Agrobacterium-mediated transfer of the Fusarium graminearum Tri6 gene into barley using mature seed-derived shoot tips as explants.

KEY MESSAGE: We transferred the Tri6 gene into the elite barley GemCraft via new transformation method through shoot organogenesis and identified the rearrangements of transgenes and phenotypic variations in the transgenic plants. Despite its agronomic and economic importance, barley transformation is still very challenging for many elite varieties. In this study, we used direct shoot organogenesis to transform the elite barley cultivar GemCraft with the RNAi constructs containing Tri6 gene of Fusarium graminearum, which causes fusarium head blight (FHB). We isolated 4432 shoot tips and co-cultured these explants with Agrobacterium tumefaciens. A total of 25 independent T0 transgenic plants were generated including 15 events for which transgene-specific PCR amplicons were observed. To further determine the presence of transgenes, the T1 progenies of all 15 T0 plants were analyzed, and the expected PCR products were obtained in 10 T1 lines. Droplet digital (dd) PCR analysis revealed various copy numbers of transgenes in the transgenic plants. We determined the insertion site of transgenes using long-read sequencing data and observed the rearrangements of transgenes. We found phenotypic variations in both T1 and T2 generation plants. FHB disease was evaluated under growth chamber conditions, but no significant differences in disease severity or deoxynivalenol accumulation were observed between two Tri6 transgenic lines and the wildtype. Our results demonstrate the feasibility of the shoot tip transformation and may open the door for applying this system for genetic improvement and gene function research in other barley genotypes.

Incidence and Predictors of Subsidence Using Modular, Tapered, Fluted Titanium Femoral Stems in Aseptic Revision Total Hip Arthroplasty.

BACKGROUND: Tapered, fluted titanium (TFT) femoral stems have become the gold standard in revision total hip arthroplasty (rTHA). However, there is a paucity of data on TFT stem subsidence rates following aseptic rTHA. Subsidence can lead to instability, mechanical failure, leg-length discrepancy, and may require revision surgery. This study evaluated the incidences and predictors of TFT subsidence in aseptic rTHA. METHODS: A total of 102 TFT femoral stems of 4 designs were retrospectively reviewed. Stem subsidence was measured on digital radiographs taken immediately after surgery and at standard clinical follow-up. Patient characteristics, risk factors for subsidence, revision etiologies, and implant characteristics were recorded. Patient-reported outcome measures were also evaluated for a subset of cases. RESULTS: Overall, 12% of stems subsided >1 cm, and subsidence was minimal (<3 mm) in ≥64% of cases. From immediate postoperative to 1-month radiographic follow-up, 79% of stems subsided a mean of 2.9 mm (range, 0.1 to 12 mm). Beyond 1 month, subsidence was minimal for ≥77% of cases. In multivariate analyses, women and less femoral implant canal fill were associated with greater subsidence (P ≤ .034). The TFT stem design was not associated with early subsidence (P = .816). There were no modular junction fractures. There were 2 fractures and 2 subsidence-related revisions for aseptic loosening that occurred postoperatively. CONCLUSIONS: The amount of subsidence in TFT stems was low and was detectable in the early (less than 1 year) postoperative period. Maximizing TFT stem fill within the femoral canal appears to reduce the risk of subsidence without increasing femoral fracture rates and should be the goal with implantation of these devices. LEVEL OF EVIDENCE: IV-Case Series, No Control Group.

Fungal biotechnology: From yesterday to tomorrow.

Fungi have been used to better the lives of everyday people and unravel the mysteries of higher eukaryotic organisms for decades. However, comparing progress and development stemming from fungal research to that of human, plant, and bacterial research, fungi remain largely understudied and underutilized. Recent commercial ventures have begun to gain popularity in society, providing a new surge of interest in fungi, mycelia, and potential new applications of these organisms to various aspects of research. Biotechnological advancements in fungal research cannot occur without intensive amounts of time, investments, and research tool development. In this review, we highlight past breakthroughs in fungal biotechnology, discuss requirements to advance fungal biotechnology even further, and touch on the horizon of new breakthroughs with the highest potential to positively impact both research and society.

DNA Markers, Pathogenicity Test, and Multilocus Sequence Analysis to Differentiate and Characterize Cereal-Specific Xanthomonas translucens Strains.

Xanthomonas translucens contains a group of bacterial pathogens that are closely related and have been divided into several pathovars based on their host range. X. translucens pv. undulosa (Xtu) and X. translucens pv. translucens (Xtt) are two important pathovars that cause bacterial leaf streak disease on wheat and barley, respectively. In this study, DNA markers were developed to differentiate Xtu and Xtt and were then used to characterize a collection of X. translucens strains with diverse origins, followed by confirmation and characterization with pathogenicity tests and multilocus sequence analysis/typing (MLSA/MLST). We first developed cleaved amplified polymorphic sequence markers based on the single-nucleotide polymorphisms within a cereal pathovar-specific DNA sequence. In addition, two Xtt-specific markers, designated Xtt-XopM and Xtt-SP1, were developed from comparative genomics among the sequenced Xtt/Xtu genomes. Using the developed markers, a collection of X. translucens strains were successfully identified as Xtu or Xtt. Pathogenicity tests on wheat and barley plants and MLSA of four housekeeping genes validated the pathovar assignation of those strains. Furthermore, MLSA revealed distinct subclades within both Xtu and Xtt groups. Seven and three sequence types were identified from MLST for Xtu and Xtt strains, respectively. The establishment of efficient Xtt/Xtu differentiation methods and characterization of those strains will be useful in studying disease epidemiology and host-pathogen interactions and breeding programs when screening for sources of resistance for these two important bacterial pathogens.

Genetic and physical localization of a major susceptibility gene to Pyrenophora teres f. maculata in barley.

KEY MESSAGE: Genetic characterization of a major spot form net blotch susceptibility locus to using linkage mapping to identify a candidate gene and user-friendly markers in barley. Spot form net blotch (SFNB), caused by the necrotrophic fungal pathogen Pyrenophora teres f. maculata (Ptm), is an economically important foliar diseases in barley. Although various resistance loci have been identified, breeding for SFNB-resistant varieties has been hampered due to the complex virulence profile of Ptm populations. One resistance locus in the host may be effective against one specific isolate, but it may confer susceptibility to other isolates. A major susceptibility QTL on chromosome 7H, named Sptm1, was consistently identified in many studies. In the present study, we conduct fine mapping to localize Sptm1 with high resolution. A segregating population was developed from selected F2 progenies of the cross Tradition (S) × PI 67381 (R), in which the disease phenotype was determined by the Sptm1 locus alone. Disease phenotypes of critical recombinants were confirmed in the following two consecutive generations. Genetic mapping anchored the Sptm1 gene to an ⁓400 kb region on chromosome 7H. Gene prediction and annotation identified six protein-coding genes in the delimited Sptm1 region, and the gene encoding a putative cold-responsive protein kinase was selected as a strong candidate. Therefore, providing fine localization and candidate of Sptm1 for functional validation, our study will facilitate the understanding of susceptibility mechanism underlying the barley-Ptm interaction and offers a potential target for gene editing to develop valuable materials with broad-spectrum resistance to SFNB.

Comparative early histologic healing quality of magnetic versus stapled small bowel anastomosis.

BACKGROUND: Successful anastomotic healing is critical to preventing complications after intestinal surgery. We aimed to compare the early healing of end-to-end small bowel anastomosis by self-forming magnets with surgical stapling in a porcine model. METHOD: Six Yorkshire pigs underwent 2 simultaneous small bowel anastomoses using a circular stapler and self-forming magnet technique. The primary outcome was healing quality, measured by 4 histologic features: inflammatory cell infiltration, collagen formation, grade of inflammation, and bacterial infiltration at the anastomosis. The samples were evaluated at days 1, 3, and 7. Gross evaluation of anastomotic integrity was a secondary outcome. RESULTS: The self-forming magnet group displayed significant differences at each time point. On day 1, the stapled group displayed dense inflammatory cell infiltration and extensively ulcerated intestinal layers with significant edema. The self-forming magnet group showed less inflammatory infiltrate, and all intestinal layers remained compressed in direct apposition. By day 3, the self-forming magnet group already exhibited neovascularization with scant bacterial colonies. By contrast, stapled anastomoses had large areas of inflammation separating collagen fibers with prevalent bacterial infiltrations. On day 7, self-forming magnet anastomoses were characterized by robust neovascularization, maturing granulation tissue, and mucosal re-epithelization without significant inflammation. Meanwhile, stapled samples had persisting dense inflammation, tissue cavities with hemorrhage, and immature fibrous tissue. Grossly, the self-forming magnet created a patent lumen without defect, whereas stapled anastomoses demonstrated focal areas of serosal separation. CONCLUSION: Bowel anastomosis by self-forming magnets is associated with superior early histologic healing metrics, including early seal generation through mechanical compression, decreased inflammation, early neovascularization, lower bacterial infiltration, and faster re-epithelization.

Why Do Plant-Pathogenic Fungi Produce Mycotoxins? Potential Roles for Mycotoxins in the Plant Ecosystem.

For many plant-pathogenic or endophytic fungi, production of mycotoxins, which are toxic to humans, may present a fitness gain. However, associations between mycotoxin production and plant pathogenicity or virulence is inconsistent and difficult due to the complexity of these host-pathogen interactions and the influences of environmental and insect factors. Aflatoxin receives a lot of attention due to its potent toxicity and carcinogenicity but the connection between aflatoxin production and pathogenicity is complicated by the pathogenic ability and prevalence of nonaflatoxigenic isolates in crops. Other toxins directly aid fungi in planta, trichothecenes are important virulence factors, and ergot alkaloids limit herbivory and fungal consumption due to insect toxicity. We review a panel discussion at the American Phytopathological Society's Plant Health 2021 conference, which gathered diverse experts representing different research sectors, career stages, ethnicities, and genders to discuss the diverse roles of mycotoxins in the lifestyles of filamentous fungi of the families Clavicipitaceae, Trichocomaceae (Eurotiales), and Nectriaceae (Hypocreales).

Complete Genomes of Two Xanthomonas translucens pv. translucens Strains Isolated from Barley in North Dakota.

Xanthomonas translucens causes the disease bacterial leaf streak in several cereal crops and grasses. Here, we report the complete genome sequences of two isolates of X. translucens pv. translucens that were isolated from barley in an important cereal crop production region.

Evaluation of sutureless anastomosis after ileostomy takedown using the self-forming magnet anastomosis system in a porcine model.

BACKGROUND: Current standard of care for creation of small bowel anastomoses after a loop ileostomy reversal includes the use of stapler devices and sutures. Compression anastomosis devices have been used for decades, aimed toward improved outcomes with a "staple free" & "suture free" anastomosis. The self-forming magnet (SFM) device is a type of compression anastomosis device used to safely and effectively create an end-to-end small bowel anastomosis without the localized inflammatory response seen with sutures or staples, as no foreign bodies are left behind. METHODS: A Good Laboratory Practice preclinical study using a porcine model to evaluate creating an in vivo anastomosis via magnetic compression between two segments of small bowel (jejunum or ileum) was performed. Magnetic anastomoses were compared to stapled and handsewn anastomoses. Six animals were used for the magnetic anastomosis and eight for the two control groups for a total of 14 subjects. RESULTS: Mean creation times were 17.1 min (SD 6.06) for the SFM group, 10.3 min (SD 6.55, CI 95%) for the stapled anastomosis group, and 28.3 min (SD 2.63, CI 95%) for the suture anastomosis group, with a statistically significant difference among groups (p < 0.0021). All evaluated SFM anastomosis, stapled anastomosis, and handsewn anastomosis underwent a burst test with a pressure of 1.3 PSI. All six magnets used for anastomoses were naturally expelled. The range of days to expel magnets was 10-17 days. Intestinal anastomoses using magnets had considerably less residual scarring and intestinal distortion than anastomoses done with either suture or staples. CONCLUSION: This preclinical study documents the safety and efficacy of creating end-to-end small bowel anastomoses after ileostomy takedown using a magnetic compression device. The result is an anastomosis free of foreign objects with less inflammation, scarring, distortion, and mural thickening than seen in sutured or stapled anastomoses.

Detection of asinine gammaherpesviruses in association with pulmonary fibrosis in free-ranging donkeys.

A mortality event among recently captured feral donkeys (Equus asinus) occurred in south-central Utah in 2016. The deaths were sporadic, and clinical signs were indicative of respiratory disease, likely associated with an infectious etiology. Ten of 13 donkeys autopsied had moderate-to-severe interstitial fibrosing pneumonia, and one had pyogranulomatous pneumonia. Consensus PCRs directed toward the DNA polymerase and DNA packaging terminase subunit 1 for herpesviruses were performed followed by sequencing of the PCR amplicons and phylogenetic analysis. Asinine herpesvirus 4 (AsHV4) and 5 (AsHV5) were consistently identified in lung tissues of affected donkeys. No other herpesviruses were identified, and herpesviral DNA was not detected in lung tissues of 2 donkeys without evidence of respiratory disease. The detection of asinine gammaherpesviruses may have been associated with the lesions described. AsHV4 and AsHV5 have been reported in previous studies as novel gammaherpesviruses based on sequences obtained from donkeys with interstitial pneumonia and marked syncytial cell formation. Our findings suggest that the association of asinine gammaherpesviruses with respiratory conditions in equids deserves further attention.

An outbreak of SARS-CoV-2 with high mortality in mink (Neovison vison) on multiple Utah farms.

The breadth of animal hosts that are susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and may serve as reservoirs for continued viral transmission are not known entirely. In August 2020, an outbreak of SARS-CoV-2 occurred on five mink farms in Utah and was associated with high mink mortality (35-55% of adult mink) and rapid viral transmission between animals. The premise and clinical disease information, pathology, molecular characterization, and tissue distribution of virus within infected mink during the early phase of the outbreak are provided. Infection spread rapidly between independently housed animals and farms, and caused severe respiratory disease and death. Disease indicators were most notably sudden death, anorexia, and increased respiratory effort. Gross pathology examination revealed severe pulmonary congestion and edema. Microscopically there was pulmonary edema with moderate vasculitis, perivasculitis, and fibrinous interstitial pneumonia. Reverse transcriptase polymerase chain reaction (RT-PCR) of tissues collected at necropsy demonstrated the presence of SARS-CoV-2 viral RNA in multiple organs including nasal turbinates, lung, tracheobronchial lymph node, epithelial surfaces, and others. Localization of viral RNA by in situ hybridization revealed a more localized infection, particularly of the upper respiratory tract. Whole genome sequencing from multiple mink was consistent with published SARS-CoV-2 genomes with few polymorphisms. The Utah mink SARS-CoV-2 strains fell into Clade GH, which is unique among mink and other animal strains sequenced to date. While sharing the N501T mutation which is common in mink, the Utah strains did not share other spike RBD mutations Y453F and F486L found in nearly all mink from the United States. Mink in the outbreak reported herein had high levels of SARS-CoV-2 in the upper respiratory tract associated with symptomatic respiratory disease and death.

Genetic mapping of host resistance to the Pyrenophora teres f. maculata isolate 13IM8.3.

Spot form net blotch (SFNB), caused by the necrotrophic fungal pathogen Pyrenophora teres f. maculata (Ptm), is a foliar disease of barley that results in significant yield losses in major growing regions worldwide. Understanding the host-parasite interactions between pathogen virulence/avirulence genes and the corresponding host susceptibility/resistance genes is important for the deployment of genetic resistance against SFNB. Two recombinant inbred mapping populations were developed to characterize genetic resistance/susceptibility to the Ptm isolate 13IM8.3, which was collected from Idaho (ID). An Illumina Infinium array was used to produce a genome-wide marker set. Quantitative trait loci (QTL) analysis identified ten significant resistance/susceptibility loci, with two of the QTL being common to both populations. One of the QTL on 5H appears to be novel, while the remaining loci have been reported previously. Single nucleotide polymorphisms (SNPs) closely linked to or delimiting the significant QTL have been converted to user-friendly markers. Loci and associated molecular markers identified in this study will be useful in genetic mapping and deployment of the genetic resistance to SFNB in barley.

Peritoneal Mesothelioma in a Free-Ranging American Black Bear (Ursus americanus).

We necropsied an American black bear (Ursus americanus) from central Utah, US and found several liters of cloudy fluid and multiple white nodules in the peritoneal cavity. Histopathologic examination and staining with pancytokeratin and vimentin markers identified a peritoneal mesothelioma. Mesothelioma has not been reported previously in black bears.

Deletion of the benzoxazinoid detoxification gene NAT1 in Fusarium graminearum reduces deoxynivalenol in spring wheat.

Benzoxazinoid (Bx) metabolites produced by wheat and other members of the Poaceae have activity against Fusarium sp. that cause cereal diseases including Fusarium head blight (FHB) on wheat and barley. Certain Bx metabolites can be detoxified by Fusarium sp. with the arylamine N-acetyltransferase NAT1. Investigation of this pathway may reveal strategies for increasing FHB resistance, such as selection for higher levels of Bx metabolites within existing germplasm and/or engineering fungal susceptibility via host induced silencing of NAT1. We assessed the reactions of fifteen wheat cultivars or breeding lines adapted to the Northwestern United States to infection with F. graminearum Δnat1 mutants that should be sensitive to Bx metabolites. Significant differences were noted in disease severity and deoxynivalenol (DON) among the cultivars 21 d after inoculation with either mutant or wildtype (PH1) strains. Mutant vs. wildtype strains did not result in significant variation for infection severity (as measured by % infected florets), but inoculation with Δnat1 mutants vs. wildtype resulted in significantly lower DON concentrations in mature kernels (p < 0.0001). Of the cultivars tested, HRS3419 was the most resistant cultivar to PH1 (severity = 62%, DON = 45 ppm) and Δnat1 mutants (severity = 61%, DON = 30 ppm). The cultivar most susceptible to infection was Kelse with PH1 (severity = 100%, DON = 292 ppm) and Δnat1 mutants (severity = 100%, DON = 158 ppm). We hypothesized that sub-lethal Bx metabolite levels may suppress DON production in F. graminearum Δnat1 mutants. In vitro assays of Bx metabolites BOA, MBOA, and DIMBOA at 30 µM did not affect growth, but did reduce DON production by Δnat1 and PH1. Although the levels of Bx metabolites are likely too low in the wheat cultivars we tested to suppress FHB, higher levels of Bx metabolites may contribute towards reductions in DON and FHB.

Silencing efficiency of dsRNA fragments targeting Fusarium graminearum TRI6 and patterns of small interfering RNA associated with reduced virulence and mycotoxin production.

Deoxynivalenol (DON) contamination of cereal grains caused by Fusarium head blight may be addressed by future RNA interference (RNAi)-based gene silencing approaches. However, utilizing these approaches will require a greater understanding of the principles that govern RNAi effectiveness in the pathogen Fusarium graminearum. RNAi in higher eukaryotes, including fungi, involves processing double stranded RNA (dsRNA) into small interfering RNA (siRNA) that silence gene expression based on base pair complementarity. This study examined virulence, DON production, and the small RNA (sRNA) populations in response to RNAi-based silencing of TRI6, a transcription factor that positively regulates DON synthesis via control of TRI5 expression. Silencing was accomplished via the expression of transgenes encoding inverted repeats targeting various regions of TRI6 (RNAi vectors). Transgene expression was associated with novel, TRI6-specific siRNAs. For RNAi vectors targeting the majority of TRI6 sequence (~600 bp), a discontinuous, repeatable pattern was observed in which most siRNAs mapped to specific regions of TRI6. Targeting shorter regions (250-350 bp) did not alter the siRNA populations corresponding to that region of TRI6. No phased processing was observed. The 5' base of ~83% of siRNAs was uracil, consistent with DICER processing and ARGONAUTE binding preferences for siRNA. Mutant lines showed TRI6 siRNA-associated reductions of TRI5 expression on toxin inducing media and DON in infected wheat and barley spikes. Shorter RNAi vectors resulted in variable levels of silencing that were less than for the ~600 bp RNAi vector, with a 343 bp RNAi vector targeting the 5' end of TRI6 having the best silencing efficiency. This work identifies efficient shorter region for silencing of TRI6 and describes the patterns of siRNA corresponding to those regions.

The Art of Science Communication-A Novel Approach to Science Communication Training.

Effective communication is a requisite skill for scientists. However, formalized training in this area is often unavailable for members of the scientific community. As one approach to combat this problem, the American Society for Biochemistry and Molecular Biology (ASBMB) developed The Art of Science Communication, an eight-week-long online course that provides facilitated instruction on how to communicate science in an oral format. The course is offered three times a year, and as of December 2017, nearly 200 individuals from all career stages have taken part in it. The course completion rate is currently 60%, a rate three to five times as high as the average for similar Massive Open Online Courses (MOOCs). Participants have indicated that taking the course has improved their ability to communicate about their research, and that the skills and lessons learned have benefited them professionally. Moving forward, we are examining approaches that will help us improve the course and expand its reach throughout the scientific community. This article details the development of the course and examines the role and potential of such training within the larger scientific community.

HAEMOPROTEUS ( PARAHAEMOPROTEUS) SPP. IN CAPTIVE-BRED BOBWHITE QUAIL ( COLINUS VIRGINIANUS) IN SOUTHERN UTAH, USA.

A captive-bred Bobwhite Quail ( Colinus virginianus) ranch in southern Utah, US experienced high mortality rates in the late summer and fall of 2012. Nine juvenile birds were necropsied at the Utah Veterinary Diagnostic Laboratory. Gross lesions included pale skeletal muscle with multifocal hemorrhages and petechiae in the air sacs and serosal surfaces of most organs. Histologically there was moderate to severe, multifocal, degenerative myositis with intramyofiber schizonts and minimal lymphoplasmacytic infiltrates in the proventriculus, ventriculus, heart, and skeletal muscle. There was also moderate fibrinoid to heterophilic vasculitis in multiple organs with vascular intraendothelial or intravascular merozoites and scattered thrombosis. In the liver and spleen there were multiple degenerative schizonts that had ruptured. Blood smears from three of the birds were stained with Wright-Giemsa stain and examined at a referral laboratory. Although the blood cells were deteriorated (postmortem artifact), life stages (exact stages not specified) consistent with Haemoproteus spp. were identified in erythrocytes. Polymerase chain reaction done on pooled tissues from two birds produced an amplicon in both pooled samples, and direct sequencing confirmed the presence of 533 base pairs of a Haemoproteus sp. in the subgenus Parahaemoproteus. The identification of Parahaemoproteus spp. in quail in southern Utah implies that appropriate Culicoides spp. vectors are present in the state and that there is potential risk to other birds such as zoo and aviary populations, wild turkeys, and other game birds.

Sharing mutants and experimental information prepublication using FgMutantDb (https://scabusa.org/FgMutantDb).

There is no comprehensive storage for generated mutants of Fusarium graminearum or data associated with these mutants. Instead, researchers relied on several independent and non-integrated databases. FgMutantDb was designed as a simple spreadsheet that is accessible globally on the web that will function as a centralized source of information on F. graminearum mutants. FgMutantDb aids in the maintenance and sharing of mutants within a research community. It will serve also as a platform for disseminating prepublication results as well as negative results that often go unreported. Additionally, the highly curated information on mutants in FgMutantDb will be shared with other databases (FungiDB, Ensembl, PhytoPath, and PHI-base) through updating reports. Here we describe the creation and potential usefulness of FgMutantDb to the F. graminearum research community, and provide a tutorial on its use. This type of database could be easily emulated for other fungal species.

Association of prospective lower extremity musculoskeletal injury and musculoskeletal, balance, and physiological characteristics in Special Operations Forces.

OBJECTIVES: Previous research has examined lower extremity (LE) musculoskeletal injury (MSI) patterns and risk factors in Special Operations Forces (SOF) trainees, conventional military personnel, and athletes; however, it is unclear if SOF have the same patterns/risk factors. This study aimed to determine the association of musculoskeletal, balance, and physiological characteristics with LE MSI in SOF. DESIGN: Cohort study. METHODS: A total of 726 Air Force (N=140), Navy Sea, Air, and Land (N=301), and Special Warfare Combatant Crewmen (N=285) SOF (age=25.72±4.77years, height=178.34±6.63cm, weight=84.28±9.03kg) participated in laboratory testing, including: LE muscular strength and flexibility; balance; body composition; anaerobic power/capacity; and aerobic capacity. Medical charts were reviewed for LE MSI 365days following laboratory testing. Participants were assigned by injury status and laboratory data stratified by tertile. Chi-square statistics were calculated to determine the frequency of LE MSI across tertiles for each characteristic. RESULTS: There was a significant association between LE MSI and: ankle inversion strength (weaker side: Χ(2)=17.703; stronger side: Χ(2)=18.911; p≤0.001); ankle eversion/inversion strength ratio (lower side: Χ(2)=13.456; higher side: Χ(2)=16.885; p≤0.001); hamstring flexibility (less flexible: Χ(2)=19.930; more flexible Χ(2)=15.185; p≤0.001); gastrocnemius-soleus flexibility (less flexible: Χ(2)=7.889, p=0.019); dynamic balance asymmetry (Χ(2)=7.444, p=0.024); Vestibular and Preference ratios (Χ(2)=9.124, p=0.010 and Χ(2)=6.572, p=0.037, respectively); and aerobic capacity (Χ(2)=13.935, p=0.001). CONCLUSIONS: Characteristics associated with LE MSI are unique in SOF. Human performance program initiatives should include efforts to optimize ankle strength and flexibility, maintain moderate hamstring flexibility, expand dynamic balance strategies, and maximize aerobic capacity to reduce LE MSI risk.