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The early identification of aggressive forms of cancer is of high importance in treating papillary thyroid cancer (PTC). Disease dissemination is a major factor influencing patient survival. Mutation status of BRAF oncogene, BRAF V600E, is proposed to be an indicator of disease recurrence; however, its influence on PTC dissemination has not been deciphered. This study aimed to explore the association of the frequency of BRAF V600E alleles in PTC with disease dissemination. In this study, 173 PTC samples were analyzed, measuring the proportion of BRAF V600E alleles by qPCR, which was then normalized against the proportion of tumor cells. Semiquantitative analysis of BRAF V600E mutant protein was performed by immunohistochemistry. The BRAF V600E mutation was present in 60% of samples, while the normalized frequency of mutated BRAF alleles ranged from 1.55% to 92.06%. There was no significant association between the presence and/or proportion of the BRAF V600E mutation with the degree of PTC dissemination. However, the presence of the BRAF mutation was significantly linked with angioinvasion. This study's results suggest that there is a heterogeneous distribution of the BRAF mutation and the presence of oligoclonal forms of PTC. It is likely that the BRAF mutation alone does not significantly contribute to PTC aggressiveness.

Purpose: : colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, and detection of new prognostic markers is mandatory for patients to receive optimal oncological treatment. The aim of the study was to assess clinical and prognostic value of red cell distribution width (RDW) in patients with CRC., Methods: : RDW values in 90 patients with CRC undergoing surgery for primary disease were analyzed in pre- and postoperative setting, and correlated with clinical and hematological parameters., Results: : Both pre- and postoperative RDW measurements were found to be associated with features of iron deficiency anemia, inflammatory response to tumor, advanced age and depth of tumor invasion. Optimal cutoff points were calculated to be 14% for preoperative and 13.6% for postoperative RDW measurements. Elevations in both pre- and postoperative RDW values had significant effects on survival in univariate and multivariate analyses. Effects were found to be independent of tumor related features, stage of the disease, development of anemia and aberrant inflammatory response to tumor., Conclusions: : RDW is an integrative parameter reflecting tumor specific features and shows significant association with overall survival in patients with CRC. This is especially important in patients with stage 2 disease where elevation in preoperative RDW values can contribute to recognition of higher risk patients., (© The Author 2017. Published by Oxford University Press on behalf of the Association of Physicians. All rights reserved. For Permissions, please email: journals.permissions@oup.com)

Quality of life (QoL) is an important outcome in assessment of breast cancer treatment. Data comparing QoL after different adjuvant treatments and QoL data on long-term survivors are modest. The aim of this study was to compare QoL scores of patients receiving adjuvant treatment with long-term breast cancer survivors, and to correlate QoL scores with clinical data. Sixty patients were recruited for the study: 20 during adjuvant radiotherapy, 20 during adjuvant chemotherapy, and 20 long-term breast cancer survivors. QoL was assessed using the self-administered EORTC core questionnaire QLQ-C30 and breast cancer-specific module QLQ-BR23. QoL scores between groups were compared using Kruskal-Wallis test and effects of clinical factors on QoL domains were tested using multiple regression analysis. No differences between three groups were observed in terms of all QoL scores. As measured by QLQ-C30, least affected QoL scales were cognitive functioning, social functioning, and physical functioning in all three patients group, while insomnia and pain scales were the most detrimentally affected. Among the groups, the highest scores of global health status and other functional scales were in adjuvant chemotherapy group. Measured by QLQ-BR23, body image scale was most affected, while sexual functioning scale was minimally affected, in all three groups. Multiple regression analysis has shown that the patient age were the only statistically significant predictor for global health status scale, and constipation scale. Our results demonstrated similar and favorable QoL in all three groups of patients and provided basic information on QoL in Croatian breast cancer patients.

The aim of the study was to determine the diagnostic value of reverse transcriptase polymerase chain reaction (RT-PCR) analysis of galectin-3 and CD44v6 as markers for preoperative diagnosis of malignancy in lesions of the thyroid. RT-PCR analysis of galectin-3 and CD44v6 expression was performed on RNA isolated from fine-needle aspirates of thyroid lesions from 428 patients. The results were evaluated against the postoperative histopathological diagnosis or definitive cytological diagnosis in cases of nodular goiter and Hashimoto thyroiditis. A total of 57 (13%) samples were inadequate for RT-PCR. Galectin-3 and CD44v6 were positive in 167 (45%) and 158 (43%) out of 371 adequate samples, respectively. Galectin-3 and CD44v6 were positive in 56 (86%) and 54 (83%) out of 65 papillary carcinomas, in 16 (29%) and 18 (32%) out of 56 Hashimoto's thyroiditis, in 61 (34%) and 52 (29%) out of 181 nodular goiters, in 23 (43%) and 23 (43%) out of 53 follicular adenomas, in 3 (100%) and 3 (100%) out of 3 follicular carcinomas, and in 8 (62%) and 8 (62%) out of 13 Hurthle cell adenomas, respectively. Specificity, sensitivity, and positive and negative predictive values in discriminating between malignant and benign thyroid nodules were 64, 87, and 35 and 96% for galectin-3; 67, 84, and 36 and 95% for CD44v6; and 79, 82, and 47 and 95% for the analysis of both markers (considered positive only if both galectin-3 and CD44v6 were positive), respectively. Owing to relatively low specificity, the clinical value of galectin-3 and CD44v6 analysis by RT-PCR as a marker for preoperative diagnosis of malignancy in thyroid lesions is limited.

The aim of this study was to analyze microphthalmia-associated transcription factor (MITF) as a marker for the detection of circulating melanoma cells, determine its prognostic value in melanoma patients, and compare it with tyrosinase. Blood samples from 201 melanoma patients in all stages of the disease and 40 healthy volunteers were analyzed. RNA was isolated from mononuclear cell fraction of the blood and assayed by reverse transcription-PCR for the expression of MITF and tyrosinase. All samples from healthy volunteers were negative for both MITF and tyrosinase. Out of 201 blood samples from melanoma patients 32 were positive for MITF, 20 for tyrosinase, and four for both MITF and tyrosinase. Analysis of MITF as an additional marker to tyrosinase allowed for detection of circulating melanoma cells in a larger number of melanoma patients in comparison to tyrosinase analysis alone (48 vs. 20 positive). A positive value of MITF was associated with shorter progression-free (P=0.005) and overall survival (P=0.042). A positive value of tyrosinase was associated with shorter overall survival (P=0.012), whereas there was no significant association between the value of tyrosinase and progression-free survival. The value of MITF was selected with multivariate analysis as the independent prognostic factor for progression-free survival, whereas the only independent prognostic factor for overall survival was the stage of disease. This study has shown that MITF is a specific marker for detection of circulating melanoma cells that has a prognostic value in melanoma patients. Determination of MITF in addition to tyrosinase improved the detection of circulating melanoma cells in melanoma patients.

We investigated if the use of two tumor markers, galectin-3 and CD44v6, could improve diagnostic accuracy of thyroid fine needle aspiration biopsy (FNAB) in cytologically indeterminate lesions (CIL). 351 patients with CIL [cellular follicular lesion/suspicious follicular neoplasm/suspicious Hürthle cell neoplasm (CFL/sFN/sHCN), Hürthle cell neoplasm (HCN), and follicular neoplasm (FN)] and surgical follow-up were investigated. 251 patients had FNAB diagnoses made without help of tumor markers and the rest of 100 patients had FNAB diagnoses made with a known expression of tumor markers determined by the reverse transcription (RT)-PCR. Risk of malignancy in all 351 patients with CIL was 6.8%. In the group with FNAB made without RT-PCR, there were 140 CFL/sFN/sHCN with the risk of malignancy of 4.2%, 92 FN with the risk of malignancy of 13.0%, and 19 HCN with the risk of malignancy of 5.2%. In the group with FNAB made with RT-PCR, there were 49 CFL/sFN/sHCN with the risk of malignancy of 2.0%, 40 FN with the risk of malignancy of 7.5%, and 11 HCN with the risk of malignancy of 9.0%. In the group with at least one positive tumor marker (N = 69), the risk of malignancy was 3.1% for CFL/sFN/sHCN, 11.1% for FN, and 10.0% for HCN. In the group with negative tumor markers (N = 31) there were no malignancies. The use of tumor markers, galectin-3 and CD44v6, determined by RT-PCR improves only sensitivity of thyroid FNAB in CIL. In most patients with CIL, and negative both tumor markers, conservative approach is advisable.

Allogeneic hematopoietic stem cell transplantation (alloHSCT) is the only potentially curative treatment available for patients with B-cell chronic lymphocytic leukemia (B-CLL). Here, we show that post-alloHSCT antibody repertoires can be mined for the discovery of fully human monoclonal antibodies to B-CLL cell-surface antigens. Sera collected from B-CLL patients at defined times after alloHSCT showed selective binding to primary B-CLL cells. Pre-alloHSCT sera, donor sera, and control sera were negative. To identify post-alloHSCT serum antibodies and subsequently B-CLL cell-surface antigens they recognize, we generated a human antibody-binding fragment (Fab) library from post-alloHSCT peripheral blood mononuclear cells and selected it on primary B-CLL cells by phage display. A panel of Fab with B-CLL cell-surface reactivity was strongly enriched. Selection was dominated by highly homologous Fab predicted to bind the same antigen. One Fab was converted to immunoglobulin G1 and analyzed for reactivity with peripheral blood mononuclear cells from B-CLL patients and healthy volunteers. Cell-surface antigen expression was restricted to primary B cells and up-regulated in primary B-CLL cells. Mining post-alloHSCT antibody repertoires offers a novel route to discover fully human monoclonal antibodies and identify antigens of potential therapeutic relevance to B-CLL and possibly other cancers. Trials described herein were registered at www.clinicaltrials.gov as nos. NCT00055744 and NCT00003838.

Background: Analysis of different tumor markers by reverse transcriptase-polymerase chain reaction (RT-PCR) in fine-needle aspiration samples of thyroid nodules has been studied with the objective of improving the accuracy of the preoperative diagnosis of thyroid lesions. The aim of the current study was to investigate thyroid fine-needle aspiration samples inadequate for RT-PCR analysis and to determine whether there is a correlation between their proportion and the method of sampling used or the greatest dimension of the nodules., Methods: A total of 350 fine-needle aspiration samples from patients with thyroid nodules were analyzed. After the aspirate was smeared for conventional cytology, the leftover material in the needle was used for RT-PCR analysis in 1 group of 175 patients. In another group of 175 patients, a separate puncture was performed to obtain material for RT-PCR analysis only. Samples were considered adequate for RT-PCR analysis if the expression of both glyceraldehyde-3-phosphate dehydrogenase and thyroglobulin was found by RT-PCR., Results: In total, 61 (17.4%) samples inadequate for RT-PCR were detected. All 12 samples that were inadequate for cytologic diagnosis were also found to be inadequate for RT-PCR analysis. The proportion of inadequate samples for RT-PCR was found to be significantly higher in samples taken from leftover material in the needle (21.7%) then in samples from a separate puncture (13.1%) (P = .049). No statistically significant correlation between the adequacy of samples for RT-PCR and the largest dimension of the nodule was found., Conclusions: The proportion of samples inadequate for RT-PCR was found to be higher in samples taken from leftover material in the needle than in samples obtained from a separate puncture., ((c) 2008 American Cancer Society.)

Detection of circulating melanoma cells by reverse transcriptase - polymerase chain reaction (RT-PCR) is a molecular diagnostic procedure which is used to predict prognosis in melanoma patients. The most widely used specific marker for detection of circulating melanoma cells by RT-PCR is expression of tyrosinase gene. This procedure has shown high specificity and low threshold for detection of melanoma cells. Most of the studies have shown that prognosis is worse in patients in which circulating melanoma cells were detected. Detection of circulating melanoma cells has been studied also as a marker for predicting response to therapy. The clinical value of this procedure is limited by the proportion of patients with clinically confirmed distant metastases being tyrosinase negative in almost all the studies. Studies have shown that analysis of additional markers to tyrosinase enables detection of circulating melanoma cells in a higher percentage of melanoma patients. RT-PCR has shown a lower threshold for detection than other methods (immunohistochemistry) in detection of melanoma metastases in lymph nodes.

Objective: To investigate whether the presence of macrophages and Hürthle cells (HC) in benign thyroid lesions could explain the false-positive expression of galectin-3 and CD44v6 detected by reverse transcriptase-polymerase chain reaction (RT-PCR)., Methods: For galectin-3 and CD44v6, RT-PCR was performed on RNA isolated from aspirates obtained by ultrasound guided fine needle aspiration cytology (FNAC) from 123 patients with benign thyroid lesions. The results of RT-PCR analysis were evaluated against the definitive FNAC diagnosis., Results: Galectin-3 expression was found in 29% follicular adenoma (FA), 26% Hashimoto thyroiditis (HT), and in 24% nodular goitre (NG). We found a statistically significant relationship between the presence of macrophages and galectin-3 positivity in NG and HT samples (P < 0.0001 and P = 0.0087 respectively). We found a statistically significant (P = 0.0219) relationship between the presence of HC and galectin-3 positivity in HT and a tendency of such a relationship (P = 0.0838) in NG. CD44v6 expression was found in 29% FA, 33% HT and in 18% NG. We found a statistically significant relationship between the presence of HC and positive expression of CD44v6 in NG (P = 0.0003) and a strong tendency of such a relationship in HT (P = 0.0571). We did not find a statistically significant relationship between the presence of macrophages and CD44v6 positivity. In FA, we did not find a statistically significant relationship between the presence of macrophages or HC and galectin-3 or CD44v6 positivity., Conclusion: Our results suggest that the presence of macrophages and/or HC may explain the positive expression of galectin-3 and CD44v6 detected by RT-PCR in HT and NG cytological samples.

Sclerosing epitheloid fibrosarcoma is a rare, histologically well-defined member of adult fibrosarcoma group of soft tissue tumors. Its main histological features are nests and cords of rounded tumor cells surrounded by hyalinized collagenous stroma. Epitheloid appearance with marked sclerosis and infiltrating growth pattern, along with occasional immunohistochemical positivity for epithelial markers may be highly suggestive of infiltrating carcinoma. Despite of bland cytological features clinical course is often protracted with a high local recurrence rate and late metastases. In this report, we present histopathological characteristics of two cases of sclerosing epitheloid fibrosarcoma, together with their clinical presentation, follow-up information and differential diagnosis.

Aim: To investigate whether analysis of microphthalmia-associated transcription factor (MITF) as an additional marker to tyrosinase in melanoma patients can improve the detection of circulating melanoma cells by reverse-transcription-polymerase chain reaction (RT-PCR)., Methods: Blood samples were taken from 33 patients with metastatic melanoma. RNA was isolated from mononuclear cell fraction of the blood and reversely transcribed into the complementary DNA (cDNA). The cDNA was assayed by PCR for the expression of tyrosinase and MITF. Peripheral blood samples from 15 healthy subjects were used as controls., Results: The threshold for detection of both tyrosinase and MITF was set low enough to detect 50 melanoma cells in 10 mL of healthy volunteer blood in the relative ratio of one melanoma cell in 0.82 x 106 peripheral blood leukocytes. Out of 33 blood samples from metastatic melanoma patients, 5 were positive for both tyrosinase and MITF, 7 for tyrosinase only, and 5 for MITF only. All samples from healthy volunteers were negative for both tyrosinase and MITF., Conclusion: Determination of MITF marker in addition to tyrosinase improved the detection of circulating melanoma cells in patients with metastatic melanoma.

Aim: To detect the expression of genes encoding tyrosinase, gp100, MART-1/Melan A, and tyrosinase-related protein-2 (TRP-2) in peripheral blood of melanoma patients by reverse transcription-polymerase chain reaction (RT-PCR)., Methods: Nineteen peripheral blood samples were obtained from 17 melanoma patients. When tested, 15 of them presented with clinically detectable metastatic disease. Samples of peripheral blood (7 mL) were collected from each patient into vacutainer cell preparation tubes. Mononuclear cells were isolated, total cellular RNA extracted, and then used as a template for reverse transcription to complementary DNA (cDNA). The cDNA was thereafter assayed by PCR for the expression of melanocyte-associated transcripts of tyrosinase, gp100, MART1/Melan-A, and TRP-2 genes., Results: Gp100 gene expression was detected in 13 out of 19 samples. In 4 of them, TRP-2 gene expression was also detectable. Expression of tyrosinase and Melan-A/MART-1 genes could not be observed. Interestingly, gp100 and TRP-2 gene transcripts were detected in patients having recurrent and/or metastatic disease at the time of testing., Conclusion: The results we obtained support the use of RT-PCR assay for indirect detection of melanoma cells in peripheral blood of melanoma patients. As the transcripts for the tyrosinase gene and MART-1/Melan A gene were not detected, additional optimization experiments of RT-PCR assay are required.