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Postoperative pain management continues to be suboptimal because of the lack of effective nonopioid therapies and absence of understanding of sex-driven differences. Here, we asked how the NLRP3 inflammasome contributes to postoperative pain. Inflammasomes are mediators of the innate immune system that are responsible for activation and secretion of IL-1β upon stimulation by specific molecular signals. Peripheral IL-1β is known to contribute to the mechanical sensitization induced by surgical incision. However, it is not known which inflammasome mediates the IL-1β release after surgical incision. Among the 9 known inflammasomes, the NLRP3 inflammasome is ideally positioned to drive postoperative pain through IL-1β production because NLRP3 can be activated by factors that are released by incision. Here, we show that male mice that lack NLRP3 (NLRP3) recover from surgery-induced behavioral and neuronal mechanical sensitization faster and display less surgical site inflammation than mice expressing NLRP3 (wild-type). By contrast, female NLRP3 mice exhibit minimal attenuation of the postoperative mechanical hypersensitivity and no change in postoperative inflammation compared with wild-type controls. Sensory neuron-specific deletion of NLRP3 revealed that in males, NLRP3 expressed in non-neuronal cells and potentially sensory neurons drives postoperative pain. However, in females, only the NLRP3 that may be expressed in sensory neurons contributes to postoperative pain where the non-neuronal cell contribution is NLRP3 independent. This is the first evidence of a key role for NLRP3 in postoperative pain and reveals immune-mediated sex differences in postoperative pain.

Objectives Bone void fillers are increasingly being used for dead space management in arthroplasty revision surgery. The aim of this study was to investigate the influence of calcium sulphate bone void filler (CS-BVF) on the damage and wear of total knee arthroplasty using experimental wear simulation. Methods A total of 18 fixed-bearing U2 total knee arthroplasty system implants (United Orthopedic Corp., Hsinchu, Taiwan) were used. Implants challenged with CS-BVF were compared with new implants (negative controls) and those intentionally scratched with a diamond stylus (positive controls) representative of severe surface damage (n = 6 for each experimental group). Three million cycles (MC) of experimental simulation were carried out to simulate a walking gait cycle. Wear of the ultra-high-molecular-weight polyethylene (UHMWPE) tibial inserts was measured gravimetrically, and damage to articulating surfaces was assessed using profilometry. Results There was no significant difference (p > 0.05) between the wear rate of implants challenged with CS-BVF (3.3 mm3/MC (95% confidence interval (CI) 1.8 to 4.8)) and the wear rate of those not challenged (2.8 mm3/MC (95% CI 1.3 to 4.3)). However, scratching the cobalt-chrome (CoCr) significantly (p < 0.001) increased the wear rate (20.6 mm3/MC (95% CI 15.5 to 25.7)). The mean surface roughness of implants challenged with CS-BVF was equivalent to negative controls both after damage simulation (p = 0.98) and at the conclusion of the study (p = 0.28). Conclusion When used close to articulating surfaces, a low-hardness, high-purity CS-BVF had no influence on wear. When trapped between the articulating surfaces of a total knee arthroplasty, CS-BVF did not scratch the surface of CoCr femoral components, nor did it increase the wear of UHMWPE tibial inserts compared with undamaged negative controls. Cite this article: R. M. Cowie, S. S. Aiken, J. J. Cooper, L. M. Jennings. The influence of a calcium sulphate bone void filler on the third-body damage and polyethylene wear of total knee arthroplasty. Bone Joint Res 2019;8:65–72. DOI: 10.1302/2046-3758.82.BJR-2018-0146.R1.

A range of surgical techniques and osteochondral interventions have been developed for early stage chondral/osteochondral repair interventions in the knee however, methods for functional, pre-clinical assessment of these therapies are limited. In this study, a method for simulating physiological loading and motion in the porcine patellofemoral joint was developed using a 6-axis simulator. As an example of how the method can be used, the influence of surgical positioning of osteochondral allografts in the patella on cartilage wear, deformation and damage and graft stability was investigated in this porcine patellofemoral joint model. The functional performance of allografts implanted either optimally (flush with the cartilage surface) or 1 mm proud of the cartilage surface was compared to a positive control (stainless steel pin implanted 1 mm proud of the cartilage surface), a negative control (no intervention) and a defect model. Allografts implanted flush with the surrounding cartilage could restore the articulating surface of the patella resulting in low wear, damage and deformation of the opposing cartilage surface, similar to that of the negative control group. Implanting the graft proud of the patella surface resulted in cartilage lesions on the femoral trochlea (ICRS grade 2) and a cartilage volume difference of 2.0 ± 3.9 mm3; the positive controls resulted in more severe lesions, a higher volume difference (14.2 ± 7.4 mm3) which in some cases exposed subchondral bone (ICRS grade 4). Defects in the patella caused deformation of the opposing cartilage surface. All grafts implanted in the patella subsided over the duration of the study. This study demonstrated a method that can be used to evaluate osteochondral repair strategies in the patellofemoral joint applying physiological loading and motions.

This study investigated the fixation of a cemented PEEK femoral TKA component. PEEK and CoCr implants were subjected to a walking gait cycle for 10 million cycles (MC), 100,000 cycles or 0 cycles (unloaded control). A method was developed to assess the fixation at the cement&ndash, implant interface, which exposed the implants to a fluorescent penetrant dye solution. The lateral condyles of the implants were then sectioned and viewed under fluorescence to investigate bonding at the cement&ndash, implant interface and cracking of the cement mantle. When tested for 100,000 cycles, debonding of the cement&ndash, implant interface occurred in both PEEK (61%) and CoCr (13%) implants. When the duration of testing was extended (10 MC), the percentage debonding was further increased for both materials to 88% and 61% for PEEK and CoCr, respectively. The unloaded PEEK specimens were 79% debonded, which suggests that, when PEEK femoral components are cemented, complete bonding may never occur. Analysis of cracks in the cement mantle showed an absence of full-thickness cracks in the unloaded control group. For the 100,000-cycle samples, on average, 1.3 and 0.7 cracks were observed for PEEK and CoCr specimens, respectively. After 10 MC, these increased to 24 for PEEK and 19 for CoCr. This was a preliminary study with a limited number of samples investigated, but shows that, after 10 MC under a walking gait, substantial debonding was visible for both PEEK and CoCr implants at the cement&ndash, implant interface and no significant difference in the number of cement cracks was found between the two materials.

PEEK-OPTIMA&trade, is being considered as an alternative to cobalt chrome (CoCr) in the femoral component of total knee replacements. To date, investigations of ultra-high molecular weight polyethylene (UHMWPE)-on-PEEK have shown an equivalent wear rate to conventional implant materials under standard conditions. In this study, the third body wear performance of UHMWPE-on-PEEK was directly compared to UHMWPE-on-CoCr in a series of pin-on-plate studies using two approaches for third body damage. Damage simulation with particles of bone cement showed a significant (p &lt, 0.001), four-fold increase in the mean surface roughness of PEEK plates compared to CoCr. However, wear simulation against the damaged plates showed no significant difference in the wear of UHMWPE pins against the different materials (p = 0.59), and a polishing effect by the pin against the PEEK plates was observed. Scratching PEEK and CoCr counterfaces with a diamond stylus to create scratches representative of severe third body damage (4 &micro, m lip height) resulted in a significantly higher (p = 0.01) wear of UHMWPE against CoCr compared to PEEK and again, against PEEK plates, polishing by the UHMWPE pin led to a reduction in scratch lip height. This study shows that in terms of its wear performance under third body wear/damage conditions, UHMWPE-on-PEEK differs from conventional knee replacement materials.

Osteochondral grafts are used clinically to repair cartilage and bone defects and to restore the congruent articulating surfaces of the knee joint following cartilage damage or injury. The clinical success of such osteochondral grafts is heavily reliant on the biomechanical and tribological properties of the surgical repair; however, a limited number of studies have investigated these factors. The aim of this study was to evaluate the influence of graft harvesting and implantation technique as well as bone properties on the primary stability of press-fit implanted osteochondral grafts using a series of uniaxial experimental push-in and push-out tests. Animal (porcine and bovine) knees were used to deliver models of different bone properties (elastic modulus and yield stress). The study showed the graft harvesting method using either a chisel or drill-aided trephine to have no influence on primary graft stability; however, the preparation technique for the graft recipient site was shown to influence the force required to push the graft into the host tissue. For example, when the length of the graft was equal to the recipient site (bottomed), the graft was more stable and dilation of the recipient site was shown to reduce short-term graft stability especially in immature or less dense bone tissue. The push-out tests which compared tissue of different skeletal maturities demonstrated that the maturity of both the graft and host bone tissue to influence the stability of the graft. A higher force was required to push out more skeletally mature grafts from mature bone tissue. The study demonstrates the importance of surgical technique and bone quality/properties on the primary stability and ultimately, the success of osteochondral grafts in the knee.

Autotaxin (ATX) is an adipokine that generates the bioactive lipid, lysophosphatidic acid (LPA). ATX-LPA signaling has been implicated in diet-induced obesity and systemic insulin resistance. However, it remains unclear whether the ATX-LPA pathway influences insulin function and energy metabolism in target tissues, particularly skeletal muscle, the major site of insulin-stimulated glucose disposal. The objective of this study was to test whether the ATX-LPA pathway impacts tissue insulin signaling and mitochondrial metabolism in skeletal muscle during obesity. Male mice with heterozygous ATX deficiency (ATX(+/−)) were protected from obesity, systemic insulin resistance, and cardiomyocyte dysfunction following high-fat high-sucrose (HFHS) feeding. HFHS-fed ATX(+/−) mice also had improved insulin-stimulated AKT phosphorylation in white adipose tissue, liver, heart, and skeletal muscle. Preserved insulin-stimulated glucose transport in muscle from HFHS-fed ATX(+/−) mice was associated with improved mitochondrial pyruvate oxidation in the absence of changes in fat oxidation and ectopic lipid accumulation. Similarly, incubation with LPA decreased insulin-stimulated AKT phosphorylation and mitochondrial energy metabolism in C2C12 myotubes at baseline and following palmitate-induced insulin resistance. Taken together, our results suggest that the ATX-LPA pathway contributes to obesity-induced insulin resistance in metabolically relevant tissues. Our data also suggest that LPA directly impairs skeletal muscle insulin signaling and mitochondrial function.

Quantitative proteins analysis was carried out in the hypothalamus of zebrafish following dietary exposure to the legacy pesticide dieldrin. Data were collected using iTRAQ labeling methodology and data were acquired using a hybrid quadrupole Orbitrap (Q Exactive) MS system (Thermo Fisher Scientific, Bremen, Germany). There were 3941 proteins identified in the hypothalamus of zebrafish, and these proteins comprised 23 unique expression patterns for proteins based on the three doses of dieldrin. There were 226 proteins that were regulated in one or more doses of dieldrin and 3715 proteins that were not affected. Thus, 5.7% of the proteins detected responded to the treatment. Many proteins that were differentially expressed were those found in, or associated with, the mitochondria. The proteomics data described in this article is associated with a research article, "Transcriptomic and proteomic analysis implicates the immune system and mitochondria as molecular targets of dieldrin in the zebrafish (Danio rerio) central nervous system" (A.M. Cowie, K.I. Sarty, A. Mercer, J. Koh, K.A. Kidd, C.J. Martyniuk, submitted) [1], and serves as a resource for researchers working in the field of pesticide exposures and protein biomarkers.

The objectives of this study were to determine the behavioral and molecular responses in the adult zebrafish (Danio rerio) central nervous system (CNS) following a dietary exposure to the pesticide dieldrin. Zebrafish were fed pellets spiked with 0.03, 0.15, or 1.8 μg/g dieldrin for 21 days. Behavioral analysis revealed no difference in exploratory behaviors or those related to anxiety. Transcriptional networks for T-cell aggregation and selection were decreased in expression suggesting an immunosuppressive effect of dieldrin, consistent with other studies investigating organochlorine pesticides. Processes related to oxidative phosphorylation were also differentially affected by dieldrin. Quantitative proteomics (iTRAQ) using a hybrid quadrupole-Orbitrap identified 226 proteins that were different following one or more doses. These proteins included ATP synthase subunits (mitochondrial) and hypoxia up-regulated protein 1 which were decreased and NADH dehydrogenases (mitochondrial) and signal recognition particle 9 which were up-regulated. Thus, proteins affected were functionally associated with the mitochondria and a protein network analysis implicated Parkinson's disease (PD) and Huntington's disease as diseases associated with altered proteins. Molecular networks related to mitochondrial dysfunction and T-cell regulation are hypothesized to underlie the association between dieldrin and PD. These data contribute to a comprehensive transcriptomic and proteomic biomarker framework for pesticide exposures and neurodegenerative diseases. Biological significance Dieldrin is a persistent organochlorine pesticide that has been associated with human neurodegenerative disease such as Parkinson's disease. Dieldrin is ranked 18th on the 2015 U.S. Agency for Toxic Substances and Disease Registry and continues to be a pesticide of concern for human health. Transcriptomics and quantitative proteomics (ITRAQ) were employed to characterize the molecular networks in the central nervous system that are altered with dietary exposure to dieldrin. We found that transcriptional and protein networks related to the immune system, mitochondria, and Parkinson's disease were preferentially affected by dieldrin. The study provides new insight into the mechanisms of dieldrin neurotoxicity that may explain, in part, the association between this pesticide and increased risks to neurodegeneration. These data contribute in a significant way to developing a molecular framework for pesticide induced neurotoxicity.

Methylmercury (MeHg) exposure and adverse health effects in fishes have been documented, but the molecular mechanisms involved in toxicity have not been fully characterized. The objectives of the current study were to (1) determine whether total Hg (THg) in the muscle was predictive of MeHg concentrations in the brain of wild female yellow perch (Perca flavescens) collected from four lakes in Kejimkujik National Park, a known biological mercury (Hg) hotspot in Nova Scotia, Canada and (2) to determine whether transcripts involved in the oxidative stress response were altered in abundance in fish collected across five lakes representing a MeHg gradient. In female yellow perch, MeHg in whole brain (0.38 to 2.00 μg/g wet weight) was positively associated with THg in muscle (0.18 to 2.13 μg/g wet weight) (R2 = 0.61, p

Postoperative pain is highly debilitating and hinders recovery. Opioids are the main pain medication used for acute postoperative pain. Given the devastating opioid addiction and overdose epidemic across the US, non-opioid pain therapeutics are desperately needed. In order to develop novel, non-opioid therapies for the treatment of postoperative pain and identify the mechanisms underlying this pain, rodent models of incisional pain have been established. The protocol herein describes in detail how to create a mouse model of postoperative pain that was adapted from established protocols. This model of postoperative pain is frequently-used, highly reproducible, and results in peripheral and central nervous system alterations.

A footbridge was built in 2013 in rural Morocco using lightweight polyester rope, spanning 64 meters (210 feet) across a deep ravine. The area is prone to perennial flash flooding, cutting off access to schools, the local clinic, and the market for weeks at a time. After five years of service, the design team returned to inspect the structural condition, and replace one of the main ropes. The decommissioned rope was subjected to load testing and dissection, and was found to be in excellent condition.The novel use of synthetic rope offered some advantages over steel wire rope typically used for this type of project, and the team developed strategies to work efficiently with this unusual material. Particularly, its light weight makes it substantially less costly and simpler to transport to the construction site (the rope was procured in the U.S. and shipped to Morocco in backpacks). As it is less sensitive to the effects of twisting, the rope does not require the level of care typical of wire rope.The paper will discuss unique aspects of construction, and challenges related to ongoing maintenance of this type of infrastructure in a developing country.

The organochlorine dieldrin (DLD) bioaccumulates in lipid-rich tissues and is associated with immunosuppression, altered metabolism, and cancer. The objective of this study was to determine the effect of DLD on the hepatic proteome in zebrafish following dietary treatment as the liver is central to metabolism. Females were fed a control dose or one of three doses of DLD-contaminated food pellets over 21 days. Both label-free and iTRAQ proteomics were conducted as two complementary methods to expand coverage of the proteome. Label-free proteomics quantified 1563 proteins: 6 proteins showed a linear dose-response with DLD. iTRAQ quantified >3500 proteins; 5 proteins were decreased and 34 proteins were increased in abundance within the liver with all three doses. Overall, DLD reduced the abundance of proteins associated with glucose and cholesterol metabolism, lipid oxidation, liver function, and immune-related processes. Few proteins were identified by both methods as being altered (~1%), suggesting that each method detected different subsets of proteins. Protein responses in the liver were largely dependent on dose, however proteins related to liver and organ function, centrosome separation, glucose/energy metabolism, and immune-related pathways were confirmed by each independent technique and were suppressed with DLD exposure. This study identifies proteomic responses that are associated with organochlorine-induced hepatotoxicity. Biological significance Environmental contaminants cause hepatotoxicity because the liver is the major organ for detoxification. The legacy pesticide dieldrin significantly bioaccumulates in tissues, and can affect molecular processes that can lead to liver pathology. LC MS/MS proteomics identified protein networks related to tumors, energy homeostasis, and chromosomal separation as those affected by dietary exposure to dieldrin. We applied two orthogonal mass spectrometry-based methods to more completely survey the liver proteome, strengthening data interpretation. These data improve understanding as to the effects of organochlorine pesticide toxicity in the liver and the study identifies proteome networks that can contribute to adverse outcome pathways for pesticide exposure.

Fipronil is a phenylpyrazole pesticide that is used in both residential and agricultural applications. Fipronil is detected in run-off and water systems that are near areas in which the pesticide has been applied. The pesticide acts to antagonize gamma aminobutyric acid receptors, leading to over-excitation in the central nervous system. Fipronil has relatively high toxicity to fish, but the mechanisms underlying the toxicity are not well understood in embryonic stages. Zebrafish embryos were exposed to a single concentration of fipronil for 48 h at ∼3-4 h-post-fertilization. Following a 7-day depuration phase, transcriptome and behavioral analyses were conducted. Transcriptomics identified neural processes as those differentially expressed with different doses of fipronil (0.2 µg, 200 µg and 2 mg fipronil/L). Gene networks associated with astrocyte differentiation, myelination, neural tube development, brain stem response, innervation, nerve regeneration, astrocyte differentiation, among other pathways were altered with exposure. In addition, miRNA-related events are disrupted by fipronil exposure and genes associated with primary or pri-miRNA processing were increased in larval fish exposed to the pesticide. These data present putative mechanisms associated with neurological impacts at later ages of zebrafish. This is important because it is not clear how early exposure to pesticides like fipronil affect central nervous system function and organisms later in life.

PEEK-OPTIMA™ (Invibio Ltd, UK) has been considered as an alternative joint arthroplasty bearing material due to its favourable mechanical properties and the biocompatibility of its wear debris. In this study, the potential to use injection moulded PEEK-OPTIMA™ as an alternative to cobalt chrome in the femoral component of a total knee replacement was investigated in terms of its wear performance. Experimental wear simulation of three cobalt chrome and three PEEK-OPTIMA™ femoral components articulating against all-polyethylene tibial components was carried out under two kinematic conditions: 3 million cycles under intermediate kinematics (maximum anterior-posterior displacement of 5 mm) followed by 3 million cycles under high kinematic conditions (anterior-posterior displacement 10 mm). The wear of the GUR1020 ultra-high-molecular-weight polyethylene tibial components was assessed by gravimetric analysis; for both material combinations under each kinematic condition, the mean wear rates were low, that is, below 5 mm3/million cycles. Specifically, under intermediate kinematic conditions, the wear rate of the ultra-high-molecular-weight polyethylene tibial components was 0.96 ± 2.26 mm3/million cycles and 2.44 ± 0.78 mm3/million cycle against cobalt chrome and PEEK-OPTIMA™ implants, respectively (p = 0.06); under high kinematic conditions, the wear rates were 2.23 ± 1.85 mm3/million cycles and 4.44 ± 2.35 mm3/million cycles, respectively (p = 0.03). Following wear simulation, scratches were apparent on the surface of the PEEK-OPTIMA™ femoral components. The surface topography of the femoral components was assessed using contacting profilometry and showed a statistically significant increase in measured surface roughness of the PEEK-OPTIMA™ femoral components compared to the cobalt chrome implants. However, this did not appear to influence the wear rate, which remained linear over the duration of the study. These preliminary findings showed that PEEK-OPTIMA™ gives promise as an alternative bearing material to cobalt chrome alloy in the femoral component of a total knee replacement with respect to wear performance.

Calcium sulfate bone void fillers are increasingly being used for dead space management in infected arthroplasty revision surgery. The presence of these materials as loose beads close to the bearing surfaces of joint replacements gives the potential for them to enter the joint becoming trapped between the articulating surfaces; the resulting damage to cobalt chrome counterfaces and the subsequent wear of ultra-high-molecular-weight polyethylene is unknown. In this study, third-body damage to cobalt chrome counterfaces was simulated using particles of the calcium sulfate bone void fillers Stimulan®(Biocomposites Ltd., Keele, UK) and Osteoset®(Wright Medical Technology, TN, USA) using a bespoke rig. Scratches on the cobalt chrome plates were quantified in terms of their density and mean lip height, and the damage caused by the bone void fillers was compared to that caused by particles of SmartSet GMV PMMA bone cement (DePuy Synthes, IN, USA). The surface damage from Stimulan®was below the resolution of the analysis technique used; SmartSet GMV caused 0.19 scratches/mm with a mean lip height of 0.03 µm; Osteoset®led to a significantly higher number (1.62 scratches/mm) of scratches with a higher mean lip height (0.04 µm). Wear tests of ultra-high-molecular-weight polyethylene were carried out in a six-station multi-axial pin on plate reciprocating rig against the damaged plates and compared to negative (highly polished) and positive control plates damaged with a diamond stylus (2 µm lip height). The wear of ultra-high-molecular-weight polyethylene was shown to be similar against the negative control plates and those damaged with third-body particles; there was a significantly higher ( p

Mechanotransduction refers to the processes whereby mechanical stimuli are converted into electrochemical signals that allow for the sensation of our surrounding environment through touch. Despite its fundamental role in our daily lives, the molecular and cellular mechanisms of mechanotransduction are not yet well-defined. Previous data suggest that keratinocytes may release factors that activate or modulate cutaneous sensory neuron terminals, including small molecules, lipids, peptides, proteins, and oligosaccharides. This study presents a first step towards identifying soluble mediators of keratinocyte-sensory neuron communication by evaluating the potential for top down mass spectrometry to identify proteoforms released during one minute of mechanical stimulation of mouse skin from naïve animals. Overall, this study identified 47 proteoforms in the secretome of mouse hind paw skin, of which 14 were differentially released during mechanical stimulation, and includes proteins with known and previously unknown relevance to mechanotransduction. Finally, this study outlines a bioinformatic workflow that merges output from two complementary analysis platforms for top down data and demonstrates the utility of this workflow for integrating quantitative and qualitative data.

In acute heart failure (AHF) syndromes significant respiratory failure (RF) is essentially seen in patients with acute cardiogenic pulmonary oedema (ACPE) or cardiogenic shock (CS). Non-invasive ventilation (NIV), the application of positive intrathoracic pressure through an interface, has shown to be useful in the treatment of moderate to severe RF in several scenarios. There are two main modalities of NIV: continuous positive airway pressure (CPAP) and pressure support ventilation (NIPSV) with positive end expiratory pressure. Appropriate equipment and experience is needed for NIPSV, whereas CPAP may be administered without a ventilator, not requiring special training. Both modalities have shown to be effective in ACPE, by a reduction of respiratory distress and the endotracheal intubation rate compared to conventional oxygen therapy, but the impact on mortality is less conclusive. Non-invasive ventilation is also indicated in patients with AHF associated to pulmonary disease and may be considered, after haemodynamic stabilization, in some patients with CS. There are no differences in the outcomes in the studies comparing both techniques, but CPAP is a simpler technique that may be preferred in low-equipped areas like the pre-hospital setting, while NIPSV may be preferable in patients with significant hypercapnia. The new modality â €high-flow nasal cannula' seems promising in cases of AHF with less severe RF. The correct selection of patients and interfaces, early application of the technique, the achievement of a good synchrony between patients and the ventilator avoiding excessive leakage, close monitoring, proactive management, and in some cases mild sedation, may warrant the success of the technique. © The Author 2017.

Background: The 69th World Health Assembly approved the Global Health Sector Strategy to eliminate viral hepatitis by 2030. Although no virological cure exists for hepatitis B virus (HBV) infection, existing therapies to control viral replication and prophylaxis to minimise mother-to-child transmission make elimination of HBV feasible. We aimed to estimate the national, regional, and global prevalence of HBsAg in the general population and in the population aged 5 years in 2016, as well as coverage of prophylaxis, diagnosis, and treatment. Methods: In this modelling study, we used a Delphi process that included a literature review in PubMed and Embase, followed by interviews with experts, to quantify the historical epidemiology of HBV infection. We then used a dynamic HBV transmission and progression model to estimate the country-level and regional-level prevalence of HBsAg in 2016 and the effect of prophylaxis and treatment on disease burden. Findings: We developed models for 120 countries, 78 of which were populated with data approved by experts. Using these models, we estimated that the global prevalence of HBsAg in 2016 was 3·9% (95% uncertainty interval [UI] 3·4–4·6), corresponding to 291 992 000 (251 513 000–341 114 000) infections. Of these infections, around 29 million (10%) were diagnosed, and only 4·8 million (5%) of 94 million individuals eligible for treatment actually received antiviral therapy. Around 1·8 (1·6–2·2) million infections were in children aged 5 years, with a prevalence of 1·4% (1·2–1·6). We estimated that 87% of infants had received the three-dose HBV vaccination in the first year of life, 46% had received timely birth-dose vaccination, and 13% had received hepatitis B immunoglobulin along with the full vaccination regimen. Less than 1% of mothers with a high viral load had received antiviral therapy to reduce mother-to-child transmission. Interpretation: Our estimate of HBV prevalence in 2016 differs from previous studies, potentially because we took into account the effect of infant prophylaxis and early childhood vaccination, as well as changing prevalence over time. Although some regions are well on their way to meeting prophylaxis and prevalence targets, all regions must substantially scale-up access to diagnosis and treatment to meet the global targets. Funding: John C Martin Foundation. © 2018 Elsevier Ltd

The first point of our body’s contact with tactile stimuli (innocuous and noxious) is the epidermis, the outermost layer of skin that is largely composed of keratinocytes. Here, we sought to define the role that keratinocytes play in touch sensation in vivo and ex vivo. We show that optogenetic inhibition of keratinocytes decreases behavioral and cellular mechanosensitivity. These processes are inherently mediated by ATP signaling, as demonstrated by complementary cutaneous ATP release and degradation experiments. Specific deletion of P2X4 receptors in sensory neurons markedly decreases behavioral and primary afferent mechanical sensitivity, thus positioning keratinocyte-released ATP to sensory neuron P2X4 signaling as a critical component of baseline mammalian tactile sensation. These experiments lay a vital foundation for subsequent studies into the dysfunctional signaling that occurs in cutaneous pain and itch disorders, and ultimately, the development of novel topical therapeutics for these conditions., eLife digest The skin is the largest sensory organ of the body, and the first point of contact with the outside world. Whether it is being pinched or caressed, the skin’s sense of touch informs organisms about their surroundings and allows them to react appropriately. Nerve cells present in the skin capture information about touch and transmit it to the brain where it is decoded. However, there are many other types of cells in the skin besides nerve cells. The role that these other skin cells play in perceiving non-painful and painful touch is still unclear. Moehring et al. now report how the skin cells that form 95% of the most outer layer of the skin are involved in detecting touch. In mutant mice whose cells can be ‘switched off’ by a certain light, artificially deactivating these cells makes the animals less able to respond to tactile stimuli. Further experiments show that when pressure is applied onto the skin, the surface skin cells release a chemical messenger, which then binds specifically to the nerve cells. When the messaging molecule is experimentally destroyed or prevented from attaching to the nerve cell, the mice react less to non-painful and painful touch. This means the cells at the surface of the skin detect tactile signals from the environment and then communicate this information to the nerve cells, where it is taken to the brain. Disrupted communication between the cells in the outer layer of the skin and the nerve cells is found in painful and itchy skin conditions such as eczema and psoriasis. Knowing how these two types of cells normally work together may help with finding new pain and itch treatments for these skin disorders.

Inflammasomes are cytosolic receptors of the innate immune system that are responsible for activation and secretion of the proinflammatory cytokine interleukin-1β (IL-1β) where IL-1β is known to cause pain by directly acting on sensory neurons. Specifically, IL-1β is known to cause peripheral sensitization to mechanical stimuli in models of inflammatory and neuropathic pain. Of the multiple inflammasomes, the NOD-like receptor protein 3 (NLRP3) inflammasome is activated by endogenous molecules that are released as a result of tissue injury and thus is ideally positioned to produce the majority of postoperative IL-1β. Therefore, we aimed to uncover the mechanism by which NLRP3 modifies postoperative pain in males and females. Male mice lacking NLRP3 recovered from incisional surgery significantly faster at the behavioral and primary afferent levels and exhibited less peri-incisional inflammation than mice expressing NLRP3 (WT). In contrast, female mice lacking NLRP3 displayed only attenuated postoperative mechanical hypersensitivity and peri-incisional inflammation was unchanged compared to WT females. Furthermore, sensory neuron-specific deletion of NLRP3 revealed that, in males, NLRP3 expressed in both sensory neurons and non-neuronal cells drives postoperative pain whereas, in females, only sensory neuron NLRP3 contributes. This is the first evidence for a direct role for NLRP3 and inflammasome-mediated sex differences in postoperative pain.

Fundamental studies characterizing transcript variability in teleost tissues are needed if molecular endpoints are to be useful for regulatory ecotoxicology. The objectives of this study were to (1) measure transcript variability of steroidogenic enzymes and steroid receptors in the fathead minnow (FHM; Pimephales promelas ) ovary to better determine normal variability and the sample sizes needed to detect specific effect sizes and to (2) determine how expression patterns related to higher level endpoints used in some regulatory ecotoxicology programs (e.g. relative gonad size). Estrogen receptor 2b ( esr2b ) and 5α-reductase a3 ( srd5a3 ) showed high variability in the ovary (CV > 1.0) while progesterone receptor ( pgr ), androgen receptor ( ar ), and esr2a showed comparatively low variability (CV = ~ 0.5–−0.7). Using these estimates, a power analysis revealed that sample sizes for real-time PCR experiments would need to be >20 to detect a 2-fold change for 7 of the transcripts examined; thus many molecular studies conducted in the fish ovary may have insufficient power to detect smaller effects. Two transcripts were correlated to steroid production in the ovary; cyp19a1 levels were positively correlated to in vitro E2 production, while ar levels were negatively correlated to in vitro T production. Thus, these transcripts may be informative molecular surrogates for ovarian steroid production. No transcript investigated showed any correlation to GSI, condition, or body weight/length. Molecular approaches in fish are increasingly used to assess biological impacts of chemical stressors; however additional studies are required that determine how molecular variability relates to higher level biological endpoints.

Considerable concern has been expressed by both domestic users and importers of Australian sorghum over the low protein content of some of the grain being offered for sale. Some grain samples have been found to contain as little as 5% crude protein, compared with the 9.2% minimum required for export sales. There has been a tendency both in Australia and overseas to associate the problem of low protein levels with increased use of hybrid sorghum varieties.This study examines, under closely controlled conditions of nitrogen supply, the effects of nitrogen supply at particular stages of growth, on both grain yield and protein concentration in the high yielding hybrid variety, Texas 610.A method was developed to control the nitrogen status of sorghum plants grown in solution culture by the use of small frequent additions of nitrogen, the size and frequency depending on the desired growth curves of the plants.In these solution culture experiments it was found that nitrogen stress could cause substantial reductions in the number of florets initiated, and the number of florets which survived to anthesis. Nitrogen stress also caused substantial reductions in the number of viable pollen grains per flower.Contrary to what has been reported for many other crops, the sorghum plant retained the capacity to absorb large amounts of nitrogen throughout the post-flowering period and grain nitrogen concentration depended almost entirely on the level of nitrogen supply during this period.Results are presented which indicate that in plants grown under non-limiting nitrogen supply conditions, grain nitrogen content is restricted by sink size. When grain size vas increased by reducing the number of grains, both grain nitrogen content and concentration increased considerably. It would appear that changes in grain structure and in the capacity of the endosperm to store nitrogen are at least partly responsible.A large field trial was conducted to determine if the results observed in solution culture under glasshouse conditions held true in the field situation. Once again it was found that a shortage of available nitrogen during the pre-flowering period had major effects on yield, and that grain protein concentration depended completely on the level of available nitrogen during the grain filling period. Side dressings of nitrogen at the boot stage or at anthesis were found to cause substantial increases in grain protein concentrations even when 300 lb of nitrogen per acre had been applied at planting.Results of the present study strongly suggest that low grain protein contents in high yielding grain sorghum varieties result from inadequate nitrogen nutrition during the grain filling period. For high yields of high protein grain, the fertilizer programme must be adjusted to ensure that the crop has an adequate supply of nitrogen up to and including the grain filling period.

Complications of patellofemoral arthroplasty often occur soon after implantation and, as well as other factors, can be due to the design of the implant or its surgical positioning. A number of studies have previously considered the wear of ultra-high-molecular-weight polyethylene patellae following suboptimal implantation; however, studies have primarily been carried out under a limited number of degrees of freedom. The aim of this study was to develop a protocol to assess the wear of patellae under a malaligned condition in a six-axis patellofemoral joint simulator. The malalignment protocol hindered the tracking of the patella centrally in the trochlear groove and imparted a constant 5° external rotation (tilt) on the patella button. Following 3 million cycles of wear simulation, this condition had no influence on the wear of ultra-high-molecular-weight polyethylene patellae aged for 4 years compared to well-positioned non-aged implants (p > 0.05). However, under the malaligned condition, ultra-high-molecular-weight polyethylene patellae aged 8–10 years after unpacking (following sterilisation by gamma irradiation in an inert atmosphere) and worn ultra-high-molecular-weight polyethylene components also aged 4 years after unpacking (following the same sterilisation process) exhibited a high rate of wear. Fatigue failure due to elevated contact stress led to delamination of the ultra-high-molecular-weight polyethylene and in some cases complete failure of the patellae. The results suggest that suboptimal tracking of the patella in the trochlear groove and tilt of the patella button could have a significant effect on the wear of ultra-high-molecular-weight polyethylene and could lead to implant failure.

BACKGROUND This report provides data on the use of social media advertising as a clinical trial recruitment strategy targeting healthy volunteers aged 60 years and older. The social media advertising campaign focused on enrollment for a Phase 1 clinical trial. Traditional means of recruiting—billboards, newspaper advertising, word of mouth, personal referrals, and direct mail—were not producing enough qualified participants. OBJECTIVE To demonstrate the effectiveness of using targeted advertising on the social networking site Facebook to recruit people aged 60 years and older for volunteer clinical trial participation. METHODS The trial sponsor used a proactive approach to recruit participants using advertising on social media. The sponsor placed and monitored an Institutional Review Board-approved advertising campaign on Facebook to recruit potential candidates for a Phase 1 clinical trial. The clinical trial required a 10-day residential (overnight) stay at a clinic in Michigan, with one follow-up visit. The sponsor of the clinical trial placed the advertising, which directed interested respondents to a trial-specific landing page controlled by the Contract Research Organization (CRO). The CRO provided all follow-up consenting, prescreening, screening, and enrollment procedures. The campaign was waged over an 8-week period to supplement recruiting by the CRO. RESULTS A total of 621 people responded to a Facebook advertising campaign by completing an online form or telephoning the CRO, and the clinical trial was fully enrolled at 45 subjects following an 8-week Facebook advertising campaign. CONCLUSIONS An 8-week Facebook advertising campaign contributed to 868 inquiries made regarding a Phase 1 clinical trial seeking to enroll healthy elderly subjects. Over the initial 11 weeks of recruitment, 178 inquiries were received using traditional methods of outreach. Respondents to the Facebook advertising campaign described in this report engaged with the sponsored advertising at a higher rate than is typical for social media-based clinical trial recruitment strategies. The older adults’ engagement rate of 4.92% was more than twice as high as click-through rates of younger adults engaged with social media advertising in other clinical trial recruitment studies. Advertising placed on the social media platform Facebook is effective with the healthy volunteer population aged 60 years and older. This approach can quickly and cost-effectively reach qualified candidates for clinical trial recruitment as a supplement to traditional means of recruiting. CLINICALTRIAL ClinicalTrials.gov: NCT02840279; https://clinicaltrials.gov/ct2/show/NCT02840279 (Archived by WebCite at http://www.webcitation.org/6wamIWXAt)

The global incidence of obesity has led to an increasing need for understanding the molecular mechanisms that drive this epidemic and its comorbidities. Quantitative real-time RT-PCR (RT-qPCR) is the most reliable and widely used method for gene expression analysis. The selection of suitable reference genes (RGs) is critical for obtaining accurate gene expression information. The current study aimed to identify optimal RGs to perform quantitative transcriptomic analysis based on RT-qPCR for obesity and diabetes research, employing in vitro and mouse models, and human tissue samples. Using the ReFinder program we evaluated the stability of a total of 15 RGs. The impact of choosing the most suitable RGs versus less suitable RGs on RT-qPCR results was assessed. Optimal RGs differed between tissue and cell type, species, and experimental conditions. By employing different sets of RGs to normalize the mRNA expression of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α), we show that sub-optimal RGs can markedly alter the PGC1α gene expression profile. Our study demonstrates the importance of validating RGs prior to normalizing transcriptional expression levels of target genes and identifies optimal RG pairs for reliable RT-qPCR normalization in cells and in human and murine muscle and adipose tissue for obesity/diabetes research.

Background: Spinal muscular atrophy is an autosomal recessive neuromuscular disorder that is caused by an insufficient level of survival motor neuron (SMN) protein. Nusinersen is an antisense oligonucleotide drug that modifies pre–messenger RNA splicing of the SMN2 gene and thus promotes increased production of full-length SMN protein. Methods: We conducted a randomized, double-blind, sham-controlled, phase 3 efficacy and safety trial of nusinersen in infants with spinal muscular atrophy. The primary end points were a motor-milestone response (defined according to results on the Hammersmith Infant Neurological Examination) and event-free survival (time to death or the use of permanent assisted ventilation). Secondary end points included overall survival and subgroup analyses of event-free survival according to disease duration at screening. Only the first primary end point was tested in a prespecified interim analysis. To control the overall type I error rate at 0.05, a hierarchical testing strategy was used for the second primary end point and the secondary end points in the final analysis. Results: In the interim analysis, a significantly higher percentage of infants in the nusinersen group than in the control group had a motor-milestone response (21 of 51 infants [41%] vs. 0 of 27 [0%], P Conclusions: Among infants with spinal muscular atrophy, those who received nusinersen were more likely to be alive and have improvements in motor function than those in the control group. Early treatment may be necessary to maximize the benefit of the drug.

Aims: To evaluate the impact of physicians' adherence to guideline-recommended medications for heart failure with reduced ejection fraction (HFrEF), including ≥50% prescription of recommended doses, on clinical outcomes at 6-month follow-up. Methods and results: In QUALIFY, an international, prospective, observational, longitudinal survey, 6669 outpatients with HFrEF were recruited 1–15 months after heart failure (HF) hospitalization from September 2013 to December 2014 in 36 countries and followed up at 6 months. A global adherence to guidelines score was developed for prescription of angiotensin-converting enzyme inhibitors (ACEIs), angiotensin receptor blockers (ARBs), beta-blockers (BBs), mineralocorticoid receptor antagonists (MRAs) and ivabradine and their dosages. Baseline global adherence score was good in 23% of patients, moderate in 55%, and poor in 22%. At 6-month follow-up, poor adherence was associated with significantly higher overall mortality [hazard ratio (HR) 2.21, 95% confidence interval (CI) 1.42–3.44, P=0.001], cardiovascular mortality (HR 2.27, 95% CI 1.36–3.77, P=0.003), HF mortality (HR 2.26, 95% CI 1.21–4.2, P=0.032), combined HF hospitalization or HF death (HR 1.26, 95% CI 1.08–1.71, P=0.024) and cardiovascular hospitalization or cardiovascular death (HR 1.35, 95% CI 1.08–1.69, P=0.013). There was a strong trend between poor adherence and HF hospitalization (HR 1.32, 95% CI 1.04–1.68, P=0.069). Conclusion: Good adherence to pharmacologic treatment guidelines for ACEIs, ARBs, BBs, MRAs and ivabradine, with prescription of at least 50% of recommended dosages, was associated with better clinical outcomes during 6-month follow-up. Continuing global educational initiatives are needed to emphasise the importance of guideline recommendations for optimising drug therapy and prescribing evidence-based doses in clinical practice. © 2017 The Authors. European Journal of Heart Failure © 2017 European Society of Cardiology

Antarctic sea ice provides a stable environment for the colonisation of diverse and highly specialised microbes that play a central role in the assimilation and regulation of energy through the Antarctic food web. However, little is known about how the bacterial community composition changes within the sea ice, the functional role bacteria play in the sea-ice microbial loop, and the effect of variations in the environment on these patterns and processes. Using terminal restriction fragment length polymorphism (T-RFLP) analysis of the total (16S rRNA gene) and the active (16S rRNA) community, we characterised spatial patterns in Antarctic sea-ice bacterial communities (SIBCO). In addition, bacterial enzymatic activities were identified using synthetic fluorogenic substrates. Both bacterial community structure and enzymatic activity were compared to changing environmental conditions vertically down through the sea ice. The structure of the DNA- and RNA-derived bacterial communities exhibited strong vertical zonation through the ice. There was no direct relationship between changes in chlorophyll a (chl a) and bacterial numbers; however, there were direct relationships between chl a, bacterial community structure and metabolic function, thus providing evidence for a coupling of the microbial loop. The SIBCO were involved in phosphate recycling and selectively assimilated proteins over carbohydrates in re - sponse to the N- and P-poor environment. We propose that Antarctic sea ice has an active microbial loop in which the spatial dynamics of bacterial communities are influenced by algal-derived substrates and nutrient availability

Dieldrin is a legacy organochlorine pesticide that is persistent in the environment, despite being discontinued from use in North America since the 1970s. Some epidemiologic studies suggest that exposure to dieldrin is associated with increased risks of neurodegenerative disease and breast cancer by inducing inflammatory responses in tissues as well as oxidative stress. However, the direct effects of organochlorine pesticides on the heart have not been adequately addressed to date given that these chemicals are detectable in human serum and are environmentally persistent; thus, individuals may show latent adverse effects in the cardiovascular system due to long-term, low-dose exposure over time. Our objective was to determine whether low-level exposure to dieldrin at an environmentally relevant dose results in aberrant molecular signaling in the vertebrate heart. Using transcriptomic profiling and immunoblotting, we determined the global gene and targeted protein expression response to dieldrin treatment and show that dieldrin affects gene networks in the heart that are associated with processes related to cardiovascular disease, specifically cardiac arrest and ventricular fibrillation. We report that genes regulating inflammatory responses, a significant risk factor for cardiovascular disease, are upregulated by dieldrin whereas transcripts related to lysosomal function are significantly downregulated. To verify these findings, proteins in these pathways were examined with immunoblotting, and our results demonstrate that dieldrin constitutively activates Akt/mTOR signaling and downregulates lysosomal genes, participating in autophagy. Our data demonstrate that dieldrin induces genes associated with cardiovascular dysfunction and compromised lysosomal physiology, thereby identifying a novel mechanism for pesticide-induced cardiotoxicity.

Dieldrin is an environmental contaminant that adversely affects aquatic organisms. The data presented in this study are proteomic data collected in liver of zebrafish that were exposed to the pesticide in a dietary exposure. For label free proteomics, data were collected with a quadrupole Time-of-Flight mass spectrometer and for iTRAQ proteomics, data were acquired using a hybrid quadrupole Orbitrap (Q Exactive) MS system. Using formic acid digestion and label free proteomics, 2,061 proteins were identified, and among those, 103 were differentially abundant (p

The herbicide linuron (LIN) is an endocrine disruptor with an anti-androgenic mode of action. The objectives of this study were to (1) improve knowledge of androgen and anti-androgen signaling in the teleostean ovary and to (2) assess the ability of gene networks and machine learning to classify LIN as an anti-androgen using transcriptomic data. Ovarian explants from vitellogenic fathead minnows (FHMs) were exposed to three concentrations of either 5α-dihydrotestosterone (DHT), flutamide (FLUT), or LIN for 12h. Ovaries exposed to DHT showed a significant increase in 17β-estradiol (E2) production while FLUT and LIN had no effect on E2. To improve understanding of androgen receptor signaling in the ovary, a reciprocal gene expression network was constructed for DHT and FLUT using pathway analysis and these data suggested that steroid metabolism, translation, and DNA replication are processes regulated through AR signaling in the ovary. Sub-network enrichment analysis revealed that FLUT and LIN shared more regulated gene networks in common compared to DHT. Using transcriptomic datasets from different fish species, machine learning algorithms classified LIN successfully with other anti-androgens. This study advances knowledge regarding molecular signaling cascades in the ovary that are responsive to androgens and anti-androgens and provides proof of concept that gene network analysis and machine learning can classify priority chemicals using experimental transcriptomic data collected from different fish species.