## 3 Results
### Metabolite Variance across Multiwell Fluidic System to Ascertain Sites of Generated Cell Byproducts
#### 3.8.1 Trending Relevance
(**b3—rat hepatocyte line graph**) ASA is inconsequential at upstream well 3, stimulates a special cause “A” in wells 4–9, then an upregulation special cause (A+) in downstream wells 10 and 11. (**c3—HepaRG line graph**) NEF induces a negative trending process shift across the entire \(\upmu\)channel. (**d3—rat hepatocyte line graph**) NEF creates a negative process shift, wells 3–6, normal functions wells 7–8, special cause wells 9–10, and unusual decline outcome in well 11. Values a1-d1 are represented as the mean \(\pm\) SEM (n = 9). * \(p<0.05\), ** \(p<0.01\), and **** \(p<0.0001\). wells 3-5, has normal function at well 6, stimulates a special cause \"A\" in wells 7-9, then enhanced special cause \"A+\" in wells 10-11. (**c3--HepaRG line graph**) NEF induces a trending process shift in wells 3-7, normal function at well 8, then a negative special cause in wells 9-11. (**d3--rat hepatocyte line graph**) NEF stimulates a process shift in wells 3-5, has normal function in wells 6-7, stimulates a special cause in wells 8-10, then unusual outcome in well 11. Values (**a1-d1**) are represented as the mean \(\pm\) SEM (n = 9). * \(p\) < 0.05, **\(p\) < 0.01, and ****\(p\) < 0.0001. Figure 12: GSH levels across multiwell culture locations at Day 7. GSH synthesis shown as fold changes (vertical axis) vs. DMSO vehicle control; vehicle normalized at 1. (**a1,b1**) Effect of aspirin exposure in HepaRG cells and rat hepatocytes. (**c1,d1**) Effect of nefazodone exposure in HepaRG cells and rat hepatocytes. (**a2,b2,c2,d2**) Re-expression of data using individual trending charts (i.e., X control charts) to convey DMSO controls with drug study outcomes. (**a2—HepaRG line graph**) ASA inhibits GSH activity across wells 3 (**)–9 (*). (**b2—rat hepatocyte line graph**) ASA promotes GSH synthesis in upstream wells 3 (****)–5 (****), displays normal function in well 6, then stimulates GSH functions in wells 7 (****)–11 (****). (**c2—HepaRG line graph**) NEF inhibits GSH activity across wells 3 (*)–7 (*) and wells 9 (*)–11 (*). (**d2—rat hepatocyte line graph**) NEF promotes GSH synthesis in upstream wells 3 (*)–5 (*), displays normal function in wells 6–7, again stimulates GSH functions in wells 7 (*)–10 (*). (**a3,b3,c3,d3**). Variability charts (i.e., MR control charts) to display functional fold changes amid adjacent wells. (**a3—HepaRG line graph**) ASA induces a trending process shift across most of the \(\upmu\)channel, wells 3–10. (**b3—rat Hepatocyte Line Graph**) ASA stimulates a process shift in wells 3–5, has normal function at well 6, stimulates a special cause “A” in wells 7–9, then enhanced special cause “A+” in wells 10–11. (**c3—HepaRG line graph**) NEF induces a trending process shift in wells 3–7, normal function at well 8, then a negative special cause in wells 9-11. (**d3—rat hepatocyte line graph**) NEF stimulates a process shift in wells 3–5, has normal function in wells 6–7, stimulates a special cause in wells 8–10, then unusual outcome in well 11. Values (**a1–d1**) are represented as the mean \(\pm\) SEM (n = 9). * \(p\) < 0.05, **\(p\) < 0.01, and ****\(p\) < 0.0001. CYP3A4 ASSAY--Functional Indicator (Studies 5-8)  
Cytochrome P450 3A4 activity is found in the liver and has a significant role in the biotransformation of compounds [49]. **ASA exposure on HepaRG cells (Study 5)** is displayed in Figure 11(a1). ASA impedes CYP3A4 activity in well 3 (**) with slightly upregulated activities in wells 6 (*), 7 (*), 8 (*), and 9 (*); remaining wells 4, 5, 10, and 11 register as non-significant. **ASA exposure in rat hepatocytes (Study 6)** is displayed in Figure 11(b1). In data presentation, the scale of the vertical axis increased 10-fold to accommodate for intensified CYP3A4 outputs, i.e., range 0-5 adjusted to 0-50. ASA exposure is non-significant in well 3, CYP3A4 upregulates in wells 4 (*)-9 (**) to denote 3-to-10-fold increases over DMSO controls, then maximized in wells 10 (****) and 11 (****) at 40-fold and 20-fold levels. **NEF exposure in HepaRG cells (Study 7)** is displayed in Figure 11(c1).