write me an abstract on The Effect of Different Temperatures with Vitamin A on Planaria Regeneration using these instructions: An abstract gives the essence of the project in a brief but complete form. Details and discussions should not be included in the abstract, but should be put in the longer, written research paper. An abstract includes 4 key pieces of information:

1. Purpose of the Experiment/ Hypothesis/ Rationale

• An introductory statement of the reason for investigating the topic of the project, the purpose of the study and the hypothesis being studied.

2. Procedures used

• A summarization of the key points and an overview of how the investigation was conducted. The control is identified and if there is no control, an explanation for why there is not one.

3. Observations/Data/Results

• This section should provide key results that lead directly to the conclusions you have drawn. It should not give too many details about the results nor include tables or graphs. The results of the inferential stats test should be included.

4. Conclusions

• Conclusions from the investigation should be described briefly. The summary paragraph should reflect on the reason for the results. If room allows, possibly state some applications and extensions of the investigation.

Item to remember when writing an abstract:

1. An abstract does not include a bibliography and should NOT have any citations.

2. It should not exceed 250 words.

Example of an abstract: (this is an example and can use improvement):

The purpose of this experiment was to find the effects of different fertilizers on pea plant growth. Recently, fertilizer A and fertilizer B have been used in the farming industry to increase the yield of wheat crops and these two fertilizers may have similar affects on pea plants. Pea plants were treated with either 3 g of fertilizer A or 3 g of fertilizer B. The plants were allowed to grow for 20 days and then their height was measured. The control that was used in the experiment was no fertilizer. It was hypothesized that pea plants treated with fertilizer B would produce more growth than fertilizer A and no fertilizer. The results revealed that plants treated with fertilizer A grew, on average, 2 cm more than fertilizer B and 4 cm more than plants not treated with any fertilizer. A t-test was done on the data and it revealed that the data was significant for fertilizer B vs. the control but not significant for fertilizer A vs. the control. The results did not support the research hypothesis. It is believed that the results are due to the fact that fertilizer A contains more nitrogen and water than fertilizer B which enables the pea plants to grow more. This research could lead to further studies that investigate the amount of fertilizer A for optimum pea plant growth and how fertilizer A may affect other legume plants.

Example of an abstract: The Effects of Marine Engine Exhaust Water on Algae

This project in its present form is the result of bioassay experimentation on the effects of two-cycle marine engine exhaust water on certain green algae. The initial idea was to determine the toxicity of outboard engine lubricant. Some success with lubricants eventually led to the formulation of "synthetic" exhaust water which, in turn, led to the use of actual two-cycle engine exhaust water as the test substance.

Toxicity was determined by means of the standard bottle or "batch" bioassay technique. Scenedesmus quadricauda and Ankistrodesmus sp. were used as the test organisms. Toxicity was measured in terms of a decrease in the maximum standing crop. The effective concentration - 50% (EC 50) for Scenedesmus quadricauda was found to be 3.75% exhaust water; for Ankistrodesmus sp. 3.1% exhaust water using the bottle technique.

Anomalies in growth curves raised the suspicion that evaporation was affecting the results; therefore, a flow-through system was improvised utilizing the characteristics of a device called a Biomonitor. Use of the Biomonitor lessened the influence of evaporation, and the EC 50 was found to be 1.4% exhaust water using Ankistrodesmus sp. as the test organism. Mixed populations of various algae gave an EC 50 of 1.28% exhaust water.

The contributions of this project are twofold. First, the toxicity of two-cycle marine engine exhaust was found to be considerably greater than reported in the literature (1.4% vs. 4.2%). Secondly, the benefits of a flow-through bioassay technique utilizing the Biomonitor was demonstrated. Also use this introdcution: The process of regeneration in planarians attracts researchers due to planarians’ extraordinary ability to regrow lost parts under various conditions (Seebeck et al, 2017). This current study investigates how different temperatures with Vitamin A impact planarian regeneration. Including different temperature ranges and constant levels of Vitamin A in the experimental design is chosen to simulate various environmental conditions and constant nutrient supplementation scenarios (Deochand et al, 2018). Consequently, the research explores how these variables affect the self-renewal ability of pluripotent stem cells using neoblasts (Lai et al, 2017). Planaria serve as model systems for embryonic stem cells in humans, driving the choice of this study. It contains real-world implications and explores a topic that assists planaria to become better models for human stem cells (Ermakova et al, 2009).

The potential implications of this research promise broader societal implications (Deochand et al, 2018). The findings may reveal information applicable in regenerative medicine, tissue engineering, and developmental biology (Sarkar et al, 2019). Insights from this research may prove beneficial to human health, for instance, in providing crucial knowledge for the development of regenerative therapies & regenerative medicine. Furthermore, this research holds the potential to assist advances in regenerative medicine by uncovering the fundamental mechanisms/functions managing and aiding regeneration in planarians (Ermakova et al, 2009).

The independent variable in this study encompasses understanding different temperatures and varying Vitamin A concentrations (Sarkar et al, 2019). This includes the impact of different temperatures (3, 20, & 37 degrees Celsius) on planarian survival and how Vitamin A potentially influences cellular processes, primarily around tissue and stem cell regeneration (Sarkar et al, 2019). Vitamin A, an external factor added to the planaria, is usually already present in the human body due to a daily diet containing a certain amount of Vitamin A. Vitamin A improves and increases vision, growth, cell division, reproduction and immunity. Vitamin A is used for cell differentiation and embryonic development. It also decreases the risk of heart disease, cancer, and skin diseases (Marie et al, 2021). Adding Vitamin A to planaria assists the planaria to become better models for regeneration applications in humans and the real world.

Understanding planarian regeneration, tissue regeneration, and neurogenesis becomes crucial for the dependent variable (DV) (Sarkar et al, 2019). This knowledge aids in understanding the processes involved in planarian tissue repair and the role of neoblasts in

pluripotent stem cells in regeneration. Pluripotent stem cells means that the stem cell can divide into any cell type. Neoblasts, which are adult stem cells that can divide, have the role of dividing and transforming into any cell type such as red blood cells, white blood, cells, platelets, and more (Lai et al, 2017). There are neoblasts spread all across planaria’s bodies, in fact, they make up about 25%-30% of all the cells in planaria (Ge et al, 2022). In other words, neoblasts are a type of pluripotent stem cells. Planaria also show better regeneration than other animals that exhibit regeneration such as zebrafish, axolotls, and spiny mice because they contain plenty more neoblasts and pluripotent stem cells than other organisms (Deochand et al, 2018).

The purpose of this study is to understand the impact of different temperatures with Vitamin A on planarian regeneration (Ermakova et al, 2009). By observing the regenerative capacity of planarians under varying experimental conditions, the research seeks to conclude on the possible effects on tissue organization, neurogenesis, and stem cell populations (Seebeck et

al, 2017). It is hypothesized that the variations in temperature, combined with Vitamin A supplementation, will have different effects on planarian regeneration (Ermakova et al, 2009).

The temperature of 23 degrees Celsius with a 4% concentration of Vitamin A is thought to increase the regeneration of the planaria the greatest. This is because planaria cultures, which are ideal habitats for growing planaria, are maintained at optimal temperatures around 23 degrees Celsius or room temperature. Also, many studies observe and utilize room temperature, typically around 23 degrees Celsius as the optimum temperature for planaria growth (Sarkar et al, 2019). 

Specifically, conditions of 23 degrees Celsius supplemented with Vitamin A are anticipated to be ideal for planaria and to support pluripotent stem cells' self-renewal ability using neoblasts, potentially resulting in the most effective planarian regeneration. This hypothesis is backed by previous studies highlighting the crucial role of environmental factors such as high humidity, low light, and a 6.0 - 7.0 (slightly acidic) pH (Ge et al, 2022). The hypothesis is also supported by nutrient supplementation play, such as glycogen and dextrose, provided by hard-boiled egg yolk, pieces of earthworm, or beef liver (Abel et al, 2022). These factors influence the developmental and regenerative processes in various organisms, including planarians (Deochand et al, 2018). It is also supported by studies indicating that planaria grow best around room temperature in a dark environment (Sarkar et al, 2019).

The independent variable in this study comprises the different temperature conditions (3, 20, and 37 degrees Celsius), with constant Vitamin A concentrations (4%). These temperatures were chosen to test how planarian regeneration and cellular processes would increase or decrease due to drastic differences in temperature of environment of the planaria; with the addition Vitamin A. This concentration of Vitamin A was chosen because it is known to not be too toxic to the planaria, but it was not too little to not have any effect on the planaria. The control group is set at 23 degrees Celsius because it is room temperature, and the temperature of the environment isn't changed. The dependent variable is planarian regeneration, determined through the length of regeneration of planaria, measured in millimeters (mm). Different temperatures with a constant concentration of Vitamin A can also affect tissue organization, neurogenesis, and the self-renewal ability of pluripotent stem cells since the change of temperature is an environmental factor change and the addition of Vitamin A is a nutritional/supplemental factor change. (Lai et al, 2017).

The purpose of this experiment is to determine if different temperatures with an addition of Vitamin A have an effect on planaria regeneration. By exploring several temperatures, the study aims to provide a comprehensive understanding of how these factors affect planarian regeneration, subsequently offering valuable insights into possible mechanisms or processes involved (Deochand et al, 2018). This study involves measuring planarian regeneration in millimeters, as planarians are ideal sources to explore regeneration processes (Emakova et al, 2009). There will be 3 different levels of the independent variable; planaria grown in an environment at 3 degrees Celsius with an addition of Vitamin A, planaria grown in an environment at 23 degrees Celsius with an addition of Vitamin A, and planaria grown in an environment at 37 degrees Celsius with an addition of Vitamin A. The planaria grown at 23 degrees Celsius with an addition of Vitamin A will be the control group of this experiment. These temperatures were selected because they are very different temperatures and could have a drastic effect on planaria regeneration, the self-renewal of neoblasts, tissue organization, neurogenesis (the growth and development of nerve tissue), and other cellular processes (Karge et al, 2020). 3 degrees Celsius is very close to freezing temperature, 23 degrees Celsius is very close to room temperature, and 37 degrees Celsius is very close to body temperature for humans. The dependent variable of planaria regeneration was measured in millimeters (mm). It is thought that planaria grown in an environment consisting of the temperature at 23 degrees Celsius (around room temperature) will result in the greatest growth of planaria. This is based on previous studies using a temperature close to room temperature in their experiments; room temperature is considered to be the optimal temperature for planaria regeneration (Sarkar et al, 2019). Most experiments use room temperature with another variable. Other temperatures haven’t been experimented with nearly as much, and their effect is not known with the addition of different variables on planaria regeneration.

USE THIS PROCEDURE: Safety equipment like gloves, safety goggles, and a pipette were prepared and safety regulations for Vitamin A were read, such as using safety goggles and not ingesting Vitamin A. Also, safety and care rules for planarians were read, such as wearing gloves and pointing knife away when cutting planaria. Safety equipment was used and worn, safety regulations for Vitamin A and planarian care were followed, loose clothing was secured, and hands were sanitized. Brown planaria were used in the experiment. A 4% concentration of Vitamin A was used in the experiment. This Vitamin A concentration was chosen because each location will contain one pill grinded and evenly distributed into each container (snack cup with lid), which is not too little to not affect planarian regeneration or too much to kill the planarians. A concentration of 4% of Vitamin A was diluted and added by grinding Vitamin A pills and mixing it to 25 mL of pond water. Then, 24 mL of pond water was put into each container, and 1 mL of diluted Vitamin A with pond water was put in each container. Then, the planaria were split into three equal groups of 25, with each group getting its own clean container. Each container was put in a temperature (23, 37, or 3 degrees Celsius) using an incubator, a fridge, or just placing the planaria containers in a garage. Carefully, 25 planarians were cut in half using an “X-Acto” knife. The bottom halves of the planaria were kept in a separate container; the top halves were used to measure regeneration. Carefully, 25 head halves of the planaria were each put in their respective containers. The initial length of the planaria, measured in millimeters, and when the experiment started were recorded. Then, the planaria were put in their groups of 3, 23, and 37 degrees Celsius. The temperature was maintained and kept steady for 14 days. The planaria were fed with pieces of earthworm every 7 days. The planaria were left for a month to regenerate in their own containers.

After 14 days, one planarian was slowly taken out of its container. The size of the planaria after their regeneration period was measured, in millimeters, with an electronic caliper. The process for measuring the planarians was duplicated for each planarian in each container and group. This procedure was rerun for 75 trials in total. Then, the data for the measurements of the planarians, the conditions the planarians were put in, and the finished date for the experiment was recorded. After the experiment was finished and the data was documented, the planaria were frozen for 48 hours and disposed of in a designated waste disposal plant. The Vitamin A was also disposed of in a waste disposal plant. USE THIS RESULT SECTION: This experiment explored the effect of different temperatures, combined with a constant concentration of vitamin A (4%), on planaria regeneration. The results of the statistical analysis were presented in table 2. The research hypothesis proposed that planaria placed in 23°C would regenerate the most. Quantitative data was collected, measuring the regeneration in millimeters at 3°C, 23°C (control), and 37°C. A statistical analysis of the average for each group revealed that the mean regeneration was higher at the control temperature of 23°C (3.3 mm) than at 3°C (1.0 mm) or 37°C (0.4 mm). This demonstrated that planaria regeneration is influenced by both temperature and vitamin A. Based on the results, the research hypothesis was supported, and it was found that planaria given a constant concentration of Vitamin A would regenerate most at 23°C.

The variance and standard deviation were calculated for each of the independent variable levels. The data showed that the 23°C control condition demonstrated the highest variance and standard deviation, indicating the most variation in planaria regeneration among all the independent variable groups. The results contained one outlier, which was trial 21 of the 23°C independent variable group, as a 6.2 millimeter regeneration of the planarian, found in table 1. This measurement of the regeneration of the planarian was outside of the 2 SD range of the 23°C independent variable group, which was 0.514 to 6.086, found in table 2.

A t-test was performed for the data, using a significance level of 0.001 and 48 degrees of freedom. This data had a null hypothesis, which stated that the effect of different temperatures with vitamin A had no significant difference on planaria regeneration. The calculated t-values for the 3°C vs. control (t=7.913), 37°C vs. control (t=10.233), and 3°C vs. 37°C (t=6.149) comparisons were all greater than the critical t-value found in the t-table (3.551). Consequently, the null hypothesis was rejected for all independent variable comparisons, indicating that the temperature did influence planaria regeneration when combined with vitamin A. The resulting probability values of the results being based on chance (p<0.001) helped the rejection of the null hypothesis, suggesting a less than 0.001/1 in 1000 chance of these results occurring by chance. This implied that the results were most likely due to the independent variable. Overall, this research revealed that different temperatures with a constant concentration of Vitamin A having an effect on planaria regeneration is statistically significant, and that 23°C/room temperature (control) helps planaria grow the most with a constant concentration of Vitamin A.

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