Limitations

One of the initial goals for this research was to investigate how metadata played a role in the exposure-response relationships. Specifically, we wanted to examine differential responses by gender, BMI, and GSTM1 status following the three exposures. There is a wealth of

scientific literature demonstrating variation in response to environmental exposures based on demographic data, and we wanted to use this type of data to elucidate any underlying differences in response using these three classifications (Alexis et al., 2009; Gilliland et al., 2002; Setlow et al., 1998). Unfortunately, there was not sufficient statistical power to differentiate responses based on gender or BMI. The results in Chapter 4 come close to demonstrating statistically significant differences for the cytokine responses and the blood pressure measurements based on GSTM1 status, but the analysis was limited by the number of GSTM1- volunteers.

The cytokine results for this dissertation came from circulating blood and not from pulmonary or upper airway tissue. Samples collected in these locations would presumably have given a more direct relationship between the inhalation exposures and the inflammatory

response. Exhaled breath condensate samples were collected and analyzed for the cytokines of interest as a substitute for tissue samples, but describing these exposure-response relationships is an area for future research.

5.5 Future Work

The work presented in Chapters 3 and 4 is focused on the relationship between the three exposures and cytokines in blood plasma. We demonstrated that the method was robust enough to use on human plasma, exhaled breath condensate, and urine. Knowing this, the EBC and urine samples from DEPOZ were analyzed for the 10 cytokines. Determining the exposure-

response relationships for these two additional biological media could offer a more complete explanation of the “total” biological response following the respective exposures.

This work was also primarily focused on the results from “Day 1” of the respective exposures. As described in the study design, the first day of the exposure was always followed by on the next day by an O3 exposure. One purpose of this design was to investigate if the first

day's exposure could be a "primer" for a more adverse response to a secondary exposure, essentially mimicking a real life exposure on two consecutive days. As such, the samples for these days need to be investigated to see if any of the prior day’s exposures could “pre-dispose” a person to have a greater inflammatory reaction following the O3 exposure on day 2.

5.6 Conclusions

In conclusion, the research presented in the dissertation has added to the exposure assessment field with several unique contributions. An immunochemistry method with certain improvements in raw signal interpretation has been shown to be sensitive and specific enough to investigate inflammatory markers in three different human biological media from “healthy” people. Second, the results demonstrate that specific cytokines respond following DE-only and O3-only exposures, and that a synergistic exposure-response relationship is created when

combining DE and O3 into a mixed exposure. Next, the results have described and created a link

between the three exposures, inflammatory markers, and cardiovascular health effects. The results also demonstrate that individuals have independent responses following exposure and that discounting this fact could lead to a miss-interpretation of the exposure-response relationship between the three exposure and the inflammatory response. Finally, I hope that the results from this dissertation and subsequent publications will be used in future risk analyses and decisions to help alleviate the overall burden of ambient-acquired cardiovascular disease.

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