scholarly journals Isotopic variations of copper at the protein fraction level in neuronal human cells exposed in vitro to uranium

The Analyst ◽  
2019 ◽  
Vol 144 (20) ◽  
pp. 5928-5933 ◽  
Author(s):  
Eduardo Paredes ◽  
Véronique Malard ◽  
Claude Vidaud ◽  
Emilie Avazeri ◽  
Richard Ortega ◽  
...  

Accurate isotope ratio determination was downscaled to the level of metal-containing protein fractions obtained from cell line lysates.

2000 ◽  
Vol 74 (1) ◽  
pp. 49-56 ◽  
Author(s):  
Carolyn A. Wilson ◽  
Susan Wong ◽  
Matthew VanBrocklin ◽  
Mark J. Federspiel

ABSTRACT We previously reported that mitogenic activation of porcine peripheral blood mononuclear cells resulted in production of porcine endogenous retrovirus(es) (PERV[s]) capable of productively infecting human cells (C. Wilson et al., J. Virol. 72:3082–3087, 1998). We now extend that analysis to show that additional passage of isolated virus, named here PERV-NIH, through a human cell line yielded a viral population with a higher titer of infectious virus on human cells than the initial isolate. We show that in a single additional passage on a human cell line, the increase in infectivity for human cells is accounted for by selection against variants carrying pig-tropic envelope sequences (PERV-C) as well as by enrichment for replication-competent genomes. Sequence analysis of the envelope cDNA present in virions demonstrated that the envelope sequence of PERV-NIH is related to but distinct from previously reported PERV envelopes. The in vitro host range of PERV was studied in human primary cells and cell lines, as well as in cell lines from nonhuman primate and other species. This analysis reveals three patterns of susceptibility to infection among these host cells: (i) cells are resistant to infection in our assay; (ii) cells are infected by virus, as viral RNA is detected in the supernatant by reverse transcription-PCR, but the cells are not permissive to productive replication and spread; and (iii) cells are permissive to low-level productive replication. Certain cell lines were permissive for efficient productive infection and spread. These results may prove useful in designing appropriate animal models to assess the in vivo infectivity properties of PERV.


Author(s):  
O. Pochapinskyi ◽  
◽  
G. Lavrenchuk ◽  
N. Atamaniuk ◽  
A. Chernyshov ◽  
...  

Objective: to investigate the structural and morpho-functional changes in test systems of malignant (A-549 cell line) and normal (fibroblasts of the 6th passage) human cells during incubation with gadolinium-containing photon-capture agent «Dotavist» and photosensitizer «Fotolon». Methods. The passaged (continuously interweaved) cell culture technique on normal human fibroblasts and malignant human cells; cytological, biophysical, statistical methods. Results. The cytotoxic properties of «Dotavist» gadolinium-containing photon-capturing agent and «Photolon» photosensitizer in a wide range of concentrations (5, 10, 25, 50, 100 and 200 μl/ml) were studied by the morphofunctional characteristics (growth kinetics, proliferative and mitotic activity, presence of atypical cells) in the in vitro test systems of malignant (non-small cell lung cancer cell line A-549) and normal (6th passage fibroblasts) human cells. It was found that the cytotoxic properties of «Dotavist» in test systems of malignant and normal cells are expressed under its administration in high concentrations (100 and 200 μl/ml). During incubation with «Photolon» photosensitizer the cytotoxic effect on malignant cells was determined at the lowest concentrations (5 and 10 μl/ml). Photosensitizer administration in the increasing concentrations has lead to genotoxic effects. Cytotoxic effect of photosensitizer on the normal human fibroblasts was evident in the 5-200 μl/ml concentration range. There was a moderate decrease in mitotic activity along with increasing concentration. Genotoxic properties of photosensitizer were evident at 25 μl/ml concentration and above. Conclusion. Study results of the effectiveness of neutron-capture and photon-capture technologies by the sensitivity assay in the in vitro test systems of human malignant cells (non-small cell lung cancer cell line A-549) and normal cells (transplantable human fibroblast culture, the 6th passage) to the gadolinium-containing photon-capture «Dotavist» agent and «Photolon» photosensitizer in different concentrations provide the basis for pre-clinical stage of evaluating the effectiveness of medications used in binary technologies. Key words: culture of human malignant cells, culture of human fibroblasts, neutron-capture agent, photon-capture agent, photosensitizer, proliferation, mitotic index.


Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2561-2561
Author(s):  
Marcin Wysoczynski ◽  
Katarzyna Miekus ◽  
Anna Marcinkowska ◽  
Anna Janowska-Wieczorek ◽  
Mariusz Z. Ratajczak

Abstract Rhabdomyosarcoma (RMS) and skeletal muscle-derived tumors frequently infiltrate bone marrow (BM). We have demonstrated that the stromal-derived factor (SDF)-1-CXCR4 receptor (Blood2002;100:2597) and hepatocyte growth factor (HGF)-c-Met receptor (Cancer Res. 2003;63:7926) play an important role in RMS metastasis to BM. Leukemia inhibitory factor (LIF) is a well known factor that plays an important role in skeletal muscle development/regeneration and similarly as SDF-1 and HGF is secreted by BM stroma. This prompted us to examine whether the LIF-LIF receptor (LIF-R) axis affects the biology/metastasis of RMS cells. We employed in our studies, human established RMS cell lines, as well as RMS samples isolated from patients and noticed that LIF-R was expressed not only on established human RMS cell lines (7/7) but more importantly, it was also detectable in patient samples (23/23). We also found that in RMS cells LIF stimulatesphosphorylation of MAPKp42/44, AKT and STAT3,chemotaxis and adhesion andincreases resistance to cytostatics (e.g., etoposide). These LIF-mediated effects were inhibited after downregulating the LIF-R by siRNA. To learn more on the biological significance of the LIF-LIF-R axis in vivo we employed two models. First, human RMS cells (RH-30) were exposed or not exposed to LIF-R siRNA and subsequently injected into SCID™-Beige immunodeficient mice. To estimate the number of RMS cells that seed to BM and liver in these animals, we isolated DNA and using real- time RT-PCR, amplified human a-satellite sequences and murine b-actin. The number of human cells present in murine organs was subsequently calculated from a standard curve derived from mixing varying numbers of human cells with a constant number of murine cells. We noticed that downregulation of LIF-R by siRNA significantly decreased the number of human RMS cells in murine BM and liver (x4 and x2 respectively). In a second model, the RH30 cell line was selected by repetitive chemotaxis for cells that are highly responsive to LIF (RH-30 L) and subsequently the cells from parental RH-30 cell line and RH-30 L cells were injected intramuscularly. Six weeks after tumour inoculation, we detected more metastasis in bone marrow and lungs in mice injected with RH-30L cells as compared to parental RH-30 clone (x6 and x3 respectively). In conclusion, we present evidence for the first time that the inhibition of LIF-LIF-R axis may decrease the invasive potential of human RMS both in vitro and in vivo. Hence, molecular targeting of LIF-LIF-R axis could possibly become a more effective new strategy to control the progression and metastasis of RMS.


2020 ◽  
pp. 1-20
Author(s):  
Rachel Verdon ◽  
Suzanne L. Gillies ◽  
David M. Brown ◽  
Theodore Henry ◽  
Lang Tran ◽  
...  

Author(s):  
John C. Garancis ◽  
R. A. Pattillo

Growth of cell system (BeWo-cell line) derived from human gestational choriocarcinoma has been established and continuously maintained in-vitro. Furthermore, it is evident from the previous studies that this cell line has retained the physiological function of the placental trophoblasts, namely the synthesis of human chorionic gonadotrophil(HCG).The BeWo cells were relatively small and possessed single nuclei, thus indicating that this cell line consists exclusively of cytotrophoblasts. In some instances cells appeared widely separated and their lateral surfaces were provided with numerous microvilli (Fig.1).


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