Combined Treatment With the Mood Stabilizers Essay

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JC5 Research Articl2: Draft

Marzia Bamiani

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Huntington's Disease

Combined treatment with the mood stabilizers lithium and valproate produces multiple beneficial effects in transgenic mouse models of Huntington's disease.

Brief Summary In Your Own Words: Goal, Experimental Design, Results, Conclusion

This research analyzes impacts of mixed-drug therapy on BDNF (Brain Derived Neurotrophic Factor) protein levels among transgenic and wild mice indicated under Huntington's Disease (HD's) N171-82Q mouse mutation. The mice's diet was chow, lithium and chow, valproate and chow, or a valproate-lithium mixture and chow. These test mice were surrendered following 14, 28, and 56 days of treatment in order to match BDNF protein levels in the cortex between different treatment time intervals. Brain cortex samples were acquired for assessment through the Western Blotting method of analysis. Investigation-specified mutual treatment using valproate and lithium proved most effective in cumulative BDNF protein stages during every treatment period. (1)

INTRODUCTION and BACKGROUND:

What Is Being Studied? Topic, Problem, Previous Studies, Gap in Knowledge

Why Are These Experiments Important? What is the scope of problem? How will these studies help?

What Must We Know To Understand the Experiments? Anatomy/Physiology, Subjects, Treatments, Outcomes

Huntington's Disease (HD) is a hereditary, dominantly autosomal, usually fatal neurodegenerative affliction, which usually presents later in life (ranging from mid-30s to mid-40s). The ailment is triggered by a Chromosome 4 gene malfunctioning; this gene is important as it generates the Huntington protein. Individuals afflicted with HD (because of the defective protein), exhibit a broad variety of symptoms, which may include impaired movement, psychiatric disorders, functional ability loss, and impairment of cognitive functioning. The symptoms surface due to degeneration of neuronal cell linked to the defective Huntington protein. The function of BDNF proteins is maintaining neuronal cell growth, survival and increase. In people suffering from HD, the neuronal cells gradually reduce HD symptoms and signs. Individuals with Huntington's disease depict lower BDNF protein levels. A combined treatment strategy using valproate and lithium has demonstrated a growth in HD patients' BDNF proteins, and it is thought to diminish the implication caused by it and inhibit the progress of HD and its symptoms. The monovalent cation -- Lithium -- has been typically used as a pharmacological agent for treating bipolar disorder for over half a period. It is the first treatment course alongside acute mania, whilst prophylactically being employed for persistent depressive and manic periods. Valproate, which functions as anticonvulsant, also works well in bipolar disorder treatment. The specific way these medications work is yet to be properly understood. Still, considerable attention has been given to these mood stabilizers' power in protecting against different abuses. As a validation, VPA and lithium may be used for co-treatment to generate largely reliable behavioral advantages in both of the HD models as well as, notably, prolonged N171-82Q mice survival time. Additionally, HDAC and GSK-3b hyperactivity was suppressed constantly using this combined treatment program, also linked to up-regulation of a couple of key neuronal protection and growth proteins - heat shock protein and the BDNF, or neurotrophic factor derived from the brain. Valproate and lithium have already been approved by the Food and Drug Administration and can be used by doctors for the treatment of this disease, even though, this is not a curable disease; the doctors frequently use these drugs to stall the progress of this fatal disease. The Gap in knowledge is that the cause of this disease is unknown and the cure is not yet discovered. Hence, scientists and health professionals are working and investigating by the use of animal models to find out the best treatment and absolute cure for Huntington's Disease. (1) (2)

EXPERIMENTAL DESIGN and DATA ANALYSIS:

What Was Done? Procedures, Treatments, Sample Collection and Method. Include Relevant Times.

How Was Data Analyzed? Statistical Tests, Descriptive Statistics, N, Significance Level

In this experiment, Transgenic mice model was used to express HD N171-82Q mouse model mutation for analyzing the impacts of mood-stabilizing medications valproate and lithium upon BDNF protein levels within brain cortex. The test mice's diet was one of the following combinations: lithium/lithium-carbonate and control chow, sodium valproate and chow, or a mixture of chow, sodium valproate and lithium carbonate. These test mice were surrendered following 14, 28, and 56 days of treatment, and this was followed by dissection of their cerebral cortex for examination (western blot analysis). Homogenization of cortex samples was conducted in T-PER protein (tissue protein) obtained from solution. A ten-minute centrifugation process was performed, following which the supernatants' western blotting commenced.

Loading control used was beta-actin, and protein level detection was performed by employing LI-COR (Odyssey infrared imaging system).

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Outcomes were standardized to the beta-actin. Cortex was examined for ascertaining if mice treatment using valproate, lithium, or both in combination, increased BDNF protein levels in the animals' brains, as well as for quantifying the drug treatment that effected greatest BNDF protein increase.

Statistical Analysis: All of the statistical analyses for this research were carried out by employing Graph Pad Prism (Graph Pad, San Diego, CA). The data of female and male mice was combined, as gender did not prove to have any significant impact on their behavioral tests. This data is expressed in the mean ± SEM (standard error of mean). Data analysis is carried out through one-way ANOVA (analysis of variance) for one outcome. Where needed, assessment of treatment or genotype comparisons at distinct time points was done using post-hoc Student t-test. When assessment of transgenic HD and wildtype mice phenotypes was done across ages, the research used two-way ANOVA (genotype age), after which data was treated using Bonferroni correction. Kaplan -- Meier analysis was utilized for assessing drug treatment's significant impacts on survival. (1)

METHOD:

What Must We Know To Understand ONE Method: How does it work? Required Equipment, Reagents, Description.

Western Blot analysis is an evaluation technique employed for identifying and quantifying protein presence in any particular sample for the specific experiment. In this experiment, Western Blot has been utilized for quantifying BDNF protein in target mice's brain cortex samples. Firstly, all proteins are detached via lysing from cortex tissues. They are subsequently separated through electrophoresis, a process by which proteins get organized on the basis of size. The first step in this process protein treatment with detergent for initiating denaturation, their unfolding, and creation of electrical charge. In the succeeding step, the proteins will be processed with electric current and injected into the gel "Nupage Bis-Tris." This enables proteins movement through the Electrophoreses gel. Proteins that are smaller in size move more rapidly through this gel, and hence, sample proteins effectively separate depending on size. Results emerge in ladder formation, and are subsequently transported to nitrocellulose membrane, which is then subject to secondary anti-rabbit or goat anti-mouse antibodies containing fluorescent molecules, whose function is BDNF protein binding. Lastly, researchers subject this nitrocellulose membrane to a photographic film, and at this stage, light is emitted by the antibody. The light takes the form of a dark area of rectangular shape on the film. Western blot interpretation is done on the basis of visible bands (lighter-darker) produced. Darker areas imply higher protein concentration. Western Blotting is one of the best methods that researchers and scientist mostly use, because it gives an accurate result. Each of the bands shows a very clear and dark enough shade under it to figure out the main result, They give the best result while doing DNA or any other kinds of experiment in the Laboratory. (3) (4)

RESULTS:

How Can You Best Sum Up the Results? Write a Short, New Title

What Was Done? Purpose of Experiment, Subjects, N, Outcome with Units

What Does Data Analysis Show? Significant Difference(s) and p-value(s), Relevant Comparisons that are the Same.

What Do Results Mean? How does outcome relate to the overall goal of the study?

After two weeks, though level of BDNF proteins in the brain cortex was found to increase above control levels following monotherapy treatment using valproate and lithium, the increase wasn't significant. Combined valproate and lithium treatment brought about a more significant rise in level of cortex BDNF proteins compared to treatment using only valproate or lithium. After four weeks, though level of BDNF proteins in the brain cortex was found to increase above control levels following monotherapy treatment using lithium, the change wasn't significant. Individual valproate treatment and combined treatment using valproate and lithium brought about a significant rise in levels of BDNF proteins compared to control.

After eight weeks, though level of BDNF proteins in the brain cortex was found to increase above control levels following monotherapy treatment using lithium, the change wasn't significant. Individual valproate treatment and combined treatment using valproate and lithium brought about a significant rise in levels of BDNF proteins compared to control. (1)

Valproate-Lithium Co-Treatment is successful in alleviating natural locomotor disorientation of HD mice. HD motor symptoms include weakened coordination of both involuntary and voluntary movements. For identifying drug treatment impacts on muscle movement and behavioral changes, YAC128 and N171-82Q mice were tested longitudinally in their motor tests,….....

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