SYNTHROID MEDICATION WITH CALCIUM ON HEART

AND BONEBy: Zaib Momin

Purpose

• Synthroid is used by many children and also adults whose thyroid gland isn’t supplying enough hormones to keep overall metabolism in working order. • The negative impacts of synthroid medication are bone loss, increase in heartbeat, and possibly heart failure.• We know that calcium can improve bone strength and assist with other necessary functions. Can calcium help reduce/balance the negative effect of synthroid medication?

Hypothesis

• The research hypothesis states that if the high concentration of calcium solution was mixed with synthroid medication, then the heart cell and bone cell viability will be higher than the control or the group with just synthroid. • If high concentration of synthroid medication was exposed to daphnia, then it would result in higher heart beats per minute. • The null hypothesis states that combining synthroid medication and calcium will have no effect on cells or daphnia.

Experimental Design Diagram

Material

• Pipette tips• Pipettes• Calcium Chloride• PBS solution

(Phosphate Buffered Saline)

• Ethanol 70%• Typan blue• Trypsin• Well plates• Depression slide

• Cell culturing media• Microfuge tubes• Synthroid medication •  Rat Heart Cells (ATCC#

CRL-1446)• Mouse Bone Cells (CRL-

12424)• Daphnia Magna• Container with spring water for daphnia replacement

• Depression slide

ProcedureTreatments were prepared to use on heart cells, bone cells, and daphnia.

Day 1: For cell line

Add 400mcL of growth media and 100mcL of cell solution to each cell plate

Incubate for 24 hours

Day 2: For cell line

Add 55mcL of treatment to appropriate wells

Procedure

Day 3: For cell line

Remove old media Add trypsin and

growth mediaCount via hemacytometer

Daphnia Procedure

Data Analysis

Heart CellMean Cell Count for HeartSample Cell

Count (cells/uL)

Trials StandardDev.

Control 236 4 29Low calcium (0.00819 mg/mL)

176 4 150

Medium calcium (0.0819 mg/mL)

294 4 51

High calcium (0.819 mg/mL)

248 4 66

Synthroid only (0.666 mg/mL)

183 4 67

Synthroid with low calcium

176 4 53

Synthroid with medium calcium

150 4 60

Synthroid with high calcium

491 4 381

Heart Cell

Heart Cell Trend

ANOVA analysis showed (f=2.10, df=7, p=0.084); therefore there was no significant difference among the mean, as p=0.084>0.01.

The presence of calcium at three concentrations showed about the some viability as the control cells.

Synthroid alone, with low calcium and medium calcium were slightly lower than the control.

Highest levels of calcium appeared to modify the negative effect.

Large standard deviation value indicate a large variation among data within several treatments.

Bone Cell

Mean Cell Count for Bone Cells

Sample Cell Count (cells/uL)

StandardDev.

Control 481 78

Low calcium (0.00819 mg/mL)

514 73

Medium calcium (0.0819 mg/mL)

576 91

High calcium (0.819 mg/mL) 775 285

Synthroid only (0.666 mg/mL)

576 248

Synthroid with low calcium 314 108

Synthroid with medium calcium

490 57

Synthroid with high calcium 498 85

Bone Cell

Trend for Bone Cells

ANOVA analysis showed no significant differences among the mean. ( P = 0.025. DF = 7. F value = 2.88)

High variation for several treatments could be a contributory factor.

Overall calcium levels increased cell viability when applied individually.

Synthroid medication caused slightly increased cell counts.

As calcium was added to synthroid treated cells there was a gradual increase in viability but none were higher than the synthroid only cell counts.

Daphnia Mean Heart Beat by Daphnia per minuteSample Heart Beat per

minuteTrials Standard

Dev.

Control 148 4 32Low calcium (0.00819 mg/mL) 225 4 9

Medium calcium (0.0819 mg/mL) 221 4 47

High calcium (0.819 mg/mL) 245 4 11

Synthroid only (0.666 mg/mL) 228 4 41

Synthroid with low calcium 226 4 26

Synthroid with medium calcium 253 4 12

Synthroid with high calcium 262 4 18

Daphnia

Daphnia Trend

ANOVA statistics showed that there was a significant difference among the mean counts for heart beats per minute for Daphnia.

F=6.25, DF=7, P=0.000. All treatments, calcium/synthroid

treatment combination showed an increase in cell counts (approximately 50% higher than the control).

Acknowledgment

I would like to acknowledge Ms. Amanda Baskett and Samah Khan for helping me come up with the topic.

I would also like to acknowledge Ms. Mary Mansell for helping me conduct the experiment and helping me finish and improve my research paper.

I would also like to acknowledge my parents for supporting me throughout the project.

I'd like to thank the Rockdale Magnet Fund and the Magnet School for providing me with the proper instruments and money to conduct the experiment.

Any Questions?

Synthroid Medication with Calcium on the Heart and Bone

By: Zaib Momin

My Achievement

C. J. Hicks Community Service

I have helped first graders in Ms. Barron’s class in C.J.Hicks during 2nd period.

I have learned how to be patient with students when teaching a new concept to them.

Driver’s License

I got my driver’s license this summer and my parents got me a car.