This text provides an introduction to multiple hypothesis testing. It covers various error criteria and testing procedures, and includes references to relevant R packages. An earlier version of this text served as the lecture notes for a PhD-level course on multiple testing.

MaxT is a highly popular resampling-based multiple testing procedure, which controls the familywise error rate (FWER) and is powerful under dependence. This paper generalizes maxT to what we term ``multi-resolution'' False Discovery eXceedance (FDX) control. FDX control means ensuring that the FDP -- the proportion of false discoveries among all rejections -- is at most $γ$ with probability at least $1-α$. Here $γ$ and $α$ are prespecified, small values between 0 and 1. If we take $γ=0$, FDX control is the same as FWER control. When $γ=0$, the new procedure reduces to maxT. For $γ>0$, our method has much higher power. Our method is then strongly connected to maxT as well. For example, if our method rejects fewer than $γ^{-1}$ hypotheses, then maxT rejects the same. Further, the implied global tests of the two methods are the same, although the implied closed testing procedures differ. Finally, our method provides an easy-to-use simultaneous, multi-resolution guarantee: the procedure outputs a single rejection threshold $q$, but ensures that with probability $1-α$, simultaneously over all stricter thresholds, the corresponding FDPs are also below $γ$.

Despite the versatility of generalized linear mixed models in handling complex experimental designs, they often suffer from misspecification and convergence problems. This makes inference on the values of coefficients problematic. In addition, the researcher's choice of random and fixed effects directly affects statistical inference correctness. To address these challenges, we propose a robust extension of the "two-stage summary statistics" approach using sign-flipping transformations of the score statistic in the second stage. Our approach efficiently handles within-variance structure and heteroscedasticity, ensuring accurate regression coefficient testing for 2-level hierarchical data structures. The approach is illustrated by analyzing the reduction of health issues over time for newly adopted children. The model is characterized by a binomial response with unbalanced frequencies and several categorical and continuous predictors. The proposed approach efficiently deals with critical problems related to longitudinal nonlinear models, surpassing common statistical approaches such as generalized estimating equations and generalized linear mixed models.

When testing many hypotheses, often we do not have strong expectations about the directions of the effects. In some situations however, the alternative hypotheses are that the parameters lie in a certain direction or interval, and it is in fact expected that most hypotheses are false. This is often the case when researchers perform multiple noninferiority or equivalence tests, e.g. when testing food safety with metabolite data. The goal is then to use data to corroborate the expectation that most hypotheses are false. We propose a nonparametric multiple testing approach that is powerful in such situations. If the user's expectations are wrong, our approach will still be valid but have low power. Of course all multiple testing methods become more powerful when appropriate one-sided instead of two-sided tests are used, but our approach often has superior power then. The proposed methods are not at all limited to safety testing and can be used for testing hypotheses about various kinds of parameters, such as coefficients of a model. The methods in this paper control the median of the false discovery proportion (FDP), which is the fraction of false discoveries among the rejected hypotheses. This approach is comparable to false discovery rate control, where one ensures that the mean rather than the median of the FDP is small. Our procedures make use of a symmetry property of the test statistics, do not require independence and have finite-sample properties.

Generalized linear models usually assume a common dispersion parameter, an assumption that is seldom true in practice. Consequently, standard parametric methods may suffer appreciable loss of type I error control. As an alternative, we present a semi-parametric group-invariance method based on sign flipping of score contributions. Our method requires only the correct specification of the mean model, but is robust against any misspecification of the variance. We present tests for single as well as multiple regression coefficients. The test is asymptotically valid but shows excellent performance in small samples. We illustrate the method using RNA sequencing count data, for which it is difficult to model the overdispersion correctly. The method is available in the R library flipscores.

Magnetic resonance imaging (MRI) has become a key technique in the study of multiple sclerosis. In a typical analysis pipeline binary lesion maps are typically modelled voxelwise using either standard linear models or logistic regression, in order to detect relationships with variables of interest. Fitting linear models does not respect the binary nature of the data and, as we shall show, this can lead to false positives. Instead voxelwise logistic regression correctly models the data however it suffers from a lack of power. Smoothing is a key means of increasing the signal to noise ratio in neuroimaging data but has previously been thought not to be applicable to lesion based logistic regression as, when applied to the 0/1 maps, it destroys the binary nature of the data [9]. In this work we show how to combine smoothing with logistic regression modelling: increasing the power of inference whilst maintaining validity. In particular we develop a test based on sign-flipping the smooth effective score contributions and control for multiple testing using the sign-flipped distribution of the maximum smoothed score test-statistic.

In many applied sciences a popular analysis strategy for high-dimensional data is to fit many multivariate generalized linear models in parallel. This paper presents a novel approach to address the resulting multiple testing problem by combining a recently developed sign-flip test with permutation-based multiple-testing procedures. Our method builds upon the univariate Flipscores test which offers robustness against misspecified variances in generalized linear models, a crucial feature in high-dimensional settings where comprehensive model validation is particularly challenging. We extend this approach to the multivariate setting, enabling adaptation to unknown response correlation structures. This approach yields relevant power improvements over conventional multiple testing methods when correlation is present.

Despite the versatility of generalized linear mixed models in handling complex experimental designs, they often suffer from misspecification and convergence problems. This makes inference on the values of coefficients problematic. To address these challenges, we propose a robust extension of the score-based statistical test using sign-flipping transformations. Our approach efficiently handles within-variance structure and heteroscedasticity, ensuring accurate regression coefficient testing. The approach is illustrated by analyzing the reduction of health issues over time for newly adopted children. The model is characterized by a binomial response with unbalanced frequencies and several categorical and continuous predictors. The proposed approach efficiently deals with critical problems related to longitudinal nonlinear models, surpassing common statistical approaches such as generalized estimating equations and generalized linear mixed models.

We introduce a multiple testing procedure that controls the median of the proportion of false discoveries (FDP) in a flexible way. The procedure only requires a vector of p-values as input and is comparable to the Benjamini-Hochberg method, which controls the mean of the FDP. Our method allows freely choosing one or several values of alpha after seeing the data -- unlike Benjamini-Hochberg, which can be very liberal when alpha is chosen post hoc. We prove these claims and illustrate them with simulations. Our procedure is inspired by a popular estimator of the total number of true hypotheses. We adapt this estimator to provide simultaneously median unbiased estimators of the FDP, valid for finite samples. This simultaneity allows for the claimed flexibility. Our approach does not assume independence. The time complexity of our method is linear in the number of hypotheses, after sorting the p-values.

There exists no consensus on the meaning of the term "randomization test". Contradicting uses of the term are leading to confusion, misunderstandings and indeed invalid data analyses. As we point out, a main source of the confusion is that the term was not explicitly defined when it was first used in the 1930's. Later authors made clear proposals to reach a consensus regarding the term. This resulted in some level of agreement around the 1970's. However, in the last few decades, the term has often been used in ways that contradict these proposals. This paper provides an overview of the history of the term per se, for the first time tracing it back to 1937. This will hopefully lead to more agreement on terminology and less confusion on the related fundamental concepts.

Equivalence testing is an important component of safety assessments, used for example by the European Food Safety Authority, to allow new food or feed products on the market. The aim of such tests is to demonstrate equivalence of characteristics of test and reference crops. Equivalence tests are typically univariate and applied to each measured analyte (characteristic) separately without multiplicity correction. This increases the probability of making false claims of equivalence (type I errors) when evaluating multiple analytes simultaneously. To solve this problem, familywise error rate (FWER) control using Hochberg's method has been proposed. This paper demonstrates that, in the context of equivalence testing, other FWER-controlling methods are more powerful than Hochberg's. Particularly, it is shown that Hommel's method is guaranteed to perform at least as well as Hochberg's and that an "adaptive" version of Bonferroni's method, which uses an estimator of the proportion of non-equivalent characteristics, often substantially outperforms Hommel's method. Adaptive Bonferroni takes better advantage of the particular context of food safety where a large proportion of true equivalences is expected, a situation where other methods are particularly conservative. The different methods are illustrated by their application to two compositional datasets and further assessed and compared using simulated data.

Summary We consider testing invariance of a distribution under an algebraic group of transformations, such as permutations or sign flips. As such groups are typically huge, tests based on the full group are often computationally infeasible. Hence, it is standard practice to use a random subset of transformations. We improve upon this by replacing the random subset with a strategically chosen, fixed subgroup of transformations. In a generalized location model, we show that the resulting tests are often consistent for lower signal-to-noise ratios. Moreover, we establish an analogy between the power improvement and switching from a t-test to a Z-test under normality. Importantly, in permutation-based multiple testing, the efficiency gain with our approach can be huge, since we attain the same power with many fewer permutations.

We propose a permutation-based method for testing a large collection of hypotheses simultaneously. Our method provides lower bounds for the number of true discoveries in any selected subset of hypotheses. These bounds are simultaneously valid with high confidence. The methodology is particularly useful in functional Magnetic Resonance Imaging cluster analysis, where it provides a confidence statement on the percentage of truly activated voxels within clusters of voxels, avoiding the well-known spatial specificity paradox. We offer a user-friendly tool to estimate the percentage of true discoveries for each cluster while controlling the family-wise error rate for multiple testing and taking into account that the cluster was chosen in a data-driven way. The method adapts to the spatial correlation structure that characterizes functional Magnetic Resonance Imaging data, gaining power over parametric approaches.

Mediation analysis in high-dimensional settings often involves identifying potential mediators among a large number of measured variables. For this purpose, a two-step familywise error rate procedure called ScreenMin has been recently proposed. In ScreenMin, variables are first screened and only those that pass the screening are tested. The proposed data-independent threshold for selection has been shown to guarantee asymptotic familywise error rate. In this work, we investigate the impact of the threshold on the finite-sample familywise error rate. We derive a power maximizing threshold and show that it is well approximated by an adaptive threshold of Wang et al. (2016, arXiv preprint arXiv:1610.03330). We illustrate the investigated procedures on a case-control study examining the effect of fish intake on the risk of colorectal adenoma. We also apply our procedure in the context of replicability analysis to identify single nucleotide polymorphisms (SNP) associated with crop yield in two distinct environments.

Non-parametric tests based on permutation, rotation or sign-flipping are examples of group-invariance tests. These tests test invariance of the null distribution under a set of transformations that has a group structure, in the algebraic sense. Such groups are often huge, which makes it computationally infeasible to test using the entire group. Hence, it is standard practice to test using a randomly sampled set of transformations from the group. This random sample still needs to be substantial to obtain good power and replicability. We improve upon this standard practice by using a well-designed subgroup of transformations instead of a random sample. The resulting subgroup-invariance test is still exact, as invariance under a group implies invariance under its subgroups. We illustrate this in a generalized location model and obtain more powerful tests based on the same number of transformations. In particular, we show that a subgroup-invariance test is consistent for lower signal-to-noise ratios than a test based on a random sample. For the special case of a normal location model and a particular design of the subgroup, we show that the power improvement is equivalent to the power difference between a Monte Carlo Z-test and a Monte Carlo t-test.

We consider the class of all multiple testing methods controlling tail probabilities of the false discovery proportion, either for one random set or simultaneously for many such sets. This class encompasses methods controlling familywise error rate, generalized familywise error rate, false discovery exceedance, joint error rate, simultaneous control of all false discovery proportions, and others, as well as seemingly unrelated methods such as gene set testing in genomics and cluster inference methods in neuroimaging. We show that all such methods are either equivalent to a closed testing method, or are uniformly improved by one. Moreover, we show that a closed testing method is admissible as a method controlling tail probabilities of false discovery proportions if and only if all its local tests are admissible. This implies that, when designing such methods, it is sufficient to restrict attention to closed testing methods only. We demonstrate the practical usefulness of this design principle by constructing a uniform improvement of a recently proposed method.

Permutation testing in linear models, where the number of nuisance coefficients is smaller than the sample size, is a well-studied topic. The common approach of such tests is to permute residuals after regressing on the nuisance covariates. Permutation-based tests are valuable in particular because they can be highly robust to violations of the standard linear model, such as non-normality and heteroscedasticity. Moreover, in some cases they can be combined with existing, powerful permutation-based multiple testing methods. Here, we propose permutation tests for models where the number of nuisance coefficients exceeds the sample size. The performance of the novel tests is investigated with simulations. In a wide range of simulation scenarios our proposed permutation methods provided appropriate type I error rate control, unlike some competing tests, while having good power.

The statistical literature is known to be inconsistent in the use of the terms 'permutation test' and 'randomisation test'. Several authors successfully argue that these terms should be used to refer to two distinct classes of tests and that there are major conceptual differences between these classes. The present paper explains an important difference in mathematical reasoning between these classes: a permutation test fundamentally requires that the set of permutations has a group structure, in the algebraic sense; the reasoning behind a randomisation test is not based on such a group structure, and it is possible to use an experimental design that does not correspond to a group. In particular, we can use a randomisation scheme where the number of possible treatment patterns is larger than in standard experimental designs. This leads to exact p values of improved resolution, providing increased power for very small significance levels, at the cost of decreased power for larger significance levels. We discuss applications in randomised trials and elsewhere. Further, we explain that Fisher's famous Lady Tasting Tea experiment, which is commonly referred to as the first permutation test, is in fact a randomisation test. This distinction is important to avoid confusion and invalid tests.

Generalized linear models are often misspecified due to overdispersion, heteroscedasticity and ignored nuisance variables. Existing quasi-likelihood methods for testing in misspecified models often do not provide satisfactory type-I error rate control. We provide a novel semi-parametric test, based on sign-flipping individual score contributions. The tested parameter is allowed to be multi-dimensional and even high-dimensional. Our test is often robust against the mentioned forms of misspecification and provides better type-I error control than its competitors. When nuisance parameters are estimated, our basic test becomes conservative. We show how to take nuisance estimation into account to obtain an asymptotically exact test. Our proposed test is asymptotically equivalent to its parametric counterpart.