Stephanie Saade, J.L. Ilsley High School

Photograph Of Human Fibroblast Cells Taken During My Experiment.





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Introduction

Turner Syndrome is a chromosomal disorder that effects approximately 1 in every 2,500 female births. Dr. Henry Turner discovered Turner Syndrome in 1938 after studying a pattern of short stature and other characteristics in what otherwise appeared to be healthy females. A syndrome is a set of features in relation to each other which stem from a root cause. Although Dr. Turner identified the syndrome, the root cause was not discovered until 1959 by Dr. C. E. Ford. There are three types of Turner Syndrome which all have to do with the total or partial loss of an X chromosome in some or all of the cells in a female’s body. This “loss” of an X chromosome occurs during cell division. Since males cannot survive without all of their x and y chromosomes present this syndrome affects only females. There are many characteristics of this syndrome which lead to a diagnosis, one of the most prominent being short stature. If Turner Syndrome were suspected in a patient, the diagnosis would be made with a test called a karyotype where a scientist would separate the chromosomes in the white blood cells.

Turner Syndrome is a condition that I have personally dealt with since I was diagnosed at 7 years of age. It is a condition a female lives with all her life. My diagnosis followed many trips to the hospital and visits with doctors. As a result of this diagnosis and the many medical tests that followed, I am now very familiar with the numerous medical conditions that could potentially exist with Turner Syndrome.

In this project I will focus on the slow growth and short stature of girls with Turner Syndrome. The average adult height of a female with Turner Syndrome is 1.43m (4 feet,8 inches). This is currently treated with injections of Human Growth Hormone (HGH). However, many families are unsure of this treatment and are weary of the potential side effects. The possible side effects include: increased weight and muscle mass, diabetes, intercranial hypertension (increase of fluid surrounding the brain), slipped femoral emphasis (increased risked of dislocating hips) and scoliosis. Parents are apprehensive to put their daughter through injections 6 nights a week for a prolonged period of time that could last 8 or 9 years. Also, the medication is very expensive and often not covered by insurance. The benefit of using HGH is that the average adult height of a patient, based on ongoing studies in several countries, will increase from 5 to 17cm (2-7 inches). Many families wish there could be an easier and more natural way to help increase the height of a female with Turner Syndrome. This is what I have experimented on in this project.

For further information on Turner Syndrome, please click here for my literature review.

For further information on Growth Hormone, please click here for my literature review.

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Objective

The objective of this project is to discover the impact of fruits and vegetables on cell growth to determine if they could be used in the future as an alternative to Human Growth Hormone (HGH) injections currently used by female patients with Turner Syndrome to increase their ultimate height.

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Apparatus

  1. Test Tubes
  2. Electronic Scale
  3. Pasture Pipettes
  4. Blender
  5. Sterile O.22 micron filters
  6. Microscope
  7. Human Fibroblast cells
  8. 4x4 well LabTek slides
  9. Cell medium (DMEM + 10% FBC)
  10. Digital Camera
  11. Organic apples, organic potatoes and organic blueberries

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Methods

Experiment #1

Organic Red Delicious apples, organic blueberries and organic potatoes were gathered to be used. The apples and potatoes were then peeled. Making a ratio of grams to distilled water created two different concentrations of all three types of produce used. 10g of the flesh of Red Delicious apple was weighed and put into a blender with 20ml of distilled water to create a 50% solution of Red Delicious apple which was put in a test tube. 12g of Red Delicious apple was then weighed and blended with 4ml of distilled water to create a 300 % solution which was put in a test tube as well. The blender was then rinsed thoroughly with distilled water. Next 10g of potatoes were weighed and blended with 20ml of distilled water to create a 50 % solution of potato and was poured into a test tube. Another 10g of potato was then weighed and blended with 5ml to create a 400% solution of potato and was put in a test tube. The blender was again thoroughly rinsed. Then, two 10g samples of blueberries were weighed and one 10g sample was added to 20ml of distilled water and another 10g sample was added to 5ml of distilled water which created a 50% solution and 200% solution of blueberries respectively. 50 microlitres (mL) of these 6 solutions of extracts were added to 6 different cell cultures of human fibroblast cells along with 50mL of medium and 50mL of Fetal Bovine Serum (FBS) after they were put through a .22 sterile micron filter. Another sample had 50mL of distilled water, 50mL of medium and 50mL of FBS. The control was fibroblast cell in 50mL of medium and 50mL of FBS. Each cell culture started with 2000 human fibroblast cells. Pictures were taken a week later. See pictures of cells for observations that were made.

Experiment #2

Organic Red Delicious, Organic Braeburn, and Organic Fuji apples were gathered and then peeled and the flesh of the apples were used. 15g of Braeburn apple were weighed. 15ml of distilled water was blended with this sample to create 100% a solution of Braeburn apple and was put in a test tube. The blender was then rinsed in distilled water. 15g of Fuji apple were weighed and blended with 15ml of distilled to create a 100% solution of Fuji apple and was put in a test tube. The blender was again rinsed in distilled water. 15g of Red Delicious apple was weighed and another 15ml of distilled water were blended together to create a 100% solution which was put in a test tube. These three samples were put through a .22 micron filter at the end of a syringe and 50mL of each were added to three cultures of 3000 human fibroblast cells with 50mL of medium and 50mL of FBS. These three samples were then added to 3000 fibroblast cells with 50mL of medium but no FBS. Also 25mL of all three samples were added to the cells with 50mL of medium and 50mL of FBS. Another 25mL of all three extracts was added to the cells with 50mL of medium and no FBS. A week later pictures of these samples were taken. See pictures for observations in this set of experiments.

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Observations

Key- Experiment 1

A-50mL of medium, 50mL of FBS and 50mL of Red Delicious apple extract at 50 % concentration added to fibroblast cells.

B-50mL of medium, 50mL of FBS and 50mL of Red Delicious extract at 300% concentration added to fibroblast cells.

C-50mL of medium, 50mL of FBS and 50mL of potato extract at 50 % concentration added to fibroblast cells.

D-50mL of medium, 50mL of FBS and 50mL of Potato extract at 200% concentration added to fibroblast cells.

E-50mL of medium, 50mL of FBS and 50mL of Blueberry extract at 50% concentration added to fibroblast cells.

F-50mL of medium, 50mL of FBS and 50mL of Blueberry extract at 200% concentration added to fibroblast cells.

G-50mL of medium and 50mL of FBS added to fibroblast cells.

H-50mL of medium, 50mL of FBS and 30mL of distilled water added to fibroblast cells.

 

Key- Experiment 2

A-50mL of medium

B-50mL of medium and 25 mL of Red Delicious extract added

C-50mL of medium and 50 mL of Red Delicious extract added

D-50mL of medium and 50 mL of Braeburn extract added

E-50mL of medium and 50mL of FBS added

F-50mL medium, 50mL of FBS and 25mL of Red Delicious apple extract added

G-50mL medium, 50mL of FBS and 50mL of Red Delicious apple extract added

H-50mL medium, 50mL of FBS and 50mL of Braeburn apple extract added

I-50mL medium and 25mL of Fuji extract added

J-50mL medium and 50mL of Fuji extract added

K-50mL medium and 25mL of Golden Delicious extract added

L-50mL medium and 50mL of Golden Delicious extract

M-50mL medium, 50mL of FBS and 25mL of Fuji apple extract added

N-50mL medium, 50mL of FBS and 50mL of Fuji apple extract added

O-50mL medium, 50mL of FBS and 25mL of Golden Delicious apple extract added

P-50mL medium, 50mL of FBS and 50mL of Golden Delicious apple extract added

CLICK HERE FOR PHOTOGRAPHS OF MY RESULTS

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Analysis

Experiment #1

This experiment was analyzed based on the density of the surviving fibroblast cells in the cell cultures. After all samples were carefully observed under a microscope, both of the Red Delicious Apple solutions had the most positive impact on the cells compared to the control which was human fibroblast cells with 50 microlitres of medium and 50 microlitres of FBS.

Experiment #2

This experiment was analyzed in much the same way as Experiment 1. All of the cell cultures grew very well and were very healthy except for the cultures with Fuji apples. The Golden Delicious apples and Braeburn had the best impact on the health and growth of the cells and should be further tested for long term effects and results.

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Applications

This project has shown great promise and the results can be used in many applications. One application is that, by just doing this project, that there will be an increased awareness of Turner Syndrome. I also see this project as being a catalyst for further experimentation and research into Turner Syndrome. I hope this project will increase awareness of HGH therapy and help Turner Syndrome patients and their families get some solid, clear facts about HGH. This project could lead to finding a much safer and natural alternative to HGH therapy to increase the height of these patients. This could either partially or totally eliminate the need to take HGH injections and as a result, lessen the possibility of side effects of HGH therapy. This diet of fruits and vegetables could also be used with patients who have a HGH deficiency.

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Conclusion

In conclusion, there were many things learned through performing these experiments. One interesting point that I noted was that human fibroblast cells could tolerate 10% water and could survive several days with no serum (FBS). The apples seemed to have an amazing effect on the morphology of the cells during the second experiment. The cells became much more elongated and were structured together in a spiral-like pattern. The cells were larger and stronger with the apples and therefore they had a similar effect on cells as HGH which is meant to stimulate the growth of cells. This means that, if further research is done and the results continue to be favorable, apples could be the missing piece of the puzzle for growth in Turner Syndrome patients and apples, in some form, could possibly be substituted for HGH therapy.

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Acknowledgements

I would like to thank the following people for their help and support with this project:

  • Mr. Greg DeLaLis, Science Teacher at J. L. Ilsley High School
  • Dr. David Byers and Robert Zwicker, Researchers at the Atlantic Research Council
  • Dr. Elizabeth Cummings, Endocrinologist at the IWK Health Centre
  • Ronelda Gillis, RN, Nurse in the Endocrine Clinic of the IWK Health Centre
  • Paula Canning BSc, PDt, CDE, Diabetes/Endocrinology Dietitian at the IWK Health Centre

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