scholarly journals Measles Virus Attenuation Associated with Transcriptional Impediment and a Few Amino Acid Changes in the Polymerase and Accessory Proteins

1998 ◽  
Vol 72 (11) ◽  
pp. 8690-8696 ◽  
Author(s):  
Makoto Takeda ◽  
Atsushi Kato ◽  
Fumio Kobune ◽  
Hiroko Sakata ◽  
Yan Li ◽  
...  
Keyword(s):  
Amino Acid ◽  
Vero Cell ◽  
Cell Line ◽  
Measles Virus ◽  
Vero Cells ◽  
Functional Difference ◽  
Pathogenic Potential ◽  
P Proteins

ABSTRACT Measles virus (MV) isolated in B95a cells, a marmoset B-cell line, retains full pathogenicity for cynomolgus monkeys, while its derivative obtained by adaptation to the growth in Vero cells, a monkey kidney cell line, loses the pathogenic potential (F. Kobune, H. Sakata, and A. Sugiura, J. Virol. 64:700–705, 1990). Here, we show with a pair of strains, a fresh isolate (9301B) in B95a cells and its Vero cell-adapted form (9301V), that the in vivo attenuation parallels the decrease of replication and syncytium-inducing capabilities in the original B95a cells and that these in vitro phenotypes are attributable to impediment of transcription, which is already obvious at the level of primary transcription catalyzed by the virion-associated RNA polymerase. On the other hand, cell fusion assays detected no functional difference between the glycoproteins of the two viruses. Essentially the same transcriptional impediment with reduced syncytium induction following Vero cell adaptation was found with two other pairs of strains that had been similarly prepared. Nucleotide sequence comparison between the 9301B and 9301V viruses revealed that a few (at most five) amino acid changes, which sporadically took place in the polymerase (L and P proteins) and/or accessory V and C proteins, were responsible for the in vitro and in vivo attenuation through adaptation to growth in Vero cells.

scholarly journals The glycoprotein of measles virus. External radioactive labelling of its carbohydrate and partial characterization of the glycopeptide

1980 ◽  
Vol 185 (1) ◽  
pp. 189-194 ◽  
Author(s):  
O Anttonen ◽  
M Jokinen ◽  
A Salmi ◽  
R Vainionpää ◽  
C G Gahmberg
Keyword(s):  
Sialic Acid ◽  
Measles Virus ◽  
Vero Cells ◽  
Galactose Oxidase ◽  
Culture Supernatants ◽  
Surface Carbohydrates ◽  
Specific Labelling

Measles virus was propagated in VERO cells and purified from the culture supernatants by two successive tartrate-density-gradient centrifugations. Surface carbohydrates were labelled both in vitro and in vivo with 3H after treatment with galactose oxidase/NaB3H4 or with [3H]glucosamine. The major labelled glycoprotein in measles virions had a mol.wt. of 79 000. After labelling with periodate/NaB3H4, which would result in specific labelling of sialic acid residues, the 79 000-mol.wt. glycoprotein was very weakly labelled. This suggests that there is no or a very low amount of sialic acid in the virions. Further analysis of the glycoprotein showed that galactose is the terminal carbohydrate unit in the oligosaccharide, and the molecular weight of the glycopeptide obtained after Pronase digestion is about 3000. The oligosaccharide is attached to the polypeptide through an alkali-stable bond, indicating a N-glycosidic asparagine linkage.

scholarly journals Pathogenicity of Entamoeba dispar under xenic and monoxenic cultivation compared to a virulent E. histolytica

2006 ◽  
Vol 48 (5) ◽  
pp. 245-250 ◽  
Author(s):  
Adriana Oliveira Costa ◽  
Maria Aparecida Gomes ◽  
Orivaldo Alves Rocha ◽  
Edward Felix Silva
Keyword(s):  
Vero Cells ◽  
Invasive Disease ◽  
In Vitro Tests ◽  
Entamoeba Dispar ◽  
Experimental Conditions ◽  
Lower Growth ◽  
Pathogenic Potential ◽  
Organism Growth

Two xenic isolates and cloned cultures of Entamoeba dispar were submitted to monoxenization using Crithidia fasciculata as the associated organism. Growth in monoxenic cultivation and ability of xenic and monoxenic trophozoites to destroy VERO cells and produce lesions in hamster livers were compared to those of a virulent E. histolytica. Parental and cloned E. dispar under monoxenic cultivation showed a remarkable lower growth than the monoxenic E. histolytica and were avirulent in both in vivo and in vitro tests. When xenically cultured, trophozoites of E. dispar showed a moderate lytic activity against VERO cells (1.5 to 41.8% of destruction) but caused severe hepatic lesions in hamsters as those caused by the virulent E. histolytica (29 to 100% in prevalence and 0.86 to 4.00 in lesion degree). Although E. dispar has not been associated with invasive disease in men, the ability of xenic trophozoites to produce prominent tissue damage in experimental conditions has indicated that some strains have a considerable pathogenic potential when in presence of bacteria.

scholarly journals RTH-149 Cell Line, a Useful Tool to Decipher Molecular Mechanisms Related to Fish Nutrition

Cells ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1754
Author(s):  
Guillaume Morin ◽  
Karine Pinel ◽  
Karine Dias ◽  
Iban Seiliez ◽  
Florian Beaumatin
Keyword(s):  
Amino Acid ◽  
Cell Line ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Fish Nutrition ◽  
Multiple Strategies ◽  
First Time ◽  
Interesting Alternative

Nowadays, aquaculture provides more than 50% of fish consumed worldwide but faces new issues that challenge its sustainability. One of them relies on the replacement of fish meal (FM) in aquaculture feeds by other protein sources without deeply affecting the whole organism’s homeostasis. Multiple strategies have already been tested using in vivo approaches, but they hardly managed to cope with the multifactorial problems related to the complexities of fish biology together with new feed formulations. In this context, rainbow trout (RT) is particularly concerned by these problems, since, as a carnivorous fish, dietary proteins provide the amino acids required to supply most of its energetic metabolism. Surprisingly, we noticed that in vitro approaches considering RT cell lines as models to study RT amino acid metabolism were never previously used. Therefore, we decided to investigate if, and how, three major pathways described, in other species, to be regulated by amino acid and to control cellular homeostasis were functional in a RT cell line called RTH-149—namely, the mechanistic Target Of Rapamycin (mTOR), autophagy and the general control nonderepressible 2 (GCN2) pathways. Our results not only demonstrated that these three pathways were functional in RTH-149 cells, but they also highlighted some RT specificities with respect to the time response, amino acid dependencies and the activation levels of their downstream targets. Altogether, this article demonstrated, for the first time, that RT cell lines could represent an interesting alternative of in vivo experimentations for the study of fish nutrition-related questions.

scholarly journals Recovery of Pathogenic Measles Virus from Cloned cDNA

2000 ◽  
Vol 74 (14) ◽  
pp. 6643-6647 ◽  
Author(s):  
Makoto Takeda ◽  
Kaoru Takeuchi ◽  
Naoko Miyajima ◽  
Fumio Kobune ◽  
Yasushi Ami ◽  
...  
Keyword(s):  
Cell Line ◽  
Measles Virus ◽  
Reverse Genetics ◽  
Recombinant Vaccinia Virus ◽  
Viral Gene ◽  
Barr Virus ◽  
Epstein Barr ◽  
Cloned Cdna ◽  
P Proteins

ABSTRACT Reverse genetics technology so far established for measles virus (MeV) is based on the Edmonston strain, which was isolated several decades ago, has been passaged in nonlymphoid cell lines, and is no longer pathogenic in monkey models. On the other hand, MeVs isolated and passaged in the Epstein-Barr virus-transformed marmoset B-lymphoblastoid cell line B95a would retain their original pathogenicity (F. Kobune et al., J. Virol. 64:700–705, 1990). Here we have developed MeV reverse genetics systems based on the highly pathogenic IC-B strain isolated in B95a cells. Infectious viruses were successfully recovered from the cloned cDNA of IC-B strain by two different approaches. One was simple cotransfection of B95a cells, with three plasmids each encoding the nucleocapsid (N), phospho (P), or large (L) protein, respectively, and their expression was driven by the bacteriophage T7 RNA polymerase supplied by coinfecting recombinant vaccinia virus vTF7-3. The second approach was transfection with the L-encoding plasmid of a helper cell line constitutively expressing the MeV N and P proteins and the T7 polymerase (F. Radecke et al., EMBO J. 14:5773–5784, 1995) on which B95a cells were overlaid. Virus clones recovered by both methods possessed RNA genomes identical to that of the parental IC-B strain and were indistinguishable from the IC-B strain with respect to growth phenotypes in vitro and the clinical course and histopathology of experimentally infected cynomolgus monkeys. Thus, the systems developed here could be useful for studying viral gene functions in the context of the natural course of MeV pathogenesis.

scholarly journals In Vitro Assessment of Gentamicin Cytotoxicity on the Selected Mammalian Cell Line (Vero cells)

2017 ◽  
Vol 1 (1) ◽  
pp. 111-116 ◽  
Author(s):  
Anton Kovacik ◽  
Eva Tvrda ◽  
Diana Fulopova ◽  
Peter Cupka ◽  
Eva Kovacikova ◽  
...  
Keyword(s):  
Cell Viability ◽  
Vero Cell ◽  
Cell Line ◽  
Mammalian Cell ◽  
Vero Cells ◽  
Mammalian Cell Line ◽  
Total Proteins ◽  
African Green Monkey Kidney ◽  
Vero Cell Line

Abstract The aim of this study was to evaluate the in vitro cytotoxicity of different concentrations (500-7500 μg/mL) of gentamicin - GENT (aminoglycoside antibiotic) on the selected mammalian cell line (Vero - cell line from African green monkey kidney). Analysis of the cell morphological changes was microscopically evaluated (magnification × 400). Quantification of Ca, Mg and total proteins was performed using spectrophotometry on device Rx Monza (Randox). Quantification of Na, K and Cl was performed on the automatic analyzer EasyLyte. The cell viability was assessed using the metabolic mitochondrial MTT test. Vero cells were able to survive at concentrations of 500 (89.21 %), 1000 (79.54 %) and 2000 μg/mL (34.59 %). We observed statistically significant decrease of vital cell content at concentrations of 2000, 4500, 7500 μg/mL against control group. Vero cell line slightly reacted to the presence of GENT but total proteins and mineral parameters were not significantly affected. Vero cells were highly sensitive to GENT with a significant decrease of viability at concentrations of 2000 and 4500 μg/mL (P < 0.001). Our data reveal that GENT has a significant cytotoxic and adverse effect on the cell viability.

A Novel Triazole Derivative Drug Presenting In Vitro and In Vivo Anticancer Properties

2018 ◽  
Vol 18 (17) ◽  
pp. 1483-1493
Author(s):  
Ricardo Imbroisi Filho ◽  
Daniel T.G. Gonzaga ◽  
Thainá M. Demaria ◽  
João G.B. Leandro ◽  
Dora C.S. Costa ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cell Line ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Hepatic Function ◽  
Anticancer Properties ◽  
Mcf 7

Background: Cancer is a major cause of death worldwide, despite many different drugs available to treat the disease. This high mortality rate is largely due to the complexity of the disease, which results from several genetic and epigenetic changes. Therefore, researchers are constantly searching for novel drugs that can target different and multiple aspects of cancer. Experimental: After a screening, we selected one novel molecule, out of ninety-four triazole derivatives, that strongly affects the viability and proliferation of the human breast cancer cell line MCF-7, with minimal effects on non-cancer cells. The drug, named DAN94, induced a dose-dependent decrease in MCF-7 cells viability, with an IC50 of 3.2 ± 0.2 µM. Additionally, DAN94 interfered with mitochondria metabolism promoting reactive oxygen species production, triggering apoptosis and arresting the cancer cells on G1/G0 phase of cell cycle, inhibiting cell proliferation. These effects are not observed when the drug was tested in the non-cancer cell line MCF10A. Using a mouse model with xenograft tumor implants, the drug preventing tumor growth presented no toxicity for the animal and without altering biochemical markers of hepatic function. Results and Conclusion: The novel drug DAN94 is selective for cancer cells, targeting the mitochondrial metabolism, which culminates in the cancer cell death. In the end, DAN94 has been shown to be a promising drug for controlling breast cancer with minimal undesirable effects.

Oxytocin analogues with non-coded amino acid residues in position 8: [8-Neopentylglycine]oxytocin and [8-cycloleucine]oxytocin

1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
The Other ◽  
Amino Acid Residues ◽  
Phase Synthesis ◽  
Other Hand ◽  
Fragment Condensation

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.

Synthesis of Fragments of the Vasoactive Intestinal Peptide (VIP) and Analysis of Their Residual Biological Activities

1995 ◽  
Vol 60 (7) ◽  
pp. 1229-1235 ◽  
Author(s):  
Ivana Zoulíková ◽  
Ivan Svoboda ◽  
Jiří Velek ◽  
Václav Kašička ◽  
Jiřina Slaninová ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biological Activities ◽  
Residual Activity ◽  
Detailed Examination ◽  
Amino Acid Residues ◽  
Linear Peptide ◽  
Zone Electrophoresis ◽  
Capillary Zone

The vasoactive intestinal (poly)peptide (VIP) is a linear peptide containing 28 amino acid residues, whose primary structure indicates a low metabolic stability. The following VIP fragments, as potential metabolites, and their analogues were prepared by synthesis on a solid: [His(Dnp)1]VIP(1-10), VIP(11-14), [D-Arg12]VIP(11-14), [Lys(Pac)15,21,Arg20]VIP(15-22), and VIP(23-28). After purification, the peptides were characterized by amino acid analysis, mass spectrometry, RP HPLC, and capillary zone electrophoresis. In some tests, detailed examination of the biological activity of the substances in vivo and in vitro gave evidence of a low, residual activity of some fragments, viz. a depressoric activity in vivo for [His(Dnp)1]VIP(1-10) and a stimulating activity for the release of α-amylase in vitro and in vivo for [Lys(Pac)15,21,Arg20]VIP(15-22) and VIP(23-28).

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