By: Stephen Pfau Pittsburgh Central Catholic High School Grade 9

By: Stephen Pfau Pittsburgh Central Catholic High School Grade 9

Everyday thousands of people take cold/flu medicine and pain relief medicine Two common medicines for cold/flu and pain are DayQuil (cold) and Ibuprofen (pain relief). Some of these people take both of these medicines at the same time

The brand used in this experiment was Top Care Ibuprofen (liquid form) NSAID used to treat arthritis, oral pain, and fever reduction Inhibits cyclooxygenase- enzyme in prostaglandin synthesis; inhibits inflammation, pain, and fever. Non selective of the isoforms of cyclooxygenase it inhibits. Side effects of this drug include: headache, nervousness; chest pain, weakness of heart; nausea; fever; bruising, muscle weakness; The recommended dose for an adult is 30 ml (600 mg ibuprofen)

Produced by Vicks and intended to relieve symptoms of the common cold and flu Active ingredients are: acetaminophen (pain reliever/fever reducer), phenylephrine (decongestant), and dextromethorphan (cough suppressant) Used in liquid form The recommended dose for an adult is 30mL: [650mg-actetaminophern, 20mg- dextromethorphan, 10mgphenylephrine]every four hours

Unicellular fungal eukaryote that has been used in food production (baking, fermentation) The most studied eukaryotic cell Has many characteristics that are similar to those of human cells like nutrition and cell growth Also used due to its characteristic rapid growth and ease of experimental manipulation

To determine the individual effects of Ibuprofen and DayQuil on yeast cell survivorship To determine the synergistic effects of Ibuprofen and DayQuil on yeast cell survivorship

Ibuprofen and DayQuil will not have a significant effect on yeast cell survivorship, either individually and synergisticly 1. Ibuprofen and DayQuil will have each have negative effects on yeast survivorship 2. synergistic effects of the drugs will reduce yeast survivorship. Null Hypothesis Alternative Hypothesis

Yeast culture 45 YEPD agar plates (1% yeast extract, 2% glucose, 2% peptone) Top Care Ibuprofen Vicks DayQuil Cold and Flu SDF (sterile dilution fluid) (10 mm KH2PO4, 10 mm K2HPO4, 1 mm MgSO4, 0.1 mm CaCl2, 100 mm NaCl) Macro pipette and sterile tips Micropipette and sterile tips Sterile test tubes Sharpie Ethanol Burner Spreader bar Incubator Test tube racks Vortex Klett Spectrophotometer Sidearm flask

1. Saccharomyces cerevisiae was grown overnight in sterile YEPD media. 2. A sample of the overnight culture was added to fresh media in a sterile sidearm flask. 3. The culture was incubated at 30 C until a density of 50-60 Klett spectrophotometer units was reached. (Cell density approximately 10 7 cells/ml). 4. The culture was diluted in a sterile dilution fluid to a concentration of approximately 10 5 cells/ml. 5. Variables and yeast were added to SDF to generate the desired exposures:

SDF (ml) Ibu (ml) DQ (ml) Yeast (ml) 9.9 9.85 9.4 9.85 9.4 9.8 8.9 9.35 9.35 0 0 0 0.05 0.5 0.05 0.5 0.05 0.5 0 0.05 0.5 0 0 0.05 0.5 0.5 0.05 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 resulting Concentration (0) control 0.5Xdq 5Xdq 0.5X ibu 5Xibu 0.5X syn 5X Syn 0.5Xib u, 5Xdq 5xibu, 0.5Xdq The total volume of all tubes was 10mL A 100x stock each of drug was created. Where x equals the calculated concentration in the blood

6. 0.1 ml. of cell culture was then added to the test tubes, yielding a final volume of 10 ml. and a cell density of approximately 10 3 cells/ml. 7. The solution was mixed by vortexing and allowed to sit at room temperature for 15 minutes. 8. After vortexing to evenly suspend cells, 0.1 ml aliquots were removed from the tubes and spread on YEPD agar plates. 9. The plates were incubated at 30 C for 48 hours. 10. The resulting colonies were counted. Each colony is assumed to have risen from one cell.

Number of colonies 180 Average number of colonies 160 140 120 100 80 60 40 20 0 control 0.5xibu 5xibu 0.5xdq 5xdq 0.5xibu 5xibu 5xsyn 0.5xsyn 5xdq 0.5xdq [Drug]

number of colonies # of colonies Control vs. Ibuprofen 200 p value= 0.155 150 100 50 0 control 0.5xibu 5xibu [Drug] Control vs. DayQuil 160 155 150 145 140 135 130 125 120 p value = 0.088 control 0.5xdq 5xdq [Drug]

number of colonies 180 Control vs. Synergistic 160 140 p values = 0.055, 0.007 120 100 80 60 40 20 0 control 0.5xibu 5xdq 5xibu 0.5xdq 5xsyn 0.5xsyn concentrations

2 factor ANOVA Source of Variation SS df MS F P-value F crit Sample 3669.556 2 1834.778 7.050059 0.00344 3.354131 Columns 4571.056 2 2285.528 8.782047 0.001154 3.354131 Interaction 2144.278 4 536.0694 2.059825 0.114094 2.727765 Within 7026.75 27 260.25 Total 17411.64 35

The null hypothesis was supported for all concentrations of the drugs There did not appear to be in any reduction in the survivorship of the yeast There did not appear to be any synergistic effects of the drugs

Limitations: The timing of the spreading for the plates varied slightly Only two concentrations Multiple inactive ingredients Only one exposure time Only one type of exposure (liquid pulse) Extensions: Test on other models such as E. coli or other micro flora Use more concentrations Test with the tablet form of each medicine Agar infusion

http://www.vicks.com/products/dayquil/cold-fluliquid-medicine/ http://en.wikipedia.org/wiki/ibuprofen http://en.wikipedia.org/wiki/yeast http://en.wikipedia.org/wiki/dextromethorphan