## 4 Discussion
A follow-up study with adiponectin sampling at different time points would be warranted to understand if the observed adiponectin result is simply an artefact of the variation present in a single measurement. PPAR\(\alpha\) was significantly upregulated in a dose-dependant manner. PPAR\(\alpha\) is a key gene involved in fatty-acid metabolism, lipolysis and fatty-acid oxidation. Flavonoids from grapefruit have been demonstrated to cause such effect [39]. However, unlike the response to grapefruit, the PPAR\(\alpha\) co-activator PGC1\(\alpha\) mRNA expression remained unchanged in the present study. These results suggest that PRSE may have a role in reducing the development of obesity via lipolysis and fatty acid oxidation. Fatty-acid synthase gave significant results in both liver and plasma. However, in the liver only the 4% PRSE treatment was significant and in plasma it was only the 2% PRSE treatment that had significant effects. Therefore, it is unlikely that weight loss would act through this pathway in a dose-dependant manner with feeding of PRSE in the diet. The reason for the significant difference at these specific dosages requires further investigation. UCP2 levels also returned contrasting results. There were no significant differences in adipose tissue UCP levels; however, in the liver a dose-dependant and highly significant relationship was observed. Compounds such as resveratrol have been suggested to have anti-obesity effects due to upregulation of UCP2 in the liver, amongst other things [40]. It is interesting that a rich source of carbohydrates such as sugarcane also contains compounds that are gaining evidence as efficient regulators of carbohydrate metabolism. Individually, some of the compounds identified in PRSE such as tricin, luteolin and apigenin have been demonstrated to have anti-obesity effects [41; 42; 43]. However, PRSE provides a novel source of these compounds. This trial has demonstrated a range of potential mechanisms for these anti-obesity effects in an animal model. Based on the data collected in this study, delivering these compounds at an optimal dosage may be a useful strategy in the prevention of obesity. The present study has identified mechanisms such as reduced absorption of carbohydrates and changes to pathways, especially those related to leptin, PPARs and liver UCP2 and to some extent adiponectin gene expression as likely candidates for weight loss in the presence of a high-carbohydrate, high-fat diet. There are other potential mechanisms such as the role of the microbiome or other cellular pathways that have not been covered in this trial and further studies that targeting these factors directly may provide valuable insights. These results demonstrate that PRSE is a bioavailable source of phytochemicals that has mitigated the damage of a high-fat, high-carbohydrate diet. Overall, further work will be required to translate these results to clinical trials. However, these initial results suggest that PRSE and sugarcane may have a positive role to play in the reduction of global obesity rates. **Supplementary Materials:** The following are available online at [[https://www.mdpi.com/article/10.3390/app11115163/s1](https://www.mdpi.com/article/10.3390/app11115163/s1)]([https://www.mdpi.com/article/10.3390/app11115163/s1](https://www.mdpi.com/article/10.3390/app11115163/s1)), Table S1: Diet Composition. **Author Contributions:** [PERSON], [PERSON], [PERSON], [PERSON], [PERSON], [PERSON]. & [PERSON] were all involved in the initial planning and completion of the trial. [PERSON]. & B.K. drafted the manuscript. All authors were involved with data interpretation. All authors have read and agreed to the published version of the manuscript.