A low-cost high-throughput phenotyping system for automatically quantifying foliar area and greenness.

Premise: With modern advances in genetic sequencing technology, plant phenotyping has become a substantial bottleneck in crop improvement programs. Traditionally, researchers have manually measured phenotypic traits to help determine genotype-phenotype relationships, but manual measurements can be time consuming and expensive. Recently, automated phenotyping systems have increased the spatial and temporal density of measurements, but most of these systems are extremely expensive and require specialized expertise. In the present paper, we develop and validate a low-cost, scalable, high-throughput phenotyping (HTP) system for automating the measurement of foliar area and greenness. Methods: During a greenhouse experiment on the effects of abiotic stress on Brassica rapa, we collected images of hundreds of plants every hour for over a month with a system that cost approximately US$1000. Results: In comparison with manually acquired images, this HTP system was able to produce similar estimates of foliar area and greenness, developmental trends, and treatment effects. Foliar area was correlated between the two image sets, but greenness was not. Discussion: These findings highlight the potential of HTP systems built from low-cost hardware and freely available software. Future work can use this system to investigate genotype-environment interactions and the genetic loci underlying morphological changes resulting from abiotic stress.

Comparing Functional Motor Control Exercises With Therapeutic Exercise in Wrestlers With Iliotibial Band Syndrome.

CONTEXT: Iliotibial band syndrome (ITBS) is a common overuse injury in runners with parallels to our findings of overuse in Greco Roman wrestlers. Despite research indicating coordination and movement-based factors about the hip, no studies were found using functional motor control (FMC) in runners or wrestlers with ITBS. Thus, we compared FMC exercises and therapeutic exercises (TEs) on pain, function, muscle strength, and range of motion (ROM) in national-level Greco Roman wrestlers with ITBS. DESIGN: Controlled laboratory study. METHODS: Sixty national-level Greco Roman wrestlers diagnosed with ITBS were randomly assigned to 8 weeks of FMC exercises, TE, and a control group (20 individuals for each group). Pain (visual analog scale), function (triple hop test for distance, single-leg vertical jump test, and agility T test), muscle strength (handheld dynamometer), and ROM (goniometer) were measured at baseline and 8 weeks after intervention as posttest. RESULTS: Although both interventions significantly reduced pain (P < .001, η2 = .87), improved function (triple hop test P = .004, η2 = .94; single-leg vertical jump P = .002, η2 = .93; and T test P < .001, η2 = .93) and strength (hip abduction (P < .001, η2 = .52), hip external rotation (P = .02, η2 = .95), knee flexion (P ≤ .001, η2 = .94), and knee extension (P < .001, η2 = .91) compared with the control group, FMC showed more significant improvements in comparison with TE. Significant differences (P = .001) were observed between FMC and TE compared with the control group in ROM outcome. However, TE was more effective than FMC in improving ROM hip abduction (P < .001, η2 = .93), hip adduction (P = .000, η2 = .92), hip internal rotation (P < .001, η2 = .92), and hip external rotation (P < .001, η2 = .93). CONCLUSION: FMC exercises were superior to TE in terms of pain, function, and muscle strength, whereas TE was more effective for improving ROM. FMC exercise is suggested as an effective intervention for improvement of the outcomes related to ITBS in national-level Greco Roman wrestlers.

Algae to angiosperms: Autofluorescence for rapid visualization of plant anatomy among diverse taxa.

PREMISE: Fluorescence microscopy is an effective tool for viewing plant internal anatomy. However, using fluorescent antibodies or labels hinders throughput. We present a minimal protocol that takes advantage of inherent autofluorescence and aldehyde-induced fluorescence in plant cellular and subcellular structures to markedly increase throughput in cellular and ultrastructural visualization. METHODS AND RESULTS: Twelve species distributed across the plant phylogeny were each subjected to five fixative treatments: 1% paraformaldehyde and 2% glutaraldehyde, 2% paraformaldehyde, 2% glutaraldehyde, formalin-acid-alcohol (FAA), and 70% ethanol. Samples were prepared by embedding and mechanically sectioning or via whole mount. A confocal laser scanning system was used to collect micrographs. We evaluated and compared fixative influence on sample structural preservation and tissue autofluorescence. CONCLUSIONS: Formaldehyde fixation of Viridiplantae taxa samples generates useful structural data while requiring no additional histological staining or clearing. In addition, a fluorescence-capable microscope is the only specialized equipment required for image acquisition. The minimal protocol developed in this experiment enables high-throughput sample processing by eliminating the need for multi-day preparations.

Analysis of CACTA transposase genes unveils the mechanism of intron loss and distinct small RNA silencing pathways underlying divergent evolution of Brassica genomes.

In comparison with retrotransposons, DNA transposons make up a smaller proportion of most plant genomes. However, these elements are often proximal to genes to affect gene expression depending on the activity of the transposons, which is largely reflected by the activity of the transposase genes. Here, we show that three AT-rich introns were retained in the TNP2-like transposase genes of the Bot1 (Brassica oleracea transposon 1) CACTA transposable elements in Brassica oleracea, but were lost in the majority of the Bot1 elements in Brassica rapa. A recent burst of transposition of Bot1 was observed in B. oleracea, but not in B. rapa. This burst of transposition is likely related to the activity of the TNP2-like transposase genes as the expression values of the transposase genes were higher in B. oleracea than in B. rapa. In addition, distinct populations of small RNAs (21, 22 and 24 nt) were detected from the Bot1 elements in B. oleracea, but the vast majority of the small RNAs from the Bot1 elements in B. rapa are 24 nt in length. We hypothesize that the different activity of the TNP2-like transposase genes is likely associated with the three introns, and intron loss is likely reverse transcriptase mediated. Furthermore, we propose that the Bot1 family is currently undergoing silencing in B. oleracea, but has already been silenced in B. rapa. Taken together, our data provide new insights into the differentiation of transposons and their role in the asymmetric evolution of these two closely related Brassica species.

Correction: Integrating transcriptomic network reconstruction and eQTL analyses reveals mechanistic connections between genomic architecture and Brassica rapa development.

[This corrects the article DOI: 10.1371/journal.pgen.1008367.].

Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

PREMISE: Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of Brassica rapa. METHODS: FAA (formaldehyde, glacial acetic acid, and ethanol)-fixed samples were stored in ethanol. Without performing any additional clearing or staining, we tested two methods of image acquisition at three locations along the proximal-distal axis of the laminae and estimated the patterns of venation using the program phenoVein. We developed and made available an R script to handle the phenoVein output and then analyzed our data using linear mixed-effects models. RESULTS: Beyond fixation and storage, staining and clearing are not necessary to estimate leaf venation using phenoVein if the images are acquired using a stereomicroscope. All estimates of venation required some manual adjustment. We found a significant effect of location within the laminae for all aspects of venation. DISCUSSION: By removing the clearing and staining steps and utilizing the semi-automated program phenoVein, we quickly and cheaply acquired leaf venation data. Venation may be an important target for crop breeding efforts, particularly if intralaminar variation correlates with variation in physiological processes, which remains an open question.

Integrating transcriptomic network reconstruction and eQTL analyses reveals mechanistic connections between genomic architecture and Brassica rapa development.

Plant developmental dynamics can be heritable, genetically correlated with fitness and yield, and undergo selection. Therefore, characterizing the mechanistic connections between the genetic architecture governing plant development and the resulting ontogenetic dynamics of plants in field settings is critically important for agricultural production and evolutionary ecology. We use hierarchical Bayesian Function-Valued Trait (FVT) models to estimate Brassica rapa growth curves throughout ontogeny, across two treatments, and in two growing seasons. We find genetic variation for plasticity of growth rates and final sizes, but not the inflection point (transition from accelerating to decelerating growth) of growth curves. There are trade-offs between growth rate and duration, indicating that selection for maximum yields at early harvest dates may come at the expense of late harvest yields and vice versa. We generate eigengene modules and determine which are co-expressed with FVT traits using a Weighted Gene Co-expression Analysis. Independently, we seed a Mutual Rank co-expression network model with FVT traits to identify specific genes and gene networks related to FVT. GO-analyses of eigengene modules indicate roles for actin/cytoskeletal genes, herbivore resistance/wounding responses, and cell division, while MR networks demonstrate a close association between metabolic regulation and plant growth. We determine that combining FVT Quantitative Trait Loci (QTL) and MR genes/WGCNA eigengene expression profiles better characterizes phenotypic variation than any single data type (i.e. QTL, gene, or eigengene alone). Our network analysis allows us to employ a targeted eQTL analysis, which we use to identify regulatory hotspots for FVT. We examine cis vs. trans eQTL that mechanistically link FVT QTL with structural trait variation. Colocalization of FVT, gene, and eigengene eQTL provide strong evidence for candidate genes influencing plant height. The study is the first to explore eQTL for FVT, and specifically do so in agroecologically relevant field settings.

A framework for genomics-informed ecophysiological modeling in plants.

Dynamic process-based plant models capture complex physiological response across time, carrying the potential to extend simulations out to novel environments and lend mechanistic insight to observed phenotypes. Despite the translational opportunities for varietal crop improvement that could be unlocked by linking natural genetic variation to first principles-based modeling, these models are challenging to apply to large populations of related individuals. Here we use a combination of model development, experimental evaluation, and genomic prediction in Brassica rapa L. to set the stage for future large-scale process-based modeling of intraspecific variation. We develop a new canopy growth submodel for B. rapa within the process-based model Terrestrial Regional Ecosystem Exchange Simulator (TREES), test input parameters for feasibility of direct estimation with observed phenotypes across cultivated morphotypes and indirect estimation using genomic prediction on a recombinant inbred line population, and explore model performance on an in silico population under non-stressed and mild water-stressed conditions. We find evidence that the updated whole-plant model has the capacity to distill genotype by environment interaction (G×E) into tractable components. The framework presented offers a means to link genetic variation with environment-modulated plant response and serves as a stepping stone towards large-scale prediction of unphenotyped, genetically related individuals under untested environmental scenarios.

Differences in Knee and Hip Adduction and Hip Muscle Activation in Runners With and Without Iliotibial Band Syndrome.

BACKGROUND: Iliotibial band syndrome has been associated with altered hip and knee kinematics in runners. Previous studies have recommended further research on neuromuscular factors at the hip. The frontal plane hip muscles have been a strong focus in strength comparison but not for electromyography investigation. OBJECTIVE: To compare hip surface electromyography, and frontal plane hip and knee kinematics, in runners with and without iliotibial band syndrome. DESIGN: Observational cross-sectional study. SETTING: Biomechanics research laboratory within a university. PARTICIPANTS: Thirty subjects were recruited consisting of 15 injured runners with iliotibial band syndrome and 15 gender-, age-, and body mass index-matched controls. In each group, 8 were male runners and 7 were female runners. Inclusion criteria for the injured group were pain within 2 months related to iliotibial band syndrome and a positive Noble compression test. Participants were excluded if they reported other lower extremity diagnoses within the last year or active lower extremity or low back pain not related to iliotibial band syndrome. Controls were excluded if they reported a history of iliotibial band syndrome. Convenience sampling was used based on referrals from local running clinics and orthopedic clinics. METHODS: Three-dimensional motion capture was performed with 10 high-speed cameras synchronized with wireless surface electromyography during a 30-minute run. The first data point was at 3 minutes, using a constant speed of 2.74 meters per second. A second data point was at 30 minutes, using a self-selected pace by the participant to allow for a challenging run until completion at 30 minutes. MAIN OUTCOME MEASUREMENTS: Motion capture was reported as peak kinematic values from heel strike to peak knee flexion for hip adduction and knee adduction. Surface electromyography was reported as a percentage of maximal voluntary contraction for the gluteus maximus, gluteus medius and tensor fascia latae muscles. RESULTS: Injured runners demonstrated increased knee adduction compared with control runners at 30 minutes (P = .002, control = -1.48°, injured = 3.74°). Tensor fasciae latae muscle activation in injured runners was increased compared with control runners at 3 minutes (P = .017, control = 7% maximal voluntary isometric contraction, injured = 11% maximal voluntary isometric contraction). CONCLUSION: The results of this study suggest that lateral knee pain in runners localized to the distal iliotibial band is associated with increased knee adduction at 30 minutes. Increased tensor fasciae latae muscle activation at 3 minutes is noted, but more investigation is needed to better understand the clinical meaning. These findings are consistent with but not conclusive evidence supporting the theory that neuromuscular factors of the hip muscles may contribute to increased knee adduction in runners with iliotibial band syndrome. We advise caution using these findings to support treatments intended to modify tensor fasciae latae activation, given the small differences of 4% in muscle activation. Increased knee adduction in runners at 30 minutes was over 5° and beyond the minimal detectable difference. Additional research is needed to confirm whether the degree of knee adduction changes earlier versus later in a run and whether fatigue is a clinically relevant factor. LEVEL OF EVIDENCE: III.

Circadian rhythms are associated with shoot architecture in natural settings.

Circadian rhythms are key regulators of diverse biological processes under controlled settings. Yet, the phenotypic and fitness consequences of quantitative variation in circadian rhythms remain largely unexplored in the field. As with other pathways, phenotypic characterization of circadian outputs in the field may reveal novel clock functions. Across consecutive growing seasons, we test for associations between clock variation and flowering phenology, plant size, shoot architecture, and fruit set in clock mutants and segregating progenies of Arabidopsis thaliana expressing quantitative variation in circadian rhythms. Using structural equation modeling, we find that genotypic variation in circadian rhythms within a growing season is associated directly with branching, which in turn affects fruit production. Consistent with direct associations between the clock and branching in segregating progenies, cauline branch number is lower and rosette branch number higher in a short-period mutant relative to wild-type and long-period genotypes, independent of flowering time. Differences in branching arise from variation in meristem fate as well as leaf production rate before flowering and attendant increases in meristem number. Our results suggest that clock variation directly affects shoot architecture in the field, suggesting a novel clock function and means by which the clock affects performance.

Bayesian estimation and use of high-throughput remote sensing indices for quantitative genetic analyses of leaf growth.

KEY MESSAGE: We develop Bayesian function-valued trait models that mathematically isolate genetic mechanisms underlying leaf growth trajectories by factoring out genotype-specific differences in photosynthesis. Remote sensing data can be used instead of leaf-level physiological measurements. Characterizing the genetic basis of traits that vary during ontogeny and affect plant performance is a major goal in evolutionary biology and agronomy. Describing genetic programs that specifically regulate morphological traits can be complicated by genotypic differences in physiological traits. We describe the growth trajectories of leaves using novel Bayesian function-valued trait (FVT) modeling approaches in Brassica rapa recombinant inbred lines raised in heterogeneous field settings. While frequentist approaches estimate parameter values by treating each experimental replicate discretely, Bayesian models can utilize information in the global dataset, potentially leading to more robust trait estimation. We illustrate this principle by estimating growth asymptotes in the face of missing data and comparing heritabilities of growth trajectory parameters estimated by Bayesian and frequentist approaches. Using pseudo-Bayes factors, we compare the performance of an initial Bayesian logistic growth model and a model that incorporates carbon assimilation (A max) as a cofactor, thus statistically accounting for genotypic differences in carbon resources. We further evaluate two remotely sensed spectroradiometric indices, photochemical reflectance (pri2) and MERIS Terrestrial Chlorophyll Index (mtci) as covariates in lieu of A max, because these two indices were genetically correlated with A max across years and treatments yet allow much higher throughput compared to direct leaf-level gas-exchange measurements. For leaf lengths in uncrowded settings, including A max improves model fit over the initial model. The mtci and pri2 indices also outperform direct A max measurements. Of particular importance for evolutionary biologists and plant breeders, hierarchical Bayesian models estimating FVT parameters improve heritabilities compared to frequentist approaches.

Polyploidy and the relationship between leaf structure and function: implications for correlated evolution of anatomy, morphology, and physiology in Brassica.

BACKGROUND: Polyploidy is well studied from a genetic and genomic perspective, but the morphological, anatomical, and physiological consequences of polyploidy remain relatively uncharacterized. Whether these potential changes bear on functional integration or are idiosyncratic remains an open question. Repeated allotetraploid events and multiple genomic combinations as well as overlapping targets of artificial selection make the Brassica triangle an excellent system for exploring variation in the connection between plant structure (anatomy and morphology) and function (physiology). We examine phenotypic integration among structural aspects of leaves including external morphology and internal anatomy with leaf-level physiology among several species of Brassica. We compare diploid and allotetraploid species to ascertain patterns of phenotypic correlations among structural and functional traits and test the hypothesis that allotetraploidy results in trait disintegration allowing for transgressive phenotypes and additional evolutionary and crop improvement potential. RESULTS: Among six Brassica species, we found significant effects of species and ploidy level for morphological, anatomical and physiological traits. We identified three suites of intercorrelated traits in both diploid parents and allotetraploids: Morphological traits (such as leaf area and perimeter) anatomic traits (including ab- and ad- axial epidermis) and aspects of physiology. In general, there were more correlations between structural and functional traits for allotetraploid hybrids than diploid parents. Parents and hybrids did not have any significant structure-function correlations in common. Of particular note, there were no significant correlations between morphological structure and physiological function in the diploid parents. Increased phenotypic integration in the allotetraploid hybrids may be due, in part, to increased trait ranges or simply different structure-function relationships. CONCLUSIONS: Genomic and chromosomal instability in early generation allotetraploids may allow Brassica species to explore new trait space and potentially reach higher adaptive peaks than their progenitor species could, despite temporary fitness costs associated with unstable genomes. The trait correlations that disappear after hybridization as well as the novel trait correlations observed in allotetraploid hybrids may represent relatively evolutionarily labile associations and therefore could be ideal targets for artificial selection and crop improvement.

Dietary antioxidants enhance immunocompetence in larval amphibians.

Dietary antioxidants have been shown to confer a variety of benefits through their ability to counter oxidative stress, including increased immunocompetence and reduced susceptibility to both infectious and non-infectious diseases. However, little is known about the effects of dietary antioxidants on immune function in larval amphibians, a group experiencing worldwide declines driven by factors that likely involve altered immunocompetence. We investigated the effects of dietary antioxidants (quercetin, vitamin E, and ß-carotene) on two components of the immune system, as well as development and growth. Lithobates pipiens tadpoles fed diets with supplemental ß-carotene or vitamin E exhibited an enhanced swelling response as measured with a phytohemagglutinin assay (PHA), but there was no induced antibody response. Effects were often dose-dependent, with higher antioxidant levels generally conferring stronger swelling that possibly corresponds to the innate immune response. Our results indicate that the antioxidant content of the larval amphibian diets not only had a detectable effect on their immune response capability, but also promoted tadpole growth (mass gain), although developmental stage was not affected. Given that many environmental perturbations may cause oxidative stress or reduce immunocompetence, it is critical to understand how nutrition may counter these effects.

Iliotibial Band Syndrome in Runners: Biomechanical Implications and Exercise Interventions.

Iliotibial band syndrome (ITBS) has known biomechanical factors with an unclear explanation based on only strength and flexibility deficits. Neuromuscular coordination has emerged as a likely reason for kinematic faults guiding research toward motor control. This article discusses ITBS in relation to muscle performance factors, fascial considerations, epidemiology, functional anatomy, strength deficits, kinematics, iliotibial strain and strain rate, and biomechanical considerations. Evidence-based exercise approaches are reviewed for ITBS, including related methods used to train the posterior hip muscles.

Selection during crop diversification involves correlated evolution of the circadian clock and ecophysiological traits in Brassica rapa.

Crop selection often leads to dramatic morphological diversification, in which allocation to the harvestable component increases. Shifts in allocation are predicted to impact (as well as rely on) physiological traits; yet, little is known about the evolution of gas exchange and related anatomical features during crop diversification. In Brassica rapa, we tested for physiological differentiation among three crop morphotypes (leaf, turnip, and oilseed) and for correlated evolution of circadian, gas exchange, and phenological traits. We also examined internal and surficial leaf anatomical features and biochemical limits to photosynthesis. Crop types differed in gas exchange; oilseed varieties had higher net carbon assimilation and stomatal conductance relative to vegetable types. Phylogenetically independent contrasts indicated correlated evolution between circadian traits and both gas exchange and biomass accumulation; shifts to shorter circadian period (closer to 24 h) between phylogenetic nodes are associated with higher stomatal conductance, lower photosynthetic rate (when CO2 supply is factored out), and lower biomass accumulation. Crop type differences in gas exchange are also associated with stomatal density, epidermal thickness, numbers of palisade layers, and biochemical limits to photosynthesis. Brassica crop diversification involves correlated evolution of circadian and physiological traits, which is potentially relevant to understanding mechanistic targets for crop improvement.

Modeling development and quantitative trait mapping reveal independent genetic modules for leaf size and shape.

Improved predictions of fitness and yield may be obtained by characterizing the genetic controls and environmental dependencies of organismal ontogeny. Elucidating the shape of growth curves may reveal novel genetic controls that single-time-point (STP) analyses do not because, in theory, infinite numbers of growth curves can result in the same final measurement. We measured leaf lengths and widths in Brassica rapa recombinant inbred lines (RILs) throughout ontogeny. We modeled leaf growth and allometry as function valued traits (FVT), and examined genetic correlations between these traits and aspects of phenology, physiology, circadian rhythms and fitness. We used RNA-seq to construct a SNP linkage map and mapped trait quantitative trait loci (QTL). We found genetic trade-offs between leaf size and growth rate FVT and uncovered differences in genotypic and QTL correlations involving FVT vs STPs. We identified leaf shape (allometry) as a genetic module independent of length and width and identified selection on FVT parameters of development. Leaf shape is associated with venation features that affect desiccation resistance. The genetic independence of leaf shape from other leaf traits may therefore enable crop optimization in leaf shape without negative effects on traits such as size, growth rate, duration or gas exchange.

Patterns of shoot architecture in locally adapted populations are linked to intraspecific differences in gene regulation.

• Shoot architecture, including the number and location of branches, is a crucial aspect of plant function, morphological diversification, life history evolution and crop domestication. • Genes controlling shoot architecture are well characterized in, and largely conserved across, model flowering plant species. The role of these genes in the evolution of morphological diversity in natural populations, however, has not been explored. • We identify axillary meristem outgrowth as a primary driver of divergent branch number and life histories in two locally adapted populations of the monkeyflower, Mimulus guttatus. • Furthermore, we show that MORE AXILLARY GROWTH (MAX) gene expression strongly correlates with natural variation in branch outgrowth in this species, linking modification of the MAX-dependent pathway to the evolutionary diversification of shoot architecture.

Node-specific branching and heterochronic changes underlie population-level differences in Mimulus guttatus (Phrymaceae) shoot architecture.

PREMISE OF THE STUDY: Shoot architecture is a fundamentally developmental aspect of plant biology with implications for plant form, function, reproduction, and life history evolution. Mimulus guttatus is morphologically diverse and becoming a model for evolutionary biology. Shoot architecture, however, has never been studied from a developmental perspective in M. guttatus. METHODS: We examined the development of branches and flowers in plants from two locally adapted populations of M. guttatus with contrasting flowering times, life histories, and branch numbers. We planted second-generation seed in growth chambers to control for maternal and environmental effects. KEY RESULTS: Most branches occurred at nodes one and two of the main axis. Onset of branching occurred earlier and at a greater frequency in perennials than in annuals. In perennials, almost all flowers occurred at the fifth or more distal nodes. In annuals, most flowers occurred at the third and more distal nodes. Accessory axillary meristems and higher-order branching did not influence shoot architecture. CONCLUSIONS: We found no evidence for trade-offs between flowers and branches because axillary meristem number was not limiting: a large number of meristems remained quiescent. If, however, quiescence is a component of meristem allocation strategy, then meristems may be limited despite presence of quiescent meristems. At the two basalmost nodes, branch number was determined by mechanisms governing either meristem initiation or outgrowth, rather than flowering vs. branching. At the third and more distal nodes, heterochronic processes contributed to flowering time and branch number differences between populations.

Iliotibial band syndrome: soft tissue and biomechanical factors in evaluation and treatment.

Muscle performance factors and altered loading mechanics have been linked to a variety of lower extremity musculoskeletal disorders. In this article, biomechanical risk factors associated with iliotibial band syndrome (ITBS) are described, and a strategy for incorporating these factors into the clinical evaluation of and treatment for that disorder is presented. Abnormal movement patterns in runners and cyclists with ITBS are discussed, and the pathophysiological characteristics of this syndrome are considered in light of prior and current studies in anatomy. Differential diagnoses and the use of imaging, medications, and injections in the treatment of ITBS are reviewed. The roles of hip muscle strength, kinematics, and kinetics are detailed, and the assessment and treatment of muscle performance factors are discussed, with emphasis on identifying and treating movement dysfunction. Various stages of rehabilitation, including strengthening progressions to reduce soft-tissue injury, are described in detail. ITBS is an extremely common orthopedic condition that presents with consistent dysfunctional patterns in muscle performance and movement deviation. Through careful assessment of lower quarter function, the clinician can properly identify individuals and initiate treatment.