A simple air-liquid interface exposure system for exposing cultured human 3D epidermis and cornea to PM2.5 collected through cyclonic separation.

Particulate matter (PM) is among the major air pollutants suspended in the atmosphere. PM2.5 has a particle size of 2.5 µm; it is known to cause inflammation, especially in the respiratory tract and skin. Since the skin acts a primary barrier against harmful environmental substances that may enter the body, it is highly exposed to PM2.5 present in the environment. However, the adverse health effects of PM2.5 exposure on human skin have not been accurately examined due to the lack of a system that exposes human epidermal tissue to the actual environmental concentration of PM2.5. In this study, we developed an air-liquid interface exposure system for exposing cultured human 3D epidermis and cornea to PM2.5 collected through cyclonic separation. PM2.5 suspension was nebulized in an acrylic chamber, and the resulting mist was pumped through a diffusion dryer into a glass exposure chamber. A particle counter was connected to the exposure chamber to continuously measure the spatial mass concentration of PM. Human 3D epidermis was cultured in the exposure chamber. Exposure of the human 3D epidermis to PM aerosol increased interleukin-8 release into the media around 50 µg/m3. Mass concentrations above 100 µg/m3 caused cell death. Furthermore, a human corneal model showed similar responses against PM2.5 exposure as 3D epidermis. The air-liquid interface exposure system developed in this study is considered useful for evaluating the health effects induced by environmental PM2.5 and can be used as an alternative to experiments involving actual human or animals.

Ca2+-dependent inhibition of branched-chain α-ketoacid dehydrogenase kinase by thiamine pyrophosphate.

Catabolism of the branched-chain amino acids (BCAAs: leucine, isoleucine, and valine) is regulated by the branched-chain α-ketoacid dehydrogenase (BCKDH) complex, which in turn is regulated by phosphorylation catalyzed by BCKDH kinase (BDK). Thiamine pyrophosphate (TPP) is required as a coenzyme for the E1 component of the BCKDH complex and can also bring about activation of the complex by inhibiting BDK. The present study shows that free Ca2+ in the physiological range greatly increases the sensitivity of BDK to inhibition by TPP (IC50 of 2.5 µM in the presence of 1 µM free Ca2+). This novel mechanism may be responsible for the stimulation of BCAA oxidation by conditions that increase mitochondrial free Ca2+ levels, e.g. in skeletal muscle during exercise.