45 degrees C) was studied. Propranolol (P) (15 mg/kg i.p.) decreased the tolerance to heat. The survival time of propranolol-treated (PT) rats was 30 min shorter. The temperature curve of control rats exposed to heat can be divided into three phases: a rapid rise in RT; a plateau (TP) and a prelethal increase. In PT animals, under identical conditions, TP disappears and RT further rises accelerating death. In the initial phase of heat exposure, HP was markedly decreased to the same extent in both experimental groups; but after 20 min HP increases in PT rats. The content of NA from SMSG both in the initial phase and in the TP phase is modified in PT rats.

[Immunohistochemical analysis of malignant tumors of the thyroid gland using 6 relevant markers]. / Imunohistohemijska analiza malignih tumora stitaste zlezde primenom sest relevantnih markera.

The results of immunohistochemical study of malignant thyroid tumours using six markers-thyroglobulin (TGB), keratin, carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), calcitonin and leucocyte common antigen (LCA)-are presented. Thirty-three cases of thyroid carcinomas were studied: 11 papillary, 8 follicular, 7 medullary and 7 anaplastic carcinomas. All cases of papillary and follicular tumours showed positive reaction with TGB and 4 papillary and 3 follicular carcinomas reacted with CEA also. 100% reactivity with calcitonin and CEA were observed in the medullary carcinoma group, while anaplastic carcinomas revealed TGB (one case), CEA (two) and EMA (three) reactivity. None of the studied tumours responded to LCA. TGB and calcitonin proved to be useful in distinguishing tumours originating in follicular cells from those originating in parafollicular cells and, along with LCA, enabled discrimination of anaplastic carcinomas from medullary carcinomas and malignant lymphomas. CEA can be only used as an auxiliary marker in cases of medullary carcinoma, while EMA and high molecular weight keratins did not prove to be reliable markers for analysis of malignant thyroid tumours.

Galectin-3 and carcinoembryonic antigen expression in medullary thyroid carcinoma: possible relation to tumour progression.

AIMS: Galectin-3 is a beta-galactoside binding protein involved in multiple biological processes through interactions with complementary glycoconjugates. We analysed the expression and coexpression of galectin-3 and carcinoembryonic antigen (CEA), one of the putative galectin-3 ligands, in medullary thyroid carcinoma (MTC). METHODS AND RESULTS: An immunohistochemical study using monoclonal antibodies was performed on paraffin sections of 20 cases of sporadic MTC comprising 10 cases without and 10 cases with lymph node metastases at the time of surgery. CEA expression was found in all tumours, distributed predominantly in the cytoplasm and occasionally at the cell surface. In the majority of cases (18/20) moderate to strong intensity of staining was found in most of the cells. Positive cytoplasmic staining for galectin-3 was found in 16/20 cases, but varied in intensity and distribution from weak/focal (7/16) to moderate (7/16) or strong (2/16). More intense staining for galectin-3 was mainly associated with MTC cases involving lymph node metastases. Eight out of these 10 cases showed moderate to strong galectin-3 expression concomitant with CEA expression throughout the tumour tissue. CONCLUSIONS: These findings suggest that galectin-3 might play a role in the pathobiology of MTC. Simultaneous expression of galectin-3 and CEA in the same tumour cells at an advanced stage of MTC indicates the possibility of their autocrine cooperation during tumour progression.

[Intercellular adhesion molecule-1 expression in renal cell carcinoma]. / Ispoljavanje intercelulne adhezivne molekule-jedan u karcinomu celija bubrega.

The presence of intercellular adhesion molecule-1 (ICAM-1) was analyzed using indirect immunoperoxidase technique in frozen sections of 27 renal cell carcinomas (RCC)--18 clear cell, 6 granular, 2 chromophobe and 1 sarcomatoid. Great variations in ICAM-1 expression were observed in clear and granular cell RCC, without correlation with mononuclear (lympho-monocytic) cell infiltration. Sarcomatoid type displayed widespread ICAM-1 distribution and intense mononuclear infiltrate, while chromophobic RCC, despite better prognosis, expressed ICAM-1 weakly and focally, almost without mononuclear infiltrate. With such results, no relation between ICAM-1 expression on tumor cells and behavior of various RCC types could be established and possible explanations are: different histogenesis of some RCC (chromophobe), nonspecific upregulation of ICAM-1 as a result of ischaemic-necrotic processes, or negative effect of ICAM-1 expression on complement action.

[Localised adenomyomatosis of the gall bladder]. / Lokalizovana adenomiomatoza zucne kese.

Adenomyomatosis of the gall bladder is a rare benign disease in which hypertrophic mucosa makes deep sinuses into hypetrophic muscular layer making, the so called Rokitansky-Aschoff's sinuses. It appears as a localised, segmental or diffuse form of the disease. Localised adnomyomatosis leads to the formation of "tumorous" lesions. Two patients, a 44-year old man and a 67-year old woman in whom "tumorous" lesions of the gall bladder were diagnosed preoperatively, are presented. Both patients were operated on. Cholecystectomy was carried out. "Tumorous" changes were, in fact, a localised form of adenomyomatosis, what was histologically confirmed.

[Anaplastic carcinomas of the thyroid gland]. / Anaplasticni karcinomi stitaste zlezde.

Anaplastic carcinoma of the thyroid gland is a tumor of great malignant potential. The majority of the patients die in the period of 6-24 months after diagnosis is made. A review of 90 cases of anaplastic carcinoma of the thyroid gland surgically treated in the Department for Endocrine Surgery of The Second Surgical Clinic and Center for Endocrine Surgery of the Clinical Centre of Serbia revealed that anaplastic carcinomas were 11.3% of all the thyroid carcinomas (796) diagnosed in the cited period. The patient's age varied from 40 to 80 (mean 63.7 years). The incidence in women is twice as large as in men (2.1:1). Intraoperative histopathologic diagnosis was made in all the patients. The variations in histological structure were noticed. Dominant histopathologic features included pleomorphic giant and spindle cells, with irregular hyperchromatic nuclei and many aberrant mitotic figures with necrosis and infiltration of blood vessels by neoplastic tissue in most of the cases.