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When scientists perform replicates of the same experiment, the effect size of each replicate often varies, which complicates interpretation of the results. This vignette documents how dabestr is able to compute the meta-analyzed weighted effect size given multiple replicates of the same experiment. This can help resolve differences between replicates and simplify interpretation.

This function uses the generic inverse-variance method to calculate the effect size, as follows:

\[\theta_{weighted} = \frac{\sum{\hat{\theta_i}}w_i}{\sum{w_i}}\] where:

\[\hat{\theta_i}=\text{Mean difference for replicate } i\] \[w_i=\text{Weight for replicate } i = \frac{1}{s_i^2}\] \[s_i^2=\text{Pooled variance for replicate } i = \frac{(n_{test}-1)s_{test}^2 + (n_{control}-1)s_{control}^2} {n_{test}+n_{control}-2}\] \[n = \text{sample size and } s^2 = \text{variance for control/test}\] Note that this uses the fixed-effects model of meta-analysis, as opposed to the random-effects model; that is to say, all variation between the results of each replicate is assumed to be due solely to sampling error. We thus recommend that this function only be used for replications of the same experiment, i.e. situations where it can be safely assumed that each replicate estimates the same population mean \(\mu\).

DABEST can only compute weighted effect size for mean difference only, and not standardized measures such as Cohen’s d.

For more information on meta-analysis, please refer to Chapter 10 of the Cochrane handbook: https://training.cochrane.org/handbook/current/chapter-10

Create dataset for demo

set.seed(12345) # Fix the seed so the results are replicable.
# pop_size = 10000 # Size of each population.
N <- 20 # The number of samples taken from each population

# Create samples
c1 <- rnorm(N, mean = 3, sd = 0.4)
c2 <- rnorm(N, mean = 3.5, sd = 0.75)
c3 <- rnorm(N, mean = 3.25, sd = 0.4)

t1 <- rnorm(N, mean = 3.5, sd = 0.5)
t2 <- rnorm(N, mean = 2.5, sd = 0.6)
t3 <- rnorm(N, mean = 3, sd = 0.75)

# Add a `gender` column for coloring the data.
gender <- c(rep("Male", N / 2), rep("Female", N / 2))

# Add an `id` column for paired data plotting.
id <- 1:N

# Combine samples and gender into a DataFrame.
df <- tibble::tibble(
  `Control 1` = c1, `Control 2` = c2, `Control 3` = c3,
  `Test 1` = t1, `Test 2` = t2, `Test 3` = t3,
  Gender = gender, ID = id
)

df <- df %>%
  tidyr::gather(key = Group, value = Measurement, -ID, -Gender)

We now have 3 Control and 3 Test groups, simulating 3 replicates of the same experiment. Our dataset also has a non-numerical column indicating gender, and another column indicating the identity of each observation.

This is known as a ‘long’ dataset. See this writeup for more details.

knitr::kable(head(df))
Gender ID Group Measurement
Male 1 Control 1 3.234211
Male 2 Control 1 3.283786
Male 3 Control 1 2.956279
Male 4 Control 1 2.818601
Male 5 Control 1 3.242355
Male 6 Control 1 2.272818

Loading Data

Next, we load data as we would normally using load(). This time, however, we also specify the argument minimeta = TRUE As we are loading three experiments’ worth of data, idx is passed as a list of vectors, as follows:

unpaired <- load(df,
  x = Group, y = Measurement,
  idx = list(
    c("Control 1", "Test 1"),
    c("Control 2", "Test 2"),
    c("Control 3", "Test 3")
  ),
  minimeta = TRUE
)

When this dabest object is printed, it should show that effect sizes will be calculated for each group, as well as the weighted delta. Note once again that weighted delta will only be calculated for mean difference.

print(unpaired)
#> DABESTR v2023.9.12
#> ==================
#> 
#> Good morning!
#> The current time is 06:04 AM on Tuesday December 12, 2023.
#> 
#> Effect size(s) with 95% confidence intervals will be computed for:
#> 1. Test 1 minus Control 1
#> 2. Test 2 minus Control 2
#> 3. Test 3 minus Control 3
#> 4. weighted delta (only for mean difference)
#> 
#> 5000 resamples will be used to generate the effect size bootstraps.

After applying the mean_diff() function to the dabest object, you can view the mean differences for each group as well as the weighted delta by printing the dabest_effectsize_obj.

unpaired.mean_diff <- mean_diff(unpaired)

print(unpaired.mean_diff)
#> DABESTR v2023.9.12
#> ==================
#> 
#> Good morning!
#> The current time is 06:04 AM on Tuesday December 12, 2023.
#> 
#> The unpaired mean difference between Test 1 and Control 1 is 0.585 [95%CI 0.307, 0.869].
#> The p-value of the two-sided permutation t-test is 0.0022, calculated for legacy purposes only.
#> 
#> The unpaired mean difference between Test 2 and Control 2 is -1.058 [95%CI -1.52, -0.577].
#> The p-value of the two-sided permutation t-test is 0.0001, calculated for legacy purposes only.
#> 
#> The unpaired mean difference between Test 3 and Control 3 is -0.254 [95%CI -0.626, 0.169].
#> The p-value of the two-sided permutation t-test is 0.1081, calculated for legacy purposes only.
#> 
#> 5000 bootstrap samples were taken; the confidence interval is bias-corrected and accelerated.
#> Any p-value reported is the probability of observing the effect size (or greater),
#> assuming the null hypothesis of zero difference is true.
#> For each p-value, 5000 reshuffles of the control and test labels were performed.

You can view the details of each experiment by accessing dabest_effectsize_obj$boot_results, as follows. This also contains details of the weighted delta.

unpaired.mean_diff$boot_result
#> # A tibble: 4 × 11
#>   control_group   test_group bootstraps nboots bca_ci_low bca_ci_high pct_ci_low
#>   <chr>           <chr>      <list>      <int>      <dbl>       <dbl>      <dbl>
#> 1 Control 1       Test 1     <dbl>        5000      0.307       0.869     0.302 
#> 2 Control 2       Test 2     <dbl>        5000     -1.52       -0.577    -1.52  
#> 3 Control 3       Test 3     <dbl>        5000     -0.626       0.169    -0.645 
#> 4 Minimeta Overa… Minimeta … <dbl>        5000     -0.186       0.238     0.0355
#> # ℹ 4 more variables: pct_ci_high <dbl>, ci <dbl>, difference <dbl>,
#> #   weight <dbl>

Unpaired Data

Simply using the dabest_plot() function will produce a Cumming estimation plot showing the data for each experimental replicate as well as the calculated weighted delta.

dabest_plot(unpaired.mean_diff)

You can also hide the weighted delta by passing the argument show_mini_meta = FALSE. In this case, the resulting graph would be identical to a multiple two-groups plot:

dabest_plot(unpaired.mean_diff, show_mini_meta = FALSE)

Paired Data

The tutorial up to this point has dealt with unpaired data. If your data is paired data, the process for loading, plotting and accessing the data is the same as for unpaired data, except the argument paired = "sequential" or paired = "baseline" and an appropriate id_col are passed during the load() step, as follows:

paired.mean_diff <- load(df,
  x = Group, y = Measurement,
  idx = list(
    c("Control 1", "Test 1"),
    c("Control 2", "Test 2"),
    c("Control 3", "Test 3")
  ),
  paired = "baseline", id_col = ID,
  minimeta = TRUE
) %>%
  mean_diff()

dabest_plot(paired.mean_diff, raw_marker_size = 0.5, raw_marker_alpha = 0.3)