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EN
To downregulate expression of the β1 integrin subunit in endothelial cells, plasmid encoding the ribozyme cassette containing hammerhead ribozyme flanked at the 5' terminus by tRNAVal and at the 3' terminus by constitutive transport element sequences was constructed. When used to transfect immortalized human endothelial cell line EA.hy 926, it selectively blocked the synthesis of the β1 integrin subunit and thus inhibited migration and proliferation of the cells. Thus, this construct may be a valuable tool to control the proangiogenic phenotype of stimulated endothelial cells.
EN
Isothiocyanate is a transient receptor potential ankyrin 1 (TRPA1) agonist and also an inhibitor of ion transporters such as anion exchanger (AE) and Na+/HCO3- co-transporter (NBC). We examined the expression of TRPA1 and ion transporters in monolayers of the rat gastric epithelial cell line RGM1 and investigated the involvement of these factors in the inhibitory action of isothiocyanate on epithelial wound healing. After obtaining a confluent monolayer, a round artificial wound of constant size was induced in the center of the cell monolayer using a pencil-type mixer with a rotating silicon tip. Immediately after the wound induction, cells at the edge of the wound started to form lamellipodia, migrating towards the center of wound, and the cell-free area decreased with time. Addition of allyl isothiocyanate to standard buffer suppressed the recovery of the wound in a concentration-dependent manner without affecting the viability of the RGM1 cells. Icilin, another TRPA1 agonist, dose-dependently inhibited wound repair. Likewise, 4,4’-diisothio- cyanatostilbene-2,2’-disulfonic acid (DIDS), a stilbene compound containing an isothiocyanate group, also inhibited the recovery of epithelial wounds. In addition, the repair of epithelial wounds was suppressed when the cells were incubated in Na+, Cl- or HCO3- free buffer. The RGM1 cells expressed the mRNAs of AE2a and NBC1 but not TRPA1. These results suggested that isothiocyanate impairs the repair of epithelial wounds in RGM1 cells, probably through the inhibition of ion transporters such as AE2a and NBC1 and not the activation of the TRPA1 channel. It is assumed that the process of epithelial repair is associated with the regulation of cell volume and intracellular pH (pHi) by these ion transporters.
EN
The results of several experimental studies have shown that ascorbic acid inhibits tumor growth and metastasis. Ascorbic acid is an antioxidant that acts as a scavenger for a wide range of reactive oxygen species (ROS). Both tumour metastasis and cell migration have been correlated with the intracellular ROS level, so it was postulated that the inhibitory effect of ascorbic acid derivatives on cell motility may be caused by scavenging of ROS. Time-lapse analyses of Walker 256 carcinosarcoma cell migration showed that both the speed of movement and the cell displacement were inhibited by ascorbic acid applied in concentrations ranging from 10 to 250 μM. This effect correlated with a reduction in the intracellular ROS level in WC 256 cells, suggesting that ROS scavenging may be a mechanism responsible for the inhibition of WC 256 cell migration. However, another potent antioxidant, N-acetyl-L-cysteine, also efficiently decreased the intracellular ROS level in WC 256 cells, but did not affect the migration of the investigated cells. These results demonstrate that intact, unmodified ascorbic acid applied in physiologically relevant and nontoxicconcentrations exerts an inhibitory effect on the migration of WC 256 carcinosarcoma cells, and that this may be one of the factors responsible for the anti-metastatic activity of vitamin C. However, our data does not support the hypothesis that the scavenging of intracellular ROS is the main mechanism in the inhibition of cancer cell migration by ascorbic acid.
EN
To downregulate expression of the β1 integrin subunit in endothelial cells, plasmid encoding the ribozyme cassette containing hammerhead ribozyme flanked at the 5' terminus by tRNAVal and at the 3' terminus by constitutive transport element sequences was constructed. When used to transfect immortalized human endothelial cell line EA.hy 926, it selectively blocked the synthesis of the β1 integrin subunit and thus inhibited migration and proliferation of the cells. Thus, this construct may be a valuable tool to control the proangiogenic phenotype of stimulated endothelial cells.
EN
The effect of triethyllead (TriEL) on the morphology and motile activity of Walker 256 carcinosarcoma cells was investigated. It was found that both 2 and 5 μM TriEL affected the cellular motility in a dose- and time- dependent manner. Initially, 2 μM TriEL caused the formation of blebs instead of lamellipodia at the front of some cells and stimulated the migration of Walker cells, but after 2 hours of 2 μM TriEL treatment, a reduction of cellular motility was observed. In the presence of 5 μM TriEL, Walker 256 carcinosarcoma cells rounded up, and their rate of movement was reduced. Moreover, the treatment of Walker carcinosarcoma cells with TriEL caused the disruption of microtubules and affected the F-actin distribution at both concentrations. At a concentration of 2 μM TriEL, the actin staining intensity was greatest in the tail of front-tail polarised blebbing cells and the actin layer was very thin at the leading edge. The control cells showed linear cortical F-actin distribution and somewhat less intense cytoplasmic staining at the same TriEL concentration. Cells treated with 5 μM TriEL showed an under-membrane pattern of actin distribution.
EN
Chemokines are relatively recently characterized and growing fast family of low molecular weight cytokines, which stimulate migration of cells in vitro and in vivo. Together with adhesion molecules chemokines are involved in the complex process of migration of leukocytes outside of blood vessels. They also direct their migration within inflamed peripheral tissues. Chemokines are divided into four subfamilies. The main criterion of this division is the localization of pairs of cysteines in the NH2 region. The major chemokine subfamilies are CXC (α) and CC (β) chemokines. In CC subfamily the first cysteines are adjacent, in CXC group they are separated by a single aminoacid. The exemptions are Lymphotactins α and β which posses only one pair of cysteines (C or γ subfamily) and Fractalkine, in which the first cysteines are separated by three aminoacids (CX3C or δ subfamily). Functionally chemokines can be divided into proinflammatory and lymphoid. Chemokines influence their target cells through the specific seven transmembrane domain receptors. They are the important mediators of migration of inflammatory cells to the central nervous system (CNS) during different pathological processes that’s why they became the target of interest in the studies on multiple sclerosis (MS). Analysis of MS brains showed significantly increased expression of chemokines CCL4 and CCL5 at mRNA level. In the cerebrospinal fluid of MS patients during relapse increased level of chemokines CCL5, CXCL9 and CXCL10 was detected. Within chronic active demyelinating plaques in MS brains expression of CCR2, CCR3 and CCR5 was observed in macrophages and microglia.
PL
Chemokiny stanowią stosunkowo niedawno wyodrębnioną i szybko rozrastającą się rodzinę cytokin charakteryzujących się małym ciężarem cząsteczkowym oraz zdolnością stymulowania migracji komórek in vitro oraz in vivo. Chemokiny wspólnie z molekułami adhezyjnymi biorą udział w złożonym procesie przemieszczania się leukocytów poza łożysko naczyniowe. Ukierunkowują one również migrację leukocytów w obrębie tkanek obwodowych, gdzie rozwija się zapalenie. Chemokiny podzielono na cztery grupy. Decydującym kryterium podziału jest położenie względem siebie par cystein w okolicy końca NH2. Dwie największe grupy che-mokin to chemokiny CXC (α) i CC (β). W grupie CC dwie pierwsze następujące po sobie cysteiny pozostają nierozdzielone, podczas gdy w grupie CXC rozdziela je pojedynczy aminokwas. Wyjątkami są limfotaktyny α i β posiadające tylko jedną parę cystein (grupa chemokin C lub γ) oraz fraktalkina, u której dwie pary cystein oddzielają trzy inne aminokwasy (grupa CX3C lub δ). Ze względu na funkcję chemokiny można podzielić na prozapalne oraz limfoidalne. Chemokiny oddziałują na komórki docelowe za pośrednictwem swoistych receptorów charakteryzujących się obecnością siedmiu domen przezbłonowych. Będąc jednym z głównych czynników odpowiedzialnych za migrację komórek zapalnych do ośrodkowego układu nerwowego (OUN) w różnych procesach patologicznych, chemokiny stały się obiektem zainteresowania również w badaniach nad stwardnieniem rozsianym (SM). Analiza mózgów pacjentów z SM wykazała istotny wzrost ekspresji chemokin CCL4 i CCL5 na poziomie mRNA. W płynie mózgowo-rdzeniowym w okresie rzutu choroby stwierdzono podwyższony poziom chemokin CCL5, CXCL9 oraz CXCL10. U chorych z SM w obrębie przewlekłych aktywnych ognisk demielinizacyjnych stwierdzano komórki barwiące się w kierunku receptorów CCR2, CCR3 i CCR5, które odpowiadały morfologią makrofagom i mikroglejowi.
EN
The purpose of the present study was to determine the role of PI 3 kinase in tumor cell migration. The migration of ovary carcinoma cells (OVP10) was strongly inhibited by wortmannin to the same extent as by TPA treatment. The strongest additive inhibitory effect was noted after cell treatment with wortmannin and TPA together. Western blotting studies showed activation / partial down regulation of PKC after TPA treatment and decreased amount of the enzyme following wortmannin exposure. Altogether, it looks like wortmannin inhibits migration of studied cells.
EN
Bioremediation is the effective remediation technology for soils contaminated by biodegradable contamination. However, bioremediation of soils contaminated by hydrophobic compounds still remains a major challenge for the scientific and industrial world. There is still the need to develop techniques which allow an increase in bioremediation efficiency. A possible solution seems to be the stimulation of bacteria migration through the subsurface while using bioremediation. In this study a weak electric field in combination with the following bacterial strains: Pseudomonas putida, Bacillus subtilis and Klebsiella pneumoniae was used to stimulate bacterial cell migration, as well as the biodegradation of crude oil contamination in soil samples. Bacterial cell migration under the influence of the weak electric field and crude oil biodegradation were estimated during the experiments. The effect of changes in electrode polarization were also included in this study. Results show that weak electric field application has a great influence on the speed and direction of bacterial migration in soil samples and biodegradation of the pollution. From the study of the application of the electric field in soil bacteria migration can be forced in the desired direction and consequently stimulate biodegradation of contamination in selected areas.
EN
Gastrointestinal ulcer healing is a complex process, involving cell migration, proliferation, angiogenesis and extracellular matrix deposition, all ultimately leading to reconstruction of tissue architecture within the ulcer scar. These processes are controlled by growth factors, cytokines and hormones. Transforming growth factor-ß (TGF-ß ), one of the multifunctional peptide growth factors, has been reported to positively regulate gastrointestinal ulcer healing. Although TGF-ß inhibits cell proliferation in a variety of cells, it induces cell migration, angiogenesis, and enhances extracellular matrix production necessary for gastrointestinal ulcer healing. TGF-ß exerts its action by binding to its transmembrane serine/threonine kinase receptors, which in turn triggers activation of various intracellular signaling pathways. Smads are intermediate effector proteins that play key roles in biological activities of TGF-ß by transmitting the signals from the cell surface directly into the nucleus and initiating transcription. New insight into the mechanisms underlying TGF-ß-Smad modulation of gastrointestinal ulcer healing will likely enhance our understanding of the mechanisms controlling the healing processes of gastrointestinal ulcers.
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