Local inflammatory response in colorectal cancer.

Type and intensity of tumor-infiltrating lymphocytes (TILs) in close proximity to the primary tumor are prognostically significant in postoperative patients. High intensity of TILs is considered to be a prognostically beneficial factor. The research included 66 postoperative colorectal cancer patients. The control group comprised 20 colon segments. Monoclonal antibodies LCA, CD3, CD4, CD5, CD8, CD20, CD23 and CD138 were used to differentiate between T and B lymphocytes. Types of cells in the infiltrate were defined. We found greater numbers of T and B lymphocytes located in close proximity to the cancerous tissue when compared to the control group. T lymphocyte intensity in the inflammatory infiltrations was directly correlated with the size of resected tumors, presence of regional lymphatic node metastases and histological grade of malignancy. Lymphocytic infiltrations of greater intensity located in close proximity to the primary tumor were found in subjects with less advanced colorectal cancer. The research presented here proves direct dependence between the immune system and colorectal cancer. The presence of lymphocytes in the inflammatory infiltrations located in close proximity to the cancerous tissue has been proved to be prognostically beneficial. The obtained results support the application of immunotherapy in colorectal cancer treatment.

Prognostic significance of ligands belonging to tumour necrosis factor superfamily in acute lymphoblastic leukaemia.

Altered activities of ligands belonging to tumour necrosis factor (TNF) superfamily, namely B-cell activating factor (BAFF), a proliferation-inducing ligand (APRIL) and apoptosis inducing ligand (TRAIL) were demonstrated in several haematological diseases including acute lymphoblastic leukaemia (ALL). BAFF, APRIL and TRAIL provide crucial survival signals to immature, naive and activated B cells. These ligands are capable of activating a broad spectrum of intracellular signalling cascades that can either induce apoptosis or protect from programmed cell death. BAFF and APRIL, which can directly activate the NF-κB pathway, have been identified as crucial survival factors for ALL cells. Here, we have analyzed serum BAFF, APRIL and TRAIL concentrations in 48 patients with newly diagnosed ALL and 44 healthy volunteers. The levels of APRIL and BAFF were significantly higher in ALL patients as compared to healthy volunteers. In contrast, concentrations of TRAIL were significantly lower in ALL patients. Moreover, following induction, the levels of APRIL, but not BAFF or TRAIL, were significantly lower in a group of patients with complete remission (CR) as compared to non-respondent (NR) ALL patients. Furthermore, we demonstrated statistically significant differences in concentrations of APRIL between CR MRD-negative and CR, MRD-positive ALL patients. Notably detection of higher concentrations of APRIL was associated with shorter leukaemia-free survival and overall survival. Altogether, our data indicate that APRIL can play an important role in the pathogenesis of ALL and the measurement of APRIL levels can improve prognostication in ALL patients.

The impact of TNF superfamily molecules on overall survival in acute myeloid leukaemia: correlation with biological and clinical features.

B cell-activating factor (BAFF), a proliferation-inducing ligand (APRIL) and apoptosis-inducing ligand (TRAIL) were demonstrated in several haematological diseases including acute myeloid leukemia (AML). Those cytokines are capable of activating a broad spectrum of intracellular signalling cascades that can either induce apoptosis or protect from programmed cell death. We have analysed BAFF, APRIL and TRAIL serum concentrations in 76 patients with newly diagnosed AML and 40 healthy volunteers. The values were significantly higher for APRIL and BAFF but lower for TRAIL compared to healthy volunteers. Induction therapy significantly reduced the values for BAFF and increased them for TRAIL. Moreover, the concentration of BAFF and APRIL was significantly lower and the concentration of TRAIL higher in a group of patients with complete remission compared to non-respondent AML patients. In addition, higher concentrations of BAFF and lower of TRAIL predicted a shorter overall survival, suggesting thereby an important prognostic marker and possible therapeutic target in AML.

BAFF and APRIL as TNF superfamily molecules and angiogenesis parallel progression of human multiple myeloma.

Tumour necrosis factor alpha (TNF-α) is an inflammatory cytokine with a wide spectrum of biological activity, including angiogenesis. B cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are members of the TNF-α family. Vascular endothelial growth factor (VEGF), on the other hand, is one of the most characteristic pro-angiogenic cytokines produced by multiple cell types in multiple myeloma (MM). We have analysed BAFF and APRIL concentrations in parallel with pro-angiogenic cytokines in serum and trephine biopsy, and the bone marrow microvascular density (MVD) in 50 patients with newly diagnosed IgG MM and 24 healthy volunteers. The study showed statistically higher concentrations of BAFF, APRIL and TNF-α, as well as VEGF and its receptor, in MM patients compared to healthy volunteers and patients in advanced stages of the disease. A statistically positive correlation between the concentration of TNF-α and the expression of VEGF was demonstrated, and so was a positive link between BAFF, APRIL, MVD and lactate dehydrogenase (LDH). Furthermore, we observed a significant decrease in all studied cytokines after anti-angiogenic therapy, with meaningful differences between responders (at least partial remission) and patients with stable disease. It was also established that APRIL, but not BAFF, correlated with pro-angiogenic cytokines such as VEGF with its receptor, MVD and syndecan-1. Finally, our results showed that serum BAFF and APRIL levels could be useful biomarkers of MM disease activity and its progression which suggests that APRIL could be a possible novel therapeutic target in MM.

Imidazole-induced contractility of vascular smooth muscle cells in the presence of U-73122, ODQ, indomethacin and 7-nitroindazole.

The aim of the study was to assess the impact of modulating factors on vascular smooth muscle cells reactivity. Vascular resistance was induced by the administration of increasing concentrations of imidazole. The experiments were performed on isolated and perfused tail artery of Wistar rats (weight 250 g - 350 g). Rats were been narcotized by urethane (intraperitoneal injection) at a dose of 120 mg/kg, stunned and then sacrificed by cervical dislocation. In the following investigation classical pharmacometric methods were used. Relationships between concentration-response curves (CRCs) for imidazole observed in the presence of ODQ [(1H-(1,2,4)oxadiazolo-[4,3-a]quinoxalin-1-one)], 7-nitroindazole and indomethacin were analyzed. Imidazole-induced contractility of vascular smooth muscle cells was independent from alpha-adrenergic receptors and PLC activity. Reactivity of VSMCsinduced by imidazole, was significantly changed in the presence of ODQ and 7-nitroindazole.

Endothelial dysfunction in Graves' disease.

PURPOSE: Graves' disease (GD) is an organ-specific autoimmune thyroid disease, characterized by hyperthyroidism due to excessive production of thyroid hormone induced by thyrotropin receptor-specific stimulatory autoantibodies. In this study, we determined serum levels of the soluble forms of ICAM-1, VCAM-1, vWF, IL-6, IL-12, IL-18, fibrinogen and CRP in patients with subclinical (SH) and overt hyperthyroidism (OH) caused by GD to elucidate a possible role of those parameters as markers of endothelium dysfunction (ED). MATERIAL/METHODS: The study included 96 patients: 52 with GD and 44 euthyroid controls, divided into 3 groups according to their thyroid function tests: SH, OH and controls (CG). RESULTS: The values of IL-6, IL-12 and IL-18 were significantly higher in GD than in CG patients (p < 0.0001, p < 0.0001; p < 0.00001, respectively). Significant difference of sVCAM-1 values were found in the patients with GD compared to CG (p < 0.0001). Patients with GD had significantly higher levels of PAI-1 (p < 0.00001), vWF (p < 0.0001), fibrinogen (p < 0.0001) in comparison to CG. In patients with OH, we observed statistically higher values of fibrinogen compared to SH group (p < 0.05). There were no significant differences in serum concentration of other study parameters in patients with SH compared to the OH. CONCLUSIONS: ED occurs during subclinical and overt hyperthyroidism causing decreased fibrinolytic activity, hypercoagulability and increased levels of IL-6, Il-12 and IL-18. These results support the notion that serum cytokines could be used as a marker of GD activity. Results of this study support the opinion that GD might require treatment as early as in the phase of SH.

The evaluation of angiogenesis and matrix metalloproteinase-2 secretion in bone marrow of multiple myeloma patients before and after the treatment.

PURPOSE: Angiogenesis appears to be a prominent feature of many hematological disorders, particularly in multiple myeloma (MM). Progression in MM also involves secretion of the metaloproteinases (MMPs). In this study, the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and its receptor, in bone marrow trephine biopsy (TB) of thirty six MM patients before and after the treatment or during progression was examined. The MMP-2 secretion was assessed from the same patients. MATERIAL/METHODS: Immunohistochemical staining of bone marrow specimens for angiogenic factors and microvessel density (MVD) and bone marrow aspirates for Western blot analysis of MMP-2 expression was performed. RESULTS: In active, untreated MM patients, we found statistically significant differences in the expression of angiogenic factors according to the patients after the anti-angiogenic treatment. We found statistical differences of the expression of angiogenic factors between the group of patients with a response after the treatment and the patients who had progression during the treatment. The data showed statistically significant decreased MVD after the treatment. The results showed statistically significant differences between initial secretion of MMP-2 in active, untreated MM patients and patients with a response after the treatment and patients with progression during the treatment. CONCLUSIONS: We showed that not only decreased expression of angiogenic cytokines is present after the anti-angiogenic treatment but also activity of MMP-2 in MM patients who responded to the treatment. Combination therapy with the inhibition of the activity of MMPs could represent an interesting therapeutical approach in MM.

Lung mass in a 28-year-old male: a case report of a rare tumor.

A twenty eight-year-old male presented with a two week history of dyspnea, cough, hemoptysis, chest pain, and fever 38-39°C. He also complained of loss of appetite, general weakness and left leg pain for two months preceding admission. He was referred with suspicion of lung tumor to our institution. Chest X-ray showed almost total atelectasis of the right lung with compensatory overinflation of the contralateral lung. Using computed tomography (CT), a lesion of diameter of 19.3 x 14.1 x 19.1 cm in the right lung, pleuritis, Th3 osteolysis, and compensatory overinflation of the left lung was seen. Bronchoscopy revealed a total obstruction of the right main bronchus due to submucosal infiltration and compression of the right main bronchus with negative histology of bronchial biopsy specimens. Transthoracic fine needle aspiration revealed celullae suspectae probabiliter neoplasmaticae suggesting tumor fusocellularis. USG of the abdomen revealed liver with numerous heterogeneous, solid areas hypo- and hyperechogenic, some of them with features of liquid or the disintegration up to diameter of 74 mm. Subsequent fine needle aspirations of the thorax and liver revealed fibrolamellar hepatocarcinoma and carcinoma adenoides of the lung. Patient underwent chemotherapy with 5-FU/DDP/VCR with no response. This report presents a case of a rare lung metastasis from FL-HCC.

Serum sICAM, sVCAM and sE-selectin levels in colorectal cancer patients.

Colorectal cancer (CRC) is one of the most common cancers of the gastrointestinal tract and the fourth cause of cancers death in the world. Soluble adhesion molecules (CAMs) are thought to have an important role in host defense against carcinogenesis. They are biomarkers of inflammation and indicators of the immune response to tumors. The study included 40 CRC patients without remote metastases and 24 control subjects. Serum concentrations of sE-selectin, sICAM and sVCAM in patients with CRC were investigated by ELISA method. The level of the sCAMs decreased significantly after radical tumor resection. Preoperative serum concentrations of sICAM and sVCAM in CRC patients were significantly higher compared to the control group, whereas there were no differences regarding serum sE-selectin. Serum levels of sE-selectin, sICAM and sVCAM correlated significantly with each other. There was a significant correlation of serum levels of sICAM-1 and sVCAM-1, but not sE-selectin, with TNM stage and lymph node involvement. No significant relationship was found between serum concentrations of sICAM-1, sVCAM-1 and sE-selectin in CRC patients and patients' age or gender. Our findings suggest that an improved understanding of the mechanisms of membrane shedding of sICAM, sVCAM and sE-selectin is required to delineate their role in tumor progression.

Cannabinoid receptors expression in bone marrow trephine biopsy of chronic lymphocytic leukaemia patients treated with purine analogues.

BACKGROUND: Cannabinoid receptors CB1 and CB2 are part the endocannabinoid system that plays an important role in the process of proliferation and apoptosis of different neoplastic cells. B-cell chronic lymphocytic leukaemia is one of the diseases in which these processes are altered. AIM: The aim of our study was the assessment of cannabinoid receptor expression on the B-lymphocytes in bone marrow trephine biopsy from leukaemic patients at diagnosis and after purine analogue treatment. METHODS: The biopsy was taken routinely and standard immunohistochemical staining procedure for paraffin embedded sections was applied. The cannabinoid receptors were detected using specific primary polyclonal antibody anti-CB1 and anti-CB2. Additionally, an existence of cannabinoid receptors was confirmed by flow cytometry. RESULTS: The results showed that the expression of CB1 receptor on the surface of neoplastic cells was lower than that of CB2 (17.0+/-3.1% and 92.1+/-1.7% respectively, p<0.001). Nine of the patients responded to applied treatment with a reduction in leukaemic infiltration (77.2+/-6.9% to 30.2+/-6.5%, p=0.007) and CB1 receptor expression (24.4+/-4.8% to 8.6+/-2.9%, p=0.01), but there was no change in CB2 expression (91.7+/-2.7% vs 90.9+/-2.8%, p=0.69). Four patients without remission expressed even greater number of the receptors. In all of the cases both cannabinoid receptor types antibodies gave positive reaction. Furthermore, the existence of cannabinoid receptors on neoplastic lymphocytes was confirmed by flow cytometry. CONCLUSION: The study provides original evidence for the existence of cannabinoid receptors on B-lymphocytes in chronic lymphocytic leukaemia patients. The receptors are thought to be a new structure that can modify the course of the disease and may be considered as a new target in leukaemia treatment.

Fine needle aspiration cytology of benign breast disease. Markers of apoptosis and proliferation.

Aim of the study was to compare the fine needle aspiration cytology findings of benign breast lesions with incidence of proliferation markers and apoptosis. This study included 37 patients with palpable breast lumps, referred for USG guided FNA. FNAC were prospectively classified as C2-benign, C4-suspicious of malignancy, and C5-malignant. The specimens were simultaneously stained for Ki-67, MPM2, Bcl2 and P53. The diagnoses in group-C2 were following: simple cyst, multiple cysts, simple cyst with apocrine metaplasia, inflammatory cyst, benign dysplasia (BD) and benign solid tumors. The final diagnoses, after histopathological verification, in cases of primary classification as C4 and C5 were as follow: proliferative fibroadenoma (FAp) and breas cancer, respectively. Great majority of C2/BD aspirates were negative for proliferative antigens Ki-67 and PCNA. These antigens were detected in part of benign solid tumors, as anticipated in suspicious solid tumor, and in all of cancer aspirates. Bcl-2 immunopositive cells were detected approximately in one quarter of C2/BD, nearly in half of C2 solid tumors and in one C4/FAp. Most of diagnosed specimens were P53-negative. Immunocytodetection of Ki67, MPM2, Bcl2, P53 might be promising, supportive method in the classification of benign breast lesions. FNAC increases the reliability of diagnosis when complemented by immunocytochemical staining. It could be helpful procedure of establishing more accurately the biology of these lesions and possibly serve as an essential factor in clinical follow-up. Nevertheless, further study on larger group of patients comparing cytological and histopathological diagnosis is required to estimate reliability of its predictive value.

Proliferative activity of chosen central nervous system (CNS) neoplasms.

Gliomas are the most common neoplastic tumours of the central nervous system. The aim of the study was to evaluate the proliferative activity of chosen types of gliomas and to analyse their correlation with histological type, malignancy grade, location, size and clinical symptoms. The study involved patients with astrocytoma, anaplastic astrocytoma, glioblastoma, oligodendroglioma, anaplastic oligodendroglioma. The proliferative activity (the labelling index--LI) of glial cells was estimated, using immunohistochemistry. In studied groups, a positive correlation was noted between the proliferative activity and tumour size, but not between the proliferative activity and tumour location. The clinical symptoms were conditioned mainly by tumour location and, to a smaller extent, by its size.

Influence of thalidomide on megakaryocytes in multiple myeloma.

The aim of the study was to assess the influence of thalidomide on megakaryocytes (MK) in patients with multiple myeloma (MM). The study was based on bone marrow trephine biopsies from 12 patients with MM before initiation of thalidomide administration and after three months of its duration. The morphometric examinations were done, using image analysis (DP 12). Quantitative assessment of MK and the analysis of the morphological parameters of MK were performed. MK with features of dysplasia were more frequently observed before the treatment. Additionally, a greater number of the so-called 'naked nuclei' was noticed then. Due to the effect of thalidomide, the mean number of MK increased and so did their area. During the treatment, a more frequent presence of emperipolesis was observed. The observations confirm the fact that thalidomide may cause changes in MK.

Apoptosis phenomenon in the selected neoplasms of the glial origin.

Gliomas cause a therapeutic problem because of their localization and asymptomatic growth in the initial phase. Neoplastic growth is connected with disturbance between proliferation and apoptosis. In the study, we assessed the Bcl-2 family proteins involved in apoptosis in gliomas. The study comprised 61 patients with gliomas and based on tissue material sent for the diagnosis. Apoptosis was assessed in various types of gliomas and was defined by the apoptotic index (IA) and shown immunohistochemically with using Bcl-2, Bax and Bcl-x antibodies. The data were statistically analyzed. We found an increased percentage of the Bax (+) cells in less matured gliomas. A reverse dependence was revealed for Bcl-x. It was found that, probably in gliomas, the assessment of the Bcl-2 family proteins may serve only as an additional parameter for the evaluation of the disease course and the therapeutic success.

Expression of PCNA and Ki-67 in posterior uveal melanomas in adults.

The aim of the study was to evaluate the expression of cell proliferation markers (PCNA and Ki-67) in posterior uveal melanomas in adults. Thirty-six enucleated eyes (without prior treatment) were included in histopatological study. A series of 15 cases of spindle cell type melanomas, 12 epithelioid and 9 of a mixed type were assessed, using the immunohistochemical method with monoclonal PCNA and Ki-67 antibodies. PCNA expression was observed in 75% and Ki-67 in 13.8% of all the examined tumours. The mean score of PCNA and Ki-67 index were the highest in tumours, which contained epithelioid cells and in large, more advanced (pT3, pT4) uveal melanomas. The results showed that the evaluation of expression of PCNA and Ki-67 may provide an additional infonnation about the progression of tumor process.

Expression of Ki-67, PCNA and MPM2 antigens in follicular cells of the thyroid gland after iodotherapy.

Hyperfunctional nodular goitre is the most common thyroid non-neoplastic condition in endemic areas. Iodotherapy is the basic method in thyroid gland hyperfunction treatment. The aim of the study was to evaluate proliferation of thyroid follicular cells in nodular goitre after iodotherapy. The study was carried out on 32 women, 30-76 years old. Cytological and immunohistochemical evaluations were based on the material, obtained by Fine-Needle Aspiration Biopsy (FNAB). Proliferative activity was immunohistochemically assessed. The influence of radioiodine on thyroid follicular cells was evaluated as a difference between the proliferation of follicular cells before and after its application. It was concluded that the proliferative activity of thyroid follicular cells decreased considerably after radioiodine therapy.

Histomorphometry of marrow megakaryocytes in experimental uraemia in rats.

Uraemic patients frequently demonstrate tendencies towards life-threatening bleeding. Reduced platelet counts and their functional immaturity seem to be caused, among other things, by disorders in the system of marrow megakaryocytes. The aim of the study was a histomorphometric evaluation of marrow megakaryocytes in the course of experimental uraemia in rats. Uraemia was induced by means of right nephrectomy and a partial removal of the left kidney cortex in rats. Morphometric analysis was conducted, using the Microlmage program set. The number of MK, MK area, N/C, CDMK, CDNMK and MK cluster formation were analysed. Uraemic rats showed a reduction in the MK count and their area and an increase in the N/C ratio, CDMK, CDNMK and in the incidence of MK clusters. The results indicate that platelet disorders, observed in uraemia, can also be conditioned by disturbed maturation of MK.

A preliminary study of the submandibular gland of the rat after one-year cadmium intoxication. Part II. Pathomorphology and ultrastructure.

The aim of the present study was to establish to what degree a one-year exposure of rat females to 5, 50 and 100 mg of Cd/l affects cell morphology of the submandibular glands. After one-year cadmium exposure of female rats, at doses of 5, 50 and 100 mg Cd/l, a pathomorphological examination revealed periductal fibrosis in the submandibular glands of rats in all the three experimental groups, which increased with cadmium dose. We also found foamish cytoplasm in the cells of the submandibular glands in all the experimental groups, the intensity of that phenomenon also increasing with Cd dose. The ultrastructural examination revealed no abnormalities in Group I. However, in Groups II and III, we observed numerous granules with a secretion, varying in shape and size that filled up the cytoplasm both in the mucous and serous cells.

Expression of PCNA and Ki-67 in the rat submandibular gland after one year cadmium intoxication--a preliminary study.

The aim of our study was to determine to what degree one-year exposure of female rats to cadmium at a dose of 5, 50 and 100 mg Cd/l affects cell proliferation in the submandibular gland, shown by PCNA and Ki-67 expression. In the present study, we found a positive nuclear reaction for PCNA in single cells in microscopic preparations of control rats. In Group I, an increase was observed in the number of PCNA-positive cells, compared to that in the control. In Group II, the number of PCNA-positive cells was markedly higher than that in Group I and in the control. In the submandibular glands of rats in Group III, the number of PCNA-positive cells was similar to that, found in Group II. However, Ki-67 expression was sporadically observed in control submandibular glands, but not in Groups I, II and III.

A preliminary study of the submandibular gland of the rat after one-year cadmium intoxication. Part I. Cadmium concentration.

The aim of the present study was to establish to what degree a one-year exposure of rat females to 5, 50 and 100 mg Cd/l affects the weight of the submandibular glands and their cadmium levels. We observed a decrease in the weight of the submandibular glands in the rat females from Groups I, II and III, compared to the control rats. We also found an increase in cadmium levels in the submandibular glands in Groups I, II and III, in comparison to the control. The highest cadmium concentration was noted in the submandibular glands in Group III, which was accompanied by the greatest weight reduction, the correlation being negative. The present experiment indicates that one-year administration of cadmium to rat females at a dose of 5, 50 and 100 mg Cd/l leads to a cadmium dose-dependent decrease in the weight of the submandibular glands.