Clinical, diagnostic and therapeutic features of keratocystic odontogenic tumors: a review.

Keratocystic odontogenic tumors (KCOTs) are benign but locally aggressive lesions of the gnathic skeleton with high propensity to recur following surgical treatment. High proliferative activity of KCOTs epithelial cells is considered as one of the factors contributing to their aggressive clinical behavior. Aggressive growth within the jaws, tendency to invade surrounding anatomical structures and occasional malignant alteration are the features that distinguish KCOTs from other types of odontogenic tumors. Due to their unique clinical and biological features, KCOTs still present an important problem in oral and maxillofacial surgery. This is especially true when a choice of the most appropriate treatment modality should be made. Establishing balance between effective reduction of recurrence risk and selection of a less aggressive surgical procedure is an issue that should be carefully considered for each individual patient.

The CD34-microvascular density in colorectal cancer patients.

PURPOSE: To investigate the influence of the angiogenesis parameter CD34 microvascular density (MVD) on overall survival of colorectal cancer (CRC) patients. METHODS: Thirty-one CRC patients were followed-up for 72 months after curative colorectal operation. Blood vessels measurement was done using the CD34-MVD immunochemistry method, and light microscopy. RESULTS: MVD was inversely correlated with patients' survival. MVD value < 35 proved as independent good prognostic factor, and patients with this value lived during the 72-month follow up after surgery, while a MVD value > 65 was an independent poor prognostic factor and such patients died within 11 months after radical surgery for CRC (p<0.01). CONCLUSION: According to these results, the CD34-MVD seems to be a significant prognosticator of overall survival in CRC patients.

Association of decreased NK cell activity and IFNγ expression with pSTAT dysregulation in breast cancer patients.

PURPOSE: Impaired IFNγ production in peripheral blood lymphocytes (PBL) and their subsets reflects immunosuppression and inadequate antitumor immune response in cancer patients. Decreased function of natural killer (NK) cells has not been investigated in breast cancer with respect to altered pSTAT signaling pathways. METHODS: PBL of breast cancer patients and healthy controls were analyzed for IFNγ and pSTAT1 expression and NK cell activity using flow cytometry and (51)Cr-release assay, respectively. The level of pSTAT1, 3 and 5 was investigated by Western blotting. RESULTS: Our results indicated that PBL and CD3(-) CD16(+) NK cells of patients had significantly lower level of IFNγ. The patients had a significantly decreased NK cell cytotoxicity compared to controls, with the decrease being dependent on the stage of disease. Positive correlation between IFNγ level in PBL and NK cytotoxicity in controls and patients was also shown. The PBL of patients, compared to controls, expressed lower level of pSTAT1, 3 and 5. The patients' T and NK cell subsets had lower pSTAT1 level. CONCLUSION: This study indicates that pSTAT1 in PBL of breast cancer patients could be a biomarker of decreased NK cell cytotoxicity and IFNγlevel that are associated with progression of this disease.

Ultrasonographic examination of the liver circulation in patients with liver metastasis from colorectal cancer.

PURPOSE: Conventional imaging modalities are presently recommended for the detection of liver metastases. However, the presence of liver micrometastases is a major diagnostic problem. It has been known that micrometastases could be associated with changes in the liver blood flow. METHODS: We examined several parameters by color Doppler ultrasound to estimate hepatic artery flow in 30 patients without and 17 patients with liver metastases from colon cancer. RESULTS: Mean values of hepatic artery diameter (4.25 + or - 0.81 mm in patients with liver metastases were not statistically different from those in patients without metastases (3.98 + or - 0.81). Patients with liver metastasis had significantly higher (p=0.007) mean values of systolic speed (61.33 + or - 30.01 cm/s) in comparison to patients without metastasis (41.38 + or - 16 cm/s). CONCLUSION: Based on these results we suggest that color Doppler examination can be an additional quick noninvasive method in the detection of circulatory changes in the estimation of liver metastases.

Specific skin lesions in hairy cell leukemia at presentation: case report and review of literature.

Skin involvement in hairy cell leukemia (HCL) at presentation is a relatively rare manifestation of the disease. A 60-year-old male patient in whom cutaneous lesions were the initial manifestation of hairy cell leukemia together with leukocytosis, monocytopenia, massive splenomegaly, and leukemic maculopapulous infiltration of the almost whole skin is described. The present case is the forth mentioned in the literature with specify of leukocytosis in peripheral blood, consisting mostly of hairy cells. The patient was treated with two courses of 2-chlorodeoxiadenosine (2-CdA, Cladribine) and splenectomy and after this cutaneous lesion disappeared and general condition is improved.

[Pathophysiological mechanism of the developing radicular cyst of the jaw].

The radicular cysts are result of inflammatory process in the periapical tissues associated with necrotic and infected pulps. Humoral and cellular immune responses play a central role in the pathogenesis of these lesions. The most important role in the growth of these lesion have proinflammatore cytokine TNF-alpha, IL-1 and IL-6. Cytokine can be secreted by macrophages, monocytes and other cells of the immune system and can participate in skeletal homeostasis including osteoclastic formation, and bone resorption in maxillofacial region. The aim of this study is to give a consise rewiew for mechanism of growth of maxilofacial radicular cysts, indicated of clinical aspect, as well as expalined role of cytokine in this pathophysiology process.

The concentration of TNF-alpha correlate with number of inflammatory cells and degree of vascularization in radicular cysts.

OBJECTIVE: To correlate values of tumor necrosis factor-alpha (TNF-alpha) depending on the count of inflammatory cells with degree of vascularization in cystic fluid of radicular cysts. MATERIAL AND METHODS: We investigated TNF-alpha concentration in 43 radicular cysts obtained from patients undergoing surgery, under local anaesthesia, and after aspiration of cystic fluid from non-ruptured cysts by enzyme-linked immunosorbent assay assay in respect of different clinical parameters as well as by histomorphometric analyses. RESULTS: Significantly higher concentration of TNF-alpha is associated with smaller radicular cysts, higher protein concentration in cystic fluid as well as with higher presence of inflammatory cells, and increased degree of vascularization in pericystic tissues and cyst wall thickness. CONCLUSIONS: We believe that determination of TNF-alpha in cystic fluid simultaneously with other parameters can be an additional parameter for clinical diagnosis of inflammed cysts.

[Comparison of clinical data, ultrasonography, mamography and histological findings obtained by ex tempore biopsy for breast cancer diagnosis].

Breast cancer is really one from tumor with highest mortality in this country as well as in the word. For early detection of the loco-regional breast cancer and disease confirmation, many diagnostic procedures were recommended with aims to help in decision for surgery treatment. This comparative study reported data by clinical analyses, ultra-sonography, mammography and histological findings obtained after ex tempore biopsy, in 59 women with breast cancer aged from 16 to 69. The highest incidence of cancer was obtained in women over 61 years of age, although high rate of breast cancer was found in aging between 21 to 50 years. The fibroadenoma were founded in 40.68%, while malignant carcinoma ductale in 13.55% of all cases. Correlation between applied analyses for fibroadenoma was found in 86.05% cases, while in malignant lesions this percentage was highest (93.33%). Apart of these methods applied for early diagnosis in general hospital and obtained statistical significance in diagnosis, new techniques based on molecular level (hormone receptor status) must be used.

Malignant histiocytosis with central nervous system involvement and hepatic mucinous cystadenoma in a single patient with review of the literature.

Malignant histiocytosis is a rare neoplasm of the reticuloendothelial system characterized by neoplastic proliferation of tissue histiocytes. We report a case of malignant histiocytosis in a 64-year-old female initially operated on for a mucinous cystadenoma of her liver. Four months after the operation, skin induration on the neck and anterior thoracic wall and systemic lymphadenopathy were noted. Histology and immunohistochemistry of the lymph node and bone marrow specimens showed extensive infiltration with atypical cells, resembling malignant histiocytes (CD45, CD45RO, CD11c, CD68, lysozyme, antitrypsin and alpha1-antichymotrypsin positive; CD1, CD35, B-cell and T-cells markers negative). She was treated with vinblastine, methotrexate and dexamethasone (3 cycles) without response. The therapy was switched to CHOP (cyclophosphamide, vincristine, doxorubicin, and prednisone) with disappearance of lymphadenopathy. Bone marrow infiltration by histiocytes was reduced to 20%. Two months after completion of 8 cycles of CHOP she experienced severe headaches, vomiting, loss of consciousness, and developed paraparesis. A CT scan of the brain was normal but the cerebrospinal fluid cytology showed presence of histiocytes. The patient was then treated with intrathecal methotrexate, prednisolone and cytosine-arabinoside and systemic chemotherapy with etoposide and cyclophosphamide. Her condition improved, she became conscious, her headache diminished, she became mobile but skin and nodal lesions reappeared along with extensive marrow histiocytic infiltration. She finally died 22 months after diagnosis.

[Doppler hemodynamic study in evaluation of liver metastases].

The color Doppler spectral analyses can be usefully in description of the liver circulation as one from non-invasive methods. Using color Doppler B mod ultrasound (Toshiba Core Vision) with duplex ultrasound we founded difference in liver circulation between patients with colon cancer and liver metastasis in comparison with those without metastases. We suggested this non invasive method for early detection of occult liver metastasis presence.

Bcl-2 expression in oral squamous cell carcinoma.

Apoptosis is a genetically regulated process involved in tissue size regulation, morphogenesis, and elimination of genetically damaged cells. A pallet of genes is involved in the control of apoptosis, such as bcl-2 family whose oncogenic potential has been demonstrated in oral tumorigenesis. Different members of bcl-2 family may promote or inhibit apoptosis by synthesizing anti- and proapoptotic proteins. One of antiapoptotic proteins, bcl-2, with a crucial role in apoptosis regulation was the object of our study. By means of immunohistochemistry we estimated the level of overexpression of bcl-2 proteins in a series of the 26 formalin fixed, paraffin-embedded samples of oral squamous cell carcinoma (OSCC). Analyzed tumors originated from different sites of oral cavity; 7/26 belonged to stage II, 14/26 to stage III, and 5/26 to stage IV. Immunoreactivity was scored according to the percentage and intensity of positive cytoplasmic bcl-2 staining. All tumors had low percentage of positively stained bcl-2 cells, with mean values for lower/higher intensity of 8.3 +/- 2.5/34.4 +/- 7, 7.5 +/- 1.1/31.9 +/- 4.3, and 8.4 +/- 5.8/31.5 +/- 5.8 within stages II, III, and IV, respectively. Low level of bcl-2 expression in our sample seems to be associated with higher survival rate: 77% for the 5-year follow-up period. Comparing clinicopathologic and risk factors data within each and between three groups of analyzed tumors (lip-tongue P = 0.58, tongue-floor of the mouth, P = 0.21, lip-floor of the mouth, P = 0.50) there was no significant difference. However, our results suggest that the level of bcl-2 expression could be a valuable predictor of tumor behavior and disease outcome.

Interferon alpha sensitisation induced fatal renal insufficiency in a patient with chronic myeloid leukaemia: case report and review of literature.

Renal dysfunction as a sequel to extended interferon alfa (IFNalpha) treatment in chronic myeloid leukaemia (CML) has been reported previously in six patients. An additional patient is presented with Philadelphia chromosome positive (Ph+) CML and nephrotic syndrome in whom initial renal insufficiency developed after only one month of low dose IFNalpha therapy. The renal biopsy showed a focal segmental mononuclear cell infiltration, basal membrane thickening, and deposits of immunoglobulins (IgG; IgAGM IC3). In spite of discontinuation of IFNalpha, renal function deteriorated and the patient died six months later. This case represents an instance of fatal kidney insufficiency as an untoward effect of sensitisation to the IFNalpha, confirmed by modified Coombs assay.

TNF-alpha induced changes in cell membrane antigen expression on K-562 cells associated with increased lactate dehydrogenase (LDH) release.

TNF-alpha is a pleiotropic cytokine, which induces death of sensitive cells, whose effect depend on cell membrane receptor expression, cell cycle phases, as well as on intracellular ratio of pro- apoptotic and anti-apoptotic molecule expression. Since determination of LDH release from cultured cells in vitro, reflects early membrane alterations, we estimated and compared LDH release from cultured cells with changes in cell membrane antigen expression on K-562 cells after TNF-alpha treatment by flow cytometry. The significant increase in LDH release activity and cytotoxicity values was associated with decrease in membrane molecule expression for CD45 and CD30 as well as for low expressed CD45RA and CD38 after TNF-alpha treatment. However, percentage of decrease of all examined molecules is not uniform, and appears to depend on the respective level of pre treatment values expression and molecule type. These results indicated the complexity of events on cell membrane, including association between increasing LDH release and decrease of antigen expression of membrane molecules following TNF-alpha mediated processes.

The comparison of spontaneous LDH release activity from cultured PBMC with sera LDH activity in non-Hodgkin's lymphoma patients.

Based on the fact that lactate dehydrogenase (LDH) enzyme is a very sensitive indicator of the cellular metabolic state, aerobic or anaerobic direction of glycolysis, activation status, and malignant transformation, in this study we compared values of the spontaneous LDH release from circulating PBMC with sera LDH activity in 53 different subtypes of non-Hodgkin's lymphoma (NHL) patients. Results shows that serum LDH was significantly (p < 0.05) elevated in comparison to the range values only in the advance clinical stage (III and IV) in all investigated subtypes of NHL according to The Working and REAL classification. On the other hand, the spontaneous LDH release from cultures PBMC is significantly (p < 0.01) elevated in early and advanced stage in all investigated forms of NHL in comparison to healthy controls. Based on consideration that an increase in spontaneous LDH release appears before elevated sera LDH activity, we conclude that determination of spontaneous LDH release by microassay from cultured cells together with other findings may help in the diagnosis of NHL patients, especially in patients with early stage of disease.

TNF-alpha induces changes in LDH isotype profile following triggering of apoptosis in PBL of non-Hodgkin's lymphomas.

Based on the possibility of tumor necrosis factor (TNF)-alpha to perform multiple and opposite biologic effects, we simultaneously investigated in vitro its effects on intracellular lactate dehydrogenase (LDH)-H and LDH-M isoenzyme activity and morphological characteristics following induction of apoptosis in peripheral blood mononuclear cells (PBMC) of non-Hodgkin's lymphoma patients (NHL) prior to and after the end of applied chemotherapy. TNF-alpha showed a significant increase ( p<0.05) of LDH-H and LDH-M activity in sonified PBMC of healthy controls after 18 h cultures accompanied with an increase of apoptotic index (AI) from 2.3 to 16.2%. Contrary to this, in PBMC of NHL patients prior to therapy TNF-alpha induced a significant decrease ( p<0.05) of LDH-H isotype activity. In patients after administration of chemotherapy, TNF-alpha in a dose of 100 U/ml induced a significant increase ( p<0.05) of LDH-M isotype activity, but not of LDH-H. In the PBMC of NHL patients prior to chemotherapy, TNF-alpha in vitro induced an increase of AI from 2.8 up to 6.8%, while in PBMC of NHL patients after applied chemotherapy AI changed from 7.2 to 14.4%. However, there was no significant difference in the increase of apoptosis in PBMC of NHL patients with high-grade malignancy and high rate response among patients who received first-line therapy, high-dose therapy, or third-line therapy regimens after in vitro TNF-alpha treatment. These results indicated different susceptibilities of PBMC of NHL to TNF-alpha when effects were analyzed by determination of intracellular LDH isotype profile and induction of apoptosis prior to and after administration of therapy in comparison to effects on healthy controls PBMC.

[Lactate dehydrogenase (LDH) in peripheral blood lymphocytes (PBL) of patients with solid tumors]. / Laktat dehidrogenaza za u limfocitima periferne krvi bolesnika sa solidnim tumorima.

Serum LDH level is a prognostic factor in different malignancies as its increase reflects tumor mass and response to therapy. Serum LDH is the consequence of the disruption of the cell membrane of a large fraction of dividing malignant cells whose metabolic hallmark is anaerobic glycolysis that leads to increased LDH enzyme activity. Moreover, as we have previously shown that spontaneous LDH release from cells represents a measure of cell membrane damage, and this parameter is used for the estimation of cell destruction in cytotoxic assays, the aim of this study was to evaluate the characteristics of LDH activity of PBMC of patients with different solid tumors (non-Hodgkin's lymphomas--NHL, n=47; Hodgkin's disease--HD, n=45; ovarian cancer--OvCa, n=6; breast cancer--BrCa, n=34; thyroid cancer--TyCa, n=3; cancer of PVU--CaPVU, n=4 and head & neck--H&N, n=6) in all clinical stages of NHL and HD and in advanced clinical stages of disease for BrCa, OvCa, CaPVU and H&N. Spontaneous LDH release from PBMC was determined by the spectrophotometric method from supernatants of 8 x 10(6)/ml PBMC cultured for 2 h in RPMI 1640 without phenol red using and LDH substrate mixture. The total LDH activity was determined after lysis of PBMC by ultrasound. The obtained results indicate that PBMC in all the investigated malignancies, compared to control PBMC, demonstrate a significant increase (p<0.01) in spontaneous LDH release act, which correlates with advanced clinical stage in all malignancies except in Hodgkin's disease, in which the spontaneous LDH release was increased in all clinical stages. Contrary to this, the total LDH activity was not increased in PBMC in all investigated tumors. However, the "percent of spontaneous LDH release" was always increased, regardless of the total LDH activity, indicating that spontaneous LDH release is the consequence of PBMC membrane damage present in advanced stages of different solid tumors.

Association of NK cell dysfunction with changes in LDH characteristics of peripheral blood lymphocytes (PBL) in breast cancer patients.

The cytotoxic activity of NK (natural killer) cells is very important in immunological surveillance against the appearance and especially the spread of malignant disease. The aim of this study was to investigate the function of this subpopulation of cells in breast cancer patients in different clinical stages of disease prior to therapy. NK cell activity was determined in breast cancer patients and healthy controls by three different methods: standard 51-chromium-release assay and by the original colorimetric uncorrected and corrected lactate dehydrogenase (LDH) release assay. A discrepancy was shown between the assays, as the uncorrected LDH assay showed, not only, much higher values, but no stage-dependent depression in NK cell activity compared to the chromium-release assay. Further analyses of separately cultured peripheral blood lymphocytes (PBL) revealed that this difference arose from an increasing, clinical stage-dependent, spontaneous LDH release from PBL of breast cancer patients. Furthermore, a stage-dependent increase in intracellular LDH activity of PBL was found, although without difference in LDH-H and LDH-M isotype ratio, compared to controls. Increased spontaneous LDH release and intracellular LDH activity was more evident in young patients, under 40 years. Correction of the original LDH-release assay for the spontaneous LDH release activity from PBL present in the assay, gave values of NK cell activity comparable to those determined by the chromium assay and indicated that breast cancer patients have a significant depression in NK cell activity which correlates with the stage-dependent increase in spontaneous LDH release. Moreover, as both assays measure the secretory, perforin-mediated, NK cell cytotoxic pathway against tumor cells, it can be concluded that the appearance of spontaneous LDH release is an indicator of cell membrane damage which not only allows the loss of LDH, but also of the components of the secretory killing pathway, resulting in NK cell dysfunction with the progression of disease. The novel findings obtained in this work reveal the association of PBL membrane damage with clinical stage of breast cancer that can, aside from reflecting NK cell depression, underlie the defect in other PBL subsets and subsequently facilitate progression of the malignant process.

Different alterations in lactate dehydrogenase activity and profile of peripheral blood mononuclear cells in Hodgkin's and non-Hodgkin's lymphomas.

The enzyme lactate dehydrogenase (LDH) activity of peripheral blood mononuclear cells (PBMC), LDH isotype H and M pattern and PBMC spontaneous LDH release activity were examined in 55 non-Hodgkin's lymphoma (NHL) patients, 46 Hodgkin's disease (HD) patients and 47 controls. The intracellular LDH and M isotype activity of PBMC, their spontaneous LDH release activity significantly increase (p < 0.01) in NHL with progressing histological grade of malignancy. Contrary to this, all classical HD patients have a significant elevation (p < 0.05) of each of these parameters. Furthermore, unlike HD, in NHL clinical stage is associated with significant (p < 0.05) increase in the level of spontaneous LDH release activity in each histological form. It is also shown that spontaneous LDH release activity of PBMC for HD and NHL patients demonstrates significant positive correlation (p < 0.005) with serum LDH level, although elevation of spontaneous LDH release precedes serum LDH increase in both diseases. The results obtained regarding alterations in intracellular, isotype and spontaneously released LDH activity of circulating PBMC show that these parameters are dependent, in NHL patients, on the grade of malignancy and tumor burden, while they are persistently present in HD patients.

A comparison of the NK cell cytotoxicity with effects of TNF-alpha against K-562 cells, determined by LDH release assay.

Effects of r h TNF-alpha as a single cytotoxic mediator against K-562 cells was examined by LDH release and compared with NK cell cytotoxicity. The mean values of the percentage of LDH release (x = 6.25 +/- 3.68%, for ten individual experiments) from K-562 cells cultured for 2 h with r h TNF-alpha 100 U/ml of culture medium did not give significant difference in comparison with mean values of percentage LDH release (x = 6.43 +/- 2.97%, for 37 individual experiments) from K-562 cells which were cultured without r h TNF-alpha (Student's t-test, P > 0.05). The results also showed, that in the presence of increasing concentrations of r h TNF-alpha there was no significant increase of LDH release through the cell membrane in these short term incubations. However, significant difference in LDH release from K-562 cells was found after 6 h between cultures treated for 30 min with or without r h TNF-alpha (Mann-Whitney test, P < 0.05). Since TNF-alpha alone shows a lower degree of K-562 cell membrane damage than NK effectors, this suggested that TNF-alpha is neither an only nor a major mediator of cell destruction, based on determination of LDH release.

The difference in NK-cell activity between patients with non-Hodgkin's lymphomas and Hodgkin's disease.

Natural killer (NK) cells play an important role in immune surveillance against malignant diseases. Considering the lymphoid origin of malignant lymphomas, as well as scarce data concerning NK-cell function in these neoplasms, we evaluated NK-cell activity in 49 patients with non-Hodgkin's lymphomas (NHL) and 47 patients with Hodgkin's disease (HD), prior to therapy. Using the recommended International Working Formulation and the Ann Arbor staging system for classification of lymphomas we found, by the LDH release cytotoxicity assay, that decreased NK-cell activity (P < 0.05) in NHL patients was essentially related to unfavourable histology (13 indolent lymphomas, 25 intermediate and 11 very aggressive lymphomas were included), but that within these categories clinical stage of the disease also contributed to the degree of NK-cell dysfunction. In contrast, in HD, NK-cell activity was persistently decreased (P< 0.05), compared to controls, irrespective of histological type and clinical stage. It is of interest also that the most profound NK-cell dysfunction that is present and persistent from the onset of HD, and which appears in very aggressive NHL was associated with the phenomenon of increased spontaneous lactate acid dehydrogenase (LDH) release activity from the separated PBMC of these patients. The difference in the level of NK-cell impairment between patients with various histological grades of malignancy in NHL and HD suggests different initial participation of innate immune reactions in these diseases.