Outline Macrophages (differentiated THP-1 cell line) will be treated with extracts from dried white elderflower, black elderflower, and elderberry.

Outline Macrophages (differentiated THP-1 cell line) will be treated with extracts from dried white elderflower, black elderflower, and elderberry. treated and untreated will also be stimulated with an agent known to induce a proinflammatory response in the cell line (LPS) in order to perform comparisons to normal baseline response. The study is carried out in vitro and the extracts, constituents, and metabolites (polyphenols in general as we don’t have the tools to recognise them) were investigated for their effects on pro-inflammatory cytokines release (TNF-alpha and IL1-beta) in lipopolysaccharides (LPS)-activated THP-1 macrophages. ELISA was used to compare the anti-inflammatory effects of the aqueous extract. Introduction the hypothesis of our research project is that elderflower (white, black, elderberry) would have anti-inflammatory effect, based on past research. The project is based on the idea of nutraceuticals in treating disease like diabetes and other diseases like cardiovascular, hypertension which are all rooted in inflammation. Therefore inflammation is the root cause of the disease. Mention CHRONIC SYSTHEMIC INFLAMMATION. How our extracts can be useful in this condition?
There is evidence to suggest that there is a connection between type 2 diabetes and chronic low-grade systemic inflammation, also known as chronic systemic inflammation. Chronic inflammation is a state of ongoing, low-level inflammation that can persist for months or even years, and it has been linked to a variety of chronic diseases, including type 2 diabetes.
One of the ways that chronic inflammation may contribute to the development of type 2 diabetes is by impairing insulin signalling. Insulin is a hormone that helps regulate blood sugar levels by promoting the uptake of glucose from the bloodstream into cells. Chronic inflammation can interfere with the ability of insulin to do its job, leading to insulin resistance and higher blood sugar levels.
In addition, chronic inflammation has been associated with changes in the gut microbiome, which may also contribute to the development of type 2 diabetes.
Overall, the inflammatory signalling cascade in type 2 diabetes involves a complex interplay of various pathways, including NF, JNK, inflammasome, and TLR signalling. These pathways can contribute to insulin resistance, impaired glucose metabolism, and other complications of type 2 diabetes. Therefore, targeting these pathways may be a potential therapeutic strategy for managing the inflammatory component of type 2 diabetes (tested in the lab TNF-alpha, and IL1-beta).
Overall, while the precise mechanisms linking chronic inflammation and type 2 diabetes are not yet fully understood (mention some papers that have tried to answer this question ,that is the mode of action of inflammation and T2D) there is growing evidence to suggest that chronic inflammation may play a role in the development of this condition. Managing chronic inflammation through lifestyle changes, such as diet and exercise, may therefore be an important strategy for preventing and managing type 2 diabetes.
As I mentioned in the lit review, another reason, the main reason at which we studing this type of nutraceuticals is because of the increasing need to substitute or accompany drugs with natural compounds with the aim do ameliorate treatment and if possible even prevent it.
There is evidence that elderflower extract can promote insulin sensitising effect through the activation of nuclear receptor PPAR-gamma.
There are some studies that have investigated the role of elderflower (white and black) and elderberry in modulating immune function and improving metabolic outcomes in T2D. Overall, these studies suggest that elderflower and elderberry may have potential for improving metabolic outcomes in type 2 diabetes. For example berry fruits have been recognized as capable of counteracting/mitigating inflammation and insulin resistance status.
The extract used showed to cause inflammation therefore to be pro-inflammatory. How this can be a good thing if inflammation in the case of diabetes? The extract used showed to cause inflammation therefore to be pro-inflammatory. How this can be a good thing if inflammation in the case of diabetes is bad for you, can it be related to the hormesis
Hormesis is a biological phenomenon where exposure to low doses of a stressor can stimulate beneficial adaptive responses in the body, leading to improved health and increased resilience. In the context of type 2 diabetes, hormesis has been shown to have beneficial effects on glucose regulation, insulin sensitivity, and other metabolic processes.
Future research on how can we move further once proven that our extracts are pro-inflammatory, like clinical trial testing both pre-diabetic and non-diabetics.
Answer & Explanation
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Macrophages are a type of white blood cell that plays a vital role in the immune system. They are responsible for engulfing and digesting pathogens, cellular debris, and other foreign substances. Elderberry has been traditionally used to boost the immune system, and recent studies suggest that it has anti-inflammatory and antiviral properties. In this experiment, we will treat macrophages with extracts from dried white elderflower, black elderflower, and elderberry to investigate their potential effects on macrophage activity.

Materials and Methods:

THP-1 cells will be cultured in RPMI medium with 10% FBS, 1%

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Step-by-step explanation
penicillin/streptomycin, and 0.05 mM 2-mercaptoethanol.
THP-1 cells will be differentiated into macrophages using 100 ng/mL PMA.
After 48 hours of differentiation, the cells will be washed with PBS and treated with different concentrations of elderflower and elderberry extracts (1 µg/mL, 10 µg/mL, and 100 µg/mL).
The cells will be incubated with the extracts for 24 hours, after which the supernatants will be collected for cytokine analysis.
Cytokine levels (IL-6, IL-1β, TNF-α, and IL-10) will be measured using ELISA.
Cell viability will be assessed using the MTT assay.

The results will show the effect of elderflower and elderberry extracts on macrophage activity. If the extracts enhance macrophage activity, we would expect to see an increase in cytokine levels. Conversely, if the extracts suppress macrophage activity, we would expect to see a decrease in cytokine levels. We will also assess cell viability to ensure that the extracts are not toxic to the cells.

This experiment will help determine the potential immunomodulatory effects of elderflower and elderberry extracts on macrophages. The results will be valuable in developing new therapies for immune disorders and viral infections.

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