Members of NADPH oxidase (Nox) enzyme family are important sources of reactive oxygen species (ROS) and are known to be involved in several physiological functions in response to various stimuli including UV irradiation. UVB-induced ROS have been associated with inflammation, cytotoxicity, cell death, or DNA damage in human keratinocytes. However, the source and the role of UVB-induced ROS remain undefined.Here, we show that Nox1 is involved in UVB-induced p38/MAPK activation and cytotoxicity via ROS generation in keratinocytes. Nox1 knockdown or inhibitor decreased UVB-induced ROS production in human keratinocytes. Nox1 knockdown impaired UVB-induced p38 activation, accompanied by reduced IL-6 levels and attenuated cell toxicity. Treatment of cells with N-acetyl-L-cysteine (NAC), a potent ROS scavenger, suppressed p38 activation as well as consequent IL-6 production and cytotoxicity in response to UVB exposure. p38 inhibitor also suppressed UVB-induced IL-6 production and cytotoxicity. Furthermore, the blockade of IL-6 production by IL-6 neutralizing antibody reduced UVB-induced cell toxicity.In vivo assay using wild-type mice, the intradermal injection of lysates from UVB-irradiated control cells, but not from UVB-irradiated Nox1 knockdown cells, induced inflammatory swelling and IL-6 production in the skin of ears. Moreover, administration of Nox1 inhibitor suppressed UVB-induced increase in IL-6 mRNA expression in mice skin.Collectively, these data suggest that Nox1-mediated ROS production is required for UVB-induced cytotoxicity and inflammation through p38 activation and inflammatory cytokine production, such as IL-6. Thus, our findings suggest Nox1 as a therapeutic target for cytotoxicity and inflammation in response to UVB exposure.

Background Wound healing is a complex biological process and complete skin regeneration is still a critical challenge. Extracellular vesicles (EVs) play essential roles in cell communication and cell regeneration, and recent studies have suggested that EVs may contribute to wound healing, though the molecular mechanisms behind this contribution remain unclear. For these reasons, we decided to use EVs isolated from human keratinocytes (HaCaT) in vitro to determine the potential mechanism of action of EV-derived wound healing. Method Scratch assays were used to determine cell migration and proliferation. Scratched cells were exposed to EVs in multiple conditions to determine how they affect wound healing. Statistical analysis between groups was carried out to using Student’s two-sided t test. A p value of <  0.05 was considered statistically significant. Result We found that proteomic analysis of purified EVs shows enrichment of proteins associated with cell communication and signal transduction, such as MAPK pathways, and keratinocyte and fibroblast cultures exposed to EVs had higher levels of proliferation, migration, and ERK1/2 and P38 activation. Moreover, we found that treatment with specific ERK1/2 and P38 signaling inhibitors PD98059 and SB239063 impaired EV-mediated cell migration, which suggests that ERK1/2 and P38 signaling is essential for EV-induced wound healing. Conclusion HaCaT cell-derived EVs accelerate the migration and proliferation of human keratinocytes and fibroblasts and may promote wound healing via the activation of MAPKinase pathways. These findings may be key in developing new methods to treat wounds and accelerate wound healing in the future.

Objective: To review the correlation between prostate specific antigen (PSA) and Gleason score and Cav-1 for diagnosing prostate adenocarcinoma. Methods: Data were collected from one hundred fifty-nine patients with prostate adenocarcinoma at the Department of Urology, Dr. Hasan Sadikin General Hospital in the period of January 2008-December 2010. The PSA levels were measured and classified into <4 ng/ml, 4-10 ng/ml, and >10 ng/ ml. The results were then analyzed and compared to the imunohistochemistry (caveolin-1) staining in the literature. The Gleason score was also noted and analyzed. Results: This study confirmed that positive caveolin-1 expression was related to the clinical markers of disease progression and was predictive of poor clinical outcome after surgery. The PSA results showed that one hundred fourty-one adenocarcinoma patients had a PSA level of >10 ng/ml with Gleason score of gleason 5-6 as the most common score. However, there was no correlation between PSA and Gleason score and caveolin-1 for diagnosing prostate adenocarcinoma. Conclusions: Caveolin-1 cannot be used to measure Gleason and PSA score due to different markers that have various advantages and disadvantages to predict carcinoma prostate. Therefore, further studies are needed.

Background: Members of NADPH oxidase (Nox) enzyme family are important sources of reactive oxygen species (ROS) and are known to be involved in several physiological functions in response to various stimuli including UV irradiation. UVB-induced ROS have been associated with inflammation, cytotoxicity, cell death, or DNA damage in human keratinocytes. However, the source and the role of UVB-induced ROS remain undefined. Method: We measured intracellular ROS, cell viability, cytotoxicity and cell signaling in response to UVB radiation (15 -100mJ/cm2) using human keratinocyte (HaCaT cell), which transfected with Nox1 or non-targeting-siRNA. We also determined ear swelling of C57BL/6 mice which administered intradermal injections of the cell lysate. Result: Nox1 knockdown decreased UVB-induced ROS production in human keratinocytes. Moreover, Nox1 knockdown impaired UVB-induced p38 activation, accompanied by reduced IL-6 levels and attenuated cell toxicity. Treatment of cells with N-acetyl-L-cysteine (NAC), a potent ROS scavenger, suppressed p38 activation as well as consequent IL-6 production and cytotoxicity in response to UVB exposure. p38 inhibitor also suppressed UVB-induced IL-6 production and cytotoxicity. Furthermore, the blockade of IL-6 production by IL-6 neutralizing antibody reduced UVB-induced cell toxicity. Coincidentally, the intradermal injection of lysates from UVB-irradiated control cells, but not from UVBirradiated Nox1 knockdown cells, induced inflammatory swelling and IL-6 production in the skin of ears in wild-type mice. Conclusion: Nox1-mediated ROS production is required for UVB-induced cytotoxicity and inflammation through p38 activation and inflammatory cytokine production, such as IL-6. Our findings suggest Nox1 as a therapeutic target for cytotoxicity and inflammation in response to UVB exposure.