Non-Alcoholic Fatty Liver Disease (NAFLD) and Bariatric/Metabolic Surgery as Its Treatment Option: A Review.

The prevalence of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) has considerably increased over the last years. NAFLD is currently the most common cause of chronic liver disease in the developing world. The diagnosis of NAFLD/NASH is often incidental, as the early-stage of disease is frequently free of symptoms. Most patients recognized with NAFLD have severe obesity and other obesity-related disease such as type 2 diabetes mellitus (T2DM), insulin-resistance, dyslipidemia and hypertension. The only proven method for NAFLD improvement and resolution is weight loss. Bariatric surgery leads to significant and long-term weight loss as well as improvement of coexisting diseases. There is a lot of evidence suggesting that metabolic/bariatric surgery is an effective method of NAFLD treatment that leads to reduction in steatosis, hepatic inflammation and fibrosis. However, there is still a need to perform long-term studies in order to determine the role of bariatric surgery as a treatment option for NAFLD and NASH. This review discusses current evidence about epidemiology, pathogenesis and treatment options for NAFLD including bariatric/metabolic surgery and its effect on improvement and resolution of NAFLD.

The effects of BAFF and APRIL signaling on non-small cell lung cancer cell proliferation and invasiveness.

Lung cancer represents the most common type of human malignancy and is the main cause of cancer-associated mortality worldwide. To improve the effectiveness of treatment strategies, a better understanding of the mechanisms of cancer progression and invasiveness is required. Recently, B-cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL), two relatively newly described cytokines belonging to the tumor necrosis factor superfamily, have been shown to play a role in cancer progression. However, at present, the effects of both cytokines on lung cancer cells remain unclear. The present study aimed therefore to understand the direct effects of BAFF and APRIL on non-small cell lung cancer (NSCLC) progression. To do so, reverse transcription quantitative PCR and western blotting were used to evaluate whether A549 and H2030 NSCLC cells express receptors for both BAFF and APRIL. The results demonstrated that both investigated cell lines expressed BAFF-R (receptor specific to BAFF only) and transmembrane activator and CAML interactor (TACI; shared receptor for both cytokines). In addition, functional experiments were performed to determine the effects of BAFF and APRIL stimulation on cancer cell viability. The results demonstrated no direct effects of BAFF and APRIL on NSCLC cell proliferation and invasiveness. In summary, the present study demonstrated that NSCLC cells possess the ability to respond directly to both BAFF and APRIL. However, activation of BAFF-R and TACI signaling in cancer cells did not increase the proliferative capacity and invasiveness. Further investigation is thus required to better understand the role of BAFF and APRIL on the progression of NSCLC.

High immunoproteasome concentration in the plasma of patients with newly diagnosed multiple myeloma treated with bortezomib is predictive of longer OS.

PURPOSE: Proteasome inhibitors (PI) bortezomib or carfilzomib among them, play a crucial role in the modern standard therapy for multiple myeloma (MM). In this study, we intended to evaluate whether immunoproteasome (IMP) concentration could act as an effective biomarker which determines the probability of response to treatment with bortezomib, in order to detect groups of patients who are more likely to respond to treatment with PI. MATERIALS AND METHODS: In our study, we evaluated IMP concentration in the plasma of 40 patients with monoclonal gammopathy of undetermined significance (MGUS) and 116 patients with newly diagnosed MM during treatment with or without PI. RESULTS: The values of all the studied parameters after the applied chemotherapy in the responders' group of patients declined considerably during the consecutive cycles of chemotherapy compared to their initial levels. On the contrary, in the group of non-responders, we observed no change in the measured IMP parameters during the consecutive cycles of therapy. We also showed that higher baseline IMP concentration might indicate longer overall survival (OS) in all patients. CONCLUSIONS: Our results indicate that assessing plasma IMP concentration can be applied as a strong biomarker for predicting clinical response to treatment and OS in patients with newly diagnosed MM.

Relationship between tumour necrosis factor-related apoptosis inducing ligand (TRAIL) and vascular endothelial growth factor in human multiple myeloma patients.

Tumour necrosis factor-alfa (TNF-α) is an inflammatory cytokine with a wide spectrum of biological activity, including angiogenesis. Tumour necrosis factor-related apoptosis inducing ligand (TRAIL), which belongs to the TNF family of proteins, plays a role in the regulation of vascular responses, but its effect on the formation of new blood vessels (angiogenesis) is unclear. We analysed TRAIL concentrations in parallel with pro-angiogenic cytokines in serum and their expression in trephine biopsy (TB) in 56 patients with newly diagnosed IgG MM and 24 healthy volunteers. The study showed statistically higher concentrations of TRAIL and TNF-α, as well as of VEGF and its receptor, in MM patients compared to healthy volunteers and patients in advanced stages of the disease. Furthermore, we observed a significant decrease in all studied pro-angiogenic cytokines and significant increase of TRAIL concentration after anti-angiogenic therapy, with meaningful differences between responders (at least partial remission) and patients with progression during the induction treatment. It was also established that TRAIL correlated statistically and negatively with pro-angiogenic cytokines such as VEGF with its receptor and expression of VEGF and syndecan-1 in TB. In summary, our data indicate that in MM patients, both clinical course and treatment responsiveness are associated with dynamic yet corresponding changes of levels of TRAIL parallel pro-angiogenic mediators such as VEGF with its receptor and expression of VEGF and syndecan-1 in TB.

Bone marrow megakaryocytes, soluble P-selectin and thrombopoietic cytokines in multiple myeloma patients.

The expression of adhesion molecules and other cell-surface molecules is substantial in the communication between plasma cells and bone marrow microenvironment, and may lead to increased proliferation of myeloma cells. Many of the cytokines involved in multiple myeloma (MM) pathogenesis, e.g. thrombopoietin (TPO) and interleukin-6 (IL-6), play a pivotal role in different developmental stages of megakaryocytopoiesis and thrombopoiesis. The principal aim of our study was to explore the relationship between thrombopoietic cytokines, megakaryocytes (MKs) and soluble P-selectin (sP-selectin) levels in MM patients before and after anti-angiogenic treatment. Forty-four patients (20 female and 24 male) with a newly diagnosed MM were examined in three groups, following a division based on the International Staging System, ISS. Plasma levels of TPO, IL-6 and soluble P-selectin (human sP-selectin) were measured by means of ELISA. Bone marrow specimens were studied to determine the number of MKs and the so-called "naked nuclei" (NN), as well as the expression of platelet-derived growth factor (PDGF). The comparison revealed a significantly higher concentration of cytokines and sP-selectin in newly diagnosed MM patients compared to healthy volunteers: for TPO, p=0.01, IL-6, p=0.0005 and sP-selectin, p=0.00008, respectively. Marked differences were observed in the concentration of sP-selectin, expression of PDGF and MKs counts between patients with MM stage I and MM stage III. Statistically meaningful correspondences were also found between MKs versus TPO, NN versus TPO, as well as MKs versus MPV, p=0.009, p=0.004 and p=0.0005, respectively. Furthermore, the analysis exhibited some statistically meaningful divergences between initial concentrations of sP-selectin in subgroups with different response after chemotherapy. The initial concentration of sP-selectin in the group of MM patients with complete or partial remission stood at 31.86 ± 6.13 ng/ml. In the remaining patients (stable disease), the concentration of sP-selectin amounted to 35.15 ± 7.23 ng/ml (p=0.048). We found a correlation between sP-selectin and IL-6 (ρ=0.57, p=0.0004), TPO and IL-6 (ρ=0.46, p=0.001) as well as sP-selectin and TPO (ρ=0.36, p=0.043), and sP-selectin and PDGF (ρ=0.36, p=0.03). Our study has eventually demonstrated that sP-selectin, as a marker of platelet activation, could be a useful marker of maximum response to therapy. Its strong association with another marker like PDGF-AB could further lead to the development of new combinational therapeutic strategies of anti-angiogenic therapy in MM patients.

Differences and similarities between LC-MS derived serum fingerprints of patients with B-cell malignancies.

Multiple myeloma (MM) and chronic lymphocytic leukaemia (CLL) are closely related B-cell non-Hodgkin's lymphomas. MM, a plasma cell malignancy, is the second most common haematopoietic cancer in Western countries, with the median survival time of 3­4 years. CLL, a lymphocyte B malignancy, is the most common leukaemia in Western countries. About 25­30% of all CLL patients do not survive the period of 5 years following diagnosis. Both malignancies are complicated, not fully understood and incurable with the current standard treatment. Biologically, MM and CLL may be preceded by associated precursor conditions, that is, monoclonal gammopathy of undetermined significance for MM and its cellular counterpart and monoclonal B-cell lymphocytosis for CLL. Similarities and differences in the biology of these malignancies prompted us to evaluate their metabolomics in stages requiring chemotherapy. Fingerprinting of serum metabolites by the use of LC-MS has never been applied in studies on MM and CLL patients. Obtained results revealed metabolites common for both malignancies (e.g. fatty acids, acylcarnitines, sphingolipids, phospholipids, phenylalanylphenylalanine and isoprene) as well as those which render them different (e.g. lysophosphatidylcholines, monoacylglycerols, aminocaproic acid, phenylacetylglutamine).

Evaluation of TNF superfamily molecules in multiple myeloma patients: correlation with biological and clinical features.

B-cell activating factor (BAFF), a proliferation-inducing ligand (APRIL) and apoptosis inducing ligand (TRAIL) are members of the tumour necrosis factor (TNF) family. They are the main survival factors for immature, naive and activated B cells. We have analysed BAFF, APRIL and TRAIL serum concentrations in 52 patients with newly diagnosed IgG multiple myeloma and 20 healthy volunteers. The values were significantly higher in the studied patients and advanced diseases, decreasing after chemotherapy, compared to the control group. It was established that BAFF as APRIL (but not TRAIL) correlated with adverse prognostic factors such as IL-6 and lactate dehydrogenase. Furthermore, higher concentrations of APRIL and BAFF (but not TRAIL) predicted a shorter progression free survival, suggesting thereby an important prognostic marker and a possible therapeutic target in myeloma.

Plasma concentration of protein Z and protein Z-dependent protease inhibitor in patients with haemophilia A.

The potential role of alterations in protein Z (PZ) concentrations in the pathogenesis of coagulation has been investigated in several studies which, however, yielded conflicting results. Protein Z deficiency may induce bleeding as well as prothrombotic tendencies and it might occur as an inherited disorder. The principal aim of the present study was to explore the concentration of protein Z and protein Z-dependent protease inhibitor (ZPI) in patients with haemophilia A. In haemophilia A patients mean plasma concentrations of PZ and ZPI were significantly higher than in healthy individuals: PZ (1.87±0.68µg/mL vs 1.49±0.54µg/mL) and ZPI (5.02±1.11µg/mL vs 4.22±0.55µg/mL), with p=0.02 and p=0.03, respectively. In the subgroup with severe haemophilia A, an in-depth analysis revealed a tendency to modulating effect of the PZ (r=-0.53; p=0.072) and a statistically significant one in the case of ZPI (rho=-0.79, p=0.002) on the bleeding rate. It simultaneously disclosed a statistically significant correlation between the number of bleeds to the joints (20.18±14.1), PZ (r=-0.72; p=0.04) and ZPI (rho=-0.88, p=0.001). With reference to this particular group of patients, the study also showed some other statistically meaningful correspondences: between PZ and ZPI (rho=0.65, p=0.02), PZ and FIX (r=-0.61, p=0.04), as well as ZPI and FVIII (rho=0.78, p=0.002). In conclusion, despite the fact that FVIII deficiency is undoubtedly the main mechanism of bleeding in haemophilia A patients, the activity of PZ/ZPI complex may play some modulating role in the matter.

The role of Interleukin-17A and Interleukin-17E in multiple myeloma patients.

BACKGROUND: Tumor growth in multiple myeloma (MM) is regulated by the cytokine networks which are produced by myeloma cells and the microenvironment of the bone marrow. Interleukin-17 (IL-17) is implicated in the increased angiogenesis in the bone marrow of MM. Recent studies reported elevated levels of interleukin 17A (IL-17A) in the sera of patients with advanced stages according to Durie-Salmon classification. MATERIAL/METHODS: We compared the concentration of IL-17A and IL-17E in the blood serum of 34 newly diagnosed MM patients with healthy subjects' sera. We also evaluated the concentration of IL-17A and IL-17E in the blood serum of MM patients and the relation to the percentage of plasma cells and other clinical parameters. The concentration of IL-17E and IL-17A of healthy subjects and patients with MM was assessed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Our data confirm that IL-17A and IL-17E serum levels were significantly higher in all MM patients and also in patients with advanced stage compared with healthy subjects. We found the correlation between serum levels of IL-17A in MM patients and percentage of plasma cells. Our results also showed that if serum levels of IL-17E were higher in MM patients, the percentage of plasma cells and beta-2-microglobulin levels were lower. CONCLUSIONS: The IL-17 family of cytokines may suppress or promote tumor growth. There seems to be some balance between the effects of IL-17A and IL-17E. The role of increased levels of IL-17E needs further investigation to understand its role in the pathobiology of MM.

The levels of sMUC-1 in patients with multiple myeloma.

Mucins have been shown to be aberrantly overexpressed in various diseases including cystic fibrosis, asthma, and cancer. Recent studies have uncovered the roles of these mucins in the pathogenesis of cancer. The presence of MUC-1 has also been detected on the cell surface of multiple myeloma (MM) cells in peripheral blood and showed direct correlation with tumor mass. In this study, we evaluated the levels of soluble MUC-1 (sMUC-1) in 50 new MM patients and correlated this with the levels of sMUC-1 after treatment. High levels of sMUC-1 were found in 20/50 (40%) MM patients, and in 2/50 (4%) healthy individuals (p = 0.001). According to the ISS, we found significant differences of mean sMUC-1 levels between the first stage of the disease (0.63 ± ± 0.26) and the third (0.93 ± 0.24; p = 0.03), but not with the second stage (0.80 ± 0.22; p = 0.08). Our study confirmed the correlation between elevated sMUC-1 and high elevated lactate dehydrogenase (p = 0.03) and the level of IgG in groups of patients with MM IgG at every stage of disease (p = 0.001). We showed for the first time that levels of sMUC-1 after treatment, in a group of patients with initially elevated levels of MUC-1, were statistically lower than in a group of patients with initially lower levels of sMUC-1 (21% vs. 42,6%; p = 0.05). At 37 months median of follow-up, we found a statistically significant difference between patients with normal versus elevated sMUC-1 in terms of progression-free survival (median 12 months vs. 8.1 months; p = 0.03).

The vitamin A family can significantly decrease the expression of ERbeta of ERs positive breast cancer cells in the presence or absence of ER ligands and paclitaxel.

Taxanes have high activity against breast cancer cells either as the single agent or in combination with other anticancer compounds. The aim of the study was to determine the effects of vitamin A compounds on the cytotoxic action of paclitaxel and on the expression of ERs in the MCF-7 breast cancer cells. Retinol and beta-carotene, but not retinoids, added to the culture exerted an effect on paclitaxel activity. However, only beta-carotene significantly reduced the percentage of proliferating cells (40.36% +/- 5.64, p < 0.01). We observed that vitamin A and its derivatives combined with paclitaxel and estradiol decreased the percentage of proliferating cells, but only in comparison to estradiol group, whereas retinol and lycopene administered together with paclitaxel and tamoxifen decrease significantly the percentage of proliferatin cells (36.85% +/- 4.71, p < 0.0001 and 37.22% +/- 1.59, p < 0.0001 respectively, compared with paclitaxel group). We have shown that paclitaxel increases the expression of ERalpha and ERbeta mRNA in MCF-7 line. The strongest effect of transcription inhibition ERalpha (2.5 times) and especially ERbeta (10 times) was observed after addition of 9-cis retinoic acid and paclitaxel. This data suggests a synergistic effect of the compounds on ERbeta down-regulation. Our results support the use of retinoid is treatment of ER positive breast cancer patients.

Vitamin A family compounds, estradiol, and docetaxel in proliferation, apoptosis and immunocytochemical profile of human ovary endometrioid cancer cell line CRL-11731.

Endometrioid carcinoma represents approximately 10% of cases of the malignant ovarian epithelial tumors. According to literature, the vitamin A (carotenoids and retinoids) plays an essential role in cell proliferation, differentiation and apoptosis in both normal and neoplastic ovarian tissues. Apart from that, the retinoids alter a cytotoxic effect of chemiotherapeutics, i.e. docetaxel, on ovarian cancer cell lines. Retinoids act on cancer cells throughout different mechanism than taxanes, so they may be the potential candidates for the new treatment strategies of ovarian cancer. The aim of the study was to determine the effects of vitamin A family compounds (retinol, beta-carotene, lycopene, all-trans -, 9-cis - and 13-cis retinoic acid) on the growth and proliferation of CRL-11731 endometrioid ovary cancer cell line and on docetaxel and estradiol activity in this culture. The assay was based on [3H] thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in CRL-11731 cells were also studied. Among vitamin A family compounds retinol and carotenoids, but not retinoids, inhibited the growth of cancer cells in dose dependent manner. Only the concentration of 100 muM of docetaxel inhibited incorporation [3H] thymidine into CRL-11731 cancer cells. Retinol (33.4%+/-8.5), carotenoids (beta-carotene 20 muM 4.7%+/-2.9, 50 muM 2.2%+/-0.9; lycopene 10 muM 7.6%+/-0.8, 20 muM 5.2%+/-2.5, 50 muM 2.9%+/-1.2), and 13-cis retinoic acid (19.7%+/-2.2) combined with docetaxel (100 muM) significantly decreased the percentage of proliferating cells (p<0.0001). The antiproliferative action of lycopene alone and in combination with docetaxel was also confirmed in immunohistochemical examination (decreased the percentage of PCNA and Ki67 positive cells). Also retinol (10 muM) and lycopene (20 and 50 muM) combined with estradiol (0.01 muM) statistically decreased the percentage of proliferating cells compared to the control (p<0.0001) and estradiol (p<0.01, p<0.0001) group (63.5%+/-14.8, 61.0%+/-20.6, 15.0%+/-5.5 respectively). In our experiments, the compounds tested induced an apoptotic effect. Docetaxel and estradiol increased the percentage of apoptotic cells (71% apoptotic cells after administration of 10 muM all-trans retinoic acid combined with 0.01 muM estradiol, p<0.0001). beta-carotene, lycopene and all-trans retinoic acid alone and in combination with docetaxel were found to influence the expression of bcl-2 and p53 antigen in the cells examined. The results of our study justified an important role of vitamin A in the pathophysiology of the ovarian endometrioid cancer.

Assessment of expression of selected Bcl-2 family proteins in lymphoid infiltration in patients with B-cell chronic lymphocytic leukaemia treated with nucleoside analogues.

B-cell chronic lymphocytic leukaemia (B-CLL) is characterized by clonal growth and accumulation of mature lymphoid cells due to disturbance in genetically regulated form of cell death called apoptosis. The intrinsic mechanism of apoptosis is controlled by Bcl-2 family proteins. Purine nucleoside analogues induce the apoptosis in cells in a state of quiescence. The aim of the study was to assess expression of selected Bcl-2 family proteins in neoplastic infiltration in bone marrow in patients with B-CLL treated with nucleoside analogues. The study comprised examination of bone marrow obtained routinely by trephine biopsy from 18 patients with B-CLL diagnosed before administration of purine nucleoside analogues treatment and after its completion. Expression of Bcl-2, Bcl-x and Bax proteins was examined. Lymphoid cells in bone marrow were present in all patients before administration of treatment. After treatment in two patients bone marrow was infiltrated in diffuse pattern, whereas other patients presented nodular pattern of infiltration. The difference between stage of infiltration before and after treatment was statistically significant (p<0.002). High percentage of infiltration cells with positive anti Bcl-2 reaction from 42.0% in one patient to 85.33+/-3.06% in four patients before treatment was observed. After treatment percentage of infiltration cells with positive anti Bcl-2 antibody reaction was from 33.0+/-18.38% in two patients to 99.0% in one patient. Positive correlation between stage of infiltration and expression of Bcl-2 protein was confirmed before and after treatment. Such correlations were not observed in case of Bax and Bcl-x. Strong staining of immunohistochemical reaction of cells in lymphoid infiltration with Bcl-2 antibody was confirmed. There was a difference between Bcl-/Bax ratio before and after treatment. Immunohistochemical assessment of expression of Bcl-2 family proteins in cells of lymphoid infiltration in bone marrow of patients with CLL is an important method in detection of minimal residual disease (MRD) after treatment.

Angiogenesis in gliomas.

Brain gliomas are characterized by invasive growth and neovascularisation potential. Angiogenesis plays a major role in the progression of gliomas and its determination has a great prognostic value. The aim of the study was to assess the vascularisation of chosen brain gliomas and to estimate how it is correlated with tumour histological type, malignancy grade, location and size, and with age and sex of patients. Tumour vascularisation analysis was based on the determination of microvascular proliferation (MVP) and microvessel density (MVD). Microvascular proliferation was measured with immunohistochemical methods using mouse monoclonal antibodies to detect cell proliferation antigens. The following antibodies were used Ki-67 and PCNA (DAKO). Identification of vessels was performed by CD31 antibody and anti-human von Willebrand factor (DAKO). The highest microvascular proliferation and microvascular density were observed in multiform glioblastomas and the lowest in oligodendrogliomas. Significant correlation was observed between the vascularisation and malignancy grade.

Can vitamin A modify the activity of docetaxel in MCF-7 breast cancer cells?

Docetaxel is one of the most effective chemotherapeutic agents in the treatment of breast cancer. On the other hand, the vitamin A family compounds play the essential roles in many biological processes in mammary gland. The aim of our study was to investigate the effect of all-trans retinol, carotenoids (beta-carotene, lycopene) and retinoids (9-cis, 13-cis and all-trans retinoic acid) on the activity of docetaxel and to compare these effects with the estradiol and tamoxifen actions on human ER(+) MCF-7 breast cancer cell line. The evaluation was based on [3H] thymidine incorporation and the proliferative activity of PCNA and Ki 67 positive cells. In our study, the incorporation of [3H] thymidine into cancer cells was inhibited to 50% by 0.2, 0.5 and 1 microM of docetaxel in the 24-hour culture and addition of estradiol (0.001 microM) didn't influence the results. However, addition of tamoxifen caused a statistically significant decrease of the percentage of the proliferating cells in the culture medium with 0.2 and 0.5 microM of docetaxel (38.99 +/- 2.84%, p<0.01 and 40.67 +/- 5.62%, p<0.01) in comparison to the docetaxel only group. The above-mentioned observations were also confirmed with the use of the immunohistochemical investigations. Among the examined vitamin A family compounds, the simultaneous application of beta-carotene (0.1 microM) and docetaxel (0.2 microM) resulted in a statistically significant reduction in the percentage of proliferating cells (40.25 +/- 14.62%, p<0.01). Lycopene (0.1 microM), which stimulates the growth of breast cancer cells in a 24-hour culture, had an inhibitory effect (42.97 +/- 9.58%, p<0.01) when combined with docetaxel (0.2 microM). Although, beta-carotene and lycopene belong to the different chemical groups, they surprisingly had a similar inhibitory influence on both growth and proliferation of MCF-7 breast cancer cells when combined with docetaxel. The application of docetaxel either with beta-carotene or lycopene had comparable inhibitory effect on breast cells growth and proliferation as tamoxifen. Therefore, it may suggest a possible important role of these carotenoids in the breast cancer therapy in women especially when docetaxel is applied.

[The effect of levothyroxine on the proliferative activity of the thyroid follicular cells]. / Wplyw lewotyroksyny na aktywnosc proliferacyjna komórekpecherzykowych tarczycy.

INTRODUCTION: Pharmacological treatment of non-toxic nodular goitre with levothyroxine has caused a discussion about its effectiveness and safety. The aim of the study was to examine the influence of levothyroxine on the proliferative activity of the thyroid follicular cells. MATERIAL AND METHODS: Cytological material was obtained from 32 euthyroid females, aged 34-69 years, by USG-guided Fine-Needle Aspiration Biopsy (FNAB), before and after 6 months treatment with levothyroxine. The patients were divided into 2 age groups: up to 45 years (14 patients) and above 45 years (18 females). Proliferative activity of the follicular cells was assessed by the proliferative index (PI), representing the percentage of follicular cells with positive reaction with the antibody Ki-67. Microscopic evaluation was carried out using morphometric computer system MicroImage InCD UDF (Olympus). RESULTS: After 6-months of treatment IP values decreased significantly in both groups studied (8.59 +/- 3.07 vs 6.51 +/- +/- 1.91; p < 0.001 in females aged < 45 years and 7.72 +/- 1.83 vs 5.09 +/- 1.51; p < 0.001 in the older group). CONCLUSION: Our results indicate that levothyroxine has an influence on the proliferative activity of the thyroid follicular cells, particularly in post-menopausal women.

The influence of N- and O-glycosylation inhibitors on the glycosylation profile of cellular membrane proteins and adhesive properties of carcinoma cell lines.

The effects of N- and O-glycosylation inhibitors on the expression of membrane proteins (MUC1 and some integrins) were evaluated in human endometrial (Ishikawa) and breast (MCF-7) cancer cells. Subconfluent cells were treated with 1-3 mg% concentration of tunicamycin and 2-10 mM of benzyl-N-acetyl-alpha-galactosaminide for 1-2 days, and used for flow cytometry, immunohistochemical staining, adhesion test and Western blotting. Benzyl-N-acetyl-alpha-galactosaminide inhibits MUC1 expression on the surface of breast more than endometrial cancer cells. Tunicamycin reduces MUC1 concentration on the cellular surface more than benzylglycoside, and greatly reduces glycosylation of glycoproteins, causing an increase in cell adhesion in both types of cancer cells. The expression of alpha2beta1 integrins on the surface of these cells was weak and decreased after treatment with inhibitors. Two different glycoforms of MUC1 proteins in endometrial cells and three in breast cancer cells were expressed and their molecular weights were reduced after treatment with glycosylation inhibitors. It was confirmed with lectin detection of carbohydrate epitopes (Tn and T) in MUC1 proteins. These observations show that glycosylation inhibitors altered the N- and O-glycan patterns in a sufficient manner, and positively modified the biological features of cancer cells.

Mast cells in neoangiogenesis.

Mast cells (MCs) always accompany connective tissue and are located in the proximity of lymphatic and blood vessels and nerve fibers. They are round or oval mononuclear cells with a diameter of 4-20 microm containing in their cytoplasm specific exocrine granules (storing neutral proteases) enclosed by a single membrane, whose presence is regarded as an index of the MC's static state. In view of their wide distribution in the organism, they play various roles in, for example, type I hypersensitivity reactions, chronic inflammatory processes, tissue reconstruction and wound healing, and pathological pulmonary fibrosis. They also play a role in angiogenesis, both in normal conditions during tissue regeneration and in pathological neoplastic states. The microcirculation provides building and nutritional substances to cancer cells and enables cancer spread via the blood. On the other hand, a tumor with good vascularization is more prone to penetration by cytostatics, which is why angiogenesis is a very important process in the course of neoplastic disease. Many authors indicate a close association between mast cells and angiogenesis. Some substances contained in the cytoplasm of these cells are potent stimulators of angiogenesis (tryptase, heparin), while others may inhibit it (protamine, platelet factor 4), and this conditions cancer growth and the development of the metastatic process. It is not known, however, what interactions occur between stimulants and inhibitors and what the proportional involvement of particular mediators in the formation of new vessels is.

The influence of the vascularisation of the follicular thyroid nodules on the proliferative activity of the follicular cells.

The enlargement of the thyroid is, in general, benign in origin and due to nodular goitre. Follicular cellular proliferation of thyroid nodules has been increasingly observed recently. With fine needle aspiration biopsy (FNAB) this is classified as a follicular tumour. These lesions present various patterns of vascularisation in ultrasound examination. The aim of the study was to establish the relation between follicular nodule vascularisation and the proliferative activity of various types of follicular cell. According to the manner of proliferation, patients were divided into groups as follows: (I). patients with hyperplastic nodules (46 cases), (II). patients with follicular adenoma (42 cases), and (III). patients with follicular cancer (9 cases). In each case B-mode sonography, Power Doppler, sonographically guided FNAB (S-FNAB), morphological examination and morphometry were performed. The proliferative activity was detected with immunohistochemical methods (PCNA, Ki 67 and MPM2) to determine the so-called "proliferative index". The study revealed increased proliferative activity in tumours of malignant origin and increased vascularisation in coexistence with increased proliferation of the follicular cells. As assessed by Power Doppler, an increased flow pattern in the centre of the nodules correlates with increased proliferative activity. The results suggest that Power Doppler examination could be helpful in selecting nodules for FNAB, especially in multinodular goitre.

The influence of chemotherapy on bone marrow in patients with chronic lymphocytic leukaemia.

In chronic lymphocytic leukaemia (CLL) bone marrow trephine biopsy (BMT) is not required for diagnosis but can have a significant prognostic value and can be used for the detection of the minimal residual disease (MRD) and for assessment of the effectiveness of the treatment applied. The aim of the study was to evaluate the morphological changes in bone marrow after treatment with purine nucleoside analogues cladribine and fludarabine. Bone marrow trephine biopsy was taken routinely from 15 patients with CLL. Bone marrow trephine biopsy was performed on every patient before as well as after chemotherapy. The number of cell elements of the marrow (the degree of atrophy), the patterns of bone marrow infiltration, the presence of reticulin and collagen fibres and the disturbances in bone marrow stroma were assessed. The infiltration of bone marrow by neoplastic cells was observed in all the patients before administration of chemotherapy. The infiltration was followed by an increase in the number of reticulin fibres. After the treatment a regression of the reticulin fibres was observed with the lessening of the infiltration. After the treatment the levels of marrow infiltrate were decreased. Increased hypoplasia of the bone marrow was observed after the chemotherapy.