Differential effects of hydrogen peroxide and ascorbic acid on the aerobic thermosensitivity of yeast cells grown under aerobic and anoxic conditions.

We have previously demonstrated that in aerobically-grown cells of the yeast Saccharomyces cerevisiae, hydrogen peroxide (H(2)O(2)) increases and ascorbic acid decreases cellular thermosensitivity, as determined by the inducibility of a heat shock (HS)-reporter gene. In this work, we reveal that the aerobic thermosensitivity of anaerobically-grown yeast cells also increases in the presence of H(2)O(2), albeit differentially between cells with two different lipid profiles. In comparison to aerobically-grown fermenting cells treated with the same H(2)O(2) concentration, both these types of anaerobically-grown cells were found to be considerably less sensitive to aerobic heat shock and considerably more thermotolerant. Paradoxically, and in contrast to ascorbate-pretreated aerobically-grown yeast cells, when anaerobically-grown cells were heat-shocked aerobically in the presence of the same ascorbic acid concentration, they exhibited increased thermosensitivity and decreased intrinsic thermotolerance with respect to their untreated counterparts. These findings are discussed with respect to what is currently known about the redox and physiological status of yeast cells grown aerobically and cells reoxygenated following anoxic growth.

The Incidence of Binocular Visual Impairment and Blindness in Children with Bilateral Retinoblastoma.

PURPOSE: The study aimed to assess the incidence of and risk factors leading to visual impairment and legal blindness in children with retinoblastoma. PROCEDURES: This is a single-center, retrospective case series of all patients with bilateral retinoblastoma presenting from 2010 to 2014. RESULTS: A total of 44 patients were included in the study. Visual impairment was present in 14 (38%) children, legal blindness was present in 7 (19%) children. Bilateral macular tumors (BMT) were associated with visual impairment (12 of 18 patients with BMT, 2 of 19 patients without BMT, p = 0.0006) and legal blindness (7 of 18 patients with BMT, 0 of 19 patients without BMT, p = 0.003). The International Intraocular Retinoblastoma Classification (IIRC) of the better eye also predicted visual impairment (16% in IIRC Group a, b, c, 75% in IIRC Group D, E, p = 0.004) and blindness (3% eye in IIRC Group a, b, c, 50% in Group D, E, p = 0.005). Various non-Snellen visual acuity measures were able to predict visual impairment in pre-verbal children, providing them with early assistance. CONCLUSIONS: The rates of visual impairment and blindness reported in this paper can be used to counsel families regarding the risk of binocular visual impairment. Early detection and support for visually impaired infants are essential as development can be affected by severe visual impairment.

Can the different heat shock response thresholds found in fermenting and respiring yeast cells be attributed to their differential redox states?

In this study we used a heat-shock (HS) reporter gene to demonstrate that respiring cells are intrinsically less sensitive (by 5 degrees C) than their fermenting counterparts to a sublethal heat shock. We also used an oxidant-sensitive fluorescent probe to demonstrate that this correlates with lower levels of sublethal reactive oxygen species (ROS) accumulation in heat-stressed respiring cells. Moreover, this relationship between HS induction of the reporter gene and ROS accumulation extends to respiring cells that have had their ROS levels modified by treatment with the anti-oxidant ascorbic acid and the pro-oxidant H(2)O(2). Thus, by demonstrating that the ROS/HSR correlation previously demonstrated in fermenting cells also holds for respiring cells (despite their greater HS insensitivity and higher level of intrinsic thermotolerance), we provide evidence that the intracellular redox state may influence both the sensitivity of the heat-shock response (HSR) and stress tolerance in the yeast Saccharomyces cerevisiae.

Reactive oxygen species may influence the heat shock response and stress tolerance in the yeast Saccharomyces cerevisiae.

Moderate levels of reactive oxygen species (ROS) have been implicated as second messengers in a number of biochemical pathways, and in animal cells have been associated with the activation of the heat shock response (HSR). Here, using an intracellular probe, we demonstrate that differential accumulation of ROS in the yeast Saccharomyces cerevisiae is strongly associated with differential induction of an HS reporter gene over a range of heat shock temperatures. There was a good correlation between cellular ROS levels and the levels of HS-induced reporter gene expression between 37 degrees C and 44 degrees C, both reaching maximal values at 41 degrees C. Furthermore, the addition of 150 microM H2O2 to the yeast cells during heat treatment resulted in a 3 degrees C decrease in the temperature required for maximal induction of the HS expression vector--an increased HS sensitivity that corresponded to a concomitant increase in ROS levels at these lower HS temperatures. Conversely, cells treated with 10 mM of the antioxidant ascorbic acid required a temperature that was 2 degrees C above that required in untreated controls for maximal induction of the HS expression vector. This decreased HS sensitivity corresponded to a decrease in ROS levels at these higher HS temperatures. Finally, cell viability assays reveal that intrinsic thermotolerance remains high in control cells despite concomitant decreases in HS-reporter gene expression and ROS accumulation between 41 degrees C and 44 degrees C. We conclude that the sensitivity of the yeast HSR is strongly associated with ROS accumulation, and suggest that ROS-mediated signalling ensures cooperation between the HS and the antioxidant responses.