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EN
Ulcerative colitis (UC) is a inflammatory disease of large bowel. The amount of people suffering from UC increases from year to year. Pathogenesis of this affection is still not entirely clear. Mechanisms of proliferation and apoptosis in colonocytes in the course of the disease are defectedThe aim of the study was to assess the rate of proliferation and intensity of apoptosis in colonocytes in patients with diagnose UC.Material and methods. Colon pathological samples taken from patients with diagnosed ulceraive colitis were examined. Patients were in both clinical and endoscopic remission and were treated with mesalazin. They were patient of Department of General and Colorectal Surgery. To estimate proliferation index dye with monoclonal antibody against Ki67. To determine apoptosis level immunohistochemistry with antybody against Bax was used.Results. Average Ki-67 in the test group was 42,13%, the largest value amounted to 57% and the lowest of 33%. Average value of Bax was 1.47 and ranged between 0-3. High index of bax appear not only in the bottom of the crypt, but also at their outlet.Conclusions. In ulecerative colitis genetic and immunological disturbances occur despite treatment. Mesalazine acting only on certain routes associated with the UC holds the remission, without, however "the molecular remission". Thus, it appears that the results of our research are another proof of the necessary caution in weaning support treatment.
EN
Helicobacter pylori (H. pylori) bacteria are human pathogens causing symptomatic gastritis, peptic ulcer or gastric cancer. Little is known about the kinetics of immune responses in H. pylori infected patients because the initial moment of infection has not been identified. Various animal models are used to investigate the immune processes related to H. pylori infection. In this study we checked whether H. pylori infection in guinea pigs, mimicking natural H. pylori infection in humans, resulted in the development of specific immune responses to H. pylori antigens by measuring the proliferation of lymphocytes localized in mesenteric lymph nodes, spleen and peripheral blood. The maturity of macrophages and cytokines, delivered by monocyte-macrophage lineage or lymphocytes, were considered as mediators, which might influence the lymphocyte blastogenic response. The obtained results showed the activation of T cells localized in mesenteric lymph nodes by H. pylori antigens in H. pylori infected guinea pigs four weeks postinfection. The blastogenic activity of lymphocytes was shaped by their interaction with antigen presenting cells, which were present in the cell cultures during the whole culture period. Moreover, the balance between cytokines derived from adherent leukocytes including interleukin 8 - IL-8 as well as interferon gamma - IFN-γ, and transforming growth factor beta - TGF-β delivered by lymphocytes, was probably important for the successful proliferation of lymphocytes. The H. pylori specific lymphocytes were not propagated in peripheral blood and spleen of H. pylori infected animals. The modulation of immunocompetent cells by H. pylori antigens or their different distribution cannot be excluded.
EN
In the present work, we have reviewed data showing that triiodothyronine and its nuclear receptors modify expression of different genes/proteins involved in cell cycle control beginning from growth factors (such as EGF and TGF-β), to cell surface receptors (EGFR), as well as proteins acting at the cell membrane (Ras), various transcription factors (c-Fos, c-Myc, E2F1), cyclins, Cip/Kip family of cdk2 inhibitors, and p53 inhibitor Mdm2 (Table 1). We have shown how TRs are also able to modify the fate of a cell, thanks to their ability to form complexes with other transcription factors such as p53 - a key regulator of apoptosis and proliferation. Available data show that the function of thyroid hormones and of their receptors on cell proliferation is not homogenous. In fact, it strongly depends on the cell type, its developmental state (progenitor or differentiated), its patho-physiological state (normal or tumor cell), and the so-called 'cellular context'. Therefore, it is not possible to uniformly recommend T3 treatment or T3 depletion to stop or initiate proliferation of all cell types. Instead, a very individual and careful action should be considered.
EN
In the search for potential therapeutics, isoxazole derivatives are still objects of interest. Previously described immunoregulatory properties of 5-amino-3-methyl-4-isoxazolecarboxylic acid (AC) benzylamides prompted us to synthesize a new class of compounds of immunotropic activity. A series of new compounds containing the isoxazole moiety were synthesized using Passerini three-component reaction. The effects on phytohemagglutinin A (PHA)-induced proliferation of human peripheral blood mononuclear cells (PBMC), production of tumor necrosis factor alpha (TNF α) in human whole blood cultures stimulated with lipopolysaccharide (LPS) and two-way mixed lymphocyte reaction (MLR) of PBMC, were investigated. Also, the effect of 1-(cyclohexylcarbamoyl)cyclohexyl 5-amino-3-methylisoxazole-4-carboxylate (PUB1) on the expression of signaling molecules associated with cell apoptosis in Jurkat cells was also determined. The results showed that the compounds inhibited to various degree mitogen-induced PBMC proliferation in a dose-dependent manner and TNF α production at 10 μg/ml. PUB1 compound, selected on the basis of its strongest antiproliferative activity, was also shown to inhibit MLR. The molecular data suggest that immunosuppressive action of PUB1 depended on induction of Fas and elevation of caspase 8 expression. In summary, we revealed immunosuppressive properties of a new class of isoxazoles and established the mechanism of action of a representative PUB1 compound.
EN
Triterpene saponins (saponosides) are found in a variety of higher plants and display a wide range of pharmacological activities, including expectorant, anti-inflamatory, vasoprotective, gastroprotective and antimicrobial properties. Recently, a potential anticancer activity of saponins has been suggested by their cytotoxic, cytostatic, pro-apoptotic and anti-invasive effects. At high concentrations (more than 100 µM) saponins exert cytotoxic and haemolytic effects via permeabilization of the cell membranes. Noteworthy, the inhibition of cancer cell proliferation, the induction of apoptosis and attenuation of cell invasiveness is observed in the presence of low saponin concentrations. Saponins might affect the expression of genes associated with malignancy. These alterations are directly related to the invasive phenotype of cancer cells and depend on "cellular context". It illustrates the relationships between the action of saponins, and the momentary genomic/proteomic status of cancer cells. Here, we discuss the hallmarks of anti-cancer activity of saponins with the particular emphasis on anti-invasive effect of diverse groups of saponins that have been investigated in relation to tumor therapy.
EN
Retinoic acid and transforming growth factor-β (TGF-β) affect differentiation, proliferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. 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 MCF-7 cells were also determined. Exogenous TGF-β1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-β1 concentrations, a marked reduction in the stimulatory action of estradiol (10-9 and 10-8 M) was observed whereas in combination with tamoxifen (10-7 and 10-6 M) only 30 ng/ml TGF-β1 caused a statistically significant reduction to aproximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-β1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 ± 19% (control =100%) by 3 ng/ml TGF-β1, and this dose was used throughout. It was found that addition of TGF-β1 and isotretinoin to the culture did not decrease proliferation, while TGF-β1 and tretinoin at low concentrations (3 × 10-8 and 3 × 10-7 M) reduced the percentage of proliferating cells by aproximately 30% (67±8% and 67±5%, P <0.05 compared to values in the tretinoin group). Both retinoids also led to a statistically significant decrease in the stimulatory effect of 10-9 M estradiol, attenuated by TGF-β1. In addition, the retinoids in combination with TGF-β1 and tamoxifen (10-6 M) caused a further reduction in the percentage of proliferating cells. Immunocytochemical analysis showed that all the examined compounds gave a statistically significant reduction in the percentage of cells with a positive reaction to PCNA and Ki 67 antigen. TGF-β1, isotretinoin and tretinoin added to the culture resulted in the lowest percentage of PCNA positive cells. However, the lowest fraction of Ki 67 positive cells was observed after addition of isotretinoin. The obtained results also confirm the fact that the well-known regulatory proteins Bcl-2 and p53 play an important role in the regulation of apoptosis in the MCF-7 cell line, with lowered Bcl-2 expression accompanying easier apoptotic induction. The majority of the examined compounds act via the p53 pathway although some bypass this important proapoptotic factor.
EN
The protein p63 plays a significant role in the development of animal epithelium. p63 is a regulator of differentiation, senescence and adhesion programs in numerous mature epithelial tissues. In patients with a healthy epidermis, p63 maintains cell progenitor potential - the ability for cellular division to occur using the delayed differentiation program. It is also responsible for the protecting the epithelial phenotype from depletion in migrating cells, thus resulting in invasion and infiltration after altering its endogenous expression.The aim of the study was to compare the number of cells with p63 protein expression and the presence of Ki67 proliferation marker in the epidermis in patients with chronic venous ulcerations versus those with properly healing wounds.Material and methods. Study materials were comprised of biopsy samples collected from healthy volunteers and patients treated for venous ulcerations. The specimens were subjected to immunohistochemical staining using available monoclonal antibodies and were analyzed with an imaging analysis program which evaluated the expression indices of both proteins in areas of intensified cellular division, i.e. wound edges.Results. The number of cells displaying protein expression in patients with chronic venous ulcerations was significantly lower in comparison to the values observed in healthy volunteers. This was determined during the intermediary phase of wound healing, the most pronounced phase of cellular response to injury.Conclusions. Decreased epidermal p63 expression in patients with venous ulcerations suggests insufficient protein production for the maintenance of autoregeneration and long-lasting division; both are required for the supplementation of migrating cells. The above-mentioned phenomenon suggests that there may be a role for p63 in regulation of the healing process and pathophysiology of chronic venous leg ulcerations.
EN
Immunosenescence is viewed as a remodeling process with the exhaustion of naïve T cells and filling up of the immunological space with memory cells. In this study some phenotypic changes of CD8+ human cells during in vivo ageing were compared with those observed in long term cultures of lymphocytes derived from cord blood or from peripheral blood from donors of different age. Both in vivo and in vitro a significant decrease of the fraction of CD8+CD28+ cells was observed. Comparing the proportions of other T cell subpopulations (the CD4/CD8+ ratio, CD56, CD57, CD27) made it possible to conclude that replicative senescence in vitro partially reflects in vivo ageing.
EN
Vitamin D (VD) and its different analogues, besides their classic role as regulators of calcium and phosphor homeostasis, have emerged as a large family of antiproliferative agents. Such properties suggested VD potential as a therapy for chronic inflammatory diseases, including nasal polyposis (NP). NP growth involves both an inflammatory process and the proliferation of fibroblast as an important factor inducing aberrations in the phenotype of the epithelium. The aim of this study was to investigate the possible influence of 1α,25-dihydroxyvitamin D3 (calcitriol) and 1α,24(R)-dihydroxyvitamin D3 (tacalcitol) in monotherapy and in combination with budesonid R (BR) on NP fibroblast proliferation. Material and methods: The study involved 26 samples of NP. NP cells were cultured on 96-well plates beginning with a concentration of 5 × 103 cells per well with RPMI 1640 medium supplemented with antibiotics and 10% foetal bovine serum. After the fourth to sixth passage the medium was replaced with a nutrient medium with calcitriol or tacalcitol in a defined concentration (from 10-9 M to 10-3 M) alone or in combination with BR in 1:1, 1:3 or 3:1 ratios, each at concentrations from 10-5 M to 10-3 M. Results: Growth inhibition of nasal fibroblasts exposed to calcitriol or tacalcitol was noted. Significant antiproliferating activity was observed at calcitriol concentrations of 10-4 M and 10-3 M after 48 h, and at a concentration of 10-3 M after 72 h with the percentage of proliferating cells reduced to 30% compared to the control samples (P < 0.05). In cells treated with tacalcitol the maximal effect was seen at 10-4 M after 48 h and at 10-3M after 72 h with a 60% inhibition with respect to the control (P < 0.05). The inhibition of fibroblast proliferation reached the maximal level when they were exposed to calcitriol: BR (1 : 1) or tacalcitol: BR (1 : 1), each at a concentration of 10-4 M, after 72 h (82% and 69%, respectively). Conclusions: The antiproliferative activity of calcitriol and tacalcitol in NP cultures was confirmed. Because of its lower toxicity and higher activity tacalcitol seems to be the more promising agent in NP therapy, both as a single medication and in treatment protocols with BR.
EN
Normal human T lymphocytes growing in culture undergo replicative senescence. Previously, we have shown that in our conditions polyclonal T cells cease proliferation after about three weeks (Radziszewska et al., 1999, Cell Biol. Int. 23, 97-103). Now we present results of a more detailed analysis of in vitro growth as well as phenotypic changes of T cells. Cell cycle analysis showed that about 20% of cells were in the S phase untill the 17th day of culture (young cells). The highest number of mitotic cells (phase G2/M; 10%) was observed during the first week of culture. All not dividing senescent cells were stopped in the G1 phase (after the 30th day of culture). The sub-G1 fraction which represents apoptotic cells did not exceed 8% during the whole period until the 30th day of culture. During in vitro T-cell growth, a rather rapid selection to CD3+CD8+ cells occurs. In the presenescent (between the 17th and 30th day) and senescent populations the majority of cells (above 90%) were CD8 positive. We also have checked the expression of α-chain interleukin-2 (IL-2) receptor (CD25). In young and presenescent cells about one third of cells was CD25 positive, but only 15% in the pool of senescent cells. Immunoblotting analysis of p16 protein recognized previously as a marker of senescent T cells, showed its highest and transient expression in presenescent cells. A critical review of the polyclonal T cell replicative senescence model is presented.
Kosmos
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2018
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vol. 67
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issue 1
131-137
PL
Białko CacyBP/SIP występuje w różnych komórkach i tkankach ssaków, a jego wysoki poziom notowany jest w mózgu, śledzionie, grasicy oraz w wielu nowotworach. CacyBP/SIP oddziałuje z wieloma białkami efektorowymi, w tym z białkami cytoszkieletu: aktyną, tubuliną, tropomiozyną. Wskazuje to, iż CacyBP/SIP bierze udział w procesach komórkowych, którym towarzyszą zmiany w organizacji cytoszkieletu zarówno w warunkach fizjologicznych jak też w różnych stanach chorobowych. W niniejszej pracy przedstawiono charakterystykę oddziaływania CacyBP/SIP z białkami cytoszkieletu oraz rolę tych interakcji w różnych procesach komórkowych.
EN
The CacyBP/SIP protein is present in different mammalian cells and tissues. Its particularly high level is observed in brain, spleen, thymus and in many cancers. CacyBP/SIP interacts with different targets including cytoskeletal proteins such as tubulin, actin, tropomyosin. This indicates that CacyBP/SIP is involved in cellular processes associated with changes in cytoskeleton organization under physiology and pathology. In the present work the characteristics of complexes formed between CacyBP/SIP and cytoskeletal proteins and the role of those interactions are presented.
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