Discriminating Between Copers and those with Chronic Ankle Instability with Clinical Outcomes

Introduction: Chronic ankle instability (CAI) is a common consequence of lateral ankle sprains and a leading cause of post-traumatic ankle osteoarthritis. Despite a history of a lateral ankle sprain, there are some individuals (copers) who appear to have a mechanism that allows them to avoid recurrent lateral ankle sprains and giving way episodes. To date, research has identified differences in perceptual (e.g. self-assessed disability), mechanical (e.g. ligament laxity), and sensorimotor (e.g. balance) outcomes between copers and those with CAI. However, the vast majority of these outcomes are considered research and/or laboratory based and thus impractical for most clinicians to use on a daily basis. The identification of clinical tools, capable of discriminating between copers and those with CAI is the first step in developing an accurate clinical battery of tests that can be used to predict patients more likely to develop CAI after an initial lateral ankle sprain. Therefore, the purpose of this investigation was to establish the ability of clinical outcome measures to discriminate between copers and those with CAI. Methods: Participants- A total of 46 participants (21 copers and 25 with CAI) were enrolled and completed the investigation. All participants read and signed the university approved informed consent document prior to participation. All subjects were between the ages of 18-30 and participated in recreational activity for at least 90 minutes each week. Copers and those with CAI had a history of at least one moderate to severe ankle sprain that required acute care. Copers resumed all pre-injury activity without limitation and without recurrent episodes of injury/giving way for at least 12 months prior to testing and scored \u3e24 on the ankle joint functional assessment tool (AJFAT). In comparison, those with CAI had at least one recurrent sprain within 6 months of testing and scored \u3c20 on the AJFAT. Protocol: Perceptual outcomes, the Foot Ankle Ability Measure (FAAM) and FAAM Sport (FAAM-S) were completed first. These questionnaires determine a subject’s self-assessed limitations during daily and physical activities and lower scores represent greater disability. Next, mechanical adaptations were assessed via the the weight bearing lunge test (WBLT) and talar glide test. The WBLT measured ankle dorsiflexion range of motion using a modified lunge technique. Maximum dorsiflexion was defined as the furthest distance the subject’s foot could be from a wall while the subject’s knee was in contact with the wall and the subject’s heel was flat on the ground. The talar glide test measures the magnitude of the talus’ posterior glide by measuring the change in passive knee flexion, from 90°, while an examiner simultaneously pushes the talus posteriorly and pulls the foot into dorsiflexion until the point of capsular restriction. Finally, sensorimotor dysfunction was assessed with the Star Excursion Balance Test (SEBT) and isometric strength testing. The SEBT is a measure of dynamic postural control and measures the distance a subject can reach with their contralateral leg without losing their balance. Reach distance was defined as the farthest point that an individual could touch without accepting weight on their reach leg and maintaining balance through the return to a bilateral stance. Reach distance was normalized to each subject’s leg length. Using a handheld dynamometer, ankle dorsiflexion, plantar flexion, eversion, and inversion strength was assessed during 3-second maximal contractions. Strength was normalized to a subject’s body weight. Statistical Analysis: Group differences in perceptual, mechanical, and sensorimotor outcomes were assessed with independent sample t-tests. If significance was achieved, the ability of an outcome to accurately discriminate between the groups was then assessed using the area under the curve (AUC) for receiver operating characteristic (ROC) curves. A ROC curve illustrates the “trade-off” between sensitivity and specificity throughout a measure’s entire range of values. Finally, clinical meaningfulness was established by the calculation of cut-off scores and likelihood ratios for outcomes that successfully discriminated between copers and those with CAI. A traditional level of significance (=0.05) was used for all analyses. RESULTS: The CAI group had significant perceptual and sensorimotor deficits relative to the coper group. Specifically, FAAM (Coper: 99.0±2.5%, CAI: 89.2±9.5%), FAAM-S (Coper: 96.6±4.1%, CAI: 79.3±16.2%), normalized posteriomedial SEBT reach distance (Coper: 91.2±8.1%, CAI: 83.5±6.5%), and normalized dorsiflexion strength (Coper: 74.5±19.9%, CAI: 62.9±12.8%) were lower in those with CAI relative to copers. The secondary analysis revealed that the FAAM (P\u3c0.01), FAAM-S (p\u3c0.01), and posteriomedial SEBT reach distance (p\u3c0.01) accurately discriminated between copers and those with CAI. The AUC scores, cut-off scores, and likelihood ratios can be seen in the table below. No differences existed with regards to talar glide, range of motion, anterior SEBT reach, posteriolateral SEBT reach, and normalized eversion, inversion, or plantar flexion strength. Outcome Measure Category AUC Cut-off Score Likelihood Ratios (95% Confidence Intervals) Positive Negative FAAM 0.93 99% 5.05 (2.92 to 10.36) 0.05 (0.01 to 0.21) FAAM-S 0.90 88% 31.00 (4.42 to 217.67) 0.33 (0.22 to 0.51) Posteriomedial SEBT Reach 0.79 89% 2.53 (1.77 to 3.79) 0.06 (0.02 to 0.27) DISCUSSION: The results of this investigation indicate that perceptual (FAAM, FAAM-S) and some sensorimotor outcomes (posteriomedial SEBT reach distance) can discriminate between copers and those with CAI. We hypothesize that these outcome measures represent part of the mechanism that 1) allows copers to function as if uninjured and 2) is absent in individuals who develop CAI. The current results, using clinical outcome measures, support previous findings that have illustrated perceptual and sensorimotor deficits using research and/or laboratory based outcomes. Further, because a variety of the current outcomes (perceptual and sensorimotor) successfully discriminated between copers and those with CAI, groups, the current findings support the theory that the causal mechanism of CAI is multi-factorial in nature. However, the most important finding of the current investigation is that the perceptual outcomes demonstrated the greatest ability to discriminate between copers and those with CAI with accuracy point estimates ranging from 0.90-0.93. Indeed, the perceptual outcomes (as a whole) demonstrated higher AUC estimates and positive likelihood ratios, as well as lower negative likelihood ratios compared with the sensorimotor outcomes (as a whole). This finding, which supports previous results, indicates that perceptual outcomes have the greatest ability to accurately predict those who became copers after initial lateral ankle sprain. Given the magnitude of the positive and negative likelihood ratios, we believe that perception based outcomes, such as the FAAM and FAAM-S, should be used in future longitudinal research investigations designed to determine if and when, post-injury, these outcomes can predict who will develop CAI after an initial lateral ankle sprain

Plasticity of a critical antigenic determinant in the West Nile virus NY99 envelope protein domain III

West Nile virus (WNV) is a mosquito-borne flavivirus that causes febrile illness, encephalitis, and occasionally death in humans. The envelope protein is the main component of the WNV virion surface, and domain III of the envelope protein (EIII) is both a putative receptor binding domain and a target of highly specific, potently neutralizing antibodies. Envelope E-332 (E-332) is known to have naturally occurring variation and to be a key determinant of neutralization for anti-EIII antibodies. A panel of viruses containing all possible amino acid substitutions at E-332 was constructed. E-332 was found to be highly tolerant of mutation, and almost all of these changes had large impacts on antigenicity of EIII but only limited effects on growth or virulence phenotypes

Recovery of West Nile Virus Envelope Protein Domain III Chimeras with Altered Antigenicity and Mouse Virulence

ABSTRACT Flaviviruses are positive-sense, single-stranded RNA viruses responsible for millions of human infections annually. The envelope (E) protein of flaviviruses comprises three structural domains, of which domain III (EIII) represents a discrete subunit. The EIII gene sequence typically encodes epitopes recognized by virus-specific, potently neutralizing antibodies, and EIII is believed to play a major role in receptor binding. In order to assess potential interactions between EIII and the remainder of the E protein and to assess the effects of EIII sequence substitutions on the antigenicity, growth, and virulence of a representative flavivirus, chimeric viruses were generated using the West Nile virus (WNV) infectious clone, into which EIIIs from nine flaviviruses with various levels of genetic diversity from WNV were substituted. Of the constructs tested, chimeras containing EIIIs from Koutango virus (KOUV), Japanese encephalitis virus (JEV), St. Louis encephalitis virus (SLEV), and Bagaza virus (BAGV) were successfully recovered. Characterization of the chimeras in vitro and in vivo revealed differences in growth and virulence between the viruses, with in vivo pathogenesis often not being correlated with in vitro growth. Taken together, the data demonstrate that substitutions of EIII can allow the generation of viable chimeric viruses with significantly altered antigenicity and virulence. IMPORTANCE The envelope (E) glycoprotein is the major protein present on the surface of flavivirus virions and is responsible for mediating virus binding and entry into target cells. Several viable West Nile virus (WNV) variants with chimeric E proteins in which the putative receptor-binding domain (EIII) sequences of other mosquito-borne flaviviruses were substituted in place of the WNV EIII were recovered, although the substitution of several more divergent EIII sequences was not tolerated. The differences in virulence and tissue tropism observed with the chimeric viruses indicate a significant role for this sequence in determining the pathogenesis of the virus within the mammalian host. Our studies demonstrate that these chimeras are viable and suggest that such recombinant viruses may be useful for investigation of domain-specific antibody responses and the more extensive definition of the contributions of EIII to the tropism and pathogenesis of WNV or other flaviviruses

Decomposability of soil organic matter over time: the Soil Incubation Database (SIDb, version 1.0) and guidance for incubation procedures

The magnitude of carbon (C) loss to the atmosphere via microbial decomposition is a function of the amount of C stored in soils, the quality of the organic matter, and physical, chemical, and biological factors that comprise the environment for decomposition. The decomposability of C is commonly assessed by laboratory soil incubation studies that measure greenhouse gases mineralized from soils under controlled conditions. Here, we introduce the Soil Incubation Database (SIDb) version 1.0, a compilation of time series data from incubations, structured into a new, publicly available, open-access database of C flux (carbon dioxide, CO2, or methane, CH4). In addition, the SIDb project also provides a platform for the development of tools for reading and analysis of incubation data as well as documentation for future use and development. In addition to introducing SIDb, we provide reporting guidance for database entry and the required variables that incubation studies need at minimum to be included in SIDb. A key application of this synthesis effort is to better characterize soil C processes in Earth system models, which will in turn reduce our uncertainty in predicting the response of soil C decomposition to a changing climate. We demonstrate a framework to fit curves to a number of incubation studies from diverse ecosystems, depths, and organic matter content using a built-in model development module that integrates SIDb with the existing SoilR package to estimate soil C pools from time series data. The database will help bridge the gap between point location measurements, which are commonly used in incubation studies, and global remote-sensed data or data products derived from models aimed at assessing global-scale rates of decomposition and C turnover. The SIDb version 1.0 is archived and publicly available at https://doi.org/10.5281/zenodo.3871263 (Sierra et al., 2020), and the database is managed under a version-controlled system and centrally stored in GitHub (https://github.com/SoilBGC-Datashare/sidb, last access: 26 June 2020)

G Protein-Coupled Estrogen Receptor (GPER) Expression in Normal and Abnormal Endometrium

Rapid estrogen effects are mediated by membrane receptors, and evidence suggests a role for both a membrane-associated form of estrogen receptor alpha (ESR1; ERα) and G-protein coupled receptor 30 (GPER; GPR30). Considering estrogen’s importance in endometrial physiology and endometriosis pathophysiology, we hypothesized that GPER could be involved in both cyclic changes in endometrial estrogen action and that aberrant expression might be seen in the eutopic endometrium of women with endometriosis. Using real-time reverse transcriptase–polymerase chain reaction (RT-PCR) and immunohistochemical analysis of normal endometrium, endometrial samples demonstrated cycle-regulated expression of GPER, with maximal expression in the proliferative phase. Eutopic and ectopic endometrium from women with endometriosis overexpressed GPER as compared to eutopic endometrium of normal participants. Ishikawa cells, an adenocarcinoma cell line, expressed GPER, with increased expression upon treatment with estrogen or an ESR1 agonist, but not with a GPER-specific agonist. Decreased expression was seen in Ishikawa cells stably transfected with progesterone receptor A. Together, these data suggest that normal endometrial GPER expression is cyclic and regulated by nuclear estrogen and progesterone receptors, while expression is dysregulated in endometriosis

SARS-like WIV1-CoV poised for human emergence

The emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome (MERS)-CoV highlights the continued risk of cross-species transmission leading to epidemic disease. This manuscript describes efforts to extend surveillance beyond sequence analysis, constructing chimeric and full-length zoonotic coronaviruses to evaluate emergence potential. Focusing on SARS-like virus sequences isolated from Chinese horseshoe bats, the results indicate a significant threat posed by WIV1-CoV. Both full-length and chimeric WIV1-CoV readily replicated efficiently in human airway cultures and in vivo, suggesting capability of direct transmission to humans. In addition, while monoclonal antibody treatments prove effective, the SARS-based vaccine approach failed to confer protection. Together, the study indicates an ongoing threat posed by WIV1-related viruses and the need for continued study and surveillance

Analyzing the Human Serum Antibody Responses to a Live Attenuated Tetravalent Dengue Vaccine Candidate

BACKGROUND: Dengue virus serotypes 1-4 (DENV-1-4) are the most common vector-borne viral pathogens of humans and the etiological agents of dengue fever and dengue hemorrhagic syndrome. A live-attenuated tetravalent dengue vaccine (TDV) developed by Takeda Vaccines has recently progressed to phase 3 safety and efficacy evaluation. METHODS: We analyzed the qualitative features of the neutralizing antibody (nAb) response induced in naive and DENV-immune individuals after TDV administration. Using DENV-specific human monoclonal antibodies (mAbs) and recombinant DENV displaying different serotype-specific Ab epitopes, we mapped the specificity of TDV-induced nAbs against DENV-1-3. RESULTS: Nearly all subjects had high levels of DENV-2-specific nAbs directed to epitopes centered on domain III of the envelope protein. In some individuals, the vaccine induced nAbs that tracked with a DENV-1-specific neutralizing epitope centered on domain I of the envelope protein. The vaccine induced binding Abs directed to a DENV-3 type-specific neutralizing epitope, but findings of mapping of DENV-3 type-specific nAbs were inconclusive. CONCLUSION: Here we provide qualitative measures of the magnitude and epitope specificity of the nAb responses to TDV. This information will be useful for understanding the performance of TDV in clinical trials and for identifying correlates of protective immunity

Immune Predictors of Mortality After Ribonucleic Acid Virus Infection

Background Virus infections result in a range of clinical outcomes for the host, from asymptomatic to severe or even lethal disease. Despite global efforts to prevent and treat virus infections to limit morbidity and mortality, the continued emergence and re-emergence of new outbreaks as well as common infections such as influenza persist as a health threat. Challenges to the prevention of severe disease after virus infection include both a paucity of protective vaccines as well as the early identification of individuals with the highest risk that may require supportive treatment. Methods We completed a screen of mice from the Collaborative Cross (CC) that we infected with influenza, severe acute respiratory syndrome-coronavirus, and West Nile virus. Results The CC mice exhibited a range of disease manifestations upon infections, and we used this natural variation to identify strains with mortality after infection and strains exhibiting no mortality. We then used comprehensive preinfection immunophenotyping to identify global baseline immune correlates of protection from mortality to virus infection. Conclusions These data suggest that immune phenotypes might be leveraged to identify humans at highest risk of adverse clinical outcomes upon infection, who may most benefit from intensive clinical interventions, in addition to providing insight for rational vaccine design

Yellow Fever: Roles of Animal Models and Arthropod Vector Studies in Understanding Epidemic Emergence

Yellow fever virus (YFV) is a mosquito-borne flavivirus circulating throughout the tropical and sub-tropical regions of Africa and South America. It is responsible for an estimated 30,000 deaths annually, and while there is a highly successful vaccine, coverage is incomplete, and there is no approved treatment for YFV infection. Despite advancements in the field, animal models for YFV infection remain scarce, and care must be taken to select an appropriate model for a given hypothesis. Small animal models require either adapted YFV strains or immunocompromised hosts. Non-human primates (NHPs) recapitulate human disease, but they require specialized facilities and training, are often in short supply and cost-prohibitive, and can present ethical concerns. The limitations in studying the mosquito vectors for YFV infection include inconsistency in the laboratory environment, the requirement for a high containment insectary, and difficulty in maintaining sylvatic mosquitoes. In this review, we discuss the roles of animal models and arthropod vector studies in understanding epidemic emergence

Morphologic and Genetic Characterization of Ilheus Virus, a Potential Emergent Flavivirus in the Americas

Ilheus virus (ILHV) is a mosquito-borne flavivirus circulating throughout Central and South America and the Caribbean. It has been detected in several mosquito genera including Aedes and Culex, and birds are thought to be its primary amplifying and reservoir host. Here, we describe the genomic and morphologic characterization of ten ILHV strains. Our analyses revealed a high conservation of both the 5′- and 3′-untranslated regions but considerable divergence within the open reading frame. We also showed that ILHV displays a typical flavivirus structural and genomic organization. Our work lays the foundation for subsequent ILHV studies to better understand its transmission cycles, pathogenicity, and emergence potential