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Considerable clinical evidence supports that increased blood–brain barrier (BBB) permeability is linked to immune extravasation of CNS parenchyma during neuroinflammation. Although BBB permeability and immune extravasation are known to be provoked by vascular endothelial growth factor-A (i.e., VEGF-A) and C-X-C motif chemokine ligand 12 (CXCL12), respectively, the mechanisms that link both processes are still elusive. The interleukin-20 (i.e., IL-20) cytokine signaling pathway was previously implicated in VEGF-mediated angiogenesis and is known to induce cellular response by way of signaling through IL-20 receptor subunit β (i.e., IL-20RB). Dysregulated IL-20 signaling is implicated in many inflammatory pathologies, but it’s contribution to neuroinflammation has yet to be reported. We hypothesize that the IL-20 cytokine, and the IL cytokine subfamily more broadly, play a key role in CNS neuroinflammation by signaling through IL-20RB, induce VEGF activity, and enhance both BBB-permeability and CXCL12-mediated immune extravasation. To address this hypothesis, we actively immunized IL-20RB–/– mice and wild-type mice to induce experimental autoimmune encephalomyelitis (EAE) and found that IL-20RB–/– mice showed amelioration of disease progression compared to wild-type mice. Similarly, we passively immunized IL-20RB–/– mice and wild-type mice with myelin-reactive Th1 cells from either IL-20RB–/– and wild-type genotype. Host IL-20RB–/– mice showed lesser disease progression than wild-type mice, regardless of the myelin-reactive Th1 cells genotype. Using multianalyte bead-based immunoassay and ELISA, we found distinctive changes in levels of pro-inflammatory cytokines between IL-20RB–/– mice and wild-type mice at peak of EAE. We also found detectable levels of all cytokines of the IL-20 subfamily within CNS tissues and specific alteration to IL-20 subfamily cytokines IL-19, IL-20, and IL-24, expression levels. Immunolabeling of CNS region-specific microvessels confirmed IL-20RB protein at the spinal cord microvasculature and upregulation during EAE. Microvessels isolated from macaques CNS tissues also expressed IL-20RB. Moreover, we identified the expression of all IL-20 receptor subunits: IL-22 receptor subunit α-1 (IL-22RA1), IL-20RB, and IL-20 receptor subunit α (IL-20RA) in human CNS microvessels. Notably, human cerebral microvasculature endothelial cells (HCMEC/D3) treated with IL-1β showed augmented expression of the IL-20 receptor. Lastly, IL-20-treated HCMEC/D3 showed alterations on CXCL12 apicobasal polarity consistent with a neuroinflammatory status. This evidence suggests that IL-20 subfamily cytokines may signal at the BBB via IL-20RB, triggering neuroinflammation.

Background: Functionality testing following anterior cruciate ligament (ACL) reconstruction can benefit clinicians and patients in determining readiness for return to sport. While a component of a multifactorial decision, inability to perform well on these tests predicts increased risk of reinjury. As of 2013, only 41% of orthopaedic surgeons report using strength or functionality testing in evaluating patients for return to sport (RTS). Indications: In the intermediate to late stages of their rehabilitation program, patients may undergo these tests to determine readiness and safety to return to sport. Technique Description: The tests described in this video include the single and triple hop for distance, triple crossover hop, single and double vertical leg jump tasks, drop jump landing task, and isokinetic and isometric strength testing. Results: Clinicians who incorporate these tests into patient rehabilitation programs may reduce patient risk of ACL reinjury by 75% to 84%. The limb symmetry index (LSI) is a reliable calculated measure for these tests, with a strong reliability for the hop tests. The limb symmetry index can be calculated for each test and represents the ratio of measured performance of the involved, or post-ACL reconstruction, leg when compared against the uninvolved leg. The commonly used limb symmetry index threshold for passing each hop test is 90%. Patients who score 90% or greater on each of these tests are less likely to experience knee reinjury. Discussion/Conclusion: The ability of knee strength and functionality tests in determining RTS following ACL reconstruction has been highlighted as a tool in potentially reducing risk of knee reinjury. Other isometric and isokinetic testing can be used in addition to the described functionality tests but may not be possible in certain practices due to equipment and funding limitations. Usage of these strength and functionality tests, in conjunction with clinician evaluation, may lead to more optimal outcomes for patients and lower rates of reinjury. Psychological assessment may aid in evaluating patient readiness for return to sport. Importantly, further sport-specific testing is still recommended and will optimize patient outcomes.

Background: Blood flow restriction (BFR) training restricts arterial inflow and venous outflow from the extremity and can produce gains in muscle strength at low loads. Low-load training reduces joint stress and decreases cardiovascular risk when compared with high-load training, thus making BFR an excellent option for many patients requiring rehabilitation. Indications: Blood flow restriction has shown clinical benefit in a variety of patient populations including healthy patients as well as those with osteoarthritis, anterior cruciate ligament reconstruction, polymyositis/dermatomyositis, and Achilles tendon rupture. Technique Description: This video demonstrates BFR training in 3 clinical areas: upper extremity resistance training, lower extremity resistance training, and low-intensity cycling. All applications of BFR first require determination of total occlusion pressure. Upper extremity training requires inflating the tourniquet to 50% of total occlusion pressure, while lower extremity exercises use 80% of total occlusion pressure. Low-load resistance training exercises follow a specific repetition scheme: 30 reps followed by a 30-second rest and then 3 sets of 15 reps with 30-seconds rest between each. During cycle training, 80% total occlusion pressure is used as the patient cycles for 15 minutes without rest. Results: Augmenting low-load resistance training with BFR increases muscle strength when compared with low-load resistance alone. In addition, low-load BFR has demonstrated an increase in muscle mass greater than low-load training alone and equivalent to high-load training absent BFR. A systematic review determined the safety of low-load training with BFR is comparable to traditional high-intensity resistance training. The most common adverse effects include exercise intolerance, discomfort, and dull pain which are also frequent in patients undergoing traditional resistance training. Severe adverse effects including deep vein thrombosis, pulmonary embolism, and rhabdomyolysis are exceedingly rare, less than 0.006% according to a national survey. Patients undergoing BFR rehabilitation experience less perceived exertion and demonstrate decreased pain scores compared with high-load resistance training. Conclusion: Blood flow restriction training is an effective alternative to high-load resistance training for patients requiring musculoskeletal rehabilitation for multiple disease processes as well as in the perioperative setting. Blood flow restriction has been shown to be a safe training modality when managed by properly trained physical therapists and athletic trainers.

Multiple Sclerosis (MS) is an immune-mediated disease that results in major locomotor deficits. However, recent studies have revealed that fatigue, slow processing speed, and memory impairment are the top variables impacting employment status for MS patients. These suggest that cognitive effects may have a greater impact on productivity, lifestyle, and quality of life than do disease-related motor deficits. However, these debilitating non-locomotive effects have been largely overlooked in rodent models of the disease, such as experimental autoimmune encephalomyelitis (EAE). We hypothesized that murine EAE can also be used to assess non-locomotive dysfunctions (mood, sociability, muscle strength, and balance), as well as potential biases in these dysfunctions due to sex and/or strain. We actively immunized male and female C57BL/6 (B6) and SJL mice for EAE and evaluated their performance on the Deacon's weight grip test, Kondziela's inverted screen test, Hall's rope grip test, manual von Frey test for somatic nociception, and a three-chamber social preference paradigm. We hypothesized that EAE progression is associated with changes in muscle strength, balance, pain, and sociability and that these variations are linked to sex and/or strain. Our results indicate that strain but not sex influenced differences in muscle strength and balance during EAE, and both sex and strain have an impact on mechanical nociception, regardless of EAE disease status. Furthermore, both sex and strain had complex effects on differences in sociability. In conclusion, testing these additional modalities during EAE helps to unveil other signs and symptoms that could be used to determine the efficacy of a drug or treatment in the modulation of a MS-like behavior.

Background: Dry needling is an increasingly popular technique for relieving musculoskeletal pain, through targeting myofascial trigger points. Existing evidence indicates that dry needling is effective in short-term management of pain, with research showing efficacy in improving functional outcomes when compared to other treatments. Indications: Myofascial trigger points may result in range of motion limitations and muscle weakness. Noninvasive treatments for musculoskeletal pain associated with myofascial trigger points, such as stretching or warm compresses, may not provide significant benefit for patients. Dry needling is a minimally invasive technique that provides significant short-term functional improvement and analgesia for musculoskeletal pain associated with myofascial trigger points, seen with reductions in visual analog scale (VAS) pain scores and decreased need for pharmacologic therapies. Technique Description: This video demonstrates dry needling at the tensor fascia latae, extensor carpi radialis longus , and gastrocnemius muscles. Myofascial trigger points and important anatomical landmarks are identified and labeled. A needle is inserted into the region of the myofascial trigger point to a depth of 10 to 100 mm and remains in situ for about 10 to 20 seconds with the use of a piston technique, although this time varies dependent upon patient response. Results: Dry needling may significantly decrease pain and improve functional outcomes in patients with short-term musculoskeletal ailments. Randomized controlled trials have examined dry needling for fibromyalgia, mechanical neck pain, myofascial pain, and following total knee arthroplasty, with these finding significant reductions in pain scores in short-term follow-up. A randomized single-blind placebo-controlled trial found dry needling with exercise to be more effective than sham dry-needling and exercise. Dry needling may provide improved long-term analgesia but also results in increased pain during the procedure and increased soreness afterwards. Pain relief may not last beyond 6 months, although little research investigating long-term outcomes has been performed. Discussion/Conclusion: The usage of dry needling in clinical settings is increasing, as it has shown strong efficacy in providing short-term pain relief and ability to improve functional outcomes. Dry needling is a simple, minimally invasive technique that is easily and quickly learned by physical therapists and may provide great benefits to patients.

Two-dimensional ultrasound transducers enable the acquisition of fully volumetric data that have been demonstrated to provide greater diagnostic information in the clinical setting and are a critical tool for emerging ultrasound methods, such as super-resolution and functional imaging. This technology, however, is not without its limitations. Due to increased fabrication complexity, some matrix probes with disjoint piezoelectric panels may require initial calibration. In this manuscript, two methods for calibrating the element positions of the Vermon 1024-channel 8 MHz matrix transducer are detailed. This calibration is a necessary step for acquiring high resolution B-mode images while minimizing transducer-based image degradation. This calibration is also necessary for eliminating vessel-doubling artifacts in super-resolution images and increasing the overall signal-to-noise ratio (SNR) of the image. Here, we show that the shape of the point spread function (PSF) can be significantly improved and PSF-doubling artifacts can be reduced by up to 10 dB via this simple calibration procedure. [ABSTRACT FROM AUTHOR]

Abstract Background The Critical Assessment of Functional Annotation (CAFA) is an ongoing, global, community-driven effort to evaluate and improve the computational annotation of protein function. Results Here, we report on the results of the third CAFA challenge, CAFA3, that featured an expanded analysis over the previous CAFA rounds, both in terms of volume of data analyzed and the types of analysis performed. In a novel and major new development, computational predictions and assessment goals drove some of the experimental assays, resulting in new functional annotations for more than 1000 genes. Specifically, we performed experimental whole-genome mutation screening in Candida albicans and Pseudomonas aureginosa genomes, which provided us with genome-wide experimental data for genes associated with biofilm formation and motility. We further performed targeted assays on selected genes in Drosophila melanogaster, which we suspected of being involved in long-term memory. Conclusion We conclude that while predictions of the molecular function and biological process annotations have slightly improved over time, those of the cellular component have not. Term-centric prediction of experimental annotations remains equally challenging; although the performance of the top methods is significantly better than the expectations set by baseline methods in C. albicans and D. melanogaster, it leaves considerable room and need for improvement. Finally, we report that the CAFA community now involves a broad range of participants with expertise in bioinformatics, biological experimentation, biocuration, and bio-ontologies, working together to improve functional annotation, computational function prediction, and our ability to manage big data in the era of large experimental screens.

Patient drug regime compliance is a major issue; sustained release implants could address this. Here, the authors report on a phase inverted in situ forming implant of PLGA for the sustained release of antiretroviral drugs and optimize and demonstrate the release of 6 different drugs over a period of up to a year.

Abstract Background Female sex workers, MSM, and transgender women—collectively referred to as key populations (KPs)—are disproportionately affected by gender-based violence (GBV) and HIV, yet little is known about the violence they face, its gender-based origins, and responses to GBV. The purpose of this study was to understand the nature and consequences of GBV experienced, to inform HIV policies and programming and to help protect KPs’ human rights. Methods Using a participatory approach, FSWs, MSM, and transgender women in Barbados, El Salvador, Trinidad and Tobago, and Haiti conducted 278 structured interviews with peers to understand their experiences of and responses to GBV. Responses to open-ended questions were coded in NVivo and analyzed using an applied thematic analysis. Results Nearly all participants experienced some form of GBV. Emotional and economic GBV were the most commonly reported but approximately three-quarters of participants reported sexual and physical GBV and other human rights violations. The most common settings for GBV were at home, locations where sex work took place such as brothels, bars and on the street; public spaces such as parks, streets and public transport, health care centers, police stations and—for transgender women and MSM—religious settings and schools. The most common perpetrators of violence included: family, friends, peers and neighbors, strangers, intimate partners, sex work clients and other sex workers, health care workers, police, religious leaders and teachers. Consequences included emotional, physical, and sexual trauma; lack of access to legal, health, and other social services; and loss of income, employment, housing, and educational opportunities. Though many participants disclosed experiences of GBV to friends, colleagues and family, they rarely sought services following violence. Furthermore, less than a quarter of participants believed that GBV put them at risk of HIV. Conclusions Our study found that across the four study countries, FSWs, MSM, and transgender women experienced GBV from state and non-state actors throughout their lives, and much of this violence was directly connected to rigid and harmful gender norms. Through coordinated interventions that address both HIV and GBV, this region has the opportunity to reduce the national burden of HIV while also promoting key populations’ human rights.

Nutritional strategies to mitigate the negative effects of heat stress on animal welfare and productivity often involve changes in ration formulation. However, cattle commonly sort their ration in favour of certain components, and it is not clear how feed sorting responds to heat stress. This study investigated the association between heat stress and feed sorting behaviour. Lactating Holstein dairy cows (n = 32; parity = 2.8±1.2; mean±SD) were housed in a free stall barn and milked 3×/day. Cows were fed individually using the Calan Broadbent Feeding System and offered ad libitum access to a total mixed ration (containing on a dry matter basis: 3.3% ryegrass hay, 16.5% ryegrass baleage, 24.7% corn silage, 11.1% brewers grains, 19.7% ground corn, 19.8% concentrate and 4.9% protein/mineral supplement), provided 1×/day. Beginning at 186±60 days in milk, cows were exposed to either: heat stress conditions (HT; n = 15) (average temperature–humidity index: 77.6), or evaporative cooling (CL; n = 17), consisting of misters and fans over the freestall and feed bunks. Data were collected during a 4-day baseline period, and two 4-day experimental periods: starting at 10 days after implementing treatments (defined as acute heat stress for HT cows), and at 62 days after implementing treatments (defined as chronic heat stress for HT cows). Daily feed intake and physiological responses to heat stress (body temperature, respiration rate) were recorded. Samples of fresh and refused feed were collected daily from individual cows for particle size analysis. The particle size separator had three screens (19, 8 and 1.18 mm) and a bottom pan, resulting in 4 fractions (long, medium, short and fine particles). Feed sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. During both heat stress periods, HT cows sorted for long particles more than CL cows (105.0% v. 100.6%; SE = 1.1). During acute heat stress, HT cows sorted to a greater extent than CL cows against medium and short particles, whereas sorting of these fractions did not differ during chronic heat stress. Body temperature and respiration rate were associated across treatments with the extent of sorting for long particles and against short particles during acute heat stress. These results suggest that feed sorting is particularly influenced during acute heat stress, and that sorting for longer particles may increase in heat stress.

Core Ideas Aerodynamic methods can be used to gap‐fill Bowen ratio energy balance micrometeorological measurements. Eddy covariance and Bowen ratio energy balance methods agree during turbulent daytime conditions. Measuring nighttime net ecosystem exchange is challenging using turbulence‐based micrometeorology. There is a need to understand the potential benefits of using the biotechnology waste by‐product from manufacturing as a fertilizer replacement in agriculture, by quantifying the economic value for the farmer and measuring the environmental impact. Measuring CO2 emissions can be used to assess environmental impact, including three widely used micrometeorological methodologies: (i) the Bowen Ratio Energy Balance (BREB), (ii) aerodynamic flux‐gradient theory, and (iii) eddy covariance (EC). As a first step in quantifying benefits of applying biotechnology waste in agriculture, a detailed examination of these three methods was conducted to understand their effectiveness in quantifying CO2 emissions for this specific circumstance. The study measured micrometeorological properties over a field planted to maize (Zea mays L. var. indentata), one plot treated with biotechnology waste applied as a nutrient amendment, and one plot treated with a typical farmer fertilizer practice. Carbon dioxide flux measurements took place over 1 yr, using both BREB and EC systems. The aerodynamic method was used to gap‐fill BREB system measurements, and those flux estimates were compared with estimates produced separately by the aerodynamic and EC methods. All methods found greater emissions over the biotechnology waste application. The aerodynamic method CO2 flux estimates were considerably greater than both the EC and a combined BREB‐aerodynamic approach. During the day, the EC and BREB methods agree. At night, the aerodynamic approach detects and accounts for buildup of CO2 at the surface during stable periods. The BREB systems combined with aerodynamic approaches provide alternate methods to EC in examining micrometeorological properties near the surface.

Core Ideas Industrial biotechnology waste as a soil amendment approximates farmer practice yields. Applying waste amendments significantly increased soil organic C compared with farmer practice. Using waste nutrients for agriculture and soil organic matter improves sustainability. Reuse of industrial biotechnology by‐products has become an important component of circular bio‐economies whereby nutrient‐rich wastes are returned to agricultural land to improve soil fertility and crop productivity. Heat‐inactivated spent microbial biomass (SMB) from the production of 1,3‐propanediol is an industrial fermentation by‐product with nutrients that could replace or supplement conventional fertilizers. Our objectives were to determine if SMB utilization as a soil amendment in agriculture could generate environmental benefits while meeting farmer yield expectations and assess the impact of SMB application on CO2 emissions. This study examined the replacement of typical farmer fertilizer practices with the application of SMB. In addition to yellow dent maize (Zea mays L. var. indentata) grain yield and aboveground biomass, soil organic carbon (SOC) was measured. The eddy covariance (EC) micrometeorological method was used to measure CO2 flux. Overall maize yields were positively correlated with increasing application rates of SMB. After two SMB applications, SOC increased by 45% on the SMB plot as compared with an increase of 11% on the farmer practice plot. The SMB‐treated plot also emitted more CO2 (794 g CO2 m−2 yr−1) compared with the farmer practice treatment (274 g CO2 m−2 yr−1). Results from this study provide information on the efficacy of waste product nutrient cycling in the soil–plant ecosystem that could improve productivity and sustainability.

Urbanization has dramatically altered habitats for local species worldwide. While some species are unable to meet the challenges that these alterations bring, others are able to persist as long as a threshold for suitable habitat is met. For reptiles, a key feature for persistence in urban areas can be access to suitable refuges from predation, high temperatures, and/or other environmental challenges. We tested for effects of local and landscape variables affecting urban occupancy in the Western Fence Lizard, Sceloporus occidentalis, in transects across an urban–rural gradient, with a specific focus on the presence of rock, tree, and shrub refuges. We found that fence lizards were much more likely to be present in areas with more rock cover, and in parks or low-density residential areas. Occupancy was also positively related to canopy cover in the general vicinity, though negatively related to number of trees along the transects. Our results highlight the importance of assessing local habitat features to successfully predict the occupancy of reptile species in urban habitats, and present directions for future research with concrete conservation and management applications.

Necrotizing soft tissue infections caused by Streptococcus pyogenes (group A streptococcus [GAS]) are characterized by rapid and extensive necrosis of fascia and muscle. Molecular epidemiological studies have demonstrated a positive correlation between GAS isolates that cause invasive infections and the production of S. pyogenes NAD+-glycohydrolase (SPN), an NADase secreted by GAS, but the effect of SPN on muscle cells has not been described. Thus, using standard βNAD+ and ATP quantification assays, we investigated the effects of SPN on cultured human skeletal muscle cell (SkMC) βNAD+ and ATP with and without streptolysin O (SLO)–a secreted cholesterol-dependent cytolysin known to act synergistically with SPN. We found that culture supernatants from GAS strains producing SLO and SPN depleted intracellular βNAD+ and ATP, while exotoxins from a GAS strain producing SLO and an enzymatically-inactive form of SPN had no effect on βNAD+ or ATP. Addition of purified, enzymatically-active SPN to NADase-negative culture supernatants or sterile media reconstituted βNAD+ depletion but had no effect ATP levels. Further, SPN-mediated βNAD+ depletion could be augmented by SLO or the homologous cholesterol-dependent cytolysin, perfringolysin O (PFO). Remarkably, SPN-mediated βNAD+ depletion was SkMC-specific, as purified SPN had minimal effect on epithelial cell βNAD+. Taken together, this study identifies a previously unrecognized role for SPN as a major disruptor of skeletal muscle βNAD+. Such activity could contribute to the rapid and widespread myonecrosis characteristic of severe GAS soft tissue infections.