Analysis of repeated measurements (KLMED8008)

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1 Analysis of repeated measurements (KLMED8008) Eirik Skogvoll, MD PhD Professor and Consultant Institute of Circulation and Medical Imaging Dept. of Anaesthesiology and Intensive Care 1

2 Repeated measurements This term is more or less equivalent with... Within subjects experiments Grouped/clustered data Hierarchical models Longitudinal data We will not consider Time series 2

3 Day 1 Welcome. Introduction. Presentation. Practical issues about the course Website: Lectures Textbook Software Excercises Exam Repeated measurements: advantages and problems One summary measure Review of variance, standard deviation, covariance and correlation Linear combination of random variables (Brief) review of linear regression (Textbook Chapter 1) 3

4 Repeated measurements Advantages The individual acts as his own control reduces total variance Effective (re-) use of recruited individuals/patients economy Problems Potential carry-over effect Spontaneous change may occur over time In general: usual statistical procedures (e.g. testing the null hypothesis) require independent observations! Special methods and software necessary 4

5 Repeated measurements Ignoring dependency between observations may lead to p-values becoming too small when doing between-patient comparisons (i.e. yield false positive results) Observations: Treatment, n=20 Control, n=20 Textbook (p. 167), Veierød et al. 7.1 (p. 231) Patients 5

6 Repeated measurements Ignoring dependency between observations may lead to p-values becoming too large when doing within-patient comparisons (i.e. yield false negative results) Observations: Treatment,n=20 Control, n=20 Textbook (p. 167), Veierød et al. 7.1 (p. 231) Individual 6

7 ONE SUMMARY MEASURE 7

8 Avoiding the problem: use one summary measure per individual Change (e.g. from baseline measurement) Mean or median Maximum Maximum change Area under the curve (AUC) Estimated slope ( ) for each individual obtained by simple linear regression These outcomes may now be analyzed as usual (with t-tests etc.), as the observations are now independent Matthews (1990), Altman (1991), Davis (2002) 8

9 Example: gastric tube to reduce postoperative nausea Double blind randomized controlled study 30 and 29 patients, without or with gastric tube, respectively Nausea registered on an ordinal 5-point ordinal scale (0 4) six times postoperatively Research question: - does gastric tube affect nausea? 9

10 Results for the first 20 patients grp t1 t2 t3 t4 t5 t6 1. No tube Tube Tube Tube No tube No tube Tube Tube No tube No tube Tube No tube Tube Tube Tube No tube Tube Tube Tube No tube Some missing observations Which summary measure to use? Mean (t1..t6) Median (t1..t6) Max (t1..t6) Does the maximum value of 4 occur at all? Do any of the values 3 or 4 occur at all? (should be clinically relevant, and decided on before results are known!) 10

11 One plot tells more than 1000 t-tests 11

12 Adding summary measures mean median max 4? 3 or 4? grp t1 t2 t3 t4 t5 t6 m md mx m4 m34 1. No tube Tube Tube Tube No tube No tube Tube Tube No tube No tube Tube No tube Tube Tube Tube No tube Tube Tube Tube No tube

13 The summary measures now independent can be analyzed as usual :. ttest m, by(grp) // slide 13 Two-sample t test with equal variances Group Obs Mean Std. Err. Std. Dev. [95% Conf. Interval] Tube No tube combined diff diff = mean(tube) - mean(no tube) t = Ho: diff = 0 degrees of freedom = 57 Ha: diff < 0 Ha: diff!= 0 Ha: diff > 0 Pr(T < t) = Pr( T > t ) = Pr(T > t) = tabulate grp m34, chi // slide 13 t1 t2 t3 t4 t5 t6 == 3 4 gtube 0 1 Total Tube No tube Total Pearson chi2(1) = Pr =

14 Review of VARIANCE AND COVARIANCE 14

15 Sample variance Definition (Rosner Def.2.7) 1 S x x n 2 2 ( i ) n 1 i 1 Comment (n -1) is used rather than n in the denominator, as we already have made use of the sample mean value. Thus one piece of information (one Degree of Freedom, DF) from the sample has been expended. This yields a slightly larger estimate of S 2. For large n, the difference is trivial. 15

16 Sample variance Properties Always positive Extreme observations contribute more (squaring) Favorable mathematical properties But cannot be interpreted on original scale (kg kg 2, mg/dl mg 2 /dl 2, cm cm 2 etc.) S 2 1 n 1 n i 1 ( x i x) 2 16

17 Sample standard deviation (S, SD) Properties S 1 n 1 Always positive Extreme observations contribute more (squaring) Favorable mathematical properties May be interpreted on original scale! n i 1 ( x i x) 2 17

18 Covariance Def. Rosner6 5.12, p. 142 Cov (X,Y) = E[(X- x )(Y- y )] More clearly : The expected product of the deviation of X and Y from their respective expected values If X og Y are independent, then cov (X,Y ) = 0 The reverse does not hold! 18

19 Covariance and correlation What about the units If X measures FEV (liters, l) and Y measures height (meters, m) then their covariance has unit l*m (compare this with the variance, which has squared units; e.g. l 2, m 2 ) The problem is solved by dividing by the product of the respective standard deviations: Corr ( X, Y ) Cov (X, Y ) X Y ( 1,1) 19

20 f(x,y) y 0 x 20

21 f(x,y) y 0,75 x 21

22 Example: Rosner6, Fig. 5.16a 22

23 Example: Rosner6, Fig. 5.16c A rather unusual situation! Unlikely with repeated measurements. 23

24 Example: Rosner6, Fig. 5.16b The usual situation with repeated measurements. 24

25 Sample covariance and sample correlation - Pearson s r n 1 S ( x x)( y y) XY, i i n i 1 r r S XY, S S X Y n n 1 ( xi x)( yi y) ( xi x)( yi y) n i 1 i n n n n n ( xi x) ( yi y) ( xi x) ( yi y) i 1 n i 1 i 1 i 1 25

26 Example Central venous PO2 measured in 12 patients during mobilisation after cardiac surgery; in two situations (x6 and x7) pnr x6 x Tasks Plot x6 and x7 Calculate by hand the following sample statistics, using only the observations on the first 3 patients : mean variance standard deviation covariance correlation coefficient 26

27 . twoway (scatter x7 x6,yline(30.75) xline(35.1)) 27

28 . cor x6 x7, cov mean (obs=12) Variable Mean Std. Dev. Min Max x x Covariance matrix x6 x7 x x pwcorr x6 x7, star(.05) sig Correlation matrix x x6 x7 x *

29 T-test for sample correlation coefficient (Rosner6 Eq , p. 496) Calculate Pearson s r Transform it to T: Example: r = 0.95, n =12, T 10 = 0.95*sqrt(10)/sqrt(1-0.95^2) = 9.62 P( T 10 >9.62) = T T n 1 n 2 x... in this case: s n r 0 r r( n 2) r 1 r 1 r n 2 n 2. pwcorr x6 x7, star(.05) sig x x6 x7 x *

30 Linear combination of random variables Rosner6, Eq. 5.12, p. 144: L n i 1 c X i i n n n 2 ( ) i ( i) 2 i j ( i, j) i 1 i 1 j 1 Var L c Var X c c Cov X X n n n 2 ci Var X i cic j i j Corr X i X j i 1 i 1 j 1 ( ) 2 (, ) i j If only two random variables X and Y, then L c X c Y Var( L) c Var( X ) c Var( Y) 2 c c Cov( X, Y)

31 Example, ctd.. pnr x6 x Calculate D = x7-x6 pnr x6 x7 D

32 Linear combination of random variables L c X c X Var( L) c Var( X ) c Var( X ) 2 c c Cov( X, X ) Example: Consider the difference ( 1) X 1 X 6 7 D X X 7 6 Var( D) ( 1) Var( X ) 1 Var( X ) 2 ( 1) 1 Cov( X, X ) Var( X ) Var( X ) 2 Cov( X, X ) Now insert estimates: Var(D) = *93.30 = considerably less than either of the original variances! 32

33 Linear combination of random variables pnr x6 x7 D Tasks: Perform the following tests By hand One sample t-test Using your favourite software One sample t-test Two sample t-test Paired samples t-test 33

34 Result (descriptive). summ D, detail D Percentiles Smallest 1% % % Obs 12 25% Sum of Wgt % -4 Mean Largest Std. Dev % % 0-2 Variance % 0 0 Skewness % 0 0 Kurtosis

35 One sample t-test. ttest D ==0 One-sample t test Variable Obs Mean Std. Err. Std. Dev. [95% Conf. Interval] D mean = mean(d) t = Ho: mean = 0 degrees of freedom = 11 Ha: mean < 0 Ha: mean!= 0 Ha: mean > 0 Pr(T < t) = Pr( T > t ) = Pr(T > t) =

36 Example, revisited now breaking the correlation x7 shuffeled pnr x6 x Shuffle x7 randomly pnr x6 x7s

37 Example, revisited Re-calculate D pnr x6 x7s D Plot x6 and x7s Calculate the following sample statistics, using all observations: mean variance standard deviation covariance correlation coefficient Re-calculate the one sample t-test 37

38 . graph twoway (scatter x7s x6, yline(30.75) xline(35.1)) 38

39 . twoway (scatter x7 x6,yline(30.75) xline(35.1)) SvO2 v/tid SvO2 v/tid 6 39

40 Results. cor x6 x7s, cov mean (obs=12) Variable Mean Std. Dev. Min Max Korrelasjonsmatrise Kovariansmatrise x x7s x6 x7s x x7s pwcorr x6 x7s, star (0.05) sig x x6 x7s x7s

41 Result (descriptive) D Percentiles Smallest 1% % % Obs 12 25% Sum of Wgt % -3 Mean Largest Std. Dev % % Variance % Skewness % Kurtosis Again, variance has changed dramatically: Var(D) = *11.16 = considerably more than the sum of the original variances! 41

42 One sample t-test. ttest D ==0 // One-sample t test Variable Obs Mean Std. Err. Std. Dev. [95% Conf. Interval] D mean = mean(d) t = Ho: mean = 0 degrees of freedom = 11 Ha: mean < 0 Ha: mean!= 0 Ha: mean > 0 Pr(T < t) = Pr( T > t ) = Pr(T > t) =

43 Conclusion Dependency changes everything! 43

44 Review of linear regression Rabe-Heskett & Skrondal, chapter 1. This material is assumed to be well known from previous courses! Points to consider ANOVA Simple linear regression Multiple linear regression Dummy variables Interaction (concept, interpretation) 44

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