Saccharomyces cerevisiae infections and antifungal susceptibility studies by colorimetric and broth macrodilution methods

Tiballi, Robert N.; Spiegel, Joan; Zarins, Lidija T.; Kauffman, Carol A.

doi:10.1016/0732-8893(95)00188-3

Cited by 32 publications

(25 citation statements)

References 24 publications

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“…Cell Viability-Cell viability after exposure to different experimental conditions was measured by the Alamar Blue assay (22). Cell media were replaced with medium containing 10% Alamar Blue and placed at 37°C in a cell incubator for 2-3 h. The media were then collected and read on a plate reader at 530 nm.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, after exposure to IL-1␤ or its vehicle on both sides of the Transwell, the cells were washed twice with wash medium (150 mM NaCl and 2 mM HEPES, pH 7.4) at 37°C and equilibrated with flux medium (140 mM NaCl, 5 mM KCl, 1 mM Na 2 HPO 4 , 1 mM MgCl 2 , 0.2 mM CaCl 2 , 10 mM glucose, and 20 mM HEPES, pH 7.4) for 10 min at 37°C. After equilibration, only the apical medium was replaced with fresh flux medium containing 22 Na ϩ at a final activity of 5 Ci/ml and ouabain (3 mM). After 6 min, cell monolayers were washed three times by overflow with cold wash medium to eliminate 22 Na ϩ not taken up by the cells and to stop further uptake.…”

Section: Methodsmentioning

confidence: 99%

“…After equilibration, only the apical medium was replaced with fresh flux medium containing 22 Na ϩ at a final activity of 5 Ci/ml and ouabain (3 mM). After 6 min, cell monolayers were washed three times by overflow with cold wash medium to eliminate 22 Na ϩ not taken up by the cells and to stop further uptake. The last wash solution was measured for 22 Na ϩ and verified not to contain any radioactivity.…”

Section: Methodsmentioning

confidence: 99%

“…Human recombinant IL-1␤ and IL-1␤ receptor antagonist (IL-1RA) were obtained from R&D Systems (Minneapolis, MN). 22 Na ϩ was obtained from PerkinElmer Life Sciences. 32 P and the chemiluminescence ECL Plus kit were obtained from Amersham Biosciences.…”

Section: Methodsmentioning

confidence: 99%

“…After 6 min, cell monolayers were washed three times by overflow with cold wash medium to eliminate 22 Na ϩ not taken up by the cells and to stop further uptake. The last wash solution was measured for 22 Na ϩ and verified not to contain any radioactivity. The cells were then lysed with 0.1% NaOH, and the radioactivity in the lysate was measured in a ␤-counter.…”

Section: Methodsmentioning

confidence: 99%

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Interleukin-1β Decreases Expression of the Epithelial Sodium Channel α-Subunit in Alveolar Epithelial Cells via a p38 MAPK-dependent Signaling Pathway

Roux

Kawakatsu²,

Gartland

et al. 2005

Journal of Biological Chemistry

157

148

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Acute lung injury (ALI) is a devastating syndrome characterized by diffuse alveolar damage, elevated airspace levels of pro-inflammatory cytokines, and flooding of the alveolar spaces with protein-rich edema fluid. Interleukin-1␤ (IL-1␤) is one of the most biologically active cytokines in the distal airspaces of patients with ALI. IL-1␤ has been shown to increase lung epithelial and endothelial permeability. In this study, we hypothesized that IL-1␤ would decrease vectorial ion and water transport across the distal lung epithelium. Therefore, we measured the effects of IL-1␤ on transepithelial current, resistance, and sodium transport in primary cultures of alveolar epithelial type II (ATII) cells. IL-1␤ significantly reduced the amiloride-sensitive fraction of the transepithelial current and sodium transport across rat ATII cell monolayers. Moreover, IL-1␤ decreased basal and dexamethasone-induced epithelial sodium channel ␣-subunit (␣ENaC) mRNA levels and total and cellsurface protein expression. The inhibitory effect of IL-1␤ on ␣ENaC expression was mediated by the activation of p38 MAPK in both rat and human ATII cells and was independent of the activation of ␣v␤6 integrin and transforming growth factor-␤. These results indicate that IL-1␤ may contribute to alveolar edema in ALI by reducing distal lung epithelial sodium absorption. This reduction in ion and water transport across the lung epithelium is in large part due to a decrease in ␣ENaC expression through p38 MAPK-dependent inhibition of ␣ENaC promoter activity and to an alteration in ENaC trafficking to the apical membrane of ATII cells. Acute lung injury (ALI)1 is a devastating clinical syndrome manifested by diffuse alveolar damage, capillary injury, and disruption of the alveolar epithelium. The acute phase of ALI is characterized by the influx of protein-rich edema fluid that impairs gas exchange, causing arterial hypoxemia and respiratory failure with an overall mortality rate of 30 -40% (1). Along with an increase in lung endothelial and epithelial permeability to protein, this syndrome is associated with abnormal surfactant production and decreased vectorial fluid transport across the lung epithelial barrier (2, 3). A number of inflammatory mediators have been found to be elevated in the alveolar space during the early phase of ALI, including interleukin (IL)-1␤, tumor necrosis factor-␣, IL-6, and IL-8 (4). IL-1␤ is one of the most biologically active cytokines in pulmonary edema and bronchoalveolar lavage fluids of patients with ALI (4 -6). Indeed, IL-1␤ increases microvascular lung epithelial permeability in in vitro and in vivo models of ALI (7). IL-1␤ also enhances alveolar epithelial repair by increasing cell spreading (8) and fibroblast activation and proliferation (5). In contrast, the role of IL-1␤ in distal lung epithelial ion transport remains unclear. In other epithelia such as the colonic epithelium, IL-1␤ inhibits aldosterone-induced electrogenic sodium absorption and attenuates aldosterone-induced up-regulation of ␤-and ␥-subunit ...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%