| 1. |
- Tågerud, Sven, et al.
(author)
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The nifk gene is widely expressed in mouse tissues and is up-regulated in denervated hind limb muscle
- 2003
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In: Cell Biology International. - 1065-6995 .- 1095-8355. ; 27:6, s. 469-475
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Journal article (peer-reviewed)abstract
- Denervation of skeletal muscle alters the expression of many genes, which may be important for establishing optimal conditions for reinnervation. Using the differential display technique we have attempted to discover neurally regulated genes in skeletal muscle. An mRNA that is up-regulated in denervated hind limb muscle was identified and cloned. The cDNA encodes an RNA-binding protein, which was discovered during the course of this work to be a nucleolar protein interacting with the fork-head associated domain of the proliferation marker protein Ki-67, and named NIFK. We show that the nifk gene is widely expressed in adult mouse tissues and that the expression is up-regulated in denervated hind limb muscle. No difference between expression in perisynaptic and extrasynaptic portions of muscle was observed. The widespread expression in adult tissues suggests that the NIFK protein has other functions in addition to its interaction with Ki-67, which is only expressed in proliferating cells.
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| 2. |
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| 3. |
- Bengtsson, Torbjörn, 1955-, et al.
(author)
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Role of the actin cytoskeleton during respiratory burst in chemoattractant-stimulated neutrophils
- 2006
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 30:2, s. 154-163
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Journal article (peer-reviewed)abstract
- The aim of this study was to clarify the role of the actin cytoskeleton during chemotactic peptide fMet-Leu-Phe (fMLF)-stimulated respiratory burst in human neutrophil granulocytes. Reactive oxygen species (ROS) was measured as luminol-amplified chemiluminescence (CL) and F-actin content as bodipy phallacidin fluorescence in neutrophils treated with latrunculin B or jasplakinolide, an inhibitor and activator of actin polymerization, respectively. Latrunculin B markedly decreased, whereas jasplakinolide increased, the F-actin content in neutrophils, unstimulated or stimulated with fMLF. Latrunculin B enhanced the fMLF-triggered ROS-production more than tenfold. Jasplakinolide initially inhibited the fMLF-induced CL-response, however, caused a potent second sustained phase (>400% of control). Both actin drugs triggered a substantial CL-response when added 5-25 min after fMLF. This was also valid for chemotactic doses of fMLF, where latrunculin B and jasplakinolide amplified the ROS-production 5-10 times. By using specific signal transduction inhibitors, we found that the NADPH oxidase activation triggered by destabilization of the actin cytoskeleton occurs downstream of phospholipase C and protein kinase C but is mediated by Rho GTPases and tyrosine phosphorylation. In conclusion, rearrangements of the actin cytoskeleton are a prerequisite in connecting ligand/receptor activation, generation of second messengers and assembly of the NADPH oxidase in neutrophil granulocytes. © 2005 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.
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| 4. |
- Bilyy, Rostyslav, et al.
(author)
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In vivo expression and characteristics of novel alpha-D-mannose-rich glycoprotein markers of apoptotic cells
- 2005
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 29:11, s. 920-928
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Journal article (peer-reviewed)abstract
- We recently established that an increased expression of alpha-D-mannose (Man)- and beta-D-galactose-rich plasma membrane glycoproteins (GPs) is characteristic for apoptotic cells in vitro [Bilyy, R.O., Stoika, R.S., 2003. Lectinocytochemical detection of apoptotic murine leukemia L1210 cells. Cytometry 56A, 89-95]. It was independent of cell line or apoptosis-inducing agent, and can therefore be considered as a selective marker for identification and isolation of apoptotic cells [Bilyy, R.O., Antonyuk, V.O., Stoika, R.S., 2004. Cytochemical study of role of alpha-D-mannose- and beta-D-galactose-containing glycoproteins in apoptosis. J. Mol. Histol. 35, 829-838]. The main goals of the present study were: (1) to determine whether an increased expression of specific GPs also takes place after apoptosis induction in vivo; and (2) to identify additional characteristics of the membrane GP markers of the apoptotic cells. To reach these goals, we studied the expression of alpha-Man-rich membrane GPs in murine leukemia L1210 cells inoculated into abdominal cavities of mice which were then subjected to the action of apoptosis inducer doxorubicin. Another experimental model used in the present work was splenocytes obtained from mice treated with dexamethasone. Lectin-affinity chromatography and PAGE electrophoresis, or PAGE electrophoresis and lectinoblot analysis were applied for isolation of plasma membrane GPs (34 kDa, and high M(W) of approximately 600 and 800 kDa) whose expressions were increased during apoptosis. Triton X-114 treatment of cell membrane samples showed that the apoptotic cell-specific GPs were localized in the peripheral and integral compartments of plasma membrane. Apoptosis in vitro and in vivo was accompanied by an increased expression of the same GP, identified by MALDI-TOF MS analysis as the microtubule-actin cross-linking factor 1. Other GPs, whose expressions were also increased at apoptosis, were similarly identified as G-protein beta-subunit like (Acc# BAA06185.1) and dystonin isoform beta.
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| 5. |
- Bolshakova, A., et al.
(author)
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Extra-cellular matrix proteins induce re-distribution of a-actinin-1 and a-actinin-4 in A431 cells
- 2007
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 31:4 SPEC. ISS., s. 360-365
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Journal article (peer-reviewed)abstract
- Alpha-actinins are actin-binding proteins of non-muscle cells, which can participate in the regulation of transcription factor activity. We describe the distribution of a-actinin-1 and -4 depending on different actin cytoskeleton formed as a result of cell adhesion to extracellular matrix proteins, such as fibronectin and laminin 2/4. Immunofluorescent studies show a difference in the distribution of a-actinin and -4. Both isoforms localise along stress-fibres, but a-actinin-1 localises in the perinuclear region more abundantly than a-actinin-4. Western blot analysis demonstrated existence of truncated forms of both isoforms. Truncated a-actinin-1 appears in cells spread on fibronectin or laminin. Cell spreading also correlated with more tight association of a-actinin-4 with chromatin. Basing on our previous finding of an interaction of a-actinin-4 with p65 subunit of the NF-?B, we checked the possible influence of immobilised ligands on its redistribution in nuclear complexes containing p65. a-Actinin-4 seems to be present in some but not all nuclear complexes containing p65. Immobilised ligands may affect the interaction of a-actinin-4/p65 complexes with chromatin. The data suggest that adhesion to extra-cellular matrix may interfere in cellular reactions mediated by a-actinin-1 and -4. © 2007 International Federation for Cell Biology.
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| 6. |
- Bolshakova, Anastasia, et al.
(author)
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Functional compartmentalisation of NF-B-associated proteins in A431 cells
- 2013
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In: Cell Biology International. - : Elsevier / Wiley-Blackwell. - 1065-6995 .- 1095-8355. ; 37:4, s. 387-396
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Journal article (peer-reviewed)abstract
- NF-B proteins belong to a family of ubiquitous transcription factors involved in a number of cellular responses. While the pathways of NF-B activation and input into the regulation of gene activity have been comprehensively investigated, its cytoplasmic functions are poorly understood. In this study we addressed effects of the compartmentalisation of NF-B proteins RelA/p65 and p50 in relation to the inhibitor IB-, using fibronectin (FN) and epidermal growth factor (EGF) for environmental stimulation of epidermoid carcinoma A431 cells. We thus assessed the presence of NF-B family proteins in the cytosol, membrane, nuclear and cytoskeletal fractions with a special attention to the cytoskeletal fraction to define whether NF-B was active or not. Sub-cellular fractionation demonstrated that the proportion of RelA/p65 differed in diverse sub-cellular fractions, and that the cytoskeleton harboured about 7% thereof. Neither the nuclear nor the cytoskeleton fraction did contain IB-. The cytoskeleton binding of RelA/p65 and p50 was further confirmed by co-localisation and electron microscopy data. During 30-min EGF stimulation similar dynamics were found for RelA/p65 and IB- in the cytosol, RelA/p65 and p50 in the nucleus and p50 and IB- in the membrane. Furthermore, EGF stimulation for 30min resulted in a threefold accumulation of RelA/p65 in cytoskeletal fraction. Our results suggest that nuclear-, membrane- and cytoskeleton-associated NF-B are dynamic and comprise active pools, whereas the cytoplasmic is more constant and likely non-active due to the presence of IB-. Moreover, we discovered the existence of a dynamic, IB--free pool of RelA/p65 associated with cytoskeletal fraction, what argues for a special regulatory role of the cytoskeleton in NF-B stimulation.
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| 7. |
- Dzanaeva, Ljubov, et al.
(author)
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The role of peroxisomes in xylose alcoholic fermentation in the engineered Saccharomyces cerevisiae
- 2020
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In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 44:8, s. 1606-1615
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Journal article (peer-reviewed)abstract
- Xylose is a second-most abounded sugar after glucose in lignocellulosic hydrolysates and should be efficiently fermented for economically viable second-generation ethanol production. Despite significant progress in metabolic and evolutionary engineering, xylose fermentation rate of recombinant Saccharomyces cerevisiae remains lower than that for glucose. Our recent study demonstrated that peroxisome-deficient cells of yeast Ogataea polymorpha showed a decrease in ethanol production from xylose. In this work, we have studied the role of peroxisomes in xylose alcoholic fermentation in the engineered xylose-utilizing strain of S. cerevisiae. It was shown that peroxisome-less pex3 Delta mutant possessed 1.5-fold decrease of ethanol production from xylose. We hypothesized that peroxisomal catalase Cta1 may have importance for hydrogen peroxide, the important component of reactive oxygen species, detoxification during xylose alcoholic fermentation. It was clearly shown that CTA1 deletion impaired ethanol production from xylose. It was found that enhancing the peroxisome population by modulation the peroxisomal biogenesis by overexpression of PEX34 activates xylose alcoholic fermentation.
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| 8. |
- Filyak, Yevhen, et al.
(author)
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Transforming growth factor beta-1 enhances cytotoxic effect of doxorubicin in human lung adenocarcinoma cells of A549 line
- 2007
-
In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 31:8, s. 851-855
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Journal article (peer-reviewed)abstract
- Transforming growth factor beta-1 (TGFbeta-1) is a regulator of cell proliferation, differentiation and apoptosis. Doxorubicin (adriamycin), an anthracycline drug causing double-strand DNA breaks, is widely used in anticancer chemotherapy. Here we demonstrated that TGFbeta-1 enhanced cytotoxic (proapoptotic) action of doxorubicin towards cultured human lung carcinoma A549 cells. Western-blot analysis and immunocytochemistry were used to show that doxorubicin induced PARP degradation in A549 cells, and TGFbeta-1 enhanced that action of the drug. The obtained results suggest a possibility of biomodulating effect of TGFbeta-1 on tumor cell treatment with doxorubicin.
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| 9. |
- Fjällström, Ann-Kristin, et al.
(author)
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Expression and phosphorylation of eukaryotic translation initiation factor 4-gamma (eIF4G) in denervated atrophic and hypertrophic mouse skeletal muscle
- 2015
-
In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 39:4, s. 496-501
-
Journal article (peer-reviewed)abstract
- The eukaryotic translation initiation factor 4-gamma (eIF4G) is important for the initiation of protein synthesis and phosphorylation on S1108 regulates this function of eIF4G. Thus, increased phosphorylation has been reported in conditions associated with increased protein synthesis such as meal feeding and insulin/IGF-1 treatment whereas decreased phosphorylation occurs following starvation, dexamethasone treatment, in sepsis and in atrophic denervated hind-limb muscle. The aim of the present study was to test the hypothesis that S1108 phosphorylation of eIF4G is differentially affected in denervated atrophic hind-limb muscles and denervated hypertrophic hemidiaphragm muscle. Protein expression and phosphorylation in innervated and 6-days denervated atrophic hind-limb muscles (pooled gastrocnemius and soleus) and hypertrophic hemidiaphragms were studied semi-quantitatively using Western blots. Total expression of eIF4G did not change in denervated hind-limb muscles but increased about 77% in denervated hemidiaphragm. S1108 phosphorylated eIF4G decreased about 64% in denervated hind-limb muscles but increased about 1.3-fold in denervated hemidiaphragm. The ratio of S1108 phosphorylated eIF4G to total eIF4G decreased about 60% in denervated hind-limb muscles but no statistically significant change was observed in denervated hemidiaphragm. The differential effect of denervation on eIF4G expression and S1108 phosphorylation in hemidiaphragm (hypertrophic) and hind-limb muscle (atrophic) may represent a regulatory mechanism that helps clarify the differential response of these muscles following denervation.
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| 10. |
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| 11. |
- Hansson Mild, Kjell, et al.
(author)
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Background ELF magnetic fields in incubators : A factor of importance in cell culture work.
- 2009
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In: Cell biology international. - : Wiley. - 1095-8355 .- 1065-6995. ; 33, s. 755-757
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Journal article (peer-reviewed)abstract
- Extremely low frequency (ELF) magnetic fields in cell culture incubators have been measured. Values of the order of tens of muT were found which is in sharp contrast to the values found in our normal environment (0.05-0.1muT). There are numerous examples of biological effects found after exposure to MF at these levels, such as changes in gene expression, blocked cell differentiation, inhibition of the effect of tamoxifen, effects on chick embryo development, etc. We therefore recommend that people working with cell culture incubators check for the background magnetic field and take this into account in performing their experiments, since this could be an unrecognised factor of importance contributing to the variability in the results from work with cell cultures.
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| 12. |
- Hegardt, Cecilia, et al.
(author)
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Changes in polyamine metabolism during glucocorticoid-induced programmed cell death in mouse thymus
- 2000
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In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 24:12, s. 871-880
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Journal article (peer-reviewed)abstract
- When mice are injected with dexamethasone, cortical thymocytes are deleted through programmed cell death (PCD). We have used this in vivo model system to investigate the kinetics of PCD and cell proliferation in relation to polyamine metabolism for 16 h after injection of dexamethasone. As a marker for PCD, we used the appearance of a sub-G(1)peak in the DNA histogram. When a sub-G(1)peak appeared at 4 h after dexamethasone treatment, the activity of the polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase (SSAT) was significantly increased and the activity of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC) was significantly decreased compared to the activities found in the thymi of control mice. Despite the significant changes in the activities of SSAT and AdoMetDC, the only change in the polyamine pool during the experimental period was that of putrescine. Presumably the complexity of this in vivo system masks changes in the spermidine and spermine pools that were expected in relation to the increased SSAT activity and decreased AdoMetDC activity.
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| 13. |
- Holst, Martina, et al.
(author)
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Novel anti-apoptotic effect of Bcl-2: Prevention of polyamine depletion-induced cell death
- 2008
-
In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 32, s. 66-74
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Journal article (peer-reviewed)abstract
- The spermine analogue N(1),N(11)-diethylnorspermine (DENSPM) efficiently depletes the polyamine pools in the breast cancer cell line L56Br-C1 and induces apoptotic cell death via the mitochondrial pathway. In this study, we have over-expressed the anti-apoptotic protein Bcl-2 in L56Br-C1 cells and investigated the effect of DENSPM treatment. DENSPM-induced cell death was significantly reduced in Bcl-2 over-expressing cells. Bcl-2 over-expression reduced DENSPM-induced release of the pro-apoptotic proteins AIF, cytochrome c, and Smac/DIABLO from the mitochondria. Bcl-2 over-expression reduced the DENSPM-induced activation of caspase-3. Bcl-2 over-expression also prevented DENSPM-induced Bax cleavage and reduction of Bcl-X(L) and survivin levels. The DENSPM-induced activation of the polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase was reduced by Bcl-2 over-expression, partly preventing polyamine depletion. Thus, Bcl-2 over-expression prevented a number of DENSPM-induced apoptotic effects
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| 14. |
- Holst, Martina, et al.
(author)
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Subcellular distribution of spermidine/spermine N(1)-acetyltransferase
- 2008
-
In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 32:1, s. 39-47
-
Journal article (peer-reviewed)abstract
- The subcellular distribution of the polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase (SSAT) was studied in L56Br-C1 cells treated with 10muM N(1),N(11)-diethylnorspermine (DENSPM) for 24h. Cells were fractioned into three subcellular fractions. A particulate fraction containing the mitochondria was denoted as the mitochondrial fraction. After DENSPM treatment, an increase in SSAT activity was mainly found in the mitochondrial fraction. Western blot analysis showed an increased level of the SSAT protein in the mitochondrial fraction compared to the cytosolic fraction. Immunofluorescence microscopy and immunogold labeling transmission electron microscopy also showed a mitochondrial association of SSAT. Transmission electron microscopy revealed that the endoplasmic reticulum was devoid of ribosomes in DENSPM-treated cells, in contrast to control cells that contained ample ribosomes. An increased SSAT activity in connection with the mitochondria may be part of the mechanism of DENSPM-induced apoptosis.
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| 15. |
- Hägerkvist, Robert, et al.
(author)
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Imatinib mesylate (Gleevec) protects against streptozotocin-induced diabetes and islet cell death in vitro
- 2006
-
In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 30:12, s. 1013-1017
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Journal article (peer-reviewed)abstract
- The tyrosine kinase inhibitor imatinib mesylate (Gleevec) has been demonstrated to protect various cell types from death by inhibition of Abelson tyrosine kinase (c-Abl). The aim of the present study was to establish whether imatinib protects the insulin producing β-cell from the different apoptosis promoting agents in vitro and whether imatinib counteracts streptozotocin-induced diabetes in NMRI mice. We observe that imatinib attenuated the actions of several different death promoting substances. In addition, mice injected with streptozotocin did not develop diabetes when given imatinib. The beneficial effects of imatinib may be related to inhibition of the pro-apoptotic MAP kinase JNK. We conclude that imatinib protects against β-cell death and that this may contribute to the previously reported anti-diabetic actions of imatinib.
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| 16. |
- Johansson Sjölander, Johanna, 1980, et al.
(author)
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The fission yeast FHIT homolog affects checkpoint control of proliferation and is regulated by mitochondrial electron transport.
- 2020
-
In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 44:2, s. 412-423
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Journal article (peer-reviewed)abstract
- Genetic analysis has strongly implicated human FHIT (Fragile Histidine Triad) as a tumor suppressor gene, being mutated in a large proportion of early-stage cancers. The functions of the FHIT protein have, however, remained elusive. Here, we investigated aph1+ , the fission yeast homolog of FHIT, for functions related to checkpoint control and oxidative metabolism. In sublethal concentrations of DNA damaging agents, aph1Δ mutants grew with a substantially shorter lag phase. In aph1Δ mutants carrying a hypomorphic allele of cds1 (the fission yeast homolog of Chk2), in addition, increased chromosome fragmentation and missegregation were found. We also found that under hypoxia or impaired electron transport function, the Aph1 protein level was strongly depressed. Previously, FHIT has been linked to regulation of the human 9-1-1 checkpoint complex constituted by Hus1, Rad1, and Rad9. In Schizosaccharomyces pombe, the levels of all three 9-1-1 proteins are all downregulated by hypoxia in similarity with Aph1. Moreover, deletion of the aph1+ gene reduced the Rad1 protein level, indicating a direct relationship between these two proteins. We conclude that the fission yeast FHIT homolog has a role in modulating DNA damage checkpoint function, possibly through an effect on the 9-1-1 complex, and that this effect may be critical under conditions of limiting oxidative metabolism and reoxygenation.
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| 17. |
- Johansson, Veronica, et al.
(author)
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Effect of polyamine deficiency on proteins involved in Okazaki fragment maturation.
- 2008
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In: Cell Biology International. - : Wiley. - 1095-8355 .- 1065-6995. ; 32, s. 1467-1477
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Journal article (peer-reviewed)abstract
- Polyamine depletion causes S phase prolongation, and earlier studies indicate that the elongation step of DNA replication is affected. This led us to investigate the effects of polyamine depletion on enzymes crucial for Okazaki fragment maturation in the two breast cancer cell lines MCF-7 and L56Br-C1. In MCF-7 cells, treatment with N(1),N(11)-diethylnorspermine (DENSPM) causes S phase prolongation. In L56Br-C1 cells the prolongation is followed by massive apoptosis. In the present study we show that L56Br-C1 cells have substantially lower basal expressions of two Okazaki fragment maturation key proteins, DNA ligase I and FEN1, than MCF-7 cells. Thus, these two proteins might be promising markers for prediction of polyamine depletion sensitivity, something that can be useful for cancer treatment with polyamine analogues. DENSPM treatment affects the cellular distribution of FEN1 in L56Br-C1 cells, but not in MCF-7 cells, implying that FEN1 is affected by or involved in DENSPM-induced apoptosis.
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| 18. |
- Jägerström, Sara, et al.
(author)
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Lipid droplets interact with mitochondria using SNAP23.
- 2009
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In: Cell biology international. - : Wiley. - 1095-8355 .- 1065-6995. ; 33:9, s. 934-40
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Journal article (peer-reviewed)abstract
- Triglyceride-containing lipid droplets (LD) are dynamic organelles stored on demand in all cells. These droplets grow through a fusion process mediated by SNARE proteins, including SNAP23. The droplets have also been shown to be highly motile and interact with other cell organelles, including peroxisomes and the endoplasmic reticulum. We have used electron and confocal microscopy to demonstrate that LD form complexes with mitochondria in NIH 3T3 fibroblasts. Using an in vitro system of purified LD and mitochondria, we also show the formation of the LD-mitochondria complex, in which cytosolic factors are involved. Moreover, the presence of LD markers in mitochondria isolated by subcellular fractionations is demonstrated. Finally, ablation of SNAP23 using siRNA reduced complex formation and beta oxidation, which suggests that the LD-mitochondria complex is functional in the cell.
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| 19. |
- Kampf, Caroline, et al.
(author)
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Mast cells accumulate in the renal capsule adjacent to transplanted pancreatic islets in rats
- 2006
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 30:12, s. 1054-1056
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Journal article (peer-reviewed)abstract
- Mast cells are important mediators of normal angiogenesis, and participate in normal would healing, i.e. processes involved in pancreatic islet engraftment. The aim of the study was to evaluate if mast cells are present in islet grafts. For this purpose, male nonnoglycaemic Wistar-Furth rats were either untreated or syngeneically implanted with 250 islets under the renal capsule. The animals were killed 1 month later, and the kidneys and endogenous pancreas were removed, fixed and embedded in paraffin. The distribution of mast cells was studied in Alcian Blue stained sections. Mast cells were rarely encountered in endogenous islets, but were frequent in the renal capsule adjacent to islet grafts. Mast cells interspersed between graft endocrine cells were as rare as in the endogenous pancreas. We conclude that mast cells may contribute to the engraftment after islet transplantation.
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| 20. |
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| 21. |
- Klemm, Anna H, et al.
(author)
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Mechano-chemical signaling in F9 cells.
- 2006
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 30:9, s. 755-9
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Journal article (peer-reviewed)abstract
- We investigated the molecular mechanism by which cells recognize and respond to physical forces in their local environment. Using a model system, to study wild type mouse F9 embryonic carcinoma cells, we examined how these cells sense mechanical stresses and translate them into biochemical responses through their cell surface receptor integrin and via the focal adhesion complex (FAC). Based on studies that show that many signal transducing molecules are immobilized on the cytoskeleton at the site of integrin binding within the focal adhesion complex, we found a time-dependent increase of focal adhesion kinase (pp125(FAK)) phosphorylation possibly due to protein kinase C (PKC) activation as well as protein kinase A (PKA) activity increase upon cell adhesion/spreading. These studies provide some insight into intracellular mechano-chemical signaling.
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| 22. |
- Pålsgård, Eva, et al.
(author)
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Effects of K+-induced depolarization and purinergic receptor activation on elemental content in insulin-producing RINm5F-cells
- 1995
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 19:1, s. 25-34
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Journal article (peer-reviewed)abstract
- X-ray microanalysis was used to detect elemental changes in the insulin-producing tumor cell-lineRINm5F. To improve discrimination between mobile ions and ions bound to macromolecules a new approach was employed, consisting of multivariate statistical analysis of correlations between the concentrations of Na, Mg, P, S, CI, K, and Ca. RINm5F cells, cultured an Formvar-coated titanium grids, were stimulated with high K+ or ATP, that are both known to stimulate insulin release. The buffers used contained Ca2+ or one of the Ca2+-analogues Sr2+ and Ba2+, to represent Ca2+ uptake inresponse to stimulation. After stimulation the cells were shock-frozen and freeze-dried overnight. Incubation for 10-20 seconds in a Ca2+-containing buffer did not significantly affect elementalcomposition, whereas cellular Mg, P and K decreased in a Sr2+-containing buffer. Depolarization with high K+ concentration caused an increase in the cellular Na content, both in Ca2+- and Sr2+-containing buffers, but not in the buffer where Ca2+ had been replaced by Ba2+. X-ray microanalysis is useful for detection of elemental changes subsequent to stimulation of cultured cells. Moreover, multivariate statistical analysis strengthens the idea that stimulation of RINm5F cells causes redistribution of ions possibly due to changes in the state of binding of some elements to cellular proteins.
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| 23. |
- Roomans, Godfried M., 1951-
(author)
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Pharmacological treatment of the basic defect in cystic fibrosis
- 2014
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In: Cell Biology International. - West Sussex, United Kingdom : John Wiley & Sons. - 1065-6995 .- 1095-8355. ; 38:11, s. 1244-1246
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Research review (peer-reviewed)abstract
- Cystic fibrosis (CF) is a genetic disease due to a mutation in the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel in epithelial cells. There are about 1900 mutations, divided in several groups, for example, stop mutations, mutations affecting the permeability of the channel, and mutations in which the mutated CFTR is recognized as abnormal and destroyed. Pharmacological treatment has become possible for stop mutations (about 10% of the patients), and for a rare mutation affecting channel permeability. For the majority of patients, however, that have a mutation in which the mutated CFTR is destroyed on its way to the cell membrane, research is still in progress, although a number of compounds have been identified that (at least partly) corrects the error in chloride transport.
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| 24. |
- Shleev, Sergey, et al.
(author)
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Simultaneous use of electrochemistry and chemiluminescence to detect reactive oxygen species produced by human neutrophils
- 2008
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In: Cell Biology International. - : Wiley. - 1065-6995 .- 1095-8355. ; 32:12, s. 1486-1496
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Journal article (peer-reviewed)abstract
- A novel approach for the simultaneous optical and electrochemical detection of biologically produced reactive oxygen species has been developed and applied. The set-up consists of a luminol-dependent chemiluminescence assay combined with two amperometric biosensors sensitive to superoxide anion radicals (O-2(center dot-))and hydrogen peroxide (H2O2), respectively. The method permits direct, real-time in vitro determination of both extra-and intracellular O-2(center dot-) and H2O2 produced by human neutrophil granulocytes. The rate of O-2(center dot-) production by stimulated neutrophils was calculated to about 10(-17) mol s(-1) per single cell. With inhibited NADPH oxidase, a distinct extracellular release of H2O2 instead of O-2(center dot-) was obtained from stimulated neutrophils with the rate of about 3 . 10(-18) mol s(-1) per single cell. When the H2O2 release was discontinued, fast H2O2 utilisation was observed. Direct interaction with and possibly attachment of neutrophils to redox protein-modified gold electrodes, resulted in a spontaneous respiratory burst in the population of cells closely associated to the electrode surface. Hence, further stimulation of human neutrophils with a potent receptor agonist (fMLF) did not significantly increase the O-2(center dot-) sensitive amperometric response. By contrast, the H2O2 sensitive biosensor, based on an HRP-modified graphite electrode, was able to reflect the bulk concentration of H2O2, produced by stimulated neutrophils and would be very useful in modestly equipped biomedical research laboratories. In summary, the system would also be appropriate for assessment of several other metabolites in different cell types, and tissues of varying complexity, with only minor electrode modifications.
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| 25. |
- Varelogianni, Georgia, 1977-, et al.
(author)
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The effect of ambroxol on chloride transport, CFTR and ENaC in cysticfibrosis airway epithelial cells
- 2013
-
In: Cell Biology International. - Hoboken, USA : Wiley-Blackwell. - 1065-6995 .- 1095-8355. ; 37:11, s. 1149-1156
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Journal article (peer-reviewed)abstract
- Ambroxol, a mucokinetic anti-inflammatory drug, has been used for treatment of cystic fibrosis (CF). The respiratoryepitheliumis covered by the airway surface liquid (ASL), the thickness and composition of which is determined by Cl− efflux viathe cystic fibrosis transmembrane conductance regulator (CFTR) and Naþ influx via the epithelial Naþ channel (ENaC). In cellsexpressing wt-CFTR, ambroxol increased the Cl- conductance, but not the bicarbonate conductance of the CFTR channels.Weinvestigated whether treatment with ambroxol enhances chloride transport and/or CFTR and ENaC expression in CF airwayepithelial cells (CFBE) cells. CFBE cells were treated with 100 mM ambroxol for 2, 4 or 8 h. mRNA expression for CFTR andENaC subunits was analysed by real-time polymerase chain reaction (RT-PCR); protein expression was measured by Westernblot. The effect of ambroxol on Cl− transport was measured by Cl− efflux measurements with a fluorescent chloride probe.Ambroxol significantly stimulated Cl− efflux from CFBE cells (a sixfold increase after 8 h treatment), and enhanced theexpression of the mRNA of CFTR and a-ENaC, and of the CFTR protein. No significant difference was observed in b-ENaCafter exposure to ambroxol, whereasmRNA expression of g-ENaC was reduced. No significant effects of ambroxol on the ENaCsubunits were observed by Western blot. Ambroxol did not significantly affect the intracellular Ca2+ concentration.Upregulation of CFTR and enhanced Cl− efflux after ambroxol treatment should promote transepithelial ion and watertransport, which may improve hydration of the mucus, and therefore be beneficial to CF-patients.
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| 26. |
- Varelogianni, Georgia, et al.
(author)
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The effect of N-acetylcysteine on chloride efflux from airway epithelial cells
- 2010
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In: Cell Biology International. - London : John Wiley & Sons. - 1065-6995 .- 1095-8355. ; 34:3, s. 245-252
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Journal article (peer-reviewed)abstract
- Defective chloride transport in epithelial cells increases mucus viscosity and leads to recurrent infections with high oxidative stress in patients with CF (cystic fibrosis). NAC (N-acetylcysteine) is a well known mucolytic and antioxidant drug, and an indirect precursor of glutathione. Since GSNO (S-nitrosoglutathione) previously has been shown to be able to promote Cl– efflux from CF airway epithelial cells, it was investigated whether NAC also could stimulate Cl– efflux from CF and non-CF epithelial cells and through which mechanisms. CFBE (CF bronchial epithelial cells) and normal bronchial epithelial cells (16HBE) were treated with 1 mM, 5 mM, 10 mM or 15 mM NAC for 4 h at 37°C. The effect of NAC on Cl– transport was measured by Cl– efflux measurements and by X-ray microanalysis. Cl– efflux from CFBE cells was stimulated by NAC in a dose-dependent manner, with 10 mM NAC causing a significant increase in Cl– efflux with nearly 80% in CFBE cells. The intracellular Cl– concentration in CFBE cells was significantly decreased up to 60% after 4 h treatment with 10 mM NAC. Moreover immunocytochemistry and Western blot experiments revealed expression of CFTR channel on CFBE cells after treatment with 10 mM NAC. The stimulation of Cl– efflux by NAC in CF airway epithelial cells may improve hydration of the mucus and thereby be beneficial for CF patients.
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| 27. |
- Nilsson, Ulrika K., et al.
(author)
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Inhibition of Ca2 +/calmodulin-dependent protein kinase or epidermal growth factor receptor tyrosine kinase abolishes lysophosphatidic acid-mediated DNA-synthesis in human myometrial smooth muscle cells
- 2003
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In: Cell Biology International. - 1065-6995. ; 27:4, s. 341-347
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Journal article (peer-reviewed)abstract
- Human myometrial smooth muscle cells (SMCs) were used to evaluate the proliferative activity of lysophosphatidic acid (LPA). This study specifically focuses on the role of Ca2+/calmodulin-dependent protein (CaM) kinase and epidermal growth factor (EGF) receptor tyrosine kinase. Myometrial SMCs were cultured from biopsies taken at Cesarean sections. The expression of LPA receptors was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and DNA-synthesis was measured by [3H]thymidine incorporation. LPA1, LPA2, and LPA3receptor subtypes were detected in the SMCs using RT-PCR. KN-62, an inhibitor of CaM kinase, and Tyrphostin AG 1478, an inhibitor of EGF receptor tyrosine kinase, dose-dependently decreased LPA-stimulated [3H]thymidine incorporation. Furthermore, BB-3103, an inhibitor of matrix metalloproteinases (MMPs), also reduced DNA-synthesis induced by LPA in these cells. The results show, for the first time, that human myometrial SMCs express all three known LPA receptor subtypes. Growth stimulatory effects of LPA on myometrial SMCs seems to be mediated by several pathways, where transactivation of EGF receptors through MMPs appears to be of importance. Furthermore, CaM kinase activity may be critical for LPA signaling since inhibition of CaM kinase totally abolish the proliferative effect of LPA.
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| 28. |
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| 29. |
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| 30. |
- Hegardt, Cecilia, et al.
(author)
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Spermine prevents cytochrome c release in glucocorticoid-induced apoptosis in mouse thymocytes.
- 2003
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In: Cell Biology International. - 1095-8355. ; 27:2, s. 115-121
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Journal article (peer-reviewed)abstract
- Apoptosis can be induced in primary cultures of mouse thymocytes using the glucocorticoid dexamethasone. Addition of the polyamine spermine simultaneously with dexamethasone reduces the induction of apoptosis compared to treatment with dexamethasone alone. We investigated the signal transduction pathway at the mitochondrial level in order to elucidate spermine's protective effect. Mitochondrial involvement is evident due to the loss of mitochondrial transmembrane potential, release of cytochrome c into the cytosol and activation of caspase-9 in dexamethasone-treated thymocytes. The addition of spermine inhibited the release of cytochrome c from the mitochondria into the cytosol, and also the activation of caspase-9. When the mitogen concanavalin A (Con A) was added to dexamethasone- plus spermine-treated thymocytes, the number of apoptotic cells in the pre-G1peak was reduced compared to thymocytes treated with only dexamethasone plus spermine. Comparing concanavalin A added to dexamethasone-treated or to dexamethasone plus spermine-treated thymocytes, showed a markedly reduced pre-G1peak in the latter. Thus, the spermine-induced inhibition of cytochrome c release confers a survival advantage on thymocytes.
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| 31. |
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| 32. |
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| 33. |
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| 34. |
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| 35. |
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| 36. |
- Rutberg, M, et al.
(author)
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Distribution of acetylated tubulin in cultured cells and tissues from the Atlantic cod (Gadus morhua). Role of acetylation in cold adaptation and drug stability.
- 1995
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In: Cell biology international. - : Wiley. - 1065-6995. ; 19:9, s. 749-58
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Journal article (peer-reviewed)abstract
- The Atlantic cod (Gadus morhua) is a poikilothermic animal living at temperatures between 2-15 degrees C. Isolated cod brain tubulin is, in contrast to mammalian brain tubulin, posttranslationally modified by acetylation to a high extent. To investigate the role of acetylation in cold adaptation, microtubules were isolated by a taxol-dependent procedure from different organs of the cod, and cells from different tissues were cultured. All cells from skin and brain were able to grow between 4 degrees C and room temperature. Microtubules in the cultured cells were sometimes severed near the periphery of the cells. Microtubules in brain cells were in general more stable to vinblastine and colchicine, when compared to skin cells. Acetylated microtubules were found only in brain cells, in peripheral nerves on scales and in nerves of the intestinal tract and in microtubules isolated from neuronal tissue. Our results show that acetylated microtubules are found both in the central and peripheral nervous system, but that there is no correlation between acetylation and cold-adaptation.
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| 37. |
- Sepehr, Arian, et al.
(author)
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Lactate damages primary hippocampal neurons in vitro.
- 2010
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In: Cell biology international. - 1095-8355. ; 34:1, s. 61-65
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Journal article (peer-reviewed)abstract
- In the present study, rat primary cultures were used to study the effect of lactate on the survival of hippocampal neurons in the presence or absence of glucose. Our results showed no extensive cell damage under glucose-free conditions compared with glucose-rich conditions. Addition of 10 and 50 mM lactate to glucose-free and glucose-rich media increased the cell damage significantly, as observed by morphology and lactate dehydrogenase activity. The results of the present study suggest that primary neurons in vitro are not sensitive to glucose deficiency and the presence of lactate damages the neurons in a concentration-dependent manner.
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| 38. |
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| 39. |
- Virtanen, Sanna S., et al.
(author)
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Inhibition of GGTase-I and FTase disrupts cytoskeletal organization of human PC-3 prostate cancer cells
- 2010
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In: Cell Biology International. - 1095-8355. ; 34:8, s. 815-826
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Journal article (peer-reviewed)abstract
- The mevalonate synthesis pathway produces intermediates for isoprenylation of small GTPases, which are involved in the regulation of actin cytoskeleton and cell motility. Here, we investigated the role of the prenylation transferases in the regulation of the cytoskeletal organization and motility of PC-3 prostate cancer cells. This was done by using FTI-277, GGTI-298 or NE-10790, the specific inhibitors of FTase (farnesyltransferase), GGTase (geranylgeranyltransferase)-I and -II, respectively. Treatment of PC-3 cells with GGTI-298 and FTI-277 inhibited migration and invasion in a time- and dose-dependent manner. This was associated with disruption of F-actin organization and decreased recovery of GFP-actin. Immunoblot analysis of various cytoskeleton-associated proteins showed that the most striking change in GGTI-298- and FTI-277-treated cells was a markedly decreased level of total and phosphorylated cofilin, whereas the level of cofilin mRNA was not decreased. The treatment of PC-3 cells with GGTI-298 also affected the dynamics of GFP-paxillin and decreased the levels of total and phosphorylated paxillin. The levels of phosphorylated FAK (focal adhesion kinase) and PAK (p-21-associated kinase)-2 were also lowered by GGTI-298, but levels of paxillin or FAK mRNAs were not affected. In addition, GGTI-298 had a minor effect on the activity of MMP-9. RNAi knockdown of GGTase-I beta inhibited invasion, disrupted F-actin organization and decreased the level of cofilin in PC-3 cells. NE-10790 did not have any effect on PC-3 prostate cancer cell motility or on the organization of the cytoskeleton. In conclusion, our results demonstrate the involvement of GGTase-I- and FTase-catalysed prenylation reactions in the regulation of cytoskeletal integrity and motility of prostate cancer cells and suggest them as interesting drug targets for development of inhibitors of prostate cancer metastasis.
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| 40. |
- Hallström, Lena, 1958-, et al.
(author)
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Breakfast habits and factors influencing food choices at breakfast in relation to socio-demographic and family factors among European adolescents : The HELENA Study
- 2011
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In: Appetite. - : Elsevier BV. - 0195-6663 .- 1095-8304. ; 56:3, s. 649-657
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Journal article (peer-reviewed)abstract
- The aim was to investigate factors associated with breakfast habits and influences on food choices at breakfast, within the framework of the EU-funded HELENA Study in 3528 adolescents from ten cities across Europe. The statement “I often skip breakfast” and personal and socio-environmental factors hypothesized to be related to food choice at breakfast were dichotomized. Logistic regression was used to investigate the association between behavioral (skipping versus consume breakfast) and individual, personal and socio-environmental factors. Half of the adolescents (fewer girls) indicated being regular breakfast consumers. Mothers’ education and family structure were associated with breakfast consumption. Adolescents with peers who gave little or no encouragement, and boys whose parents gave encouragement, were more likely to be regular breakfast consumers. Personal factors influenced the girls more than the boys in their choice of food for breakfast and socio-environmental factors influenced younger adolescents more than older adolescents. In conclusion, a broad range of (behavioral, individual, personal and socio-environmental) factors influence breakfast habits and food choices at breakfast among European adolescents. Breakfast habits were inappreciably influenced by socio-demographical factors. These factors need to be considered in discussions surrounding the development of nutritional intervention programs intended for adolescents.
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