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.

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.

Estradiol and tamoxifen differently affects the inhibitory effects of vitamin A and their metabolites on the proliferation and expression of alpha2beta1 integrins in MCF-7 breast cancer cells.

PURPOSE: Retinoids are well known inhibitors of estrogen-dependent breast cancer cell growth and differentiation. alpha2beta1 integrins are involved in the normal growth and differentiation of breast cells, they also take part in many pathological processes including malignancies. The aim of the study was to evaluate the effect of estradiol and tamoxifen on the inhibitory action of retinoids on the proliferation of MCF-7 breast cancer cells and alpha2beta1 integrin expression. MATERIALS AND METHODS: Evaluation was based on [3H]thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. Expression of alpha2beta1 was assessed through immunocytochemical analysis. RESULTS: Treatment of cancer cells with the examined compounds and tamoxifen (10 microM) revealed that only 13-cis retinoic acid (13-cis RA) and all-trans retinoic acid (ATRA) (10(-5) M) decreased cells proliferation compared to the tamoxifen group (30.84%+/-3.32, p<0.01 and 31.05%+/-4.67, p<0.01, respectively). The lowest fraction of PCNA positive cells was also observed after the simultaneous addition ATRA (10(-5) M) and tamoxifen (10 microM) (30.75%+/-0.95, p<0.01, compared to the tamoxifen group). Our results showed that the decrease of alpha2beta1 integrin expression by 13-cis RA (10(-5) M, 49.6+/-3.25%) and ATRA (10-9 M, 15.0%+/-5.0) was augmented by tamoxifen and to a lesser extent by estradiol, particularly in the case of ATRA at 10(-7) or 10(-9) M. CONCLUSIONS: This data suggest that tamoxifen augments the inhibitory effect of retinoids on proliferation and alpha2beta1 integrin expression in MCF-7 cells.

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.

Effect of brefeldin A on membrane localization of MUC1 mucin and adhesive properties of cancer cells.

Transmembrane glycoproteins play a significant role in cancer cells adhesion and metastatic process, just for that reason the glycosylation inhibitors are used to change the glycan structure and in this way the membrane expression of glycoproteins. The inhibitory effect of brefeldin A (BFA) on the expression of some glycoproteins: MUC1 mucin and alpha2beta1 integrin on cell surface of breast (MCF-7 and MDA-MB-231 lines) and endometrial (Ishikawa line) cancer cells was evaluated in our study. In MCF-7 and MDA-MB-231 cells, a decrease in MUC1 expression depended on brefeldin A concentration and equaled about 40% in cells treated with 1mg% of drug. In Ishikawa cells, a decrease in MUC1 expression was lower and amounted to about 25%. The expression of alpha2beta1 integrin was greatly inhibited in brefeldin-treated MCF-7 and Ishikawa cells, though it was unchanged in MDA-MB-231 cells. A decrease in MUC1 mucin and alpha2beta1 integrin level reduced the adhesive properties of BFA-treated cells. Adhesion to type I collagen was greatly diminished in BFA-treated MCF-7 and Ishikawa cells (above 70%), and to a lesser degree in MDA-MB-231 cells (about 50%); which was mainly caused by the inhibited integrin expression. These findings have proved that brefeldin A, by changing the surface glycoproteins level, can alter carcinoma cells adhesion to extracellular matrix proteins.

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.

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.

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.

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.

Proliferative activity of papillary carcinoma and benign papillary hyperplasia of thyroid follicular cells.

Papillary structures of follicular cells are observed in nodular goiter, cysts, hyperplastic areas of follicular tumors, Graves' disease, thyroiditis and carcinomas. The distinction of papillary carcinomas from benign lesions has important implication for clinical management. The aim of the study was to test a marker of proliferation activity (MIB-1) in the diagnosis of benign and malignant thyroid papillary proliferation. The study was carried out in 98 women with papillary carcinoma, nodular goiter. intracystic proliferation. Graves' disease and hyperplastic areas of follicular benign tumors. The formalin fixed, paraffin-embedded specimens were microscopically examined using HE staining and immunostaining with MIB-1 antibody (DAKO). The proliferative index (PI) was significantly higher in malignant than in benign papillary hyperplasia. Our results may provide additional information for differential papillary proliferation diagnosis by FNAB.

Megakaryocytes emperipolesis in bone marrow of the patients with non-Hodgkin's lymphoma.

Emperipolesis of hematopoietic cells within the cytoplasm of the megakaryocytes was most often described in association with various pathologic conditions. The aim of the research was estimation of the incidence of emperipolesis in the bone marrow of the patients with non-Hodgkin's lymphoma (NHL). 30 patients with different histological types of NHL (chronic lymphocytic leukaemia--CLL, hairy cell leukaemia--HCL, multiple myeloma--MM) in compliance with clinical stage of the disease, patient's age and sex were analyzed. Trephine biopsies of the bone marrow were carried out in fixative solution and paraffin embedding. Hematoxylin and eosin and monoclonal antibody CD 61 were applied on thin sections. Phenomenon of megakaryocytic emperipolesis in human bone marrow was found in 6 cases: in 5 cases in CLL and in 1 case in HCL. In most of them emperipolesis was related to single megakaryocytes. We observed in the cytoplasm of the megakaryocytes single hematopietic cells-most often lymphocytes, rarely eosinophilic granulocytes. We found no correlations between histological types of NHL, clinical stage of the disease, patients' age, sex and the incidence of megakaryocytic emperipolesis.

MIB-I as a proliferative activity marker of the multiform glioblastomas.

MIB-I is a proliferative activity marker of multiform glioblastomas which are the most frequent tumors of the central nervous system. They are characterizad by differential rate and prognosis. The aim of the study was to determine the proliferative activity of multiform glioblastomas and estimation of the correlation between tumors' proliferative activity and tumors' localization, size, patients' age and sex. 24 patients (18 females and 6 males) with multiform glioblastomas were analyzed. The mean patients' age was 52.1. The proliferative activity was calculated as a proliferation index: IP for MIB-I. Cells with positive reaction were determined by MIB-I which was compared to all neoplastic cells. The most frequent localization of the tumors were frontal and temporal lobes of the brain. The size of the tumors ranged from 2.5 to 5.3 cm (mean 3.9). Mean IP was 43.2 (SD+/-17.4). We found no correlation between IP MIB-I and localization of the tumor, patients' age and sex. There was a marginal statistically significant correlation between IP MIB-I and size of the tumor (p=0.005).

[Chylomicronemia syndrome complicating type 1 hyperlipoproteinemia with concomitant insulin-dependent diabetes]. / Zespól chylomikronemii w przebiegu hiperlipoproteinemii typu 1 z towarzyszaca cukrzyca insulinozalezna.

The case of chylomicronaemia syndrome complicating clinical course of type-1 hyperlipoproteinaemia and insulin-dependent diabetes is described. The patient required hypoglycaemic and lipid-lowering therapy.