Cells viability

In the present authors laboratory, in-vitro tests have been carried out on numerous alloys and pure metals in order to determine their effect on cell viability and their capacity to induce inflammatory reactions (Table 5.4). 

The viability tests consisted of the establishment of the relative plating efficiency (RPE) and subsequently, the 50% lethal concentration LCjo (or RPE50) by using the colonyforming method on human epithelial cells in culture, the LI 32 cell-line. This test measures quantitatively only one criterion of toxicity which is cell death or cell survival, and consequently is specific, liable, and easily reproducible. It makes possible the ranking of cytotoxic effects of any chemical substance by comparison with the LC50 (Puck and Marcus 1955 Frazier and Andrews 

Pure metals or alloys LQo (mg L ) Survival rates (% SD) at 400 mg L Multinucleated Giant Cells (% SD) 

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Figure 15.4 shows the linear model for (15.6.3), the loss of cell viability at various temperatures. As the temperature increases from 105 to 121 °C, the value for the slope of the line increases. This means that the number of viable cells at a fixed time of sterilisation will drastically decrease as the temperature increases by 16 °C. 

In contrast to the DNA damage checkpoint, the mitotic spindle checkpoint is essential for cell viability. Dierefore, targeting kinases of the spindle checkpoint including Bubl, BubRl, and Mpsl might be a valid strategy for anticancer treatment. 

Constitutive exocytosis/secretion takes place in all eukaryotic cells and is essential for cell viability and growth. Trafficking vesicles destined for constitutive exocytosis originate from the trans-Golgi-network and contain secretory macromolecules derived from the... 

In order to assess the effect of the corn cob xylan on the cell viability and proliferation rate, xylan solutions at concentrations of 0.1, 0.25, 0.50, 0.75, and 1 mg/ml were placed in contact with human cervical adenocarcinoma cells (HeLa cells) for 24 and 72 h. Finally, the cell viability was determined by the MTT assay. It was observed that regardless of the xylan concentration, the samples tested did not affect the viability of HeLa cells after incubation for 24 h (Figure 13) (Unpublished data). 

Besides, the statistical analysis of the results obtained confirmed that the xylan samples did not present a significant effect on the cell viability and cell proliferation rate when in direct contact with HeLa cells at the concentrations used in this study and compared to the control. 

The biological impact of starch capped copper nanoparticles on mouse embryonic fibroblast (3T3L1) cells in vitro) was also evaluated by various parameters. More than 85 % of the 3T3Llcells were found to be viable, even after 20 hours time exposure which implies minimum impact on cell viability and morphology. The study demonstrates dose dependent cytotoxic potential of SCuNPs, that is non cytotoxic in the nanogram dose and moderately cytotoxic in the microgram doses (Fig. 10). Comparison of SCuNPs with Cu ions and uncapped copper nanoparticles (UCuNPs) revealed that, ions are more cytotoxic than SCuNPs. This observation supports the theory of slow release of ions from starch coated nanoparticles. 

Fig. 10. Effect of starch capped copper nanoparticles on cell viability (MTT assay) in case of mouse embryonic fibroblast (3T3L1).

Table 3. Effect of AR concentration ICHM on the saving of E.coU recA y.lux cell viability and relative index of SOS-system induction at UV exposion (3.64 J/m ). - P<0.05 - P<0.01.

So this research reveals new mechanisms of bacterial autoregulation under extreme conditions, controlled by low weight molecules - alkyiresorcinols. It applied aspects are defined by developing of methods for DNA protection in vitro and elongators of bacterial cells viability at UV exposure. 

Drought is perhaps one of the most complex examples to choose but it illustrates well the possibilities of, and pitfalls to, progress. Drought affects almost every facet of plant function and we are faced with the paradox that yield and evapotranspiration are intimately linked. In general, increases in yield when water supply is limiting are likely to result from characteristics which increase the available water supply, increase water use efficiency or increase biomass allocation to the economically useful plant parts (Pass-ioura, 1986). Additionally, features which maintain cell viability and protect metabolism in water-stressed tissue and allow rapid recovery after dry periods will contribute yield under some circumstances. 

Cationic aPNAs are taken up into living cells in a dose-dependant manner without affecting cell viability. (4) We have shown that aPNAs are stable to the nucleases present in human semm for up to 6 h. 

Fig. 15. Effects of turbulent shear stress level and exposure time on cell viability measured by trypan blue staining. Cells were sheared in a concentric cylinder viscometer [1]...

Models based on Eqs. (47)-(50) have been used in the past to describe the disruption of unicellular micro-organisms and mammalian (hybridoma) cells [62]. The extent of cell disruption was measured in terms of loss of cell viability and was found to be dependent on both the level of stress (deformation) and the time of exposure (Fig. 25). All of the experiments were carried out in a cone and plate viscometer under laminar flow conditions by adding dextran to the solution. A critical condition for the rupture of the walls was defined in terms of shear deformation given by Eq. (44). Using micromanipulation techniques data were provided for the critical forces necessary to burst the cells (see Fig. 4)... 

The influence of mechanical forces on cell viability is of great importance when growing the cells in agitated systems. By far the greatest amount of work reported in the literature has been done on suspension cells but adherent cells also experience shear forces not only in bioreactors also in vivo. Therefore, most research has be done on endothelial cells but studies exists done on non-endothelial cells. The influence of shear forces on cell growth, morphology and productivity will be discussed as well as possibilities of making the cells more resistant. 

The main reasons for the damage to cells in a reactor are the apparent shear forces and the collision of microcarriers with themselves and with turbulent eddies. In the literature studies are mainly focused on suspension cells and there again on hybridoma cells. The work reported in the hterature can be divided into two fields studies dealing with the influence of various stirrer speeds on cell viability and those investigating the influence of defined shear forces on cells with a viscosimeter. 

A dual isotope labeling technique [85] has been used to measure membrane permeability in plant cells, based on the selective permeabiHty of the membranes of living cells to tritiated water and carbon-14 labeled mannitol. Kieran [29] showed that the results of the dual isotope labeling and Evan s Blue staining methods correlated well as indicators of cell viability however, the latter was preferable in terms of reagent cost and ease of analysis. 

Tissue Culture Assay. Kogure et al. (48) report a novel tissue culture assay for detecting several types of sodium channel blockers. The mouse neuroblastoma cell line ATCC CCL 131 is grown in RPMI 1640 supplemented with 13.5% fetal bovine serum and 100 pg/ml gentamycin, in an atmosphere of 5% C0 95% air at 37 C. Ninety-six well plates are seeded with 1 x 10 cells in 200 pi of medium containing 1 mM ouabain and 0.075 mM veratridine. Veratridine and ouabain cause neuroblastoma cells to round-up and die. In the presence of sodium channel blockers (e.g., TTXs or STXs), the lethal action of veratridine is obviated and cells retain normal morphology and viability. An important feature of this assay is that a positive test for sodium channel blockers results in normal cell viability. Since bacterial extracts can contain cytotoxic components, this assay offers an advantage over tests that use cell death as an endpoint. The minimum detectable level of TTX is approximately 3 nM, or approximately 1/1000 mouse unit. 

Studies by Hudson et al, (2000) have demonstrated the presence of eight polyphenols in rice bran by using high-pressure liquid chromatography. They are protocatechuic acid, p-coumaric acid, ferulic acid, sinapic aci vanillic acid, caffeic acid, which is a methoxycirmamic acid derivative, and tricin. The effect of these polyphenols on cell viability and on the colony-forming ability of human-derived MDA MB 468 and HBL 100 breast cells, colon-derived SW 480 and human colonic epithelial cells was assessed. These authors concluded that rice bran polyphenols have putative cancer chemopreventive properties. 

The effect of endogenous PL3 (A) and externally applied E. carotovora enzymes (B) on the cell viability of tuber tissue disks from PL-inactive (I) and PL-active (II) plant lines. 

IC50 values of cisplatin and AU55 and Figures 29a and b show the course of cell viabilities in dependence of the molar concentrations in case of melanoma cells. 

Isolated hepatocytes incubated with ionic iron rapidly undergo lipid peroxidation. Some studies have not shown a consequent decrease in viability (as indicated by uptake of trypan blue or release of enzymes). This is probably a result of short incubation times, as changes in viability lag behind increases in lipid peroxidation, and may not occur for more than 2 h after lipid peroxidation begins (Bacon and Britton, 1990). Recent studies have shown strong correlations between increased lipid peroxidation [production of thiobarbituric acid (TBA) reactants] and loss of cell viability (trypan blue staining) (Bacon and Britton, 1989). The significance of the lag between lipid peroxidation and decreases in cell viability is as yet uncertain. 

Treatment with iron chelators and a-tocopherol protect against lipid p>eroxidation and hepatocellular injury in iron-overloaded rats (Sharma etal., 1990). When hepatocytes are isolated from rats, which have been pretreated with a-tocopherol, there is a significant reduction in iron-induced lipid peroxidation and improvement in cell viability in vitro (Poli et al., 1985). Similar effects were seen when hepatocytes were incubated with iron chelators (Bacon and Britton, 1990). Treatment of moderately, but not heavily, iron-loaded rats with desferrioxamine in vivo inhibits the pro-oxidant activity of hepatic ultrafiltrates (Britton et al., 1990b). 

Grisham, M.B., Gaginella, T.S., Ritter, C.V., Tamai, H., Be, R.M. and Granger, D.N. (1990b). The effects of neutrophil-derived oxidants on ileal mucosal permeability, electrolyte transport and epithelial cell viability in the rat. Inflammation 14, 531-542. 

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