The prognostic value of E-cadherin and Ki-67 compared to standard histopathologic examination in non-muscle invasive bladder cancer.

OBJECTIVES: The objectives of this study were to determine the prognostic value of expression levels of selected biomarkers and their statistical analysis in relation to survival and standard histopathologic examination and other clinicopathologic variables in non-muscle invasive bladder cancer (NMIBC). BACKGROUND: Worldwide, bladder cancer is a frequent malignant disease with rising incidence. Characteristic invasiveness and high recurrence rates call for more diagnostic methods to obtain more accurate information. Prognosis is affected by a significant interpersonal variability of the disease. For this reason, constant search for alternative and better diagnostic methods is essential. METHODS: We analysed cancer tissue from patients with Ta and T1 bladder cancer. E-cadherin and Ki-67 expression levels were analysed using immunohistochemical staining. The expression levels quantified to a percentual amount were statistically analysed in relation to survival and their frequency distribution in the study group. RESULTS: E-cadherin and Ki-67 expression levels show high association with tumor stage and grade         (p<0.001), in contrast, the association with recurrence has proven insignificant. Patients with non-aberrant biomarker expression levels have much higher survival rates than the cases with aberrant expression. CONCLUSION: Low expression levels of Ki-67 and high expression levels of E-cadherin positively affect survival of patients, whereas aberrant expressions pose poorer prognosis (Tab. 2, Fig. 2, Ref. 33).

Bone metastases in neuroendocrine tumors.

Neuroendocrine tumors arise from various cells that form a part of the endocrine system and account for a small number of cases encountered by oncologists in clinical practice. The clinical incidence of these tumors used to be low, and newer imaging modalities have now begun to be used for detecting bone metastases at an earlier stage. Bone metastases arising from neuroendocrine tumors are a well-recognized complication. Their presence carries along a poor prognosis. Clinical symptoms are similar to those encountered in other forms of cancer that are complicated by bone metastasis. Over the last decade or so, the clinical detection, diagnostic methods and treatment strategies have changed dramatically, and new treatments are emerging slowly. The indolent course of neuroendocrine tumors and the development of bone metastasis have limited our current knowledge on how to best prevent and manage the condition. Current information available from clinical studies is marred by paucity and small sample sizes, making further clinical trials an absolute necessity. In this review, we discuss the current status in the diagnosis and management of bone metastases arising from neuroendocrine tumors (Fig. 3, Ref. 28).

[Benign solitary cecal ulcer]. / Benígny solitárny vred slepého creva.

Solitary benign cecal ulcer is a non-usual finding without characteristic clinical or radiological performance. Because ofits little frequency and its clinical symptoms often suspicious for the presence of severe diseases like are malignancies, acute abdomen or lower gastrointestinal bleeding, this finding is only rarely diagnosed preoperatively. Definitive diagnosis is almost always made pathologically. In this case report we describe clinical, radiological and pathological findings in our patient with difficulties caused by a solitary benign cecal ulcer.

[Ductal carcinoma in situ (DCIS) and biopsy of the sentinel lymph node]. / DCIS a biopsia sentinelovej uzliny.

INTRODUCTION: Ductal carcinoma in situ (DCIS) is the disease with increasing incidence. Nowadays, approximately 80% DCIS are diagnosed via mammography and represent more than 20% of all types of breast cancer. The acceptance of surgical procedures with this type of breast carcinoma is controversial as primary diagnosis of non-invasive carcinoma is often underestimated and in the end, histopathological examination reveals invasive carcinoma with biological potential to metastasize. In cases of "risk" patient groups with DCIS, several studies report lymph node metastases. The aim of the study has been to assess the incidence of sentinel lymph node metastatic involvement in high-risk patient group with DCIS and in ductal carcinoma in situ with microinvasion (DCISMI), to note the incidence of invasive carcinoma in definitive histopathology in patients with pre-operative diagnosis of DCIS and to analyze some predictors of invasivity. STUDY TYPE AND PATIENT GROUP: In retrospective analysis, we evaluated the setting of 119 patients who have been operated on at our Clinic from January, 1st 2008 until December, 31th 2010 for the diagnosis of DCIS. Prospectively, we have created the setting of 44 patients with high-risk DCIS with sentinel lymph node biopsy (SLNB) performed. METHODS AND RESULTS. Metastatic involvement of sentinel lymph node in high-risk DCIS has been found in 4 cases (9.0%)--in 1 patient (2.2%) with correct diagnosis of DCIS and in 3 patients (6.8%) with invasive carcinoma according to final histopathology. In the patient with DCIS, a micrometastasis of 0.4 mm was found in one sentinel lymph node. After complete axillary dissection, non-sentinel axillary lymph nodes metastatic involvement was not demonstrated (14/0). In 6 cases (5.0%), we identified DCISMI and did not find metastasis in sentinel lymph node. In the high-risk DCIS group, in 4 patients (9.0%) DCISMI and in 12 patients (27.2%) invasive carcinoma was found after definitive histopathologic examination. In this group, the overall ratio of invasive lesions was 36.2%. As for predictors of invasivity, high-grade carcinoma (OR 4.2; 95% CI 1,40-12,58) has more than 4-fold higher influence and lesion size

Detection of circulating tumor cells in metastatic breast cancer patients.

OBJECTIVE: The objective of this study was the detection of circulating tumor cells (CTCs) in metastatic breast cancer patients. METHODS: Since only small numbers of circulating tumor cells (CTCs) are found in peripheral blood, at first we performed immunomagnetic separation as a concentration method suitable for selecting circulating tumor cells in peripheral blood. This was followed by analysis of isolated cells with the aid of laser scanning cytometry (LSC). Twenty eight patients with metastatic breast cancer were enrolled in the study and the control group consisted of 19 clinically healthy women. Six milliliters of peripheral blood was drawn for the analyses, but only in two patients the blood has been drawn twice. Blood samples were taken when no chemotherapy was administered, but hormonal therapy has been allowed. RESULTS: The positivity for CTCs was found in 20 (50.0 %) patients with metastatic breast cancer patients, while in 6 (31.6 %) healthy controls false positive circulating epithelial-like cells were detected. Because we did not use CD45 staining, we could not distinguish these circulating epithelial-like cells from CTCs. In a majority of metastatic breast cancer patients we found a mixed population of HER-2 gene expressing CTCs. We found that HER2+ CTCs in high numbers are CK19 + CTCs, while almost all HER2-CTCs are CK19- CTCs. CONCLUSION: The described method was found promising for estimating HER2 status on CTCs from peripheral blood in metastatic breast cancer patients.

[Significance of the sentinel lymph node biopsy in early breast carcinomas]. / Význam biopsie sentinelovej uzliny pri vcasnom karcinóme prsníka.

We present our experience regarding sentinel lymph node biopsy (SLNB) at St. Elizabeth Institute of Oncology during 48 months. From January 1st, 2006 until December 31st, 2009, we had performed SLNB in 269 patients. Primary tumour size was 0.3-3.5cm including non-invasive breast carcinoma (i.e. TIS, T1 and T2 of TNM classification). Invasive carcinoma accounted for 255 (94.8%) cases, while non-invasive carcinoma for 14 (5.2%) cases. From total of 269 patients with invasive carcinoma, we used validation method in 157 (72.7%). In 255 patients with invasive carcinoma, sentinel node was not identified in 4 (1.6%) cases--in 1 patient with T1 invasive carcinoma and in 3 patients with T2 tumours. False negativity of sentinel node in T1 tumours was 4.3%. The incidence of macrometastases in sentinel nodes was confirmed using standard histopathologic examination with hematoxylin-eosin stain. In negative instances, the examination was then completed with serial sections and immunohistochemistry using cytoskeletal antibodies for confirmation of presence of micrometastases. In 6 (2.4%) cases, we found micrometastase in originally negative sentinel lymph node. Subsequent axillary dissection has not confirmed non-sentinel nodes involvement.

Chromatin texture, DNA index, and S-phase fraction in primary breast carcinoma cells analysed by laserscanning cytometry.

OBJECTIVES: Laser scanning cytometry (LSC) is a slide-based technique capable of measuring a number of biological parameters both in immobilised cell suspensions and in formalin-fixed paraffin-embedded tissue sections. BACKGROUND: High proliferation rate in surgically removed breast tumours is an unfavourable prognostic factor. In node negative cases it can help distinguish patients with higher risk for distant metastases from those with a lower risk. PATIENTS AND METHODS: In a prospective study we investigated 140 breast tumours, of which 113 were invasive ductal carcinomas, 11 were invasive lobular carcinomas, and 16 tumours were of other histological types. Cells for LSC investigations were prepared from fresh, surgically removed tumours by mechanical disintegration. After fixation the cells were stained with FITC-conjugated anti-cytokeratin (CK-FITC) to distinguish CK+ tumour cells from CK- stroma, and with propidium iodide to stain DNA. RESULTS: We identified three S-phase fraction (SPF) groups, with low (30 patients), moderate (54 patients), and high SPF (51 patients). Thirty-seven tumours were diploid, 83 were aneuploid, while 5 tumours had a bimodal distribution of DNA content. Chromatin texture values were increasing in the respective subclasses from the hypodiploid group to the tetraploid/hypertetraploid group. CONCLUSION: The measurement of DNA content and SPF of tumours by LSC completed by and correlated with other biological properties of the tumour cells may be a useful tool in assessing prognosis and clinical outcome of patients with breast cancer. (Tab. 5, Fig. 4, Ref. 18). Full Text (Free, PDF) www.bmj.sk.

Advanced detection and measurement of cells on membrane from peripheral blood by laser scanning cytometry (LSC) in early stage breast cancer patients.

BACKGROUND: The aim of our study was the potential detection of circulating tumour cells (CTCs) in early stage breast cancer patients. Our approach was cell microfiltration through polycarbonate membrane as a concentration method suitable for CTC selection in peripheral blood. The isolated cells on membrane were further analysed by laser scanning cytometry. METHODS: Sixteen patients were enrolled in the study, of which 13 had early stage breast carcinoma and 3 patients had metastatic breast carcinoma. The analyses were performed from 9 ml of peripheral blood, in one patient blood was drawn twice. Blood samples were taken after adjuvant chemotherapy but prior to adjuvant radiotherapy. The control group consisted of 12 clinically healthy subjects. CONCLUSIONS: In the control group 3 subjects out of 12 had 1 CTC, the mean CTC numbers being 0.25 +/- 0.45. In the early stage breast cancer patients 0-36 CTCs were detected (mean 13.9 +/- 12.9 CTCs. 10 patients out of 13 had more than 2 CTCs (62%). The detection and measurement of cells on membrane is a simple and reproducible method of detection of CTCs in peripheral blood. Sensitivity of the method is 88.5%. Detection of CTCs seems to be a promising method for the monitoring of adjuvant therapy in early stage breast cancer patients and for the identification of high risk patients in whom elevated numbers of CTCs are persisting following the termination of adjuvant therapy (Tab. 1, Fig. 4, Ref. 35). Full Text (Free, PDF) www.bmj.sk.

Radionavigated detection of sentinel nodes in breast carcinoma--first experiences of our department.

INTRODUCTION: Biopsy and histological evaluation of sentinel lymphatic node limits the axillary node dissection only in cases of positive histological finding and decreases the occurrence of postoperative complications related to the axillary node dissection. METHODS: We used radiotracer SentiScint, Medi-Radiopharma Ltd, Hungary and preoperatively administered blue dye--Blue Patenté V, Guebert, Aulnay-Sous-Bios, France. 11 (18%) patients were subdued to deep peritimorous application of radiotracer, 10 (16.4%) to sub/intradermal application over the lesions and n 40 (65.6%) patients the application was sub/intradermal and periareolar. The patients underwent an operation protocol of corresponding quadrantectomy, radionavigated blue-dye sentinel node biopsy and axillary dissection. RESULTS AND CONCLUSIONS: From May 2006 to June 2008, we examined 61 patients with breast carcinoma. They underwent radionavigated and blue-dye sentinel node biopsy. We detected 57 (93.4%) sentinel nodes with preoperative scintigraphy, of which only 51 (83.6%) were detected peroperatively and underwent histological evaluation. In six (9.8%) cases, the "frozen cut" histology of the primary lesion had shown a benign lesion; hence no sentinel node biopsy or axillary disection was performed. 12 (19.7%) of 51 histologically evaluated sentinel nodes had metastatic invasion. We retrospectively compared the histological fund in sentinel and axillary nodes in patients with metastatic sentinel nodes. In 6 (16.6%) cases, the sentinel node was positive of metastatic invasion but axillary nodes were histologically negative, in 6 (16.6%) cases the sentinel node and axillary nodes were positive for metastatic invasion. We observed falsely negative findings in 3 (8.3%) patients with negative histological fund in the sentinel node, but positive axillary nodes (Tab. 3, Fig. 2, Ref. 11). Full Text (Free, PDF) www.bmj.sk.

Synaptophysin negative central neurocytoma.

BACKGROUND: Central neurocytoma is a rare primary brain tumour, mostly localised in the lateral ventricles in relation to the foramen of Monro. OBJECTIVES: To report a case of a rare central neurocytoma with a complete loss of Synaptophysin expression and provide the differential diagnosis. METHODS AND RESULTS: We describe a case of a 34-year old man with a headache, unsteady gait and dim vision. MRI demonstrated a tumorous expansion localised in both lateral ventricles. The patient underwent a subtotal resection. Histology showed a picture consistent with central neurocytoma, but tumour was completely negative for Synaptophysin. We describe our approach in such a diagnostically difficult case. CONCLUSIONS: In the rare case of Synaptophysin-negative central neurocytoma, its neuronal differentiation should be substantiated by electron-microscopic examination. Unfortunately in the routine work, biopsy samples are usually fixed in formalin fixative which does not preserve ultrastructure well. In such situations, an accurate diagnosis is disputable and based on careful assessment of the histological features, exclusion of tumours with similar morphology and detailed correlation with MRI pictures (Fig. 4, Ref. 6). Full Text (Free, PDF) www.bmj.sk.

Recurrent malignant epitheloid schwannoma of the lower lip.

The authors describe a case of recurrent malignant epitheloid schwannoma of the lower lip. Histologically, the tumor was composed of fibroblast-like spindle cells in compact fascicles and areas of epitheloid growth, combined with demonstration of S-100, GFAP and NF positivity, which is characteristic for this type of tumor. The therapy consisted of a combination of surgery and radiotherapy and the patient was followed-up since the disease was diagnosed. A local re-operation had to follow the first surgical intervention consisting of a radical excision of tumor in the lower lip together with suprahyoid neck dissection six months later. After the first operation, the patient received a radiation therapy with a total dosage of 12 Gy in seven fractions to the tumor area of the lower lip. After the second operation, an external radiotherapy with total dosage of 50 Gy was applied. Despite the complex intensive therapy, the patient died of metastases into lungs, liver and spine 37 months after the initiation of the therapy (Fig. 2, Ref. 17). Full Text (Free, PDF) www.bmj.sk.

Genetic analysis of KRAS mutation status in metastatic colorectal cancer patients.

Colorectal carcinoma (CRC) represents a serious problem worldwide: in the Slovak republic are diagnosed about 2600 new CRC cases annually and its incidence is increasing. Colorectal cancer patients may succumb to the disease because of local recurrence or local formation of metastasis. Therefore, it is necessary to modulate therapeutic algorithm with new methods, leading to early diagnostic of CRC or changing the existing therapeutic procedures. Recent progresses have been made in understanding of EGFR pathway involved in CRC carcinogenesis, especially the role of Ras protein. Mutations in KRAS oncogene are frequently found in human cancers, particularly colorectal, pancreatic, billiary tract and lung tumors. The presence of the KRAS mutations in metastatic colorectal cancer patients correlates with lack of response to the certain epidemal growth factor receptor (EGFR) inhibitor therapies, such as Panitumumab and Cetuximab. Consequently, screening for KRAS mutations status may be used as a prognostic marker, because the CRC patients with KRAS positive tumors have a worse prognosis. The aim of our study was to establish the methods for rapid and sensitive detection of KRAS mutation status in formalin fixed paraffin embedded (FFPE) tissues DNA. We applied Real Time PCR analysis (TheraScreen KRAS Mutation Test Kit) and sequencing analysis (optimised for the analysis of FFPE tissues) to detect somatic mutations in codon 12 and 13 of KRAS gene. Both methods were used concurrently in the panel of DNA isolated from 25 colorectal FFPE tissues tumor. The positive or negative results from all 25 samples were identified by both methods independently. The KRAS mutations were presented in 8 of 25 patients (32%). Our results demonstrate that the Real Time PCR analysis can be used for detection of somatic KRAS mutations in FFPE clinical samples. However, we also recognize that the sequencing analysis of approximately 200bp amplicons may be used for mutations status screening, but with care of method sensitivity.

Laser scanning cytometry (LSC) in pathology--a perspective tool for the future?

Cytometry is becoming a standard method of examination not only in biology but also in various fields of experimental and clinical medicine. While in flow cytometry suspensions of cells are measured, laser scanning cytometers enable both the measurement of cells in single-cell suspensions (after immobilising the cells on a conventional glass slide) and in frozen or paraffin-embedded tissue sections. We discuss the possible fields of utilisation and future perspectives of laser scanning cytometry in medicine with special reference to clinical pathology and cytology (Fig. 3, Ref. 49). Full Text (Free, PDF) www.bmj.sk.

Thyroid tumors: histological classification and genetic factors involved in the development of thyroid cancer.

Classification of thyroid tumours and their variants is described with special respect to some recent findings on somatic mutations characteristics which are associated with individual types of malignity. Special attention is paid to the interrelations between thyroid nodules and malignity and predictive risk factors are listed.

Semi-quantitative RT-PCR assessment of molecular markers in breast large-core needle biopsies.

Large-core needle biopsies are frequently used for the preoperative evaluation of the breast lesions. In addition to initial diagnostic information, they can show the status of molecular markers with predictive and prognostic value and further contribute to an optimal selection of treatment strategy. So far, the potential use of large-core needle biopsies in assessment of marker profile of the breast lesions was studied using the immunostaining approaches. In this work, we sought to determine whether analysis of the large-core needle biopsy by semi-quantitative reverse-transcription polymerase chain reaction reveals molecular data that correspond with the marker status of the subsequently removed tumor. Five molecular markers including ER, PR, c-erbB-2/HER-2/neu, c-erbB-3/HER-3, and CD44 were assessed on mRNA isolated from 23 large-core needle biopsies and corresponding surgical breast cancer specimens using beta2- microglobulin as an internal standard. Significant or highly significant correlation between core biopsies and excised tumors was observed for each marker when the mRNA expression status was scored as positive or negative, with the concordance of the data ranging from 73.9% to 86%. Using the dichotomous scoring, majority of the biopsies (75%) displayed molecular profile that was either identical to the profile of the related tumor specimen, or with the difference in one marker. However, no significant correlation was found when the levels of the markers were expressed as continuous variables, possibly due to intratumoral cell heterogeneity. These results suggest potential usefulness and reliability of semi-quantitative RT PCR in the evaluation of large-core needle biopsies with regard to marker positivity or negativity. On the other hand, the marker-related data expressed as continuous variables cannot be accurately assessed on the large- core needle specimens using this approach, indicating the need for methodological improvements.

Hepatoprotective effect of rooibos tea (Aspalathus linearis) on CCl4-induced liver damage in rats.

Hepatoprotective properties of rooibos tea (Aspalathus linearis) were investigated in a rat model of liver injury induced by carbon tetrachloride (CCl(4)). Rooibos tea, like N-acetyl-L-cysteine which was used for the comparison, showed histological regression of steatosis and cirrhosis in the liver tissue with a significant inhibition of the increase of liver tissue concentrations of malondialdehyde, triacylglycerols and cholesterol. Simultaneously, rooibos tea significantly suppressed mainly the increase in plasma activities of aminotransferases (ALT, AST), alkaline phosphatase and billirubin concentrations, which are considered as markers of liver functional state. The antifibrotic effect in the experimental model of hepatic cirrhosis of rats suggests the use of rooibos tea as a plant hepatoprotector in the diet of patients with hepatopathies.

Heterotopic pancreas in gastric antrum with macroscopic appearance of gastric polyp.

Heterotopic pancreas is a relatively rare clinical diagnosis, not commonly involved in differential diagnostic considerations of GI symptoms. The authors report a case of heterotopic pancreas discovered endoscopically in the gastric antrum. A 60-year-old woman presented with epigastric pain. The patient took alendronate for osteoporosis. The endoscopic examination revealed Helicobacter pylori positive antral atrophic gastropathy and a well delineated hemispherical polyp, 8 mm in diameter, in the antrum of the stomach. Histology showed antral gastritis and the presence of heterotopic pancreas. After dietary measures and Helicobacter eradication, the patient was relieved of symptoms. According to the authors' opinion, the finding of heterotopic pancreas did not necessitate intervention, and was an incidental finding. The authors discuss the significance of heterotopic pancreas with the conclusion that the resection of the lesion is indicated only if consistent symptoms are present. Fig. 3, Ref. 28.)

Serum markers of liver fibrogenesis, and liver histology findings in patients with chronic liver diseases.

AIM OF THE STUDY: Investigation of the relationships between the grade and stage of chronic liver diseases irrespective of their etiology using some novel serum markers of liver fibrogenesis, the "classical" serum markers of liver necro-inflammatory injury (such as transaminases), and the histomorphological evaluation of liver biopsies. METHODS: Markers of liver fibrogenesis: serum metalloproteinase 1 (MMP-1), tissue inhibitor of MMP-1 (TIMP-1), and N-terminal propeptide of the procollagen III (PIIINP); "liver function tests" (LFTs): bilirubin, transaminases ALT, AST; ALP, GMT; and liver morphology findings: necro-inflammatory activity, fibrosis; were studied in the series of 32, 'naive', i.e. yet untreated patients (women/men--11/21) with various CLDs: chronic viral hepatitis B or C 13 (CHB 3, CHC 10), non-alcoholic steatohepatitis 9, liver steatosis 4, primary biliary cirrhosis 5, drug-induced hepatitis. The diagnoses were based on the clinical, laboratory and liver imaging (ultrasonography) findings and confirmed by the liver biopsy. CONCLUSIONS: Investigation of liver fibrogenesis serum markers (PIIINP, MMP-1, TIMP-1) in patients with various CLDs has shown statistically significant correlations of these parameters with "classical" serum markers of liver necro-inflammation (ALT, AST) and the results of histomorphological evaluation of the necro-inflammatory activity (parameters NAI, MEF) and fibrosis (parameter FI) in liver biopsies. (Tab. 4, Ref. 31.)