Correspondence of cytological and histopathological diagnoses in diagnostic category V of the Bethesda system: "suspicious for malignancy".

The progress in imaging methods enables fine needle aspiration (FNA) biopsy to be performed on smaller and smaller lesions, including malignant ones (papillary microcarcinomas). The follicular variant predominates in this group, with cytological features often not permitting an unbiased interpretation. The aim of the study was to determine the degree of reliability of the "suspicious for malignancy" (SM) diagnosis in material from the Institute of Oncology in Gliwice (IO). 290 primary SM diagnoses were established from 2010 to 2015 in the IO, including the consultations. None of the patients was treated surgically after the first FNA resulting in diagnostic category V (DC V). After the second FNA 80 patients underwent surgery, after the third 58, and after subsequent FNA 10. Together, 148 surgical resections were performed. Among 148 patients treated surgically, 111 were diagnosed with malignant lesions, which constitutes 75%. Predominantly - in 91 cases - the histopathological outcome was papillary carcinoma. The others were: 16 medullary carcinomas, 2 follicular carcinomas, and 2 poorly differentiated carcinoma cases. Moreover, 8 follicular adenomas and 28 nonneoplastic lesions were found. The high positive predictive value (PPV = 75%) of SM diagnosis established in the IO testifies to the high reliability of this test. Diagnostic category V in FNA should be an indication for surgical treatment.

The diagnosis of cancer in thyroid fine needle aspiration biopsy. Surgery, repeat biopsy or specimen consultation?

Fine needle aspiration biopsy (FNA) is the only diagnostic method that allows a preoperative diagnosis of thyroid carcinoma. An unequivocal diagnosis of a malignant change is achievable only in cases in which all cytological criteria of carcinoma are met. The aim of the study was to evaluate the necessity of repeat thyroid FNA in patients with papillary thyroid carcinoma verified on consultative examination (CE). We analyzed cytology reports of thyroid FNA and CE that resulted in the diagnosis of papillary carcinoma. Evaluation of the correlation of the cytological diagnosis with the histopathology report was based on data obtained after the surgery. Between 2010 and 2015 in the Institute of Oncology (IO) there were 184 cancers diagnosed on CE or in thyroid FNA performed primarily in IO. Additionally, 74 patients were subjected to repeat biopsy after confirmation of cancer in CE. Histopathological diagnosis of cancer was obtained in 62 (100%) cases that were doubly confirmed with cytological examination. The remaining 12 patients were operated on outside the institute. From 110 FNA primarily performed in the IO, histopathological verification was achievable in 92 cases, from which 92 (100%) provided a confirmation of cancer, and the remaining 18 patients were operated on outside the institute. High (100%) specificity of cancer diagnosis in FNA established primarily and verified on CE (second independent assessment) indicates that repeat FNA in order to confirm the diagnosis is unnecessary.

A novel quantitative method of pten expression assessment in tumor tissue.

Phosphatase and Tensin Homolog deleted on chromosome 10 (PTEN) gene is one of the most important tumor suppressor genes which is involved in the regulation of many signaling cascades (AKT/PKB and MAPK). Subtle changes in its activity lead to cancer susceptibility or aggressive tumor behaviour. Despite the diversity of mechanisms leading to PTEN inactivation, it is frequently associated with a decreased or complete loss of protein expression. About 20% decrease in PTEN expression could lead to the development of cancer. There have been no objective, quantitative methods of PTEN expression assessment that allow to measure the subtle variations of the protein concentration in a tissue-contextual manner. A new quantitative algorithm of immunostaining evaluation based on combination of color deconvolution and relative chromogen signal intensity was used in the study. The proposed algorithm was implemented in the popular ImageJ image analysis software and positively verified in cancer cell lines and tissue models as well as in the tissue samples of colorectal cancer (CRC) patients. The proposed quantitative method of PTEN expression assessment creates an alternative to currently available subjective methods and forms the basis for inter-case and inter-tissue comparisons. Using the algorithm it would be possible to identify three groups of patients with advanced colorectal cancer which could significantly differ in the overall survival. The research should be continued.

Does human papilloma virus participate in colorectal carcinogenesis?

Colorectal cancer is one of the most commonly diagnosed neoplasms still associated with relatively high mortality. Viral infections are often mentioned among the neoplasm transformation risk factors. Incidence of human papilloma virus (HPV), associated with high oncogenic risk, in the large intestine and the meaning of its presence in the colorectal carcinogenesis are still not clear. The aim of the study was to show a presence of HPV in specimens of adenomatous polyps and colorectal cancer using the Q-PCR method. Fifty patients (32 M/18W, mean age 62.8 years) were enrolled in the study, for whom tissue samples were obtained. Study material involved paraffin blocks derived from samples collected by flexible sigmoidoscopy from 10 polyps and 10 large intestine adenocarcinomas and 30 paraffin blocks with specimens of surgically removed large intestine adenocarcinomas. Presence of HPV genome was confirmed by quantitative PCR method using commercially available Abbott RealTime High Risk HPV test. The test is able to detect 14 most prevalent high oncogenic risk subtypes of human papilloma virus. Status of HPV DNA was successfully assessed in all 50 samples. No HPV DNA was discovered in any of the tested samples. Presence of high oncogenic risk HPV subtypes in large intestine adenoma and adenocarcinoma seems to be very rare, and its dominating role in the pathogenesis of colorectal cancer, even if possible, is unlikely.

Influence of local peripheral temporary ischaemia on biochemical and histological effects in small intestine and serum of rats following abdominal irradiation.

The local temporary ischaemia effect on radiation-induced lipid peroxidation, superoxide dismutase isoenzyme activities, and intestinal crypt number was estimated in male WAG-strain rats in vivo. The animals were irradiated in the abdomen area with doses of 2 Gy for ten consecutive days using a Philips 60Co source. The calculated dose rate was 0.595 Gy/min. Local temporary ischaemia was induced by clamping the tail base before each irradiation. The parameters evaluated were: TBA-RS level and enzymatic activities of CuZnSOD, MnSOD in serum and jejunum. The number of jejunum crypts was assigned as a histopathologic parameter. The results showed a clear protection by ischaemic preconditioning for crypt survival. The difference in the number of crypts in irradiated animals with and without local temporary ischaemia was statistically significant (Student's t-test P < 0.05). Also, significant enhancement of TBA-RS was observed in the serum of irradiated animals. Local temporary ischaemia application diminished the concentration of radiation- induced TBA-RS. The differences in the levels of TBA-RS in the serum were statistically significant (ANOVA P < 0.002). In contrast, there was no evident effect on the level of TBA-RS in tissue homogenates in any investigated groups. Some fluctuation of CuZnSOD isoenzyme activity in intestinal tissue was noted; however, the differences were not significant. Local temporary ischaemia had no influence on Mn- SOD activity in serum, and in both irradiated groups the behaviour of this isoenzyme was similar. Also, there were no differences in MnSOD activity measured in tissue homogenates. These findings support results of our previous in vivo studies, suggesting that local temporary ischaemia can prevent oxidative effects of fractionated radiotherapy.

Intracellular distribution of Photofrin in malignant and normal endothelial cell lines.

Compared to current treatments including surgery, radiation therapy, and chemotherapy, PDT offers the advantage of an effective and selective method of destroying diseased tissues without damaging surrounding healthy tissues. One of the aspects of antitumour effectiveness of PDT is related to the distribution of photosensitizing drugs. The localization of photosensitizers in cytoplasmic organelles during PDT plays a major role in the cell destruction; therefore, intracellular localization of Ph in malignant and normal cells was investigated. The cell lines used throughout the study were: human malignant A549, MCF-7, Me45 and normal endothelial cell line HUV-EC-C. After incubation with Ph cells were examined using fluorescence and confocal microscopy to visualize the photosensitizer accumulation. For cytoplasm and mitochondria identification, cells were stained with CellTracker Green and MitoTracker Green, respectively. Distribution of Ph was different in malignant and normal cells and dependent on the incubation time. The maximal concentration of Ph in two malignant cell lines (A549 and MCF-7) was observed after 4 hours of incubation, and the most intensive signal was observed around the nuclear envelope. Intracellular distribution of Ph in the Me45 cell line showed that the fluorescence emitted by Ph overlaid that from MitoTracker. This indicates preferential accumulation of the sensitizer in mitochondria. Our results based on the mitochondrial localization support the idea that PDT can contribute to elimination of malignant cells by inducing apoptosis, which is of physiological significance.

DNA ploidy assessment method applied to primary breast carcinoma FNA biopsies.

The goal of our study was to assess suitability of FNA biopsy material as a source of samples (cell suspension) for DNA ploidy assessment in neoplastic tumors using flow cytometry. DNA ploidy is an established prognostic factor in many types of cancers. Aneuploid breast tumors are characterized by increased aggressiveness which manifests itself through rapid local progression and metastatic spread. Investigated specimens were breast cancer FNA biopsy cell suspensions. Measurements were performed using flow cytometry. Material studied comprised 143 cases analyzed in 1999-2000. We found in this group 101 carcinoma cases with aneuploid type and 42 cases of primary breast carcinoma with diploid type of cell cycle. Immunocytochemical assesssment of estrogen receptor and progesterone receptor status was performed in group of 105 cases. DNA ploidy was compared to receptor status of the investigated cells. DNA aneuploidy correlated with weak or no reaction for the presence of estrogen and progesterone receptors. Our study demonstrates the suitability of DNA ploidy assessment method applied to cytological material from FNA biopsies.

Effect of phosphoramidon on brain tissue angiogenesis in the chronic phase of vasospasm after subarachnoid hemorrhage in the rat.

BACKGROUND: Subarachnoid hemorrhage (SAH) frequently leads to prolonged cerebral vasospasm resulting in vascular pathology due to endothelial cell ischemia and neuronal hypoxia. Posthemorrhagic vasospasm can be counteracted by the administration of phosphoramidon, which blocks the endothelin converting enzyme (ECE) responsible for the conversion of big endothelin into a fully active ET1 peptide. The aim of the study was to determine the effect of chronic vasospasm after SAH on angiogenesis and the effect on this process of endothelin-1, the main causative factor in vasospasm. MATERIAL AND METHODS: Male Wistar rats were examined. Seven days after cannulation of the cisterna magna, blood was administered to induce SAH. The ECE inhibitor phosphoramidon was administered in a dose of 40 nmol in 50 microl of cerebrospinal fluid three times: 20 min before SAH, 60 min after SAH, and 24 hours after SAH. The brains were removed 48 hours later for histological evaluation. The vascular surface density was measured in cerebral hemisphere sections (at the level of the dorsal part of the hippocampus) and brainstem sections (1/2 of the pons). CONCLUSION: Increased angiogenesis was observed in the cerebral hemispheres after SAH. The administration of phosphoramidon inhibits angiogenesis in cerebral hemispheres after SAH.

Cerebral angiogenesis after subarachnoid hemorrhage (SAH) and endothelin receptor blockage with BQ-123 antagonist in rats.

The aim of the study was to determine the effect of chronic vasospasm after SAH on angiogenesis and the effect of endothelin-1, the main causative factor in vasospasm, on this process. Male Wistar rats, 220-250 g, were examined. Seven days after cannulation of the cisterna magna (CM), a 100 microl dose of non-heparinized blood was administered to induce SAH. Sham SAH (aSAH) was induced by intracisternal injection of 100 microl of artificial cerebrospinal fluid. Endothelin receptor antagonist BQ-123 in a dose of 40 nmol in 50 microl of cerebrospinal fluid was given three times: 20 min. before SAH and aSAH, 60 min and 24 hours after SAH and aSAH. The same pattern of BQ-123 administration was used in the nonSAH group. The brains were removed 48 hours later for histological evaluation. Vascular surface density was measured in cerebral hemisphere sections (at the level of the dorsal part of the hippocampus) and brain stem sections (1/2 of the pons). An increase in angiogenesis was observed after SAH, compared to control values. The administration of BQ-123, a specific endothelin receptor blocker inhibits angiogenesis in cerebral hemispheres after SAH.