# Hypothesis testing S2

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1 Basic medical statistics for clinical and experimental research Hypothesis testing S2 Katarzyna Jóźwiak 2nd November /43

2 Introduction Point estimation: use a sample statistic to estimate a population parameter γ of interest. Interval estimation: use a 95% CI to give a 95% probability that this interval contains γ. Hypothesis testing: assume a value for γ and make a probability statement about the value of the corresponding statistic. A statistical hypothesis is an assumption about a population parameter. Hypothesis testing refers to the formal procedures used to accept or reject statistical hypotheses. 2/43

3 Introduction Example: The average person sleeps 8 hours per day. But would you say college students tend to sleep also 8 hours? Take random sample of 100 college students and ask them how long they sleep on an average day. Sample mean x = 6.5 hours. Is this difference just due to sampling variation? Or do college students really sleep different hours than the general population? Would you say students do not sleep 8 hours on average if x = 3? And if x = 8.5? Would your decision depend on something else? 3/43

4 Null and alternative hypothesis Null hypothesis (H 0 ): specifies (a) hypothesized true value/s for a parameter. Always a statement about independence, no effect or equality in populations (e.g., no difference between means; no relationship between variables). Very precise statement. Usually a statement that we wish to reject. Keep it simple: if e.g. we compare the mean effect of a new treatment with a standard treatment, a simple H 0 is that the treatments have the same effect. Alternative hypothesis (H 1 ): specifies (a) true value/s for a parameter, that will be considered if H 0 rejected. Is always a statement about dependence, an effect or differences in populations (e.g., there is a difference between means; there is a relationship between variables). Not precise statement. 4/43

5 Null and alternative hypothesis Null hypothesis: H 0 : Parameter = Parameter value Alternative hypothesis: H 1 : Parameter Parameter value or H 1 : Parameter > Parameter value or H 1 : Parameter < Parameter value Parameter is the population parameter; Parameter value is the hypothesized value of the parameter Example with number of hours college students sleep: H 0 : µ = 8 H 1 : µ < 8 5/43

6 Null and alternative hypothesis We test the null hypothesis: Assuming that the null hypothesis is correct, what is the probability of obtaining our pattern of results? We either "reject the null hypothesis" or "fail to reject the null hypothesis" If the probability of obtaining our pattern of results is high, the result is considered to be likely so we do not reject the null hypothesis. 6/43

7 Test statistic We construct a test statistic: Test statistic = Statistic Parameter value Standard error of the statistic = Effect Error where Statistic is the observed sample statistic, i.e. point estimate of the population parameter Parameter value is the hypothesized value of the parameter Test statistic measures by how many SEs the statistic differs from its hypothesized value in H 0 Test statistic has a sampling distribution Example with number of hours college students sleep where H 0 : µ = 8 and x = 6.5, SE=1: Test statistic = = /43

8 Significance level and critical value Level of significance, α, is the probability value of the criterion for rejecting the null hypothesis Usually α =.05, sometimes α =.01 or α =.1 Level of significance should be chosen before conducting any null hypothesis Rejection region is the set of values for the test statistic that leads to rejection of the null hypothesis Non-rejection region is the set of values not in the rejection region that leads to non-rejection of the null hypothesis Critical value is the value of the test statistic that separate the rejection and non-rejection regions 8/43

9 Significance level and critical value One-sided test, H 1 : Parameter > Parameter value When Test statistic (1-α) th quantile: we reject the null hypothesis Sampling distribution under H0 α (1 α)th percentile Rejection region 9/43

10 Significance level and critical value One-sided test, H 1 : Parameter < Parameter value When Test statistic α th quantile: we reject the null hypothesis Sampling distribution under H0 α Rejection region αth percentile 10/43

11 Significance level and critical value Two-sided test, H 1 : Parameter Parameter value When Test statistic α/2 th quantile or Test statistic (1-α/2) th quantile: we reject the null hypothesis Sampling distribution under H0 α/2 α/2 Rejection region α/2th percentile (1 α/2)th percentile Rejection region 11/43

12 Statistical STATISTICAL TABLES table with critical values 2 TABLE A.2 t Distribution: Critical Values of t Significance level Degrees of Two-tailed test: 10% 5% 2% 1% 0.2% 0.1% freedom One-tailed test: 5% 2.5% 1% 0.5% 0.1% 0.05% /43

13 p value Using distribution of a test statistic we find probability of obtaining value of our test statistic or more extreme values p value is the probability of obtaining a test statistic result at least as extreme as the one that is actually observed, assuming that the null hypothesis is true when p value is small the test statistic is significant 13/43

14 p value One-sided test, H 1 : Parameter > Parameter value When p value < α: we reject the null hypothesis Sampling distribution under H0 p value Sample value of test statistic More extreme test statistic values 14/43

15 Sample p value One-sided test, H 1 : Parameter < Parameter value When p value < α: we reject the null hypothesis p value Sampling distribution under H0 More extreme test statistic values value of test statistic 15/43

16 p value Two-sided test, H 1 : Parameter Parameter value When p value < α: we reject the null hypothesis Sampling distribution under H0 p value/2 p value/2 More extreme values Sample value of test statistic Sample value of test statistic More extreme values 16/43

17 p value p value lower than 0.05 (0.1): results are "significant at the 5% (10%) level" (small enough to reject H 0 ). Reject H0 at a 5% significance level. There is evidence to reject H0. p value not lower than 0.05 (0.1): results are "not significant at the 5% (10%) level". Fail to reject H0 at a 5% significance level. There is not enough evidence to reject H0. But this does NOT mean that we accept that H0 is true. The cut-off level 0.05 (0.1) is the significance level of the test. 17/43

18 Statistical table with probabilities Standard Normal Cumulative Probability Table Cumulative probabilities for NEGATIVE z-values are shown in the following table: z /43

19 Statistical table with probabilities Standard Normal Cumulative Probability Table Standard Normal Cumulative Probability Table Cumulative -2.6 probabilities for NEGATIVE z-values are shown in the following table: z /43

20 Statistical table with probabilities Example with number of hours college students sleep: H 0 : µ = 8 vs H 1 : µ < 8 x = 6.5, SE=1: Test statistic = = 1.5, p value = > α = 0.05 if we assume that the test statistic has a normal distribution; when we change the alternative hypothesis to H 1 : µ 8, then p value=2*0.0668= /43

21 Hypothesis testing step by step 1. Define the null and alternative hypotheses under study. 2. Collect relevant data from a sample of individuals. 3. Calculate the value of the test statistic. 4. Compare the value of the observed test statistic to a critical value or p value of the distribution of the test statistic under H Interpret the results. 21/43

22 Type I and Type II errors In a hypothesis test we draw a conclusion about H 0 (there is strong/weak evidence to reject/not reject) based on a sample. We can make mistakes! Decision State of nature Reject H 0 Do not reject H 0 H 0 true Type I error (α) Correct decision (1-α) H 0 false Correct decision (1 β) Type II error (β) α: significance level of the test. Probability of rejecting H0 when H 0 is true. β: Probability of not rejecting H 0 when H 0 is false. 1 β: power of the test that is probability of rejecting H0 when it is false. 22/43

23 Power Essential to know the power of a proposed test. We want to be confident that our statistical method can correctly reject the null hypothesis. It is accepted that power should be 0.8 or greater. Study with a smaller power might waste our time and resources. If the power is smaller than 0.8 we might want to replicate our study. Factors relevantly related to power: sample size power. variability of observations power. effects of interest power : A hypothesis test has a greater chance of detecting a large true effect than a small one. significance level α power 23/43

24 Power Sampling distribution associated with H0 Sampling distribution associated with H1 power reject H0 24/43

25 Power Sampling distribution associated with H0 Sampling distribution associated with H1 power µ=0 µ=5 25/43

26 Power Sampling distribution associated with H0 Sampling distribution associated with H1 power µ=0 µ=4 26/43

27 Power Sampling distribution associated with H0 Sampling distribution associated with H1 power µ=0 µ=3 27/43

28 Power Statistical significance versus Clinical relevance Obtaining significant results does not mean that the effect it measures is meaningful or important. Small underpowered studies fail to detect clinically relevant effects as statistically significant. Clinical trial comparing octreotide and sclerotherapy in patients with variceal bleeding (Sung et al., 1993): Sample of 100, 5% power (while reported calculation suggested a sample of 1800 was needed). Large sample sizes with large power might show small clinically irrelevant effects to be statistically significant. Reduction of blood pressure by two points between treatment and placebo. An expensive new psychiatric treatment technique reduces the average hospital stay from 60 days to 59 days. 28/43

29 Power Power analysis calculation should be used at the planning stage of our investigation to ensure that the sample size is big enough. For any power calculation we need to know: Type of a test (e.g., independent t-test, paired t-test, ANOVA, regression, etc.) The significance level The expected effect size The sample size Fixing the significance level, the expected effect size and sample size we can calculate power Fixing the significance level, the expected effect size and power we can calculate sample size 29/43

30 Power How large should the sample be in order to detect a specific effect size? One-sample case example: H 0 : µ = m vs H 1 : µ < m or vs H 1 : µ > m n ) 2 H 0 : µ = m vs H 1 : µ m n ( σ x z 1 α/2 +z 1 β x m Two-sample case example: H 0 : µ 1 = µ 2 vs H 1 : µ 1 < µ 2 or vs H 1 : µ 1 > µ 2 n 1 ( σ x1 2 + σ x2 2 /k ) ( ) z 1 α +z 2 1 β x 1 x 2 ( z 1 α +z 1 β σ x x m H 0 : µ 1 = µ 2 vs H 1 : µ 1 µ 2 n 1 ( σ x1 2 + σ x2 2 /k ) ( ) z 1 α/2 +z 1 β 2 x 1 x 2 x: the observed value of the sample mean σ x: the standard error of the mean k = n 1 /n 2 : ratio between the sample sizes of the two groups ) 2 30/43

31 Power z 1 α, z 1 β : critical values from the standard normal distribution α = 5% z1 α = 1.65, z 1 α/2 = 1.96 α = 1% z1 α = 2.33, z 1 α/2 = 2.58 power= 80% z1 β = 0.84 power= 85% z1 β = 1.04 power= 90% z1 β = 1.28 power= 95% z1 β = /43

32 Power Example A family sociologist is interested in the difference in marital satisfaction between working and notworking wives. How many women should be asked about their marital satisfaction to be able to say that working wives are more happy than not working wives? H 0 : working wives are as happy as notworking wives; H 1 : working wives are more happy than notworking wives expected effect size = 7.5 standard deviation = 25 equally sized groups, k = 1 n 1 ( σ x σ x 2 2 /k ) ( z 1 α +z 1 β x 1 x 2 ) 2 = ( /1) ( ) 2 = /43

33 Power Example 33/43

34 One-tailed vs. two-tailed If e.g. we wish to test parameter µ: H0 : µ = 5 vs. H 1 : µ 5 will require a two-tailed test (two-sided). We test for the possibility of deviation from H 0 in both directions. If α = 0.05, then half of it is used to test statistical significance in one direction (µ > 5) and the other half for the other direction (µ < 5). H0 : µ = 5 vs. H 1 : µ > 5 will require a one-tailed test (one-sided). We completely disregard the possibility of µ < 5. This provides more power to detect an effect in direction µ > 5 by not testing µ < 5. Hardly ever appropriate with biomedical data. 34/43

35 One-tailed vs. two-tailed When would a one-tailed test be appropriate? E.g.: new drug that we believe improves other existing drug. Should we use a one-tailed test? "Yes, because it has more power to detect an effect in that direction", "Yes, because a two-tailed test did not reject H 0 but got very close to rejecting it, so a one-tailed test will do it", THESE ARE NO GOOD ANSWERS... Why? Because we fail to test that it could be actually (much) worse than the existing drug. 35/43

36 One-tailed vs. two-tailed E.g.: we use a one-tailed test to test the efficacy of a cholesterol lowering drug, because the drug cannot raise cholesterol (so individuals taking the drug who have higher cholesterol level than controls are treated as failing to show a difference). Imagine individuals taking the drug end up with average higher levels than those of the control group. Using a one-tailed test implies we say this is just due to random variation! But what if there is an underlying cause? But, there is so much variation in medical data that nothing should be excluded. 36/43

37 Multiple hypothesis testing Example: We have three groups of participants and we want to compare every pair of groups. We carry out three separate independent tests with α = 0.05: 1 test: probability of a correct decision (given H0 is true) is =0.95, overall α = = tests: probability of a correct decision (given H0 is true) is (1-0.05)(1-0.05)=0.9025, overall α = = tests: probability of a correct decision (given H0 is true) is (1-0.05)(1-0.05)(1-0.05)=0.8573, overall α = = The probability of making a Type I error (probability of falsely rejecting the null hypothesis) increases from 5% to 14.3%! 37/43

38 Multiple hypothesis testing Familywise error rate or experimentwise error rate or overall α is the error rate across statistical tests conducted on the same data; i.e., this is the probability that at least one test erroneously rejects the null hypothesis: familywise error = 1 (0.95) number of tests E.g. We carry out 20 significance tests on a dataset. The probability that at least one of them erroneously rejects H 0 is 1 (1 α) 20 = 64%! We cannot identify which one(s), if any, are false positives... We can use some post-hoc adjustment, control the familywise error rate, to account for how many tests we perform (will be covered later this week). E.g. Bonferroni: divide α by the number of comparisons to ensure that the overall α is below For 10 tests we use as our criterion for significance 38/43

39 Non-parametric tests Hypothesis test: Parametric: based on knowledge of probability distribution the data follow. Non-parametric: replace data with their ranks (numbers describing position in the ordered dataset) and make no assumption about probability distribution of the data. Example: scores: 5, 2, 6, 8, 7, 1, 10 ordered scores: 1, 2, 5, 6, 7, 8, 10 ranks: 1, 2, 3, 4, 5, 6, 7, Useful when sample size is small and/or data are measured on a categorical scale. However: less power to detect a true effect than the equivalent parametric test (given assumptions of this are satisfied). 39/43

40 Exact vs. asymptotic tests Asymptotic method: p-values estimated assuming the data, given a sufficiently large sample size, conform to a particular parametric distribution. But if the data is small, sparse, heavily tied, unbalanced or poorly distributed then asymptotic method may not yield reliable results. Obtain instead exact p-value based on exact distribution of the test statistic. If dataset is too large for calculating exact p-value then Monte Carlo algorithms are applied. Both methods are implemented in SPSS. We will use them during Practicals 3 and 4. 40/43

41 Hypothesis tests vs. confidence intervals A CI can be used to make a decision in a similar way to a hypothesis test: In the sleeping hours example: if the hypothesized mean value 8 lies outside the corresponding 95% CI for the mean then hypothesized value is implausible and enough evidence to reject H 0 (no p-value is used): x = 6.5, SEM=1: CI=[ *1; *1]=[4.54; 8.46] 8 lies inside the 95%CI If a hypothesis test and a CI can do the same, why not use just one of them? 41/43

42 Hypothesis tests vs. confidence intervals Hypothesis testing Hypothesis testing provides no information about the probability of H0, just gives the probability of finding a specific or more extreme results when the null hypothesis is true. Statistical significance is not the same as medical importance or biological relevance. With large sample sizes we can find statistically significant small differences that are not interesting (the sample estimates fall near the parameter value in H 0 ), while in small samples effects that are clinically relevant might not be statistically significant. Hypothesis testing is used to calculate required sample size and statistical power. 42/43

43 Hypothesis tests vs. confidence intervals Confidence interval CI does not depend on a priori hypotheses. CI displays the entire set of believable values of the parameter, so we can determine whether or not the difference between the true value and the H 0 value has practical importance. 43/43

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