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Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serological tests have been suggested as an additional diagnostic tool in highly suspected cases with a negative molecular test and determination of seroprevalence in population. We compared the diagnostic performance of eight commercial serological assays for IgA, IgM, and IgG antibodies to the SARS-CoV-2 virus. Materials and methods: The comparison study was performed on a total of 76 serum samples: 30 SARS-CoV-2 polymerase chain reaction (PCR)- negative and 46 SARS-CoV-2 PCR-positive patients with asymptomatic to severe disease and symptoms duration from 3-30 days. The study included: three rapid lateral flow immunochromatographic assays (LFIC), two enzyme-linked immunosorbent assays (ELISA), and three chemiluminescence immunoassays (CLIA). Results: Agreement between IgM assays were minimal to moderate (kappa 0.26 to 0.63) and for IgG moderate to excellent (kappa 0.72 to 0.92). Sensitivities improved with > 10 days of symptoms and were: 30% to 89% for IgM; 89% to 100% for IgG; 96% for IgA; 100% for IgA/IgM combination; 96% for total antibodies. Overall specificities were: 90% to 100% for IgM; 85% to 100% for IgG; 90% for IgA; 70% for IgA/IgM combination; 100% for total antibodies. Diagnostic accuracy for IgG ELISA and CIA assays were excellent (AUC ≥ 0.90), without significant difference. IgA showed significantly better diagnostic accuracy than IgM (P < 0.001). Conclusion: There is high variability between IgM assays independently of the assay format, while IgG assays showed moderate to perfect agreement. The appropriate time for testing is crucial for the proper immunity investigation.

Anastomotic dehiscence is one of the most serious complications following surgery for colorectal cancer, and early detection of anastomotic dehiscence is critical to minimize mortality and morbidity. The aim of this study was to determine the value of peritoneal interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) as markers for preclinical detection of anastomosis dehiscence following colorectal surgery. Concentrations of IL-6 and TNF-α were measured in drain fluid obtained from 58 patients on days 1 to 4 following surgery for colorectal cancer. Five out of 58 patients developed anastomosis dehiscence. Patients who developed anastomosis dehiscence had significantly higher concentration of IL-6 on day 1 after surgery, and TNF-α on day 1, 2 and 4 after surgery. Interleukin-6 on day 1 was predictive for anastomosis dehiscence with specificity of 83%, sensitivity of 80%, positive predictive value (PPV) of 31% and negative predictive value (NPV) of 98%. TNF-α was predictive for anastomosis dehiscence on day 1 (specificity 92%, sensitivity 80%, PPV 50%, NPV 98%), day 2 (specificity 94%, sensitivity 80%, PPV 57%, NPV 98%), and day 4 (specificity 83%, sensitivity 100%, PPV 27%, NPV 100%). Our study indicates the potential use of peritoneal cytokines IL-6 and TNF-α as additional diagnostic tool for early detection of anastomosis dehiscence following colorectal surgery.

Objectives: Cytokeratin 20 (CK20) is one of the most investigated markers for the detection of circulating colorectal cancer (CRC) cells by reverse transcription polymerase chain reaction (RT-PCR). The aim of this study was to evaluate prognostic value of RT-PCR detection of circulating CRC cells using CK20 as a marker, and to compare the value of preoperative and postoperative blood sample analysis for that purpose. Methods: Ribonucleic acid (RNA) was isolated from mononuclear cell fraction of blood samples taken from 95 CRC patients before and after tumor resection and from 23 healthy volunteers and assayed by real-time RT-PCR for CK20 expression. Results: In patients positive for CK20 postoperatively both progression-free survival (PFS) and overall survival were significantly shorter than in patients negative for CK20 postoperatively, while the difference between patients positive and negative for CK20 preoperatively was not statistically significant in terms of neither PFS nor overall survival. Conclusion: Our results have shown prognostic value of circulating cancer cells detected in postoperative blood samples from CRC patients using CK20 as marker for RT-PCR, which has potential implications for treatment of these patients. In clinical practice, CK20 expression profile could be a factor in weighting treatment options in CRC patients. In cases where multiple treatment options are possible, patients with positive postoperative CK20 expression could be candidates to receive more aggressive treatment.

There is a great interest in molecular markers that would help in the preoperative diagnosis of malignant thyroid nodules in cases of indeterminate fine-needle aspiration cytology. The aim of this study was to determine the diagnostic accuracy of HMGA2 gene expression in discriminating benign from malignant thyroid nodules. In this study, 237 preoperative thyroid fine-needle aspiration samples were analyzed prospectively for the expression of the HMGA2 gene by real-time reverse transcription polymerase chain reaction. The results were evaluated against the postoperative histopathologic diagnosis or definitive cytologic diagnosis in cases of nodular goiter and Hashimoto thyroiditis. Among 237 samples from patients with thyroid nodules that were analyzed, 231 were adequate for real-time reverse transcription polymerase chain reaction analysis. With a cutoff value of 8.71 for relative gene expression, HMGA2 was positive in 19 (16.4%) of 116 nodular goiter, 1 (2.6%) of 39 Hashimoto thyroiditis, 9 (28.1%) of 32 follicular adenoma, 0 (0%) of 5 Hurthle cell adenoma, 32 (88.9%) of 36 papillary carcinoma, and 3 (100%) of 3 follicular carcinoma samples. In discriminating between malignant and benign thyroid nodules, HMGA2 has shown specificity of 84.5%, sensitivity of 91.9%, positive predictive value of 53.1%, and negative predictive value of 98.2%. High sensitivity and negative predictive value of HMGA2 for preoperative detection of malignant thyroid nodules shown in this study indicate that it may have a role as an ancillary marker in cytology in the management of patients with thyroid nodules.

Usage of complementary and alternative medicine (CAM) is steadily increasing over the last decades, gaining medical, economic and sociological importance. Th e aim of the present study was to assess the use of complementary and alternative therapies in cancer patients. A cross-sectional, descriptive survey design was used to collect data through an anonymous questionnaire. A total of 267 patients were included in the study. The prevalence of CAM use among cancer patients in this study was 60.3%. It was found that 61 heterogeneous CAM therapies were used, the most popular among patients being naturopathy/folk medicine. In multivariate logistic regression analysis, independent predictors of CAM use were high income, divorced status, female sex and younger age. In conclusion, considering the fact that a large proportion of patients used at least one CAM approach, we need to continue our efforts to improve the patient-oncologist communication in order to deliver most reliable information to patients and to better understand the possible standard medicine-CAM interactions. According to results of the latest studies, CAM therapies that help manage pain, nausea, fatigue, anxiety, and other symptoms should be integrated into the patient overall care.

Background Detection of circulating cancer cells by reverse transcription polymerase chain reaction (RT-PCR) has been studied as a prognostic marker in patients with colorectal cancer (CRC) but so far with conflicting results regarding specificity and prognostic value. In this study cytokeratin-20 (CK20) was evaluated by real-time RT-PCR as a marker for circulating CRC cell detection and the influence of surgical tumor resection on the presence of circulating CRC cells was analyzed. Methods RNA was isolated from the mononuclear cell fraction of blood samples taken from 95 CRC patients before and after tumor resection and from 23 healthy volunteers and assayed by real-time RT-PCR for CK20 expression. Results Among 23 healthy volunteers one was positive for CK20. Among 95 CRC patients, 25 were positive for CK20 before and 23 after surgery. Sixteen patients positive before surgery became negative after surgery, while 14 patients negative before surgery became positive after surgery. An increase in the proportion of CK20-positive samples with increasing stage of disease was observed for preoperative but not postoperative blood samples. Conclusions Its association with clinical stage indicates that CK20 might have prognostic value as a marker for detection of circulating CRC cells. Surgical tumor resection can both reduce and induce the presence of circulating CRC cells.

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.

Protein connexin 43 (Cx43), a part of intercellular gap junctions, is frequently down-regulated in tumors. The aim of the study was to compare Cx43 expression in primary colorectal tumors of patients with stage III and IV disease.Immunohistochemical expression of Cx43 was analyzed in 50 colorectal adenocarcinomas from surgically-treated patients of stage pT3N1-2 without metastases (M0) and 50 specimens of the same pTN stage from patients with synchronous liver metastases (M1). Association of Cx43 expression with clinicopathological factors and tumor site was also analyzed.There was no significant difference in Cx43 expression between M0 and M1 tumor specimens (p=0.817), as well as in Cx43 expression between colonic and rectal tumors (p=0.116), respectively. Stromal expression of Cx43 was higher in M1 than in M0 tumors (p=0.004).Stromal Cx43 expression is possible indicator of metastatic potential of colorectal adenocarcinoma.

Objective: To investigate whether the presence of macrophages and Hurthle 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.

Prisutnost aktivirajućih mutacija gena BRAF je prediktivni biljeg za liječenje bolesnika s metastatskim ili inoperabilnim melanomom lijekovima koji ciljano djeluju na mutirani BRAF (vemurafenib, dabrafenib). Aktivirajuće mutacije gena BRAF koje su prisutne u 50-60% melanoma mogu se odrediti različitim metodama, koje se najčešće temelje na metodi real-time PCR ili na sekvenciranju gena.

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.

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.

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-polymerase chain reaction 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). Positive value of MITF was associated with shorter progression-free (P =0.005) and overall survival (P =0.042). 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. Value of MITF was selected with multivariate analysis as independent prognostic factor for progression-free survival, whereas the only independent prognostic factor for overall survival was 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.

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. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.

Primary prevention and early detection of cancer are areas of growing interest in oncology as they present the most powerful weapon in fighting cancer and the curability rates increase with earlier disease detection. Breast cancer is one of the most often diagnosed tumors and one of the biggest public health issues and importance of high-risk individuals detection has been recognized decades ago. Such high-risk individuals are candidates for earlier and more frequent implementation of breast cancer screening methods. To recognize high-risk women, it is necessary to individually investigate the presence or absence of all risk factors known up to date and express them as a single number which will present one's risk for breast cancer development. PrOnDi (Preventive Oncology Diagnostics) questionnaire is designed to detect high risk persons for tumors of 18 different localizations. It consists of 84 questions in total and 18 of those are related to breast cancer risk factors. All the anwers to the PrOnDi questionnaire are linked to certain relative risk value, connected to a specific breast cancer risk factor. As a person completes the questionnaire, all the relative risks for breast cancer are summarized according to the carefully designed formula and a person is being classified into one of three possible risk groups (traffic-light concept): red-high risk (breast cancer risk is significantly higher compared to general population) ; yellow-moderate risk (risk is moderately higher than in general population) ; green-low risk (risk is comparable to the one in general population). In the pre-pilot testing of the PrOnDi questionnaire, 59 women with previously diagnosed breast cancer were given the questionnaire. The results were very encouraging: 98, 3% of patients were classified in the moderate (28, 81%) or high risk (69, 49%) groups, and only 1, 7% were classified in the 'green' group. According to these results, PrOnDi questionnaire could present a mighty tool in breast cancer prevention, early diagnosis and detection of high risk individuals. Of course, prior to definite conclusions, a survey on much bigger number of healthy individuals must be performed, but preliminary results are very optimistic and we might have a valuable tool for the future of oncology.