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Revision as of 15:06, 19 October 2021
Overview
Exposure to prolonged chronic stress induces heightened vulnerability to anxiety, depression, and a lot of mood disorders. Dysfunction of the medial prefrontal cortex (mPFC) in the brain has been linked to the cognitive and emotional deficiency induced by long-term stress exposure. Chronic stress-induced depression (CSID), therefore, is an imperative mental disorder. If treated improperly, upcoming psychiatric disorders, such as major depressive disorders (MDD), anxiety, and even post-traumatic stress disorder (PTSD) caused by lasting sensitivity to recurring sensors, will eventually tear the patient to shreds[1]. According to the World Health Organization (WHO), depression is already becoming the leading cause of disability around the world. It even haunts more than 264 million people worldwide[2]. However, it is only the tip of the iceberg.
To alleviate damage of stress becoming crucial before it accumulates into a greater problem. We came up with a novel solution called Menble, where the engineered stress-relieving probiotic is wrapped inside a bubble. It is a modification of Taiwan’s representative drink, bubble tea, offering destressing benefits for people of all ages, especially working adults and teenagers. By incorporating our solution in the daily lives of people, it will be more accessible and affordable for consumers from all socioeconomic backgrounds, promoting health equality and sustainable communities in the long run.
Oxidative Stress @PQ
Hydrogen peroxide (H2O2) was changed to paraquat (PQ) as the inducer for the oxidative stress sensing system for the following reasons:
>Paraquat: a better inducer for the oxidative stress sensing system
1. PQ produces superoxide radical (O2¯·) catalyzed by NADPH-cytochrome P450 reductase.[e] Then, O2¯· is converted into hydrogen peroxide (H2O2) by the SOD enzyme system [f] or into hydroxyl radical (OH¯·) by the HWR enzyme system[f][g]. The pathway is shown in (Fig.3).
2. Because chronic stress-induced depression (CSID) is related to a variety of abnormal changes in oxidative stress,[h] a single kind of oxidative stress molecule cannot specifically mimic the changes in the human body. Therefore, PQ was chosen as the inducer in our project to more closely simulate oxidative stress in the intestine.
3. In our oxidative stress assay, we compared the strength of the oxidative stress sensing system when induced by PQ compared to when it is induced by hydrogen peroxide (H2O2). As shown in figure 4, sfGFP expression is greater when induced by PQ .(Fig.4)
Microfluidics
Flow Chart
Fabrication of Microfluidic Chip
1. Place the PDMS mixture inside a vacuum chamber as long as possible. Based on our own experience with a total PDMS value of 15gr on a circular container (diameter: 9cm), we put the solution in the vacuum chamber for at least 30minutes (you do not have to pay attention very much to this progress, or even better if you leave it running for an extended duration). But your mileage might vary: a more robust vacuum pump, more extensive surface area of the container used, and less solution mixed will make this progress faster.
Microfluidic Chip Assembly
Our team was honoured to interview the general manager of Chen En Food Product Enterprise, Mr Vincent, as Chen En is one of the largest bubble suppliers in both local and global markets. Before the online interview session began, we signed a Memorandum of Understanding (MOU) with them. As a giant bubble supplier, they provided advice on preparing our bubble with the engineered bacteria inside. In addition, they also gave us valuable market data to further our business model.
References
- Saito, Y., Sato, T., Nomoto, K., & Tsuji, H. (2018). Identification of phenol- and p-Cresol-producing intestinal bacteria by using media supplemented with tyrosine and its metabolites. FEMS Microbiology Ecology, 94(9).
- Zhang, G., Brokx, S., & Weiner, J. H. (2005). Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. Nature Biotechnology, 24(1), 100–104.
- Passmore, I. J., Letertre, M., Preston, M. D., Bianconi, I., Harrison, M. A., Nasher, F., … Dawson, L. F. (2018). Para-cresol production by Clostridium difficile affects microbial diversity and membrane integrity of Gram-negative bacteria. PLoS pathogens, 14(9), e1007191.
- InterPro EMBL-EBI. “4-Hydroxy-Tetrahydrodipicolinate Synthase, DapA (IPR005263) < InterPro < EMBL-EBI.” Ebi.Ac.Uk, 2019, www.ebi.ac.uk/interpro/entry/IPR005263. Accessed 5 July 2019.
- Merlin, C., Masters, M., McAteer, S., & Coulson, A. (2003). Why Is Carbonic Anhydrase Essential to Escherichia coli? Journal of Bacteriology, 185(21), 6415–6424.
- Hashimoto, M., & Kato, J.-I. (2003). Indispensability of the Escherichia coli Carbonic Anhydrases YadF and CynT in Cell Proliferation at a Low CO2 Partial Pressure. Bioscience, Biotechnology, and Biochemistry, 67(4), 919–922.
- Coralli, C., Maja Cemazar, Chryso Kanthou, Tozer, G. M., & Dachs, G. U. (2001). Limitations of the Reporter Green Fluorescent Protein under Simulated Tumor Conditions. Cancer Research, 61(12), 4784–4790.
