14.3 – Fixed effects, Random effects

Introduction

With few exceptions (eg, repeatability and intraclass correlation calculations, Chapter 12.3), we have been discussing the Model I ANOVA or fixed-effects ANOVA– fixed implies that we select the levels for the factor. It may not be obvious — in hindsight it is — but levels may also be randomly selected, eg, nature provides the levels. Thus, levels are random and the model is a random-effects ANOVA. Beginning with our discussions of ANOVA, it becomes increasingly important to incorporate concept of models in statistics. As you have been working in R and Rcmdr with the lm() function, you have been forced to address the statistical model concept — you enter the response variable then type in both factors and create a term for the interaction.

We’ve just completed an experiment in which the response (cholesterol levels) of 36 individuals, from one of three drug treatments (placebo, Drug A, Drug B), given one of 3 types of diets (low, medium, or high carbohydrate), was observed. Thus, we say that the specific treatments of drug and diet contribute to variation in cholesterol levels. More formally, we say that the observed response of the kth individual (Yijk) is equal to the overall mean (m) plus the added effect of Drug (a) plus the added effect of Diet (b) plus the interaction between Diet and Population (ab) plus unidentified sources of variation generally called error (e). In symbols, we write

    \begin{align*} Y_{ijk} = \mu + \alpha_{j} + \beta_{2} + \left (\alpha\beta \right )_{ij} +\epsilon_{ijk} \end{align*}

where i is the number of levels of the first factor (in our example, Diet had 2 levels, so i = A or i = B), j is the number of levels of the second factor (in our example, Population had 2 levels, so j = 1, or j = 2), and k is the total number of observations in the experiment (12 in our example, so k = 1, 2, …, 11, 12). Thus, we can think of each term “adding up” to give us the observed value.

Although it is a bit confusing at first, these equations help us understand how the experiment was conducted and therefore how to analyze and interpret the results.

Model I, Model II & Model III ANOVA

Statisticians recognize that how levels of the factors were selected for an experiment impact conclusions from ANOVAs. The key: whether or not the levels of the factor were selected (1) randomly from all possible levels of the factor or (2) specifically selected by the experimenters. We introduced the concepts of fixed and random effects in Chapter 12.3. For one-way ANOVA, the distinction between fixed and random effects influences the interpretation, but not the calculation of the ANOVA components. For two or more treatment factors, both the interpretations and the calculations of ANOVA components are affected.

There are two general types of Factors that we can choose to employ in an ANOVA: Fixed Model ANOVA and Random Model ANOVA. Where two or more factors apply, by far the most common model in experimental sciences is a combination of fixed and random, so we need to add a third general type, the Mixed Model ANOVA design.

Fixed Factors. Where the levels of the factor are selected by us. In this case we would only be interested in the response of the individuals that are given those specific treatments.

Medicine – for example, where we choose a treatment given to patients with a history of coronary heart disease; compare outcomes of patients given a statin (drug used to lower serum cholesterol) drug versus a placebo. (Note that this is the same study we discussed in our lecture on about risk analysis).

Ecotoxicology – for example, compare growth rates and deformities of tadpole frogs given Aldicarb, Atrazine (both estrogen-mimicking pesticides), or a control (ie, no pesticides). If you’re interested in these topics, here’s a link to the EPA’s web site, listing pesticide sales and use in the United States. Here’s a link to a NIH National Institute of Environmental Sciences, with a nice description of estrogen mimicking pesticides.

Agriculture & Genetics – for example, monitor growth of a particular hybrid corn available from three different manufacturers. See an example of Wisconsin corn hybrid studies.

In each of these examples we might be interested in those specific treatments and no other treatments.

HO: No difference in the means among the levels of the Factor

HA: Some difference in the means among these specific levels of the Factor; the specific levels of this factor affects the response variable.

This is an example of a Model I ANOVA, also called a “fixed effects” model ANOVA.

Random Factors. We still only use a relatively few number of different levels of a particular factor. However, in this case we are interested in many different levels of the factor — we want to generalize beyond our sample. The levels that we use would be a “sample” of all possible levels that we would be interested in.

Medicine – for example, where we randomly choose a treatment level given to patients; four concentrations of a drug and a placebo. Since concentration is a ratio scale data type, concentration can range from 0 (the placebo) to 100%.

Ecotoxicology – for example, release different concentrations or mixtures of air plus components of air pollution to chambers, record the response of plants or animals.

Agriculture & Genetics – for example, grow three different varieties of a plant in three different soils or different genetic strains of animals on three different diets. In these cases, factor levels are random because we are drawing from a large pool of possible levels: genetic varieties or strains — we selected three, but it’s rare that were are specifically interested in the three chosen. More often, we want to make generalizations and the three were somehow representative (we hope) of genetic variation in the species of interest.

In each of these examples, we write the null hypothesis to reflect that the particular levels are only of interest in so far as they can be used to generalize back to the population.

HO: No difference in the variation between groups.

HA: Some difference in the variation among these groups.

This is an example of a Model II ANOVA, also called a “random effects” model ANOVA. Note that we specify the hypothesis in terms of variation, not of the means.

Your two-way ANOVA could be Model I, Model II, or it could be mixed, with one factor fixed, the other random (this later model is called a Model III, or “mixed model” ANOVA).

For the most part, the distinction between whether you have fixed or random effects is clear, but whether we use fixed or random or combinations, this design decision has consequences for testing.

The decision does not affect the Sources of Variation for the different Models. Last time, we showed the tests for a Model I ANOVA (the “fixed effects model”).

For Random Effects or Mixed Effects, we only change how we determine the statistical significance of the Factors. Here’s a summary of how the experimental design changes the calculation of the F-test statistic for the Factors.

Table 1. Calculation of F

The Critical Value for each of the different F values will be obtained by simply finding the degrees of freedom for the numerator and denominator SS. This was discussed and can be found sources of variation in 2-way ANOVA lecture.

From the formulas, we can see that the major difference is that sometimes the Factor MS is divided by the error MS and sometimes it is divided by the interaction MS.

If the interaction term is NOT statistically significant, then the Interaction MS (mean square) estimates the Error MS. In other words, if the interaction term is not statistically significant it will be similar in magnitude as the Error MS. In this case there will be no large difference in the computed outcomes if the Factor A or B is fixed or random.

However, there will be times when the interaction is not significant but the interaction MS is still larger than the Error MS. Then there could be a difference in the F value for the Factor.

If the interaction term is Significant and the interaction MS is larger than the Error MS then there will be difference in F values for the Factors A and/or B. The F values will be smaller for the Factors MS that are divided by the interaction MS.

It is possible that they will become non-significant with the interaction MS as the denominator. So it will become Harder to detect a significant Factor effect if there is also a significant Interaction effect. A graphical representation will help us understand why we use the interaction MS in some instances as the denominator.

In fact, if the interaction is found to be statistically significant, we must then interpret the effects of factors with caution. In general, if the interaction is significant, then the main factors are generally not interpreted in the 2-way ANOVA. Instead, a series of one-way ANOVA’s are conducted holding one of the factors constant. For example, evaporative water loss (EWL) in frogs in the presence of air pollution (ozone) may depend on the relative humidity (RH) — if the RH is low, the frog may lose less body water at different concentrations of ozone than if the RH is moderate. Therefore, since the interaction is significant, the best thing to do is to look at the effects of ozone concentration on EWL at each level of saturation (RH).

This is a critical point in your understanding of complex ANOVA designs. Let us examine a case where there is a mildly significant interaction effect between two factors. In the first graph below (Fig 1) we see that Genotype 2 performs better on average (combining the two density treatments). If we are only interested in these two density treatments then it might be that the Genotype Factor is significant. This would be the case if Factor A is fixed and Factor B (density) is also fixed.

The Formula for Factor A would be: F = Genotype MS / error MS

Example of an interaction; genotype 1 has higher growth rate in density 2, genotype 2 does best in density 1 conditions.

Figure 1. Interaction example. At density D1, genotype 2 (red line) has higher growth rate; at density D2, the ranking switches: now, genotype 1 (black line) has higher growth rate.

However, it is likely that both Factor A (genotype) and Factor B (densities) are actually “samples” of many other possible genotypes and densities that we could examine.

Consider more than two genotypes raised in more than 2 densities (Fig 2). The outcome might look like

Interaction example expanded for multiple genotypes over multiple densities. 

Figure 2. Interaction example expanded for multiple genotypes over multiple densities. 

The graph (Fig 2) shows that genotype 2 does not do better than genotype 1 if we have more densities. If we also have other genotypes we see that there are other genotypes that have better (higher) responses than genotype 2.

    \begin{align*} F = \frac{Genotype \ MS}{Interaction \ MS} \end{align*}

In these cases it would have been more appropriate to calculate the F value for Factor A (genotype) using the interaction MS as the denominator. In Figure 1 there was some interaction this will make it harder to reject the null hypothesis that there is no effect of Factor A (genotype). So you must be careful to think about how you plan to interpret your data before you decide how to analyze the data using a Two-Factor ANOVA.

Questions

  1. Which of the following statements regarding fixed and random factors is true?
    A. With fixed factors, the subjects are selected by the researcher
    B. With fixed factors, the treatment levels are selected by the researcher at random from all possible levels
    C. With fixed factors, the subjects are selected at random by the researcher
    D. With random factors, the treatment levels are selected by the researcher at random from all possible levels
  2. Please write the equation for the one-way ANOVA with four levels of of fixed effects treatment factor A (you may wish to review Chapter 12.2)
  3. Selecting from all possible levels of a statin drug would be an impossible and meaningless experimental design. Explain why.
  4. For a multiway ANOVA design, when will the differences in the Random versus Fixed Factor make a difference?

Quiz Chapter 14.3

Fixed effects, random effects


Chapter 14 contents

12.3 – Fixed effects, random effects, and agreement

Introduction

Within discussions of one-way ANOVA models the distinction between two general classes of models needs to be made clear by the researcher. The distinction lies in how the levels of the factor are selected. If the researcher selects the levels, then the model is a Fixed Effects Model, also called a Model I ANOVA. On the other hand, if the levels of the factor were selected by random sampling from all possible levels of the factor, then the model is a Random Effects Model, also called a Model II ANOVA. This page is about such models and I’ll introduce the intraclass correlation coefficient, abbreviated as ICC, as a way to illustrate applications of Model II ANOVA.

Here’s an example to help the distinction. Consider an experiment to see if over the counter painkillers are as good as prescription pain relievers at reducing numbers of migraines over a six week period. The researcher selects Tylenol®, Advil®, Bayer® Aspirin, and Sumatriptan (Imitrex®), the latter an example of a medicine only available by prescription. This is clearly an example of fixed effects; the researcher selected the particular medicines for use.

Random effects, in contrast, implies that the researcher draws up a list of all over the counter pain relievers and draws at random three medicines; the researcher would also randomly select from a list of all available prescription medicines.

Fixed effects are probably the more common experimental approaches. To be complete, there is a third class of ANOVA called a Mixed Model or Model III ANOVA, but this type of model only applies to multidimensional ANOVA (eg, two-way ANOVA or higher), and we reserve our discussion of the Model III until we discuss multidimensional ANOVA (Table 1).

Table 1. ANOVA models

ANOVA model Treatments are
I Fixed effects
II Random effects
III Mixed, both fixed & random effects

Although the calculations for the one-way ANOVA under Model I or Model II are the same, the interpretation of the statistical significance is different between the two.

In Model I ANOVA, any statistical difference applies to the differences among the levels selected, but cannot be generalized back to the population. In contrast, statistical significance of the Factor variable in Model II ANOVA cannot be interpreted as specific differences among the levels of the treatment factor, but instead, apply to the population of levels of the factor. In short, Model I ANOVA results apply only to the study, whereas Model II ANOVA results may be interpreted as general effects, applicable to the population.

This distinction between fixed effects and random effects can be confusing, but it has broad implications for how we interpret our results in the short-term. This conceptual distinction between how the levels of the factor are selected also has general implications for our ability to acquire generalizable knowledge by meta-analysis techniques (Hunter and Schmidt 2000). Often we wish to generalize our results: we can do so only if the levels of the factor were randomly selected in the first place from all possible levels of the factor. In reality, this may not often be the case. It is not difficult to find examples in the published literature in which the experimental design is clearly fixed effects (ie, the researcher selected the treatment levels for a reason), and yet in the discussion of the statistical results, the researcher will lapse into generalizations.

Random Effects Models and agreement

Model II ANOVA is common in settings in which individuals are measured more than once. For example, in behavioral science or in sports science, subjects are typically measured for the response variable more than once over a course of several trials. Another common setting of Model II ANOVA is where more than one raters are judging an event or even a science project. In all of these cases what we are asking is about whether or not the subjects are consistent, in other words, we are asking about the precision of the instrument or measure.

In the assessment of learning by students, for example, different approaches may be tried and the instructor may wish to investigate whether the interventions can explain changes in test scores. To assess instrument validity — an item score from a grading rubric (Goertzen and Klaus 2023), tumor budding as biomarker for colorectal cancer (Bokhorst et al 2020) — agreement or concordance among two or more raters or judges (inter-rater reliability) needs to be established (same citations and references therein). For binomial data, we would be tempted to use the phi coefficient, Is it actually reliable Cohen’s kappa (two judges) or Fleiss Kappa (two or more judges) is a better tool. Phi coefficient and both assessment scores were introduced in Chapter 9.2.

For ratio scale data, agreement among scores of  two or more scores from judges There is an enormous number of articles on reliability measures in the social sciences and you should be aware of a classical paper on reliability by Shrout and Fleiss (1979) (see also McGraw and Wong, 1996). Both the ICC and the Pearson product moment correlation, r, which we will introduce in Chapter 16, are measures of strength of linear association between two ratio scale variables (Jinyuan et al 2016). But ICC is more appropriate for association between repeat measures of the same thing, eg, repeat measures of running speed or agreement between judges. In contrast, the product moment correlation can be used to describe association between any two variables, eg, between repeat measures of running speed, but also between say running speed and maximum jumping height. The concept of repeatability of individual behavior or other characteristics is also a common theme in genetics and so you should not be surprised to learn that the concept actually traces to RA Fisher and his invention of ANOVA. Just as in the case of the sociology literature, there are many papers on the use and interpretation of repeatability in the evolutionary biology literature (eg, Lessels and Boag 1987; Boake 1989; Falconer and Mackay 1996; Dohm 2002; Wolak et al 2012).

Agreement and repeatability are Model II (random effects) because the subjects, items, or raters being studied are treated as a random sample from a larger population rather than specific, fixed levels of interest. In reliability studies, subjects are not chosen for their specific characteristics but as representatives of the population variability. The goal is to infer whether a measurement tool works for any future subject or rater, not just the ones tested in the study. These factors are designed to estimate variance components and generalize results beyond the specific sample used. Put another way, the goal of Model II is often to estimate how much variance is due to differences between subjects versus within subjects (repeatability), which requires treating the subject effect as random.

There are many ways to analyze these kinds of data but a good way is to treat this problem as a one-way ANOVA with Random Effects. Thus, the Random Effects model permits the partitioning of the variation in the study into two portions: the amount that is due to differences among the subjects or judges or intervention versus the amount that is due to variation within the subjects themselves. The Factor is the Subjects and the levels of the factor are how ever many subjects are measured twice or more for the response variable.

If the subjects performance is repeatable, then the Mean Square Between (Among) Subjects, MS_{B}, component will be greater than the Mean Square Error component, MS_{W}, of the model. There are many measures of repeatability or reliability, but the intraclass correlation coefficient or ICC is one of the most common. The ICC may be calculated from the Mean Squares gathered from a Random Effects one-way ANOVA. ICC can take any value between zero and one.

    \begin{align*} ICC = \frac{s_{B}^2}{s_{B}^2-s_{W}^2} \end{align*}

where

    \begin{align*} s_{B}^2 = MS_{B}-\frac{MS_{W}}{k} \end{align*}

and

    \begin{align*} s_{W}^2 = MS_{W} \end{align*}

B and W refers to the among group (between or among groups mean square) and the within group components of variation (error mean square), respectively, from the ANOVA, MS refers to the Mean Squares, and k is the number of repeat measures for each experimental unit. In this formulation k is assumed to be the same for each subject.

By example, when a collection of sprinters run a race, if they ran it again, would the outcome be the same, or at least predictable? If the race is run over and over again and the runners cross the finish lines at different times each race, then much of the variation in performance times will be due to race differences largely independent of any performance abilities of the runners themselves and the Mean Square Error term will be large and the Between subjects Mean Square will be small. In contrast, if the race order is preserved race after race: Jenny is 1st, Ellen is 2nd, Michael is third, and so on, race after race, then differences in performance are largely due to individual differences. In this case, the Between-subjects Mean Square will be large as will the ICC, whereas the Mean Square for Error will be small, and so will the ICC.

Can the intraclass correlation be negative?

In theory, no. Values for ICC range between zero and one. The familiar Pearson product moment correlation, Chapter 16, takes any value between -1 and +1. However, in practice, negative values for ICC will result if MS_{B} < MS_{W}.

In other words, if the within group variability is greater than the among group variability, then negative ICC are possible. The question is whether negative estimates are ever biologically relevant or, simply result of limitations of the estimation procedure. Small ICC values and few repeats increases the risk of negative ICC estimates. Thus, a negative ICC would be “simply a(n) “unfortunate” estimate (Liljequist et al 2019). However, there may be situations in which negative ICC is more than unfortunate.  For example, repeatability, a quantitative genetics concept (Falconer and Mackay 1996), can be negative (Nakagawa and Schielzeth 2010).

    \begin{align*} r=\frac{V_{G}+V_{Eg}}{V_{P}} \end{align*}

Repeatability reflects the consistency of individual traits over time (or spatially, eg, scales on on a snake), is defined as the ratio of within individual differences and among individuals differences. Thus, within individual variance is attributed to general environmental effects, V_{Eg}, whereas variance among individuals is both genetic and environmental. Scenarios for which repeated measures on the same individual may occur under environmental conditions, termed specific environmental effects or V_{Es} by Falconer and Mackay (1996), that negatively affect performance are not hard to imagine — it remains to be determined how common these conditions are and whether or not estimates of repeatability are negative.

ICC Example

I extracted 15 data points from a figure about nitrogen metabolism in kidney patients following treatment with antibiotics (Figure 1, Mitch et al 1977). I used a web application called WebPlot Digitizer (https://apps.automeris.io/wpd/), but you can also accomplish this task within R via the digitize package. I was concerned about how steady my hand was using my laptop’s small touch screen, a problem that very much can be answered by thinking statistically, and taking advantage of the ICC. So, rather than taking just one estimate of each point I repeated the protocol for extracting the points from the figure three times generating a total of three points for each of the 15 data points or 45 points in all. How consistent was I?

Let’s look at the results just for three points, #1, 2, and 3.

In the R script window enter

points = c(1,2,3,1,2,3,1,2,3)

Change points to character so that the ANOVA command will treat the numbers as factor levels.

points = as.character(points)
extracts = c(2.0478, 12.2555, 16.0489, 2.0478, 11.9637, 16.0489, 2.0478, 12.2555, 16.0489)

Make a data frame, assign to an object, eg, “digitizer”

digitizer = data.frame(points, extracts)

The dataset “digitizer” should now be attached and available to you within Rcmdr. Select digitizer data set and proceed with the one-way ANOVA.

Output from oneway ANOVA command

Model.1 <- aov(extracts ~ points, data=digitizer)

 summary(Model.1)
            Df    Sum Sq   Mean Sq  F value     Pr(>F)    
points       2 313.38895 156.69448 16562.49 5.9395e-12 ***
Residuals    6   0.05676   0.00946                        
---
Signif. codes:  0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1

numSummary(extracts , groups = points, 
+   statistics=c("mean", "sd"))
         mean           sd data:n
1  2.04780000 0.0000000000      3
2 12.15823333 0.1684708085      3
3 16.04890000 0.0000000000      3

End R output. We need to calculate the ICC

    \begin{align*} ICC = \frac{156.69448-\frac{0.00946}{3}}{156.69448-\frac{0.00946}{3}+0.00946} = 0.9999 \end{align*}

I’d say that’s pretty repeatable and highly precise measurement!

But is it accurate? You should be able to disentangle accuracy from precision based on our previous discussion (Chapter 3.5), but now in the context of a practical way to quantify precision.

ICC calculations in R

We could continue to calculate the ICC by hand, but better to have a function. Here’s a crack at the function to calculate ICC along with a 95% confidence interval.

myICC <- function(m, k, dfN, dfD) {
 testMe <- anova(m) 
 MSB <- testMe$"Mean Sq"[1]
 MSE <- testMe$"Mean Sq"[2]
 varB <- MSB - MSE/k
 ICC <- varB/(varB+MSE)
 fval <- qf(c(.025), df1=dfN, df2=dfD, lower.tail=TRUE)
 CI = (k*MSE*fval)/(MSB+MSE*(k-1)*fval)
 LCIR = ICC-CI
 UCIR = ICC+CI
 myList = c(ICC, LCIR, UCIR)
 return(myList)
}

The user supplies the ANOVA model object (eg, Model.1 from our example), k, which is the number of repeats per unit, and degrees of freedom for the among groups comparison (2 in this example), and the error mean square (6 in this case). Our example, run the function

m2ICC = myICC(Model.1, 3, 2,6); m2ICC

and R returns

[1] 0.9999396 0.9999350 0.9999442

with the ICC reported first, 0.9999396, followed by the lower limit (0.9999350) and the upper limit (0.9999442) of the 95% confidence interval.

Instead of writing your own function, several R packages can calculate the intraclass correlation coefficient (ICC) and its variants. These packages include: irr, psy, psych, and rptR.  For complex experiments involving multiple predictor variables, these packages are helpful for obtaining the correct ICC calculation (cf Shrout and Fleiss 1979; McGraw and Wong 1996). For the one-way ANOVA it is easier to just extract the information you need from the ANOVA table and run the calculation directly. We do so for a couple of examples.

Example: Are marathon runners consistent more consistent than my commute times?

Honolulu Marathon 2024 participant. Image credit: Pdubs.94, licensed under CC BY 4.0 (Creative Commons Attribution 4.0 International License), via Wikipedia.

Figure 1.Honolulu Marathon 2024 participant. Image credit: Pdubs.94, licensed under CC BY 4.0 (Creative Commons Attribution 4.0 International License), via Wikipedia.

A marathon is 26 miles, 385 yards (42.195 kilometers) long (World Athletics.org). And yet, tens of thousands of people choose to run in these events. For many, running a marathon is a one-off, the culmination of a personal fitness goal. For others, it’s a passion and a few are simply extraordinary, elite runners who can complete the courses in 2 to 3 hours (Table 3). That’s about 12.5 miles per hour. For comparison, my 20 mile commute on the H1 freeway on Oahu typically takes about 40 minutes to complete, or 27 miles per hour (Table 2, yes, I keep track of my commute times, per Galton’s famous maxim: “Whenever you can, count”).

View west along Interstate H-1 (Lunalilo Freeway) at 6AM, Honolulu, Oahu, Hawaii. Image used with permission, credit: S. Dohm.

Figure 2. View west along Interstate H-1 (Lunalilo Freeway) at 6:30AM on a weekday, Honolulu, Oahu, Hawaii. Image used with permission, credit: S. Dohm.

Table 2. A sampling of average commute speeds, miles per hour (mph),  on the H1 freeway during DrD’s morning commute (distance about 19.3 miles).

WeekMondayTuesdayWednesdayThursdayFriday
128.523.828.530.226.9
225.822.429.326.227.7
326.222.624.924.234.3
423.326.931.326.230.2

Try on your own.

Calculate the ICC for my commute speeds.

Run the one-way ANOVA to get the necessary mean squares and input the values into our ICC function. We have

require(psych)
m2ICC = myICC(AnovaModel.1, 4, 4,11); m2ICC
[1] 0.7390535 0.6061784 0.8719286

Repeatability, as estimated by the ICC, was 0.74 (95% CI 0.606, 0.872), for repeat measures of commute times.

We can ask the same about marathon runners — how consistent from race to race are these runners? The following data are race times drawn from a sample of runners who completed the Honolulu Marathon in both 2016 and 2017 in 2 to 3 hours (times recorded in minutes). In other words, are elite runners consistent?

Table 3. Honolulu marathon running times (min) for eleven repeat, elite runners.

IDTime
P1179.9
P1192.0
P2129.9
P2130.8
P3128.5
P3129.6
P4179.4
P4179.7
P5174.3
P5181.7
P6177.2
P6176.2
P7169.0
P7173.4
P8174.1
P8175.2
P9175.1
P9174.2
P10163.9
P10175.9
P11179.3
P11179.8

Stacked worksheet, P refers to person, therefore P1 is person ID 1, P2 is person ID 2, and so on.

After running a one-way ANOVA, here are results for the marathon runners

m2ICC = myICC(Model.1, 2, 10,11); m2ICC
[1] 0.9780464 0.9660059 0.9900868

Repeatability, as estimated by the ICC, was 0.98 (95% CI 0.966, 0.990), for repeat measures of marathon performance. Put more simply, knowing what a runner did in 2016 I would be able to predict their 2017 race performance with high confidence, 98%!

And now, we compare: the runners are more consistent!

Clearly this is an apples-to-oranges comparison, but it gives us a chance to think about how we might make such comparisons. The ICC will change because of differences among individuals. For example, if individuals are not variable, if everyone finishes the race at the same time, then the ICC will be low (or even zero).  Because there is no variance to be attributed to individual differences, the proportion of total variance explained by those differences will be being minimal.

Visualizing repeated measures.

Waterfall plot.

If just two measures, then a profile plot will do. We introduced profile plots in the paired t-test chapter (Ch10.3). For more than two repeated measures, a line graph will do (see question 9 below). Another graphic increasingly used is the waterfall plot (waterfall chart), particularly for before and after measures or other comparisons against a baseline. For example, it’s a natural question for our eleven marathon runners — how much did they improve on the second attempt of the same course? Figure 3 shows my effort to provide a visual.

Simple waterfall plot or race improvement for Table 3 data. Dashed horizontal line at zero.

Figure 3. Simple waterfall plot of race improvement for Table 3 data. Dashed horizontal line at zero.

R code

Data wrangling: Create a new variable, eg, race21, by subtracting the first time from the second time. Multiply by -1 so that improvement (faster time in second race) is positive. Finally, apply descending sort so that improvement is on the left and increasingly poorer times are on the right.

df <- data.frame(
ID = rep(paste0("P", 1:11), each = 2),
Time = c(179.9,192.0,
129.9,130.8,
128.5,129.6,
179.4,179.7,
174.3,181.7,
177.2,176.2,
169.0,173.4,
174.1,175.2,
175.1,174.2,
163.9,175.9,
179.3,179.8)
)

# Convert to wide format
df_wide <- reshape(df, idvar = "ID", timevar = "ID",
direction = "wide")

# Extract first and second times manually
time_matrix <- matrix(df$Time, ncol = 2, byrow = TRUE)
colnames(time_matrix) <- c("Time1", "Time2")

# Calculate improvement (second - first)
base21 <- time_matrix[,2] - time_matrix[,1]

# Sort for waterfall display
base21 <- sort(base21, decreasing = TRUE)

# Waterfall plot
barplot(base21, col="blue", border="blue", space=0.5,
ylim=c(-20,20),
ylab="Improvement from first race (min)")
abline(h=0, lty=2)

Interpretation.

From the graph (Fig 3) we see that few of the racers improved their times the second time around — a few had decreased times.

Spaghetti plots.

Spaghetti plots, which display individual trajectories over time or multiple measurements, is a specialized line plot effective for visualizing the consistency of performance. The vertical thickness or spread of a single subject’s line across measurements indicates their level of repeatability. A “flat” line indicates high repeatability, while a volatile, zig-zagging line indicates low repeatability. Spaghetti plots are useful to visualize growth, decline, or stability over time for a single performance measure.

A spaghetti plot of average commute speeds from Table 2 data. 

Figure 4. A spaghetti plot of average commute speeds from Table 2 data. 

R code

# 1. Enter the data from Table 2.
commute <- data.frame(
Week = 1:4,
Monday = c(28.5, 25.8, 26.2, 23.3),
Tuesday = c(23.8, 22.4, 22.6, 26.9),
Wednesday = c(28.5, 29.3, 24.9, 31.3),
Thursday = c(30.2, 26.2, 24.2, 26.2),
Friday = c(26.9, 27.7, 34.3, 30.2)
)

head(commute)

# 2. Set the X-axis points
days <- 1:5
day_names <- c("Mon", "Tue", "Wed", "Thu", "Fri")

# 3. Create empty plot
# Find range of Y to ensure all data fits
ymin <- min(commute[, 2:6]) - 2
ymax <- max(commute[, 2:6]) + 2

plot(NA, xlim = c(1, 5), ylim = c(ymin, ymax),
xlab = "Day of Week", ylab = "Average Speed (mph)",
main = "Weekly Commute Speeds", xaxt = "n")

# Add X-axis labels
axis(1, at = 1:5, labels = day_names)

# 4. Add lines for each week (spaghetti)
colors <- c("red", "blue", "darkgreen", "purple")
for (i in 1:nrow(commute)) {
# Plot lines
lines(days, commute[i, 2:6], col = colors[i], lwd = 2, type = "b", pch = 19)
}

# 5. Add legend
legend("topleft", legend = paste("Week", 1:4), col = colors, lwd = 2, pch = 19)

Parallel coordinates plots.

Parallel coordinates plot allows each entity to be represented by a unique, continuous line traced across several vertical axes (each representing a different measurement or variable). High repeatability is indicated when a line remains roughly horizontal across all axes, while poor repeatability is visualized as lines that zig-zag or change direction between measurements.

A parallel coordinates plot of average commute speeds from Table 2 data.

Figure 5. A parallel coordinates plot of average commute speeds from Table 2 data.

Both spaghetti plots and parallel coordinate plots can be used to visualize consistency of performance over multiple measures, but they differ significantly in their handling of data, scale, and context. Spaghetti plots are best for tracking within-subject variability over time, but quickly become a mess — like pasta tossed and sticking to a wall — when subject count increases. Parallel coordinate plots are a multivariate generalization — each line represents an individual’s profile across different tests, eg, accuracy, speed, VO2max.

R code.

# 1. Create the data frame
# see above

# 2. Extract only the numerical commute data for plotting
commute_data <- commute[, 2:6]

# 3. Setup the plot area
# Define range for y-axis
y_range <- range(commute_data)
# Define number of days (5)
num_days <- ncol(commute_data)

# Create empty plot
plot(NA, xlim = c(1, num_days), ylim = y_range, 
xaxt = "n", xlab = "Day of Week", ylab = "Average Speed (mph)",
main = "Average Commute Speed: Weekly Comparison")

# Add x-axis labels
axis(1, at = 1:num_days, labels = colnames(commute_data))

# 4. Draw vertical lines for each day
for (i in 1:num_days) {
lines(c(i, i), y_range, col = "grey", lty = 2)
}

# 5. Add lines for each week (repeated measures)
colors <- c("red", "blue", "darkgreen", "orange")
for (i in 1:nrow(commute_data)) {
lines(1:num_days, commute_data[i, ], col = colors[i], lwd = 2, type = "b", pch = 19)
}

# 6. Add legend
legend("topleft", legend = paste("Week", commute$Week), col = colors, lwd = 2, pch = 19)

Heat map.

Heat map quickly identify patterns, such as an individual’s performance improving, degrading, or remaining stable over time. For example, uniform, solid colors across rows (individuals) or columns (time/trials) indicate high consistency, while varied colors indicate poor repeatability.

A heat map of the commute speeds data set.

Figure 6. A heat map of the commute speeds data set.

R code.

# 1. Create the data
# Week is excluded from matrix to only include numerical commute times
commute_data <- matrix(c(
28.5, 23.8, 28.5, 30.2, 26.9,
25.8, 22.4, 29.3, 26.2, 27.7,
26.2, 22.6, 24.9, 24.2, 34.3,
23.3, 26.9, 31.3, 26.2, 30.2
), nrow = 4, byrow = TRUE)

# 2. Add row and column names
rownames(commute_data) <- c("Week 1", "Week 2", "Week 3", "Week 4")
colnames(commute_data) <- c("Mon", "Tue", "Wed", "Thu", "Fri")

# 3. Create the heatmap
# Use t() to transpose so weeks are rows and days are columns
# Use rev() on rows to have Week 1 at the top
image_data <- t(apply(commute_data, 2, rev))

# Set up the plotting area (margins for labels)
par(mar = c(5, 6, 4, 2))

# Plot the matrix
# Option 1
image(1:nrow(image_data), 1:ncol(image_data), image_data,
col = heat.colors(12),
axes = FALSE, xlab = "Days", ylab = "",
main = "Commute Times Heatmap")

# Add axes labels
axis(1, at = 1:5, labels = colnames(commute_data))
axis(2, at = 1:4, labels = rev(rownames(commute_data)), las = 1)
box()

Interpretation.

The graphs (Fig 4 – 6) show a lack of trends in consistency of drive speeds. The heat map suggests more consistency on Monday — low speeds (Fig 4 & Fig 5), but that’s a stretch.

An example for you to work, from our Measurement Day

If you recall, we had you calculate length and width measures on shells from samples of gastropod and bivalve species. In the table are repeated measures of shell length, by caliper in mm, for a sample of Conus shells (Fig 7 and Table 4).

Conus shells

Figure 7. Conus shells, image by M. Dohm.

Repeated measures of the same 12 Conus shells.

Table 4. Unstacked dataset of repeated length measures on 12 shells

SampleMeasure1Measure2Measure3
145.7446.4446.79
248.7949.4149.67
352.7953.4553.36
452.7453.1453.14
553.2553.4553.15
653.2553.6453.65
731.1831.5931.44
840.7341.0341.11
943.1843.2343.2
1047.147.6447.64
1149.5350.3250.24
1253.9654.554.56

Maximum length from from anterior end to apex with calipers. Repeated measures were conducted blind to shell id, sampling was random over three separate time periods.

Questions

  1. Consider data in Table 2, Table 3, and Table 4. True or False: The arithmetic mean is an appropriate measure of central tendency. Explain your answer.
  2. Enter the shell data into R; Best to copy and stack the data in your spreadsheet, then import into R or R Commander. Once imported, don’t forget to change Sample to character, otherwise R will treat Sample as ratio scale data type. Run your one-way ANOVA and calculate the intraclass correlation (ICC) for the dataset. Is the shell length measure repeatable?
  3. The marathon results in Table 2 are paired data, results of two races run by the same individuals. Make a profile plot for the data (see Ch10.3 for help).
  4. A profile plot is used to show paired data; for two or more repeat measures, use a line graph instead.
    A. Produce a line graph for data in Table 4. The line graph is an option in R Commander (Rcmdr: Graphs Line graph…), and can be generated with the following simple R code:
lineplot(Sample, Measure1, Measure2, Measure3)

B. What trends, if any, do you see? Report your observations.
C. Make another line plot for data in Table 2. Report any trend and include your observations.

Quiz Chapter 12.3

Fixed effects, random effects, and agreement


Chapter 12 contents