Infrared Thermographic Analysis of Surface Temperature of the Hands During Exposure to Normobaric Hypoxia.

Frostbite and other cold-related injuries commonly develop during prolonged exposure to the low environmental temperatures of polar and mountainous regions. Hypoxia is a potent sympathetic stimulus that causes vasoconstriction of the peripheral blood vessels, which may further compound the risk of developing a cold-related injury during high-altitude exposure. To investigate this, we utilized portable infrared thermographic technology to quantitatively measure changes in the surface temperature of the hands during exposure to increasing levels of normobaric hypoxia in a temperature-controlled high-altitude simulation. Surface temperature was assessed at four anatomical locations on both the left and right hands in a cohort of 10 healthy male participants at a series of predetermined levels of hypoxia (0.20 fraction of inspired oxygen [FIO2] [pre- and postexposure], 0.172 FIO2, 0.145 FIO2, 0.128 FIO2). Thermographic analysis revealed an overall decrease in peripheral temperature across the anatomical regions of the hands as the hypoxic stimulus increased, with statistically significant reductions observed at all four anatomical sites during exposure to 0.128 FIO2 (p < 0.05). These findings demonstrate that portable infrared thermography can be used to detect reductions in peripheral surface body temperature during exposure to normobaric hypoxia.

A role for human mitochondrial complex II in the production of reactive oxygen species in human skin.

The mitochondrial respiratory chain is a major generator of cellular oxidative stress, thought to be an underlying cause of the carcinogenic and ageing process in many tissues including skin. Previous studies of the relative contributions of the respiratory chain (RC) complexes I, II and III towards production of reactive oxygen species (ROS) have focussed on rat tissues and certainly not on human skin which is surprising as this tissue is regularly exposed to UVA in sunlight, a potent generator of cellular oxidative stress. In a novel approach we have used an array of established specific metabolic inhibitors and DHR123 fluorescence to study the relative roles of the mitochondrial RC complexes in cellular ROS production in 2 types of human skin cells. These include additional enhancement of ROS production by exposure to physiological levels of UVA. The effects within epidermal and dermal derived skin cells are compared to other tissue cell types as well as those harbouring a compromised mitochondrial status (Rho-zero A549). The results show that the complex II inhibitor, TTFA, was the only RC inhibitor to significantly increase UVA-induced ROS production in both skin cell types (P<0.05) suggesting that the role of human skin complex II in terms of influencing ROS production is more important than previously thought particularly in comparison to liver cells. Interestingly, two-fold greater maximal activity of complex II enzyme was observed in both skin cell types compared to liver (P<0.001). The activities of RC enzymes appear to decrease with increasing age and telomere length is correlated with ageing. Our study showed that the level of maximal complex II activity was higher in the MRC5/hTERT (human lung fibroblasts transfected with telomerase) cells than the corresponding wild type cells (P=0.0012) which can be considered (in terms of telomerase activity) as models of younger and older cells respectively.

Implications of using the fluorescent probes, dihydrorhodamine 123 and 2',7'-dichlorodihydrofluorescein diacetate, for the detection of UVA-induced reactive oxygen species.

During investigation of UVA-induced oxidative stress in HaCaT keratinocytes with dihydrorhodamine 123 (DHR123) and 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA), exaggerated baseline values were observed within control samples, suggesting a mechanism of probe oxidation and subsequent change in fluorescence intensity (FI) independent of cellular ROS generation. The effects of diluent, UVA pre-treatment and loading protocols upon the FI of the probes have therefore been investigated. The study confirmed the capacity of Dulbecco's Modified Eagle's Medium (DMEM) to confer fluorescence intensity changes in both probes, most notably DCF-DA. In addition, UVA pre-treatment compromises the effectiveness of DHR123 and DCF-DA to detect ROS generated in a cell-free system. In vitro data shows a greater UVA-induced FI increase in HaCaT cells loaded with probe before rather than after UVA treatment. This study has important implications for future research, the understanding of previous studies and associated confounding effects using DHR123 and DCF-DA as ROS sensitive probes.