Frontiers in Pharmacology

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Summary of Research Method Applied

In Vitro Studies: These studies give us useful information about the effects of Bryostatin-1 on the mechanisms involved in MS, such as oxidative stress. Explanation: Oxidative stress is an imbalance between the production of free radicals and the ability of the body to counteract or detoxify their harmful effects through neutralization by antioxidants.

Viability Tests: Explanation: This is an assay to determine the ability of organs, cells, or tissues to maintain or recover viability. For example, the ratio of potassium to sodium in cells can be considered as an indicator of survival.

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If the cells do not contain a high proportion of potassium and low sodium, this could indicate that (1) the cell membrane may not be intact, and/or (2) the potassium sodium pump may not be functioning well. Viability tests include a variety of assessments, from quantitative measures of physiological function to determinations of whether damage can be repaired and recovered.

Qualitative and Quantitative Analysis of Apoptotic Markers: Explanation: Apoptosis occurs through a complex series of signals. Illustrating this principle, the image below shows the key parameters of apoptosis and the approximate relative time when the signs of those events are likely to be detected.

Measurement of Neurotrophin Levels: Explanation: Neurotrophins are a group of proteins that stimulate the survival and development of neurons. They also play an important role in inducing the apoptosis of PCD neurons. Neurotrophic survival signals in neurons are mediated by the high affinity binding of these proteins to their Trk receptors. In contrast, the binding of neurotrophins to p75NTR receptors mediates a majority of neuronal apoptosis signals. PCD, which occurs during brain growth, is responsible for the loss of the majority of excess neurons. This is necessary, as, during development, there is an overproduction of neurons that must be eliminated for optimal functioning.

Mitochondrial Assays Before and After Treatment with Bryostatin-1: Explanation: Mitochondrial function has been assessed using Clark-type electrode probes for measuring oxygen consumption, luminescent ATP assays for quantifying total energy metabolism, and MTT or Alamar Blue for determining metabolic activity.

In Vivo Studies: To evaluate the in vivo effects of Bryostatin-1, the toxic model of MS is created in C57BL/6 mice using cuprizone. Explanation: The cuprizone model simulates toxic demyelination. In this model, young mice are fed the copper chelator cuprizone, which leads to oligodendrocyte death and subsequent reversible demyelination. Spontaneous remyelination can occur as early as four days after withdrawing from cuprizone.

Immunohistochemistry: Explanation: This is the most common application of immunostaining. It involves selectively identifying antigens (proteins) in cells of a tissue section by exploiting the principle of antibody binding specificity to antigens in biological tissues.

Western Blot: Explanation: This is a common method for detecting and analyzing proteins. It is based on technology that involves the transfer, or blotting, of proteins separated by electrophoresis from a gel to a membrane, where they can be precisely visualized.

Real-time PCR analyses are conducted on myelin genes and proteins. This analysis plays an important role in monitoring the increase in the product generated during the reaction in “real time.” The quantitative approach is based on the time or session in which amplification is first detected, instead of a quantitative measurement of PCR products.

Microscopic examinations will help us to investigate the effects of Bryostatin-1. Elisa kits are used to determine the reduction of inflammation. Elisa kits play an important role in particle detection or conducting ligand-linked assays. Many ELISA types use the detection of enzymes of radish peroxidase (HRP) or alkali phosphatase (AP), along with chromogenic, fluorescent, or chemiluminescent substrates.

Real-time PCR and Western blot analyses are used to evaluate the effects of Bryostatin on OPC differentiation. OPC is a subtype of glial cells in the central nervous system. Introduction to oligodendrocytes reveals that they may also be able to differentiate into neuronal astrocytes. Various oligodendrocytes support and provide electrically isolated sheaths in the form of a myelin sheath, allowing for potential proliferation of work faster and high-precision transmission without the need to increase the axial diameter. The loss of OPCs, and differentiated oligodendrocytes, is associated with impairment of neurological functions.


Bryostatin-1 administration resulted in improvement of cognition, synaptogenesis, and neurogenesis, and decline of necrosis and astrogliosis in infarcted tissue. Moreover, it has direct antitumor effects in the treatment of lymphoma, sarcoma, leukemia, melanoma, and is used for breast, colon, lung, and ovarian cancers.

Explanation: Cognition is defined as the ability of an individual to perform the various mental activities most closely associated with learning and problem solving. Synaptogenesis is the formation of synapses between neurons in the nervous system. Neurogenesis is the process by which nervous system cells are produced by neural stem cells. Astrogliosis is an abnormal increase in the number of astrocytes due to the destruction of nearby neurons from CNS trauma, infection, ischemia, stroke, autoimmune responses, and neurodegenerative disease.

It is well-tolerated and its adverse reactions are rare, mild, and reversible. Explanation: Tolerability refers to the degree to which overt adverse effects of a drug can be tolerated by a patient. Usually, it is measured by the rate of “dropouts”, or patients that forfeit participation in a study due to extreme adverse effects.

Explanation: The blood-brain barrier is a semipermeable membrane separating the blood from the cerebrospinal fluid, and constituting a barrier to the passage of cells, particles, and large molecules. When the drug crosses the BBB, it means it has an effect on brain cells.


Bryostatin-1, with its anti-inflammatory, antioxidant properties, MMPs inhibitors, and activation of neuronal composition, has additional features and advantages. These include the ability to cross BBB and oral availability, which may make it appropriate for treatment for MS. To explain further: anti-inflammatory is the property of a substance that reduces inflammation or edema, which are induced by certain chemical mediators of inflammation like bradykinin, serotonin, and prostaglandins.

When we say ‘antioxidants,’ we mean it has the power to neutralize the ROS in a process called radical scavenging, and then carry them away. Antioxidants exist in both enzymatic and non-enzymatic forms in the intracellular and extracellular environment, enhancing cellular defenses. A Matrix Metalloproteinase Inhibitor (MMPI) inhibits matrix metalloproteinases. They reduce cell migration and have anti-angiogenic effects. They may be both endogenous and exogenous. The most notorious endogenous metalloproteinase inhibitors are Tissue Inhibitors of Metalloproteinases (TIMPs).

Future Perspectives

It is necessary to focus on the molecular events occurring after treatment of the cells with Bryostatin-1 in future studies. Explanation: A molecular event is the initial interaction between a molecule and a biomolecule or biosystem that can be causally linked to an outcome via a pathway. MIEs use knowledge of the chemistry of these processes to aid AOP research and toxicity risk assessment.

Summary of the Paper

Multiple sclerosis is characterized by inflammation, high concentration of MMPs, myelin damage, and BBB damage. Bryostatin-1 has antiinflammatory effects, decreases MMPs levels, and BBB damage, along with antioxidant properties and a neuroprotective effect. Based on these properties, this paper hypothesizes that Bryostatin-1 is an effective treatment in MS. Explanation: Multiple sclerosis (MS) is a disease that can potentially disrupt the brain and spinal cord (central nervous system).

In MS, the immune system attacks the protective sheath (myelin) that covers nerve fibers, causing problems in communication between your brain and the rest of your body. In the end, the disease can cause permanent damage or deterioration of the nerves. Signs and symptoms of MS vary widely and depend on the amount of nerve damage and the nerves affected. Some people with severe MS may lose their ability to walk independently or at all, while others may experience long periods of remission without any new symptoms. Bryostatins are potent modulators of protein kinase C. They have been studied in clinical trials as anti-cancer agents, anti-AIDS/HIV agents, and in people with Alzheimer’s disease.

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Frontiers in Pharmacology. (2019, Jul 25). Retrieved from