Inactivation of lactate dehydrogenase by several chemicals: Implications for in vitro toxicology studies

Kendig, Derek M.; Tarloff, Joan B.

doi:10.1016/j.tiv.2006.08.004

Cited by 71 publications

(45 citation statements)

References 19 publications

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“…This most likely results from very low or absent cytolysis, combined with the ability of T. vaginalis to scavenge and ingest proteins and other metabolites from the surrounding medium (48,49,50,51), resulting in a lower LDH readout. A similar phenomenon of LDH values below background was observed in several previous studies (52,53).…”

Section: Resultssupporting

confidence: 89%

Trichomonas vaginalis Contact-Dependent Cytolysis of Epithelial Cells

et al. 2013

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bTrichomonas vaginalis is an extracellular protozoan parasite that binds to the epithelium of the human urogenital tract during infection. In this study, we examined the propensities of 26 T. vaginalis strains to bind to and lyse prostate (BPH-1) and ectocervical (Ect1) epithelium and to lyse red blood cells (RBCs). We found that only three of the strains had a statistically significant preference for either BPH-1 (MSA1103) or Ect1 (LA1 and MSA1123). Overall, we observed that levels of adherence are highly variable among strains, with a 12-fold range of adherence on Ect1 cells and a 45-fold range on BPH-1 cells. Cytolysis levels displayed even greater variability, from no detectable cytolysis to 80% or 90% cytolysis of Ect1 and BPH-1, respectively. Levels of adherence and cytolysis correlate for weakly adherent/cytolytic strains, and a threshold of attachment was found to be necessary to trigger cytolysis; however, this threshold can be reached without inducing cytolysis. Furthermore, cytolysis was completely blocked when we prevented attachment of the parasites to host cells while allowing soluble factors complete access. We demonstrate that hemolysis was a rare trait, with only 4 of the 26 strains capable of lysing >20% RBCs with a 1:30 parasite/RBC ratio. Hemolysis also did not correlate with adherence to or cytolysis of either male (BPH-1)-or female (Ect1)-derived epithelial cell lines. Our results reveal that despite a broad range of pathogenic properties among different T. vaginalis strains, all strains show strict contact-dependent cytolysis. T he exclusively human-infective flagellated parasitic protistTrichomonas vaginalis is the cause of the most common nonviral sexually transmitted disease, trichomoniasis. In women, the parasite resides in the vagina and colonizes the cervix; in men, it can be found in the urogenital tract and the prostate. In 2008 alone there were an estimated 276.4 million new cases in adults between 19 and 49 years of age, with a 10-fold-higher prevalence in females than in males (1). T. vaginalis in males often goes undetected, and thus the incidence is difficult to determine. However, it has been reported that in men with infected female partners, the incidence can range from 15 to 73% (2, 3, 4, 5). Additionally, men can clear infections 3 to 12 times faster than women whether the infection is treated or not (6, 7). The percentage of men who reportedly remain asymptomatic upon T. vaginalis infection is highly variable depending on the population examined and the diagnostic technique utilized. Regardless, it is thought that a higher proportion of men than women have asymptomatic infections (4,8,9).T. vaginalis infection can cause symptoms ranging from irritation and swelling in the urogenital tract to severe complications such as cervical erosion and premature birth during pregnancy (10,11,12). Trichomoniasis is also associated with infertility in both men and women (13,14). Moreover, infections are linked to cervical cancer (15,16,17,18), benign prostate hyperplasia (19), advance...

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Section: Resultssupporting

confidence: 89%

Trichomonas vaginalis Contact-Dependent Cytolysis of Epithelial Cells

et al. 2013

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“…LDH release is commonly used as a marker for necrotic/oncotic cell death (Valentovic and Ball, 1998). Most cells contain LDH and when these cells are lethally injured, loss of membrane integrity can be assessed by monitoring activity of LDH in the incubation medium (Kendig and Tarloff, 2007). This is consistent with Yang et al's report that a COEsrelated dose-dependent increase in levels of DNA damage, micronuclei rate, MDA concentration, and LDH activity (Yang et al, 2007).…”

Section: Discussionsupporting

confidence: 82%

Synergistic Increase of Oxidative Stress and Tumor Markers in PAH-Exposed Workers

Gao

Chen²,

Li³

et al. 2014

Asian Pacific Journal of Cancer Prevention

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“…In previous studies, loss of viability determined using alamarBlue paralleled loss of intracellular lactate dehydrogenase activity (Kendig and Tarloff, 2007), suggesting that loss of viability measured with alamarBlue represents cytotoxicity. LLC-PK 1 cells were seeded in 24-well plates at a density of 1.0 × 10 5 cells/ml and allowed to attach overnight.…”

Section: Methodsmentioning

confidence: 71%

Contribution of reactive oxygen species to para-aminophenol toxicity in LLC-PK1 cells

Foreman¹,

Tarloff²

2008

Toxicology and Applied Pharmacology

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Para-aminophenol (PAP) causes nephrotoxicity by biochemical mechanisms that have not been fully elucidated. PAP can undergo enzymatic or non-enzymatic oxidation to form reactive intermediates. Using modulators of reactive oxygen species (ROS), the role of ROS in PAP toxicity in LLC-PK 1 cells was investigated. ROS formation was determined using a fluorescein derivative and viability using alamarBlue. Following treatment of cells with PAP, ROS formation occurred prior to loss of cell viability. Several modulators of ROS were used to identify the pathways involved in PAP toxicity. Viability was improved with catalase treatment, while viability was decreased when cells were treated with superoxide dismutase (SOD). Both catalase and SOD exert their effects outside of cells in the incubation medium, since there was no evidence of uptake of these enzymes in LLC-PK 1 cells. In cell-free incubations, hydrogen peroxide (H 2 O 2 ) was produced when 0.5 mM PAP was included in the incubation medium. Further, SOD greatly increased and catalase greatly decreased H 2 O 2 production in these cell-free incubations. These data suggest that H 2 O 2 formed in the incubation medium contributes to loss of viability following PAP treatment. When cells were coincubated with 0.5 mM PAP and tiron, pyruvate, bathocuproine, 1, 10-phenanthroline, or dimethylthiourea (DMTU), ROS formation was decreased. However, there was minimal improvement in cell viability. Paradoxically, DMTU exacerbated PAP-induced loss of viability. These data suggest ROS are generated in cells exposed to PAP but these species are not the predominant cause of cellular injury.

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Inactivation of lactate dehydrogenase by several chemicals: Implications for in vitro toxicology studies

Cited by 71 publications

References 19 publications

Trichomonas vaginalis Contact-Dependent Cytolysis of Epithelial Cells

Trichomonas vaginalis Contact-Dependent Cytolysis of Epithelial Cells

Synergistic Increase of Oxidative Stress and Tumor Markers in PAH-Exposed Workers

Contribution of reactive oxygen species to para-aminophenol toxicity in LLC-PK1 cells

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