STA 6167, Section 1648, Fall 2007 Project #2 Due Thursday 2/07/08

RAMI SHAMSHIRI
UFID#: 9021-3353

Part1
Response: Clearance of theophyline Factor A: popular heartburn medication: Placebo/Tagment/Pepcid Factor B: Drug: Theophylline Subjects: 14 patients suffering from chronic obstructive pulmonary disorder Goal: To compare the true population mean theophylline clearances in the treatment groups Treatment groups: 1. Theophylline/Placebo (i=1) 2. Theophylline/Tagamet (i=2) 3. Theophylline/Pepcid (i=3) Answer: Here we have t=3 treatments (groups) to be compared. We also have b=14 blocks (subjects). The outcome (Response) is Clearance of theophyline and is labeled Yij which means when treatment i is assigned to block j. ai : Effect of treatment i bj : Effect of treatment j ij: Random error This problem can be modeled as below: (Block effects and random errors independent):

Yij = + i + j + ij = i + j + ij
t

i =1

=0 j ~

(0, )
2 b

ij ~

(0, )
2 e

Questions: 1- Give the sample means for each treatment: Answer: From SAS
N intagnt Obs Mean 1 14 3.0792857 2 14 3.1592857

3 14 2.2557143

From Excel: Treatment groups Theophylline/Placebo (i=1) Theophylline/Tagamet (i=2) Theophylline/Pepcid (i=3) Treatment Mean 3.079285714 3.159285714 2.255714286

2- Give the sample means for each subject: Answer: From Excel: 1
5.88 5.13 3.69

2
5.89 7.04 3.61

3
1.46 1.46 1.15

4
4.05 4.44 4.02

5
1.09 1.15 1

6
2.59 2.11 1.75

7
1.69 2.12 1.45

8
3.16 3.25 2.59

9
2.06 2.11 1.57

10
4.59 5.2 2.34

11
2.08 1.98 1.31

12
2.61 2.38 2.43

13
3.42 3.53 2.33

14
2.54 2.33 2.34

4.9

5.513 1.357 4.17 1.08 2.15 1.753

1.913 4.043 1.79 2.473 3.093 2.403

Sample means for each subject from SAS output:

subject Obs Mean 1 3 4.9000000 2 3 5.5133333 3 3 1.3566667 4 3 4.1700000 5 3 1.0800000 6 3 2.1500000 7 3 1.7533333 8 3 3.0000000 9 3 1.9133333 10 3 4.0433333 11 3 1.7900000 12 3 2.4733333 13 3 3.0933333 14 3 2.4033333

3- Obtain the Analysis of Variance Table Answer:

Source SS SST

Treatments Blocks Error Total Where:

t b

SSB SSE TSS

df t-1 b-1 (b-1)(t-1) bt-1

MS MST = SST/(t-1) MSB = SSB/(b-1) MSE = SSE/[(b-1)(t-1)]

F = MST/MSE

TSS = i =1 j =1 yij y ..

= 87.41 df Total = bt 1 = 41 df T = t 1 = 2 df B = b 1 = 13 = TSS SST SSB = 87.41 7.01 71.81 = 8.59 df E = (b 1)(t 1) = 26

( SSB = t (y SSE = (y
t b j =1

SST = bi =1 y i. y ..
.j

) y )
..

= 7.01 = 71.81

ij

y i. y . j + y ..

Source

Treatments Blocks Error Total

SS SST=7.01

SSB=71.81 SSE=8.59 TSS=87.41

df 2 13 26 41

MS MST = 3.505 MSB = 5.52 MSE = 0.33

F = 10.59

From SAS output, we can see the same results.

Sum of Squares 78.81656667 8.59874762 87.41531429

Source Model Error Corrected Total R-Square 0.901633

DF 15 26 41

Mean Square 5.25443778 0.33072106

F Value 15.89

Pr > F <.0001

Coeff Var 20.31072

Root MSE 0.575084

thcl Mean 2.831429

Source intagnt subject

DF 2 13

Type I SS 7.00518571 71.81138095

Mean Square 3.50259286 5.52395238

F Value 10.59 16.70

Pr > F 0.0004 <.0001

4- Test the null hypothesis that the 3 treatment means are all equal (which implies neither Tagamet or Pepcid interact with theophylline) versus the alternative that the true means are not all equal. (=0.05) Answer: Test for differences among treatment effects: H0: a1 = ... = at = 0 (m1 = ... = mt ) HA: Not all ai = 0 (Not all mi are equal)

T .S . : Fobs = R.R. : Fobs

MST MSE F ,t 1,(b 1)(t 1)

P val : P ( F Fobs )
F-obs= 10.59 => P-value: 0.0004 <0.05 => Reject H0 F-critical=F0.05,2,26 = (From F-tables)=3.37 F-obs= 10.59 is larger than F-critical=F0.05,2,26=3.37, or it s equivalent to say that The P-value from the Test statistic is smaller than 0.05 significant level, thus we reject the null hypothesis and conclude that Not all the 3 treatment means are equal. 5- Use Tukeys method to compare all pairs of treatment means with an experimentwise error rate of E=0.05. Answe: Since we have rejected the H0 and concluded differences exist among the treatment means, we can now use Tukeys method to determine which treatments differ significantly. t= number of treatments n= total number of observations=bt 4

ni= The number of measurements per treatment

, ,

, ,

0.33

3.514 0.33 Comparison

Placebo VS Tagamet Placebo VS Pepcid Tagamet VS Pepcid

0.54 Confidence Interval (-0.62,0.46) (-0.02,1.06) (0.05,1.133) Conclusion NSD Pl>Pe T>P

3.07-3.15= -0.08 3.07-2.55=0.52 3.15-2.557=0.593

Alpha 0.05 Error Degrees of Freedom 26 Error Mean Square 0.330721 Critical Value of Studentized Range 3.51417 Minimum Significant Difference 0.5401 Means with the same letter are not significantly different. Tukey Grouping Mean N intagnt A A A B 3.1593 3.0793 2.2557 14 14 14 2 1 3

6-

Give the relative efficiency of the RBD to CRD for this experiment. How many subjects would be needed per treatment for CRD to give treatment means estimates with the precision of this RBD? Answer: The relative efficiency of using this design as opposed to a Completely Randomized Design is obtained as:

t = 3 b = 14 MSB = 5.525 MSE = 0.33 RE ( RCB, CRD ) = (b 1) MSB + b(t 1) MSE 13(5.52) + 14(2)(0.33) 81 = = = 5.98 (bt 1) MSE (14(3) 1)(0.33) 13.53

We will need 5.98 times as many subject per treatment for CRD to give Treatment means estimate with the precision of this RBD. 14(5.98) 84 subject per treatment 3(84) = 252 total subjects 7- Qualitatively, what is the studys implications?
Answer:Here we compared the mean theophylline clearances when it is taken with each of the three drugs, Placebo, Tagment, Pepcid. Here Randomized Block Design was used and we controlled for the subject-to-subject variation when comparing the three treatments. We tested for treatment effects, and when we realized that not all of the treatments have equal effects, we used the Tukeys method to make pairwise comparisons among the three drugs. This result showed that Placebo and tagamet has a greater effect than Pepcid.

Part 2Here we have more than one set of treatments that wed like to compare simultaneously. If we wish to measure the interaction we will have to have more than one measurement (replicate) corresponding to each combination of levels of the 2 factors. Response: 85 day weight gains in the rats Factor A: Model (a=2 Levels) Male (i=1) Female (i=2) Factor B: dosing regimens (b=6 Levels) Control (0 mg/kg/day) Subcutaneous Injection (1.0 mg/kg/day) Oral Glavage (0.1 mg/kg/day) Oral Glavage (0.5 mg/kg/day) Oral Glavage (5 mg/kg/day) Oral Glavage (50 mg/kg/day) Replicates: n=5 per Treatment =>5*(2*6)=60 rats

Model:

Yijk = + i + j + ( )ij + ijk

i = 1,2 j = 1,2,3,4,5,6 k = 1,..,5

where : Yijk 85 day weight gains in the rats

Overall Mean
i Effect of gender i

j Effect of dosing regimens level j

( )ij
i

Interaction effect between gender i and dosing regimens j = ( )ij = 0

j

ijk Random error term

= = ( )
j i j i

ij

ijk ~

(0, )
2

Questions: 1- Give the sample means for the 12 treatments (2 Gender x 6 Dosing Regimens) Answer:
N gender trt Obs Mean 1 1 5 324.0020000 2 5 432.0020000 3 5 326.9980000 4 5 318.0000000 5 5 324.9980000 6 5 328.0000000 2 1 5 148.0020000 2 5 217.0000000 3 5 140.0000000 4 5 152.0000000 5 5 147.0000000 6 5 152.0000000

Analysis Variable : wtgain N gender Obs Mean 1 30 342.3333333 2 30 159.3336667

Analysis Variable : wtgain N trt Obs Mean 1 10 236.0020000 2 10 324.5010000 3 10 233.4990000 4 10 235.0000000 5 10 235.9990000 6 10 240.0000000

2- Test whether there is an interaction between Gender and Dosing Regimen. (=0.05) Answer:

Source Gender Dosing regimens Interaction Error Total

df 1 5 5 48 59

SS
502333.170 65354.856 3630.3300 69564.394 640882.7514

MS
502333.170 13070.971 726.0660 1449.2582

F
346.6 9.02 0.50

P-Value 0.0001 0.0001

0.7740

To test the interaction of factors A and B is as below: H0: ()11=...=( )ab=0 No interaction effect HA: Not all ()ij=0 Interaction effect exist = = 0.5

The P-value from this test is much greater than any acceptable significance level, thus we DO NOT reject the null hypothesis and conclude that No interaction effect exist between factor A and Factor B.
3- If there is not an interaction: a) Test whether there are gender or dosing regimen main effects. (Each at =0.05). Answer: Now that we realized no interaction effects exist, we can test for differences among the effects of the levels of factor A and for differences among the effect of the level of factor B as follows. Test for Factor A effect:

H0: 1=...=a=0 No Factor A effect HA: Not all i=0 Factor A effect exist = = 346.6 with P-value= 0.0001 The P-value from this test is smaller than any acceptable significance level, thus we REJECT the null hypothesis and conclude that Factor A (gender) effect exist.
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Test for Factor B effect: H0: 1=...=b=0 No Factor A effect HA: Not all j=0 Factor A effect exist = = 9.02 with P-value= 0.0001 The P-value from this test is smaller than any acceptable significance level, thus we REJECT the null hypothesis and conclude that Factor B (dosing regimens) effect exist.
b) Use Tukeys method to compare genders (E=0.05), and all pairs of dosing regimens (E=0.05). Answer: Post-Hoc Comparisons: Now that we realized there is no interaction, we can make pairwise comparisons among levels of Factor A and Factor B, respectively. For Factor A, we use Tukeys method, and obtain simultaneous confidence intervals of the form:
.. .

, ,

..

, ,

, ,

= 2.843 (From Table) => W 19.75

1449.25

= 2.843 6.95 = 19.75885

Gender (2 levels => 1 comparison) Male vs Female rats: .. .. = 342.3 152.3 = 190
, ,

95%CI: 190 19.75=(170.25,209.75)

For Factor B, the Tukeys method, is in the form:

. . . . .

, ,

, ,

= 4.197 (From Table)

1449.25

= 4.197 12.03 = 50.52 => W 50.5

Dosing regiments (6 levels =>C(6,2)=15 comparison)

Comparisons 1 0mg vs 1 mg 2 0mg vs 0.1 mg 3 0mg vs 0.5 mg 4 0mg vs 5 mg 5 0mg vs 50 mg 6 1mg vs 0.1 mg 7 1mg vs 0.5 mg 8 1mg vs 5 mg 9 1mg vs 50 mg 10 0.1mg vs 0.5 mg 11 0.1mg vs 5 mg 12 0.1mg vs 50 mg 13 0.5mg vs 5 mg 14 0.5mg vs 5 mg 15 5mg vs 50 mg
. . . . 236-324=-88 236-233=3 236-235=1 236-235.9=0.09 236-240=-4 324-233=91 324-235=89 324-235.9=88.1 324-240=84 233-235=-2 233-235.9=-2.9 233-240=-7 235-235.9=-0.9 235-240=-5 235.9-240=-4.1

Confidence Interval -8850.5 350.5 150.5 0.0950.5 -450.5 9150.5 8950.5 88.150.5 8450.5 -250.5 -2.950.5 -750.5 -0.950.5 -550.5 -4.150.5

Conclusion (1mg)> (0mg) NSD NSD NSD NSD (1mg)> (0.1mg) (1mg)> (0.5mg) (1mg)> (5mg) (1mg)> (50mg) NSD NSD NSD NSD NSD NSD

From SAS output:

Alpha 0.05 Error Degrees of Freedom 48 Error Mean Square 1449.258 Critical Value of Studentized Range 2.84352 Minimum Significant Difference 19.764

Means with the same letter are not significantly different. Tukey Grouping A B Mean 342.333 159.334 N 30 30 gender 1 2

Alpha 0.05 Error Degrees of Freedom 48 Error Mean Square 1449.258 Critical Value of Studentized Range 4.19724 Minimum Significant Difference 50.529 Means with the same letter are not significantly different. Tukey Grouping A B B B B B B B B B Mean 324.50 240.00 236.00 236.00 235.00 233.50 N 10 10 10 10 10 10 trt 2 6 1 5 4 3

5) Qualitatively, what is the studys implications? Answer:Here we compared more than one set of treatment simultaneously using 2-way ANOVA. Testing for interaction effect between the two factors, gender and dose regimens shows that NO interaction effect between the two factors exists, however testing for individual effects of factors revealed that gender and dose regimens have significant effects on the rats. For the first factor, gender, this effect was obviously observed between the only possible comparison, male and female groups. For the second factor, this effect was observed when the dose regimen was 1mg.

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