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  • Elsevier (CrossRef)  (15)
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  • 1
    Language: English
    In: The American Journal of Pathology, April 2012, Vol.180(4), pp.1370-1377
    Description: The influences of cytotoxic drugs on endothelial cells remain incompletely understood. Herein, we examined the effects of chemotherapeutic agents in experimental angiogenesis models and analyzed vessel densities in clinical neuroblastoma tumor samples. Cisplatin (20 to 500 ng/mL), doxorubicin (4 to 100 ng/mL), and vincristine (0.5 to 4 ng/mL), drugs commonly involved in neuroblastoma therapy protocols, induced pro-angiogenic effects in different angiogenesis models. They enhanced endothelial cell tube formation, endothelial cell sprouting from spheroids, formation of tip cells in the sprouting assay, expression of αvβ3 integrin, and vitronectin binding. All three drugs increased global cellular kinase phosphorylation levels, including the angiogenesis-relevant molecules protein kinase Cβ and Akt. Pharmacological inhibition of protein kinase Cβ or Akt upstream of phosphatidylinositol 3-kinase reduced chemotherapy-induced endothelial cell tube formation. Moreover, the investigated chemotherapeutics dose dependently induced vessel formation in the chick chorioallantoic membrane assay. Tumor samples from seven high-risk patients with neuroblastoma were analyzed for vessel density by IHC. Results revealed that neuroblastoma samples taken after chemotherapy consistently showed an enhanced microvessel density compared with the corresponding samples taken before chemotherapy. In conclusion, our data show that chemotherapy can activate endothelial cells by inducing multiple pro-angiogenic signaling pathways and exert pro-angiogenic effects and . Moreover, we report a previously unrecognized clinical phenomenon that might, in part, be explained by our experimental observations: chemotherapy-associated enhanced vessel formation in tumors from patients with neuroblastoma.
    Keywords: Medicine
    ISSN: 0002-9440
    E-ISSN: 1525-2191
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  • 2
    Language: English
    In: Biochemical Pharmacology, 15 January 2010, Vol.79(2), pp.130-136
    Description: Artemisinin derivatives are well-tolerated anti-malaria drugs that also exert anti-cancer activity. Here, we investigated artemisinin and its derivatives dihydroartemisinin and artesunate in a panel of chemosensitive and chemoresistant human neuroblastoma cells as well as in primary neuroblastoma cultures. Only dihydroartemisinin and artesunate affected neuroblastoma cell viability with artesunate being more active. Artesunate-induced apoptosis and reactive oxygen species in neuroblastoma cells. Of 16 cell lines and two primary cultures, only UKF-NB-3 CDDP showed low sensitivity to artesunate. Characteristic gene expression signatures based on a previous analysis of artesunate resistance in the NCI60 cell line panel clearly separated UKF-NB-3 CDDP from the other cell lines. -Buthionine-S,R-sulfoximine, an inhibitor of GCL (glutamate–cysteine ligase), resensitised in part UKF-NB-3 CDDP cells to artesunate. This finding together with bioinformatic analysis of expression of genes involved in glutathione metabolism showed that this pathway is involved in artesunate resistance. These data indicate that neuroblastoma represents an artesunate-sensitive cancer entity and that artesunate is also effective in chemoresistant neuroblastoma cells.
    Keywords: Neuroblastoma ; Artesunate ; Artemisinin ; Chemoresistance ; Cancer ; Chemotherapy ; Pharmacy, Therapeutics, & Pharmacology ; Chemistry
    ISSN: 0006-2952
    E-ISSN: 1873-2968
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  • 3
    Language: English
    In: International Journal of Medical Microbiology, 2002, Vol.291, pp.190-193
    Description: Altogether 112 sera from 67 patients hospitalised for tick-borne encephalitis (TBE) were tested for the presence of antibodies against human granulocytic ehrlichiosis (HGE). Indirect immunofluorescence assay and western blotting were used for the screening and confirmation respectively. A laboratory culture of HGE agent in HL-60 cells served as an antigen supply. Commercially available human sera were used as positive controls; the respective IgG and IgM cut-offs of 1/80 and 1/40 were applied. Six patients (8.9%) exhibited specific IgG antibodies against HGE at or above the cut-off level, while no serum sample was shown to be IgM-reactive at the cut-off. The mean age of the subset of positive patients was 42 (± 16.7) years which exceeded that of the total TBE set averaging 37.5 (± 17.3) years. One patient, a woman aged 32, exhibited an elevated HGE titre of 1/80 on admission 15 days after TBE onset, that still slightly increased up to 1/160 by the end of the following week. Corresponding WB-pattern showed significant MSP, p30, and p28 bands. These findings suggest a case of concurrent TBE with HGE. Overall, this study demonstrates that TBE/HGE co-infections could be encountered in central Europe.
    Keywords: Human Granulocytic Ehrlichiosis (Hge) ; Tick-Borne Encephalitis (Tbe) ; Co-Infections ; Serology ; Central Europe ; Biology
    ISSN: 1438-4221
    E-ISSN: 1618-0607
    Source: ScienceDirect Journals (Elsevier)
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  • 4
    Language: English
    In: Cancer Letters, 2007, Vol.250(1), pp.107-116
    Description: The efficacy of Onconase on the growth of a panel of chemosensitive and chemoresistant neuroblastoma cell lines was investigated. Onconase decreased cell viability of chemosensitive (IMR-32, UKF-NB-3) and chemoresistant neuroblastoma cell lines characterised by high expression of P-glycoprotein (P-gp) (UKF-NB-3 DOX ) or by high P-gp expression in combination with mutated p53 (UKF-NB-3 VCR , Be(2)-C), in a similar manner. Moreover, Onconase caused cell cycle block in G1 phase and induced caspase-independent cell death. Transmission electron microscope investigations suggested that Onconase-induced autophagy contributes to Onconase-induced cell death. Antitumour activity of Onconase against naïve and drug-resistant neuroblastoma xenografts was confirmed in animals.
    Keywords: Onconase ; Neuroblastoma ; Multi-Drug-Resistance ; P-Glycoprotein ; P53 ; Medicine
    ISSN: 0304-3835
    E-ISSN: 1872-7980
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  • 5
    Language: English
    In: Biochemical Pharmacology, 2002, Vol.64(2), pp.239-246
    Description: Continuous cultivation of T-lymphoid H9 cells in the presence of 3′-azido-2′,3′-dideoxythymidine (AZT) resulted in a cell variant cross-resistant to both thymidine and deoxycytidine analogs. Cytotoxic effects of AZT, 2′,3′-didehydro-3′-deoxythymidine as well as different deoxycytidine analogs such as 2′,3′-dideoxycytidine, 2′,2′-difluoro-2′-deoxycytidine (dFdC) and 1-ß-D-arabinofuranosylcytosine (Ara-C) were strongly reduced in H9 cells continuously exposed to AZT when compared to parental cells (〉8.3-, 〉6.6-, 〉9.1-, 5×10 4 -, 5×10 3 -fold, respectively). Moreover, anti-HIV-1 effects of AZT, d4T, ddC and 2′,3′-dideoxy-3′-thiacytidine (3TC) were significantly diminished (〉222-, 〉25-, 〉400-, 〉200-fold, respectively) in AZT-resistant H9 cells. Study of cellular mechanisms responsible for cross-resistance to pyrimidine analogs in AZT-resistant H9 cells revealed decreased mRNA levels of thymidine kinase 1 (TK1) and lack of deoxycytidine kinase (dCK) mRNA expression. The loss of dCK gene expression was confirmed by western blot analysis of dCK protein as well as dCK enzyme activity assay. Moreover, enzyme activity of TK1 and TK2 was reduced in AZT-resistant cells. In order to determine whether lack of dCK affected the formation of the active triphosphate of the deoxycytidine analog dFdC, dFdCTP accumulation and retention was measured in H9 parental and AZT-resistant cells after exposure to 1 and 10 μM dFdC. Parental H9 cells accumulated about 30 and 100 pmol dFdCTP/10 6 cells after 4 hr, whereas in AZT-resistant cells no dFdCTP accumulation was detected. These results demonstrate that continuous treatment of H9 cells in the presence of AZT selected for a thymidine analog resistant cell variant with cross-resistance to deoxycytidine analogs, due to deficiency in TK1, TK2, and dCK.
    Keywords: Cellular Resistance ; Zidovudine ; Gemcitabine ; Cytarabine ; Thymidine Kinase 1 ; Deoxycytidine Kinase ; Pharmacy, Therapeutics, & Pharmacology ; Chemistry
    ISSN: 0006-2952
    E-ISSN: 1873-2968
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  • 6
    Language: English
    In: Biologicals, 1991, Vol.19(2), pp.87-92
    Description: Mouse NCTC clone 919 L (L-929) cells were propagated continuously for 3 years as monolayers in a protein-free chemically-defined medium. These cells, designated L-929-WS, were used for quality control testing of the surfaces of commercially available cell culture plastic flasks. Differences in attachment and saturation density of L-929-WS cells in a protein-free culture medium were taken to define various levels of quality of the culture vessels tested. The rate of attachment and growth of L-929-WS cells on a surface of a given quality correlated directly with that of human embryonal fibroblasts and embryonal epithelial cells grown in a serum-free medium supplemented with growth factors and hormones. L-929-WS cells propagated continuously in a protein-free medium provide a simple and sensitive assay system for more general quality control testing of surfaces used for the culture of monolayer cells.
    Keywords: Biology
    ISSN: 1045-1056
    E-ISSN: 1095-8320
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  • 7
    Language: English
    In: Cell Biology International, September 1993, Vol.17(9), pp.885-895
    Description: A protein-free chemically defined medium designated PFEK-1 was developed for culture of VERO cells on polyvinyl formal (PVF) culture surface without serum or other macromolecular supplements. VERO cells proliferated in PFEK-1 medium on PVF surface to a similar extent as cells in serum-supplemented medium without previous adaptation from serum-containing conditions. The protein-free culture infected with coxsackievirus B4, herpes simplex virus types 1 and 2, measles virus and poliovirus types 1, 2 and 3 developed viral titers comparable to those found in conventionally grown cells. The results demonstrated that VERO cells in protein-free culture provide a sensitive substrate for the production of human pathogenic viruses which are not contaminated by serum or other protein factors usually added to a culture medium.
    Keywords: Biology
    ISSN: 1065-6995
    E-ISSN: 1095-8355
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  • 8
    Language: English
    In: Antiviral Research, 1997, Vol.33(3), pp.165-175
    Description: An l -glutamine antagonist, 6-diazo-5-oxo- l -norleucin ( l -DON), inhibits replication of vesicular stomatitis virus, poliovirus and paramyxoviruses in cultured cells. We tested the antiviral activity of l -DON against different strains of herpes simplex virus type 1 (HSV-1) in Vero cells. In the presence of a physiological plasma concentration of l -glutamine (0.5 mM) l -Don inhibited 50% production of virus plaques at concentrations ranging from 7.9 to 16 μ M. At concentrations of 40 μ M l -Don inhibited infectious virus yield by 99%. The antiviral activity of l -DON decreased with increasing l -glutamine concentrations. A concentration of 5000 μ M of l -Don had no significant effects on the viability of Vero cells. Transmission electron microscopical investigations showed that l -DON prevented mainly envelopment of viral nucleocapsids in the cytoplasm. The immunoprecipitation experiments demonstrated selective inhibition of synthesis of HSV-1 glycoproteins in l -DON treated cells. The results showed that l -DON inhibits HSV-1 replication at a late stage in the virus replication cycle, probably the cytoplasmic maturation of virions and subsequent virion egress from the cells.
    Keywords: Hsv ; Acyclovir ; 6-Diazo-5-Oxo- L-Norleucin ; Virus-Resistance ; Medicine ; Biology
    ISSN: 0166-3542
    E-ISSN: 1872-9096
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  • 9
    Language: English
    In: Cancer Letters, 1993, Vol.70(1), pp.15-24
    Description: Sodium phenylacetate (NaPA) at concentrations ranging from 2 to 6 mM stimulated morphological differentiation of two human neuroblastoma cell lines IMR-32 and UKF-NB-3. These concentrations inhibited growth and DNA synthesis of the cells in a dose dependent manner without significant effect on cell viability. The differentiated cells showed pseudoganglia formation and extension of cellular processes. The morphological differentiation in both cell lines was accompanied by decreased expression of N- myc oncoprotein. These results suggest that NaPA at concentrations, which have been achieved in humans with no significant adverse effects, promotes differentiation of cultured human neuroblastoma cells in association with the reduced expression of the malignant phenotype.
    Keywords: Neuroblastoma ; Differentiation ; Phenylacetate ; Retinoic Acid ; N- Myc ; Medicine
    ISSN: 0304-3835
    E-ISSN: 1872-7980
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  • 10
    Language: English
    In: Cell Biology International, April 1994, Vol.18(4), pp.271-278
    Description: The effects of aphidicolin, a specific inhibitor of DNA polymerase alpha, on cell growth, DNA synthesis and myogenic differentiation in the human alveolar rhabdomyosarcoma cell line KFR were studied. The treatment with aphidicolin at 5 x 10(-6) M concentration, which completely inhibited DNA synthesis and cell growth, induced morphological differentiation of small mononuclear cells to elongated, multinucleated (myotube-like) structures. The morphological differentiation was accompanied by the expression of skeletal muscle myosin; about 30% myosin-positive cells were observed after 14 days of treatment, compared to 2.3% in untreated cultures. The results showed that aphidicolin induces differentiation of human rhabdomyosarcoma cells and that multinucleated myotube-like elements may develop simply by cell fusion without cell division and DNA synthesis.
    Keywords: Biology
    ISSN: 1065-6995
    E-ISSN: 1095-8355
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