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  1. Project Summary

Project title (Maximum 20 words)

The effect of branched-chain amino acids (BCAAs) on microglia cells associated with Alzheimer’s Disease
Word Count: 14

Project Summary (Maximum 250 words)
A healthy microenvironment is maintained by the resident macrophages of the central nervous system (CNS) known as microglia to ensure neuronal survival. [1] Under pathological or physiology condition, the immune protection can be initiated by microglia to clear away the pathogens, metabolic wastes and cellular debris. [2]
A notable features of Alzheimer’s disease (AD) is the activation and proliferation of microglia in the brain which concentrated around amyloid plagues. [3] Genetic data in humans indicated that microglia have an important role in the pathogenesis of AD. [3]
One of the crucial amino acids involved in several important brain functions is branched-chain amino acid (BCAAs). [4] The brain function is affected by BCAAs which modify the large neutral amino acid (LNAA) transport at the blood-brain barrier. [5] Normal concentrations of BCAA in human cerebrospinal fluid usually has a range of 5 – 15 μM that is lower in magnitude by several level. [6]
An examined study was performed for testing a hypothesis of excess consumption of sports drinks containing BCAAs on leading to amyotrophic lateral sclerosis among professional football players as those sports drinks contain 1 – 3 grams of total BCAAs per 342 grams. [7]
The aim of the project is to investigate the possible morphological and functional changes of the microglial cells while in the low or high concentration of BCAAs which is associated with Alzheimer’s disease.
Word Count: 226

  1. Project Details
    Please provide an outline of the project and relevance of the proposed work to the subject domain postgraduate programme. Please refer to guidelines for further details. (Maximum 500 words)

A study from Bernier L.P. et al., (2020) has carried out the experiment of microglial cells being blocked out from consuming glucose to see any changes occurred. However, the microglial cells were changed to consume glutamine rapidly as alternative metabolic fuel after were flexible metabolically. [8] Also, the equipment was using use fluorescence lifetime imaging of intracellular NAD(P)H and time-lapse two-photon imaging of microglial dynamics in vivo and in situ. [8] Therefore, in this project, the experiment will be doing in the similar way but with amino acid metabolism of BCAAs and using cell culture for observe any significant changes in morphological and functional of microglial cells, and this will be the hypothesis of this project.
The aim is to investigate the possible morphological and functional changes of microglial cells under low or high concentration of BCAAs, some significant changes are expected when it is under very high or low concentration of BCAAs. The project objectives are shown below:

  1. Morphological changes under the microscope
  2. Proliferative rate of microglial cells
  3. Apoptosis rate of microglial cells
    If there are significant morphological changes of microglial cells due to low or high concentration of BCAAs, it can be supplement when it is low concentration or remove when it is under high concentration for stabling the changes of microglial cells, therefore, this can be reducing the risk of developing AD.
    As if the proliferation of microglial cells is longer or shorter due low or high concentration of BCAAs, ensuring the optimal concentration of BCAAs will provide the best environment for microglial cells to live longer for performing their function better, so the risk of developing AD can be reduced.
    If there are significant number changes of apoptosis of microglial cells due to low or high concentration of BCAAs, the optimum concentration of BCAAs can ensure that microglial cells are at the best condition for living longer to reduce the chance and risk of developing Alzheimer’s disease.
    Therefore, if any of these possible outcomes do occur, it can be very important to investigate further which would be a great approach and one of the efficient therapies for treating AD, even with the patient who are in very critical condition of AD. Patients with critical conditions are likely to show more improvement than before which may reduce mortality rate of AD or the chances of development into dementia in the future.
    Word Count: 396
  4. Programme of Work
    Provide a detailed description of the proposed work programme. Please refer to guidelines for further details. (Maximum 500 words)

The cells that will be used in the cell culture procedure is SIM-A9, which is a spontaneously immortalised microglial cell line and is isolated from the postnatal murine cerebral cortex. [9]
Cells culture: The cells will be cultured in Dulbecco’s modified Eagle’s Medium/nutrient mixture F12 (DMEM/F12) (Gibco) medium which has 17.5 mM glucose containing 10% FBS and 0.1% Penicillin/Streptomycin antibiotics or Neurobasal A medium (Gibco). Every 3 days, the 70-80% confluence with medium will be replaced as the cells passaged inside. The sub-passage was carried out by using 2X trypsin-EDTA (Gibco) in Dulbecco’s Phosphate Buffered Saline (PBS) (Thermo Fisher Scientific) for 5 minutes to detach cells. After that, manipulate composition of culture media to measure end point such as cell viability, immunological marker following stimulation. Finally, high or low concentration of BCAAs will be added.
Morphological Studies
The cells will be looked at under a microscope to observe any significant morphological changes of microglial cells under high or low concentration of BCAAs.
Functional Studies
Cells proliferation study. Reagent WST-1 kit will be used to study proliferation rates of microglial cells under low or high concentration of BCAAS. The low concentration of BCAAs will be less than 5 μM.
Apoptosis Studies. Apoptosis kit (Caspase-3) will be used to study the apoptosis of microglial cells under very low or high concentration of BCAAs. The high concentration of BCAAs will be higher 15 μM.
Project objective Possible Outcomes

  1. Morphological changes under the microscope 1.1. There are significant morphological changes of microglial cells under low concentration of BCAAs.
    1.2. There are no significant morphological changes of microglial cells under low concentration of BCAAs.
    1.3. There are significant morphological changes of microglial cells under high concentration of BCAAs.
    1.4. There are no significant morphological changes of microglial cells under high concentration of BCAAs.
  2. Proliferative rate of microglial cells 2.1. The proliferation of microglial cells will be longer under low concentration of BCAAs.
    2.2. The proliferation of microglial cells will be shorter under low concentration of BCAAs.
    2.3. The proliferation of microglial cells will be longer under high concentration of BCAAs.
    2.4. The proliferation of microglial cells will be shorter under high concentration of BCAAs.
  3. Apoptosis rate of microglial cells 3.1. There are significant number changes of apoptosis of microglial cells under low concentration of BCAAs.
    3.2. There are no significant number changes of apoptosis of microglial cells under low concentration of BCAAs.
    3.3. There are significant number changes of apoptosis of microglial cells under high concentration of BCAAs.
    3.4. There are no significant number changes of apoptosis of microglial cells under high concentration of BCAAs.
    Table 1. Project objects and possible outcomes.

Table 2. The schedule of project shown by weeks
Word Count: 449

Type of service: Academic Paper Writing
Type of assignment: Report
Subject: Biology
Pages/words: 3/825
Number of sources: 0
Academic level: HNC/ HND
Paper format: Harvard
Line spacing: Double
Language style: UK English