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Phylogenomic subsampling and upsampling for efficient evolutionary analyses of big data.

Long runtimes, high memory demands, and reliance on high-performance computing impede phylogenomic analyses. We review a scalable phylogenomic subsampling with upsampling (PSU) framework, in which small subsamples of sites from a concatenated alignment are expanded by upsampling before inference, and the resulting analyses are then aggregated to obtain evolutionary estimates. PSU harnesses the fact that the computational cost of maximum likelihood analysis is strongly influenced by the number of distinct site patterns in the concatenated alignment, whereas statistical power depends primarily on the amount of evolutionary information represented by the total number of sites and substitutions. By reducing the former while restoring the latter through upsampling, PSU can approximate many full-data analyses at substantially lower computational cost. Analysis of simulated and empirical datasets shows that PSU can accurately estimate bootstrap support values, select the optimal substitution model, test evolutionary hypotheses, and infer branch lengths, divergence times, and associated uncertainty measures, while reducing runtime and memory requirements by orders of magnitude. PSU also provides distributions of inferred clade support across independent subsamples, enabling detection of conflicting phylogenetic signals that may remain hidden in conventional bootstrap analysis. Automated tuning of subsample size, the number of subsamples, and the number of upsampling replicates make PSU practical across diverse datasets. We suggest that PSU is a general strategy for scalable phylogenomic inference using a broad range of statistical methods. By enabling analyses of genome-scale alignments on commodity hardware, PSU broadens research access and reduces environmental and infrastructural costs of big-data phylogenomics.

confidence limits↗

Bayesian hierarchical error model for analysis of gene expression data.

MOTIVATION: Analysis of genome-wide microarray data requires the estimation of a large number of genetic parameters for individual genes and their interaction expression patterns under multiple biological conditions. The sources of microarray error variability comprises various biological and experimental factors, such as biological and individual replication, sample preparation, hybridization and image processing. Moreover, the same gene often shows quite heterogeneous error variability under different biological and experimental conditions, which must be estimated separately for evaluating the statistical significance of differential expression patterns. Widely used linear modeling approaches are limited because they do not allow simultaneous modeling and inference on the large number of these genetic parameters and heterogeneous error components on different genes, different biological and experimental conditions, and varying intensity ranges in microarray data. RESULTS: We propose a Bayesian hierarchical error model (HEM) to overcome the above restrictions. HEM accounts for heterogeneous error variability in an oligonucleotide microarray experiment. The error variability is decomposed into two components (experimental and biological errors) when both biological and experimental replicates are available. Our HEM inference is based on Markov chain Monte Carlo to estimate a large number of parameters from a single-likelihood function for all genes. An F-like summary statistic is proposed to identify differentially expressed genes under multiple conditions based on the HEM estimation. The performance of HEM and its F-like statistic was examined with simulated data and two published microarray datasets-primate brain data and mouse B-cell development data. HEM was also compared with ANOVA using simulated data. AVAILABILITY: The software for the HEM is available from the authors upon request.

Algorithms↗

Exact inference for growth curves with intraclass correlation structure.

We consider repeated observations taken over time for each of several subjects. For example, one might consider the growth curve of a cohort of babies over time. We assume a simple linear growth curve model. Exact results based on sufficient statistics (exact tests of the null hypothesis that a coefficient is zero, or exact confidence intervals for coefficients) are not available to make inference on regression coefficients when an intraclass correlation structure is assumed. This paper will demonstrate that such exact inference is possible using generalized inference.

Biometry↗

Markovian domain fingerprinting: statistical segmentation of protein sequences.

MOTIVATION: Characterization of a protein family by its distinct sequence domains is crucial for functional annotation and correct classification of newly discovered proteins. Conventional Multiple Sequence Alignment (MSA) based methods find difficulties when faced with heterogeneous groups of proteins. However, even many families of proteins that do share a common domain contain instances of several other domains, without any common underlying linear ordering. Ignoring this modularity may lead to poor or even false classification results. An automated method that can analyze a group of proteins into the sequence domains it contains is therefore highly desirable. RESULTS: We apply a novel method to the problem of protein domain detection. The method takes as input an unaligned group of protein sequences. It segments them and clusters the segments into groups sharing the same underlying statistics. A Variable Memory Markov (VMM) model is built using a Prediction Suffix Tree (PST) data structure for each group of segments. Refinement is achieved by letting the PSTs compete over the segments, and a deterministic annealing framework infers the number of underlying PST models while avoiding many inferior solutions. We show that regions of similar statistics correlate well with protein sequence domains, by matching a unique signature to each domain. This is done in a fully automated manner, and does not require or attempt an MSA. Several representative cases are analyzed. We identify a protein fusion event, refine an HMM superfamily classification into the underlying families the HMM cannot separate, and detect all 12 instances of a short domain in a group of 396 sequences. CONTACT: jill@cs.huji.ac.il; tishby@cs.huji.ac.il.

Algorithms↗

Genetics and population history of Caucasus populations.

We describe aspects of genetic diversity in several ethnic populations of the Caucasus Mountains of Daghestan using mitochondrial DNA sequences and a sample of 100 polymorphic Alu insertion loci. The mitochondrial DNA (mtDNA) sequences are like those of Europe. Principal coordinates and nearest neighbor statistics show that there is little detectable structure in the distances among populations computed from mtDNA. The Alu frequencies of the Caucasus populations suggest that they have undergone more genetic drift than most other groups since the dispersal of modern humans. Genetic differences among these populations are not large; instead, they are of the same order as distances among populations of Europe. We compare two methods of inference about the demography of ancient colonizing populations from Africa, one based on conventional FST statistics and one based on mean Alu insertion frequencies. The two approaches agree reasonably well if we assume that there was demographic growth in Africa before the diaspora of ancestors of contemporary regional human groups outside Africa.

DNA, Mitochondrial↗

Fuzzy modelling for selecting headgear types.

The purpose of this study was to develop a computer-assisted inference model for selecting appropriate types of headgear appliance for orthodontic patients and to investigate its clinical versatility as a decision-making aid for inexperienced clinicians. Fuzzy rule bases were created for degrees of overjet, overbite, and mandibular plane angle variables, respectively, according to subjective criteria based on the clinical experience and knowledge of the authors. The rules were then transformed into membership functions and the geometric mean aggregation was performed to develop the inference model. The resultant fuzzy logic was then tested on 85 cases in which the patients had been diagnosed as requiring headgear appliances. Eight experienced orthodontists judged each of the cases, and decided if they 'agreed', 'accepted', or 'disagreed' with the recommendations of the computer system. Intra-examiner agreements were investigated using repeated judgements of a set of 30 orthodontic cases and the kappa statistic. All of the examiners exceeded a kappa score of 0.7, allowing them to participate in the test run of the validity of the proposed inference model. The examiners' agreement with the system's recommendations was evaluated statistically. The average satisfaction rate of the examiners was 95.6 per cent and, for 83 out of the 85 cases, 97.6 per cent. The majority of the examiners (i.e. six or more out of the eight) were satisfied with the recommendations of the system. Thus, the usefulness of the proposed inference logic was confirmed.

Adolescent↗

Molecular evolution and phylogeny of the buzzatii complex (Drosophila repleta group): a maximum-likelihood approach.

The buzzatii complex of the mulleri subgroup (Drosophila repleta group) consists of three clusters of species whose evolutionary relationships are poorly known. We analyzed 2,085 coding nucleotides from the xanthine dehydrogenase (XDH:) gene in the 10 available species of the complex and Drosophila mulleri and Drosophila hydei. We adopted a statistical model-fitting approach within the maximum-likelihood (ML) framework of phylogenetic inference. We first modeled the process of nucleotide substitution using a tree topology which was reasonably accurate. Then we used the most satisfactory description so attained to reconstruct the evolutionary relationships in the buzzatii complex. We found that a minimally realistic description of the substitution process of XDH: should allow six substitution types and different substitution rates for codon positions. Using this description we obtained a strongly supported, fully resolved tree which is congruent with the already-known (yet few) relationships. We also analyzed published data from three mitochondrial cytochrome oxidases (CO I, II, and III). In our analyses, these relatively short DNA sequences failed to discriminate statistically among alternative phylogenies. When the data of these three gene regions are combined with the XDH: sequences, the phylogenetic signal emerging from XDH: becomes reinforced. All four of the gene regions evolve faster in the buzzatii and martensis clusters than in the stalkeri cluster, paralleling the amount of chromosomal evolution.

Animals↗

Estimation and control of multiple testing error rates for microarray studies.

The analysis of microarray data often involves performing a large number of statistical tests, usually at least one test per queried gene. Each test has a certain probability of reaching an incorrect inference; therefore, it is crucial to estimate or control error rates that measure the occurrence of erroneous conclusions in reporting and interpreting the results of a microarray study. In recent years, many innovative statistical methods have been developed to estimate or control various error rates for microarray studies. Researchers need guidance choosing the appropriate statistical methods for analysing these types of data sets. This review describes a family of methods that use a set of P-values to estimate or control the false discovery rate and similar error rates. Finally, these methods are classified in a manner that suggests the appropriate method for specific applications and diagnostic procedures that can identify problems in the analysis are described.

Computational Biology↗

HTSinfer: inferring metadata from bulk Illumina RNA-Seq libraries.

SUMMARY: The Sequencing Read Archive is one of the largest and fastest-growing repositories of sequencing data, containing tens of petabytes of sequenced reads. Its data is used by a wide scientific community, often beyond the primary study that generated them. Such analyses rely on accurate metadata concerning the type of experiment and library, as well as the organism from which the sequenced reads were derived. These metadata are typically entered manually by contributors in an error-prone process, and are frequently incomplete. In addition, easy-to-use computational tools that verify the consistency and completeness of metadata describing the libraries to facilitate data reuse, are largely unavailable. Here, we introduce HTSinfer, a Python-based tool to infer metadata directly and solely from bulk RNA-sequencing data generated on Illumina platforms. HTSinfer leverages genome sequence information and diagnostic genes to rapidly and accurately infer the library source and library type, as well as the relative read orientation, 3' adapter sequence and read length statistics. HTSinfer is written in a modular manner, published under a permissible free and open-source license and encourages contributions by the community, enabling easy addition of new functionalities, e.g. for the inference of additional metrics, or the support of different experiment types or sequencing platforms. AVAILABILITY AND IMPLEMENTATION: HTSinfer is released under the Apache License 2.0. Latest code is available via GitHub at https://github.com/zavolanlab/htsinfer, while releases are published on Bioconda. A snapshot of the HTSinfer version described in this article was deposited at Zenodo at 10.5281/zenodo.13985958.

Metadata↗

Invited review: Integrating quantitative findings from multiple studies using mixed model methodology.

In animal agriculture, the need to understand complex biological, environmental, and management relationships is increasing. In addition, as knowledge increases and profit margins shrink, our ability and desire to predict responses to various management decisions also increases. Therefore, the purpose of this review is to help show how improved mathematical and statistical tools and computer technology can help us gain more accurate information from published studies and improve future research. Researchers, in several recent reviews, have gathered data from multiple published studies and attempted to formulate a quantitative model that best explains the observations. In statistics, this process has been labeled meta-analysis. Generally, there are large differences between studies: e. g., different physiological status of the experimental units, different experimental design, different measurement methods, and laboratory technicians. From a statistical standpoint, studies are blocks and their effects must be considered random because the inference being sought is to future, unknown studies. Meta-analyses in the animal sciences have generally ignored the Study effect. Because data gathered across studies are unbalanced with respect to predictor variables, ignoring the Study effect has as a consequence that the estimation of parameters (slopes and intercept) of regression models can be severely biased. Additionally, variance estimates are biased upward, resulting in large type II errors when testing the effect of independent variables. Historically, the Study effect has been considered a fixed effect not because of a strong argument that such effect is indeed fixed but because of our prior inability to efficiently solve even modest-sized mixed models (those containing both fixed and random effects). Modern statistical software has, however, overcome this limitation. Consequently, meta-analyses should now incorporate the Study effect and its interaction effects as random components of a mixed model. This would result in better prediction equations of biological systems and a more accurate description of their prediction errors.

Analysis of Variance↗

Testing statistical hypotheses about rat liver foci.

Tests of statistical hypotheses concerning treatment effect on the development of hepatocellular foci can be carried out directly on two-dimensional observations made on histologic sections or on estimates of the density and volume of foci in three dimensions. Inferences about differences in the density or size of foci from tests based on two-dimensional observations, however, can be misleading. This is because both the number of focus cross-sections observed in a tissue section and the percent area occupied by foci can be expressed in terms of the number of foci per unit volume of liver tissue and the mean focus size. As a consequence, a treatment difference may be caused by a difference in the density of foci, their average size, or both. Of more serious concern is the possibility that failure to detect a treatment effect may occur not only when there is no treatment effect but also when the density and size of foci differ between treatments in such a way that their product is unchanged. This can happen if the effect of treatment is to increase the number of foci and decrease their average size, or vice versa. A similar difficulty of interpretation is associated with hypothesis tests based on average focus cross-section area. Tests based on estimates of the number of foci per unit volume and mean focus volume allow direct inference about the quantities of interest, but these estimates are unstable because they have large variances. Empirical estimates of statistical power for the Wilcoxon rank sum test and the t-test from data on control rats suggest power may be limited in experiments with group sizes of ten and low observed numbers of focus cross-sections. If hypothesis tests based on estimates of the density and size of foci are to form the basis for a bioassay, then the power of statistical tests used to identify treatment effects should be investigated.

Animals↗

A simple local sensitivity analysis tool for nonignorable coarsening: application to dependent censoring.

Right- and interval-censored data are common special cases of coarsened data (Heitjan and Rubin, 1991, Annals of Statistics19, 2244-2253). As with missing data, standard statistical methods that ignore the random nature of the coarsening mechanism may lead to incorrect inferences. We extend a simple sensitivity analysis tool, the index of local sensitivity to nonignorability (Troxel, Ma, and Heitjan, 2004, Statistica Sinica14, 1221-1237), to the evaluation of nonignorability of the coarsening process in the general coarse-data model. By converting this index into a simple graphical display one can easily assess the sensitivity of key inferences to nonignorable coarsening. We illustrate the validity of the method with a simulated example, and apply it to right-censored data from an observational study of cardiac transplantation and to interval-censored data on time to detectable viral load from a clinical trial in HIV disease.

Biometry↗

Fundamentals of massive automatic pairwise alignments of protein sequences: theoretical significance of Z-value statistics.

MOTIVATION: Different automatic methods of sequence alignments are routinely used as a starting point for homology searches and function inference. Confidence in an alignment probability is one of the major fundamentals of massive automatic genome-scale pairwise comparisons, for clustering of putative orthologs and paralogs, sequenced genome annotation or multiple-genomic tree constructions. Extreme value distribution based on the Karlin-Altschul model, usually advised for large-scale comparisons are not always valid, particularly in the case of comparisons of non-biased with nucleotide-biased genomes (such that of Plasmodium falciparum). Z-values estimates based on Monte Carlo technics, can be calculated experimentally for any alignment output, whatever the method used. Empirically, a Z-value higher than approximately 8 is supposed reasonable to assess that an alignment score is significant, but this arbitrary figure was never theoretically justified. RESULTS: In this paper, we used the Bienaymé-Chebyshev inequality to demonstrate a theorem of the upper limit of an alignment score probability (or P-value). This theorem implies that a computed Z-value is a statistical test, a single-linkage clustering criterion and that 1/Z-value(2) is an upper limit to the probability of an alignment score whatever the actual probability law is. Therefore, this study provides the missing theoretical link between a Z-value cut-off used for an automatic clustering of putative orthologs and/or paralogs, and the corresponding statistical risk in such genome-scale comparisons (using non-biased or biased genomes).

Algorithms↗

A statistical causal model for the assessment of dysarthric speech and the utility of computer-based speech recognition.

The evaluation of the degree of speech impairment and the utility of computer recognition of impaired speech are separately and independently performed. Particular attention is paid to the question concerning whether or not there is a relationship between naive listeners' subjective judgements of impaired speech and the performance of a laboratory version of a speech recognition system. It is a difficult task to relate a speech impairment rating with speech recognition accuracy. Towards this end, a statistical causal model is proposed. This model is very appealing in its structure to support inference, and thus can be applied to perform various assessments such as the success of automatic recognition of dysarthric speech. The application of this model is illustrated with a case study of a dysarthric speaker compared against a normal speaker serving as a control.

Adult↗

Semiparametric variance-component models for linkage and association analyses of censored trait data.

Variance-component (VC) models are widely used for linkage and association mapping of quantitative trait loci in general human pedigrees. Traditional VC methods assume that the trait values within a family follow a multivariate normal distribution and are fully observed. These assumptions are violated if the trait data contain censored observations. When the trait pertains to age at onset of disease, censoring is inevitable because of loss to follow-up and limited study duration. Censoring also arises when the trait assay cannot detect values below (or above) certain thresholds. The latent trait values tend to have a complex distribution. Applying traditional VC methods to censored trait data would inflate type I error and reduce power. We present valid and powerful methods for the linkage and association analyses of censored trait data. Our methods are based on a novel class of semiparametric VC models, which allows an arbitrary distribution for the latent trait values. We construct appropriate likelihood for the observed data, which may contain left or right censored observations. The maximum likelihood estimators are approximately unbiased, normally distributed, and statistically efficient. We develop stable and efficient numerical algorithms to implement the corresponding inference procedures. Extensive simulation studies demonstrate that the proposed methods outperform the existing ones in practical situations. We provide an application to the age at onset of alcohol dependence data from the Collaborative Study on the Genetics of Alcoholism. A computer program is freely available.

Age of Onset↗

An exploratory instrumental variable analysis of the outcomes of localized breast cancer treatments in a medicare population.

This study is motivated by the potential problem of using observational data to draw inferences about treatment outcomes when experimental data are not available. We compare two statistical approaches, ordinary least-squares (OLS) and instrumental variables (IV) regression analysis, to estimate the outcomes (three-year post-treatment survival) of three treatments for early stage breast cancer in elderly women: mastectomy (MST), breast conserving surgery with radiation therapy (BCSRT), and breast conserving surgery only (BCSO). The primary data source was Medicare claims for a national random sample of 2907 women (age 67 or older) with localized breast cancer who were treated between 1992 and 1994. Contrary to randomized clinical trial (RCT) results, analysis with the observational data found highly significant differences in survival among the three treatment alternatives: 79.2% survival for BCSO, 85.3% for MST, and 93.0% for BCSRT. Using OLS to control for the effects of observable characteristics narrowed the estimated survival rate differences, which remained statistically significant. In contrast, the IV analysis estimated survival rate differences that were not significantly different from 0. However, the IV-point estimates of the treatment effects were quantitatively larger than the OLS estimates, unstable, and not significantly different from the OLS results. In addition, both sets of estimates were in the same quantitative range as the RCT results.We conclude that unadjusted observational data on health outcomes of alternative treatments for localized breast cancer should not be used for cost-effectiveness studies. Our comparisons suggest that whether one places greater confidence in the OLS or the IV results depends on at least three factors: (1) the extent of observable health information that can be used as controls in OLS estimation, (2) the outcomes of statistical tests of the validity of the instrumental variable method, and (3) the similarity of the OLS and IV estimates. In this particular analysis, the OLS estimates appear to be preferable because of the instability of the IV estimates.

Aged↗

Deduction of functional peptide motifs in scorpion toxins.

Scorpion toxins are important physiological probes for characterizing ion channels. Molecular databases have limited functional annotation of scorpion toxins. Their function can be inferred by searching for conserved motifs in sequence signature databases that are derived statistically but are not necessarily biologically relevant. Mutation studies provide biological information on residues and positions important for structure-function relationship but are not normally used for extraction of binding motifs. 3D structure analyses also aid in the extraction of peptide motifs in which non-contiguous residues are clustered spatially. Here we present new, functionally relevant peptide motifs for ion channels, derived from the analyses of scorpion toxin native and mutant peptides.

Amino Acid Motifs↗

[Juvenile bone cysts and giant cell tumors (author's transl)].

The attempt is made to demonstrate clinical, radiographical, morphological differences between juvenile bone cyst and giant cell tumor as shown in the patients of the Surgical Department of the University of Dusseldorf and literature. It is possible to derive inferences in differential diagnosis from the age of the patient, clinical course and statistically most frequent locations. Radiographically the bone cyst distinguishes itself by its central localisation in the metaphysis, where as the giant cell tumor has an excentric position in the epiphysis with a tendency of extending into the metaphysis. Giant cells occur in both diseases. While the bone cyst is an tumor-like lesion, the giant cell tumor is a real neoplasia. The therapeutical procedure varies according to different prognostical significance of the two diseases. Therefore, it is necessary to draw an exact line of demarcation between the comparetively harmless juvenile bone cyst and the semimalign giant cell tumor.

Adolescent↗