Interleukin-6 induces nascent protein synthesis in human dorsal root ganglion nociceptors primarily via MNK-eIF4E signaling.

Plasticity of dorsal root ganglion (DRG) nociceptors in the peripheral nervous system requires new protein synthesis. This plasticity is believed to be responsible for the physiological changes seen in DRG nociceptors in animal models of chronic pain. Experiments in human DRG (hDRG) neurons also support this hypothesis, but a direct observation of nascent protein synthesis in response to a pain promoting substance, like interleukin-6 (IL-6), has not been measured in these neurons. To fill this gap in knowledge, we used acutely prepared human DRG explants from organ donors. These explants provide a physiologically relevant microenvironment, closer to in vivo conditions, allowing for the examination of functional alterations in DRG neurons reflective of human neuropathophysiology. Using this newly developed assay, we demonstrate upregulation of the target of the MNK1/2 kinases, phosphorylated eIF4E (p-eIF4E), and nascently synthesized proteins in a substantial subset of hDRG neurons following exposure to IL-6. To pinpoint the specific molecular mechanisms driving this IL-6-driven increase in nascent proteins, we used the specific MNK1/2 inhibitor eFT508. Treatment with eFT508 resulted in the inhibition of IL-6-induced increases in p-eIF4E and nascent proteins. Additionally, using TRPV1 as a marker for nociceptors, we found that these effects occurred in a large number of human nociceptors. Our findings provide clear evidence that IL-6 drives nascent protein synthesis in human TRPV1+ nociceptors primarily via MNK1/2-eIF4E signaling. The work links animal findings to human nociception, creates a framework for additional hDRG signaling experiments, and substantiates the continued development of MNK inhibitors for pain.

Microbial, holobiont, and Tree of Life eDNA/eRNA for enhanced ecological assessment.

Microbial environmental DNA and RNA (collectively 'eNA') originate from a diverse and abundant array of microbes present in environmental samples. These eNA signals, largely representing whole organisms, serve as a powerful complement to signals derived from fragments or remnants of larger organisms. Integrating microbial data into the toolbox of ecosystem assessments and biotic indices therefore has the potential to transform how we use eNA data to understand biodiversity dynamics and ecosystem functions, and to inform the next generation of environmental monitoring. Incorporating holobiont and Tree of Life approaches into eNA analyses offers further holistic insight into the range of ecological interactions between microbes and other organisms, paving the way for advancing our understanding of, and ultimately manipulating ecosystem properties pertinent to environmental management, conservation, wildlife health, and food production.

Connecting blue-green infrastructure elements to reduce combined sewer overflows.

By infiltrating and retaining stormwater, Blue-Green Infrastructure (BGI) can help to reduce Combined Sewer Overflows (CSOs), one of the main causes of urban water pollution. Several studies have evaluated the ability of individual BGI types to reduce CSOs; however, the effect of combining these elements, likely to occur in reality, has not yet been thoroughly evaluated. Moreover, the CSO volume reduction potential of relevant components of the urban drainage system, such as detention ponds, has not been quantified using hydrological models. This study presents a systematic way to assess the potential of BGI combinations to mitigate CSO discharge in a catchment near Zurich (Switzerland). Sixty BGI combinations, including four BGI elements (bioretention cells, permeable pavement, green roofs, and detention ponds) and four different implementation rates (25%, 50%, 75%, and 100% of the available sewer catchment area) are evaluated for four runoff routing schemes. Results reveal that BGI combinations can provide substantial CSO volume reductions; however, combinations including detention ponds can potentially increase CSO frequency, due to runoff prolongation. When runoff from upstream areas is routed to the BGI, the CSO discharge reductions from combinations of BGI elements differ from the cumulative CSO discharge reductions achieved by individual BGI types, indicating that the sum of effects from individual BGI types cannot accurately predict CSO discharge in combined BGI scenarios. Moreover, larger BGI implementation areas are not consistently more cost-effective than small implementation areas, since the additional CSO volume reduction does not outweigh the additional costs. The best-performing BGI combination depends on the desired objective, being CSO volume reduction, CSO frequency reduction or cost-effectiveness. This study emphasizes the importance of BGI combinations and detention ponds in CSO mitigation plans, highlighting their critical factors-BGI types, implementation area, and runoff routing- and offering a novel and systematic approach to develop tailored BGI strategies for urban catchments facing CSO challenges.

The effect of green infrastructure on resilience performance in combined sewer systems under climate change.

Climate change is currently reshaping precipitation patterns, intensifying extremes, and altering runoff dynamics. Particularly susceptible to these impacts are combined sewer systems (CSS), which convey both stormwater and wastewater and can lead to combined sewer overflow (CSO) discharges during heavy rainfall. Green infrastructure (GI) can help mitigate these discharges and enhance system resilience under historical conditions; however, the quantification of its effect on resilience in a future climate remains unknown in the literature. This study employs a modified Global Resilience Analysis (GRA) framework for continuous simulation to quantify the impact of climate change on CSS resilience, particularly CSOs. The study assesses the efficacy of GI interventions (green roofs, permeable pavements, and bioretention cells) under diverse future rainfall scenarios based on EURO-CORDEX regional climate models (2085-2099) and three Representative Concentration Pathways (2.6, 4.5, 8.5 W/m2). The findings underscore a general decline in resilience indices across the future rainfall scenarios considered. Notably, the total yearly CSO discharge volume increases by a range of 145 % to 256 % in response to different rainfall scenarios. While GI proves effective in increasing resilience, it falls short of offsetting the impacts of climate change. Among the GI options assessed, green roofs routed to pervious areas exhibit the highest adaptive capacity, ranging from 9 % to 22 % at a system level, followed by permeable pavements with an adaptation capacity between 7 and 13 %. By linking the effects of future rainfall scenarios on CSO performance, this study contributes to understanding GI's potential as a strategic tool for enhancing urban resilience.

Do baseline assumptions alter the efficacy of green stormwater infrastructure to reduce combined sewer overflows?

Green stormwater infrastructure (GSI) is growing in popularity to reduce combined sewer overflows (CSOs) and hydrologic simulation models are a tool to assess their reduction potential. Given the numerous and interacting water flows that contribute to CSOs, such as evapotranspiration (ET) and groundwater (GW), these models should ideally account for them. However, due to the complexity, simplified models are often used, and it is currently unknown how these assumptions affect estimates of CSOs, GSI effectiveness, and ultimately planning guidance. This study evaluates the effect on estimates of CSOs and GSI effectiveness when different flows and hydrologic processes are neglected. We modified an existing EPA SWMM model of a combined sewer system in Switzerland to include ET, GW, and upstream inflows. Historical rainfall data over 30 years are used to assess volume and duration of CSOs with and without three types of GSI (bioretention basins, permeable pavements and green roofs). Results demonstrate that neglect of certain flows in modelling can alter CSO volumes from -15 % to 40 %. GSI effectiveness also varies considerably, resulting in differences in simulated percent of CSO volume reduced from 8 % to 35 %, depending on the GSI type and modeled flow or process. Representation of GW within models is particularly crucial when infiltrating GSI are present, as CSOs could increase in certain subcatchments due to higher GW levels from increased infiltration. When basing GSI planning decisions on modeled estimates of CSOs, all relevant hydrologic processes should be included to the extent possible, and uncertainty and assumptions should always be considered.

The effect of pre-exercise protein intake on substrate metabolism, energy expenditure, and energy intake: a dose-response study.

BACKGROUND: Pre-exercise protein consumption does not seem to influence substrate metabolism during exercise compared to fasted exercise, however it is unclear if the protein dose impacts on this effect. METHODS: In a randomized, double-blinded within-subject design trial, healthy, active males and females (n = 15, 25 ± 5 yrs, O2peak: 47.5 ± 8.8 ml/kg/min) completed 1 h of cycling exercise at 60% peak power output 30 min after having consumed either 0, 20, or 40 g of whey protein. Indirect calorimetry was used to measure substrate oxidation during exercise and baseline and post-exercise resting energy expenditure. Blood samples were taken throughout the trials to measure metabolic responses. Free-living food intake post-trial was collected using food diaries. RESULTS: Fat oxidation rates during exercise did not differ between the three conditions (p = 0.19) with small effect sizes between conditions (Cohen's dz: 0 vs. 20 g = 0.22, 0 vs. 40 g = 0.47, 20 vs. 40 g = 0.27). Serum insulin was higher in the protein groups vs. 0 g (p < 0.05), whereas non-esterified fatty acids were higher in the 0 g compared to 20 and 40 g (p < 0.05). Glucose was significantly lower after 15 min of exercise in 20 and 40 g vs. 0 g (p = 0.01). Resting energy expenditure was elevated post-exercise (p < 0.001), without an interaction for protein dose (p = 0.90). Post-trial free-living energy intake was not different between conditions (p = 0.31), but 24-h energy intake was significantly higher in 40 vs. 0 g (p = 0.04). CONCLUSION: Protein doses up to 40 g do not seem to impair fat oxidation rates during exercise compared to fasted exercise and could be considered as a nutritional strategy for exercising individuals who struggle to include fasted exercise in their training.

Which Algorithm Best Propagates the Meyer-Miller-Stock-Thoss Mapping Hamiltonian for Non-Adiabatic Dynamics?

A common strategy to simulate mixed quantum-classical dynamics is by propagating classical trajectories with mapping variables, often using the Meyer-Miller-Stock-Thoss (MMST) Hamiltonian or the related spin-mapping approach. When mapping the quantum subsystem, the coupled dynamics reduce to a set of equations of motion to integrate. Several numerical algorithms have been proposed, but a thorough performance comparison appears to be lacking. Here, we compare three time-propagation algorithms for the MMST Hamiltonian: the Momentum Integral (MInt) (J. Chem. Phys., 2018, 148, 102326), the Split-Liouvillian (SL) (Chem. Phys., 2017, 482, 124-134), and the algorithm in J. Chem. Phys., 2012, 136, 084101 that we refer to as the Degenerate Eigenvalue (DE) algorithm due to the approximation required during derivation. We analyze the accuracy of individual trajectories, correlation functions, energy conservation, symplecticity, Liouville's theorem, and the computational cost. We find that the MInt algorithm is the only rigorously symplectic algorithm. However, comparable accuracy at a lower computational cost can be obtained with the SL algorithm. The approximation implicitly made within the DE algorithm conserves energy poorly, even for small timesteps, and thus leads to slightly different results. These results should guide future mapping-variable simulations.

Global resilience analysis of combined sewer systems under continuous hydrologic simulation.

Managing and reducing combined sewer overflow (CSO) discharges is crucial for enhancing the resilience of combined sewer systems (CSS). However, the absence of a standardised resilience analysis approach poses challenges in developing effective discharge reduction strategies. To address this, our study presents a top-down method that expands the existing Global Resilience Analysis to quantify resilience performance in CSS. This approach establishes a link between threats (e.g., rainfall) and impacts (e.g., CSOs) through continuous and long-term simulation, accommodating various rainfall patterns, including extreme events. We assess CSO discharge impacts from a resilience perspective by introducing eight new metrics. We conducted a case study in Fehraltorf, Switzerland, analysing the performance of three green infrastructure (GI) types (bioretention cells, green roofs, and permeable pavements) over 38 years. The results demonstrated that GI enhanced all resilience indices, with variations observed in individual CSO performance metrics and their system locations. Notably, in Fehraltorf, green roofs emerged as the most effective GI type for improving resilience, while the downstream outfall displayed the highest resilience enhancement. Overall, our proposed method enables a shift from event-based to continuous simulation analysis, providing a standardised approach for resilience assessment. This approach informs the development of strategies for CSO discharge reduction and the enhancement of CSS resilience.

Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types.

Two multiplex immunoassays are routinely used to assess antibody responses in clinical trials of the 9-valent human papillomavirus (9vHPV) vaccine. The HPV6/11/16/18/31/33/45/52/58 competitive Luminex immunoassay (HPV-9 cLIA) and HPV6/11/16/18/31/33/45/52/58 total immunoglobulin G Luminex immunoassay are used for measurements of immunogenicity. Following their initial validation in 2010, both assays were redeveloped, and several parameters were optimized, including the coating concentration of virus-like particles, type of Luminex microspheres, serum sample and reference standard diluent, reference standard starting dilution and titration series, and vendor and concentration of the phycoerythrin-labeled antibodies. Validation studies evaluated the assay performance parameters, including the intra-assay precision (repeatability), intermediate precision, linearity, relative accuracy, and limits of quantitation. In addition, since maintaining a link to the original assays that were used in trials supporting vaccine licensure is critical, the assays were formally bridged to the previous assay versions by using individual patient sera from a 9vHPV vaccine clinical trial (n = 150 day 1 [prevaccination] samples; n = 100 month 7 [1 month post-last vaccine dose] and n = 100 month 36 [30 months post-last vaccine dose; antibody persistence] samples). The results of the validation studies indicate that both optimized assays are accurate, specific, and precise over their respective quantifiable ranges. There was a strong linear association between the new and previous versions of both assays. Assay serostatus cutoffs for the redeveloped assays were established based on the bridging studies and, for the HPV-9 cLIA, further refined, based on additional data from HPV vaccine clinical studies so as to align the seropositivity rates between assay versions. IMPORTANCE Assay modernization is a key aspect of vaccine life cycle management. Thus, new, reoptimized versions of two 9vHPV immunoassays have been developed and validated for use in ongoing and future HPV vaccine clinical trials. These assays are suitable for use in high-throughput testing for HPV antibodies in serum samples. Bridging to previous versions of the assays allows for the continuous monitoring of immune responses across assay versions, including in immunogenicity studies that involve new populations as well as long-term follow-up studies.

Environmental DNA/RNA for pathogen and parasite detection, surveillance, and ecology.

Detection of pathogens, parasites, and other symbionts in environmental samples via eDNA/eRNA (collectively eNA) is an increasingly important source of information about their occurrence and activity. There is great potential for using such detections as a proxy for infection of host organisms in connected habitats, for pathogen monitoring and surveillance, and for early warning systems for disease. However, many factors require consideration, and appropriate methods developed and verified, in order that eNA detections can be reliably interpreted and adopted for surveillance and assessment of disease risk, and potentially inclusion in international standards, such as the World Organisation for Animal Health guidelines. Disease manifestation results from host-symbiont-environment interactions between hosts, demanding a multifactorial approach to interpretation of eNA signals.

Characterization of Fragile X Mental Retardation Protein expression in human nociceptors and their axonal projections to the spinal dorsal horn.

Fragile X Mental Retardation Protein (FMRP) regulates activity-dependent RNA localization and local translation to modulate synaptic plasticity throughout the central nervous system. Mutations in the FMR1 gene that hinder or ablate FMRP function cause Fragile X Syndrome (FXS), a disorder associated with sensory processing dysfunction. FXS premutations are associated with increased FMRP expression and neurological impairments including sex dimorphic presentations of chronic pain. In mice, FMRP ablation causes dysregulated dorsal root ganglion (DRG) neuron excitability and synaptic vesicle exocytosis, spinal circuit activity, and decreased translation-dependent nociceptive sensitization. Activity-dependent, local translation is a key mechanism for enhancing primary nociceptor excitability that promotes pain in animals and humans. These works indicate that FMRP likely regulates nociception and pain at the level of the primary nociceptor or spinal cord. Therefore, we sought to better understand FMRP expression in the human DRG and spinal cord using immunostaining in organ donor tissues. We find that FMRP is highly expressed in DRG and spinal neuron subsets with substantia gelatinosa exhibiting the most abundant immunoreactivity in spinal synaptic fields. Here, it is expressed in nociceptor axons. FMRP puncta colocalized with Nav1.7 and TRPV1 receptor signals suggesting a pool of axoplasmic FMRP localizes to plasma membrane-associated loci in these branches. Interestingly, FMRP puncta exhibited notable colocalization with calcitonin gene-related peptide (CGRP) immunoreactivity selectively in female spinal cord. Our results support a regulatory role for FMRP in human nociceptor axons of the dorsal horn and implicate it in the sex dimorphic actions of CGRP signaling in nociceptive sensitization and chronic pain.

Development and Validation of a Sensitive and Robust Multiplex Antigen Capture Assay to Quantify Streptococcus pneumoniae Serotype-Specific Capsular Polysaccharides in Urine.

Streptococcus pneumoniae is a major cause of community-acquired pneumonia (CAP) in young children, older adults, and those with immunocompromised status. Since the introduction of pneumococcal vaccines, the burden of invasive pneumococcal disease caused by vaccine serotypes (STs) has decreased; however, the effect on the burden of CAP is unclear, potentially due to the lack of testing for pneumococcal STs. We describe the development, qualification, and clinical validation of a high-throughput and multiplex ST-specific urine antigen detection (SSUAD) assay to address the unmet need in CAP pneumococcal epidemiology. The SSUAD assay is sensitive and specific to the 15 STs in the licensed pneumococcal conjugate vaccine V114 (STs 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F) and uses ST-specific monoclonal antibodies for rapid and simultaneous quantification of the 15 STs using a Luminex microfluidics system. The SSUAD assay was optimized and qualified using healthy adult urine spiked with pneumococcal polysaccharides and validated using culture-positive clinical urine samples (n = 34). Key parameters measured were accuracy, precision, sensitivity, specificity, selectivity, and parallelism. The SSUAD assay met all prespecified validation acceptance criteria and is suitable for assessments of disease burden associated with the 15 pneumococcal STs included in V114. IMPORTANCE Streptococcus pneumoniae has more than 90 serotypes capable of causing a range of disease manifestations, including otitis media, pneumonia, and invasive diseases, such as bacteremia or meningitis. Only a minority (<10%) of pneumococcal diseases are bacteremic with known serotype distribution. Culture and serotyping of respiratory specimens are neither routine nor reliable. Hence, the serotype-specific disease burden of the remaining (>90%) noninvasive conditions is largely unknown without reliable laboratory techniques. To address this need, a 15-plex urine antigen detection assay was developed and validated to quantify pneumococcal serotype-specific capsular polysaccharides in urine. This assay will support surveillance to estimate the pneumococcal disease burden and serotype distribution in nonbacteremic conditions. Data obtained from this assay will be critical for understanding the impact of pneumococcal vaccines on noninvasive pneumococcal diseases and to inform the choice of pneumococcal serotypes for next-generation vaccines.

Exploring the development of professional values in an online RN-to-BSN program.

BACKGROUND: Development of professional nursing values is critical within registered nurse-to-bachelor of science in nursing programs to prepare nurses for increasingly complex and diverse work environments. The results of previous studies have been inconsistent, with few studies focusing on online registered nurse-to-bachelor of science in nursing programs. In addition, little is known regarding the effectiveness of the educational methods used to support advancement of professional values and ethical practice. OBJECTIVE: The object of this study was to gain an understanding of nursing students' attitudes and beliefs about professional values at entry and exit of an online registered nurse-to-bachelor of science in nursing program that includes a standalone ethics course and integrates American Nurses Association Code of Ethics provisions throughout the curriculum. RESEARCH DESIGN: For this one-group pretest-posttest, quasi-experimental design, longitudinal matched-pair data were gathered at program entry and exit using the Nurses Professional Values Scale-Revised. PARTICIPANTS AND RESEARCH CONTEXT: In all, 119 students of an online registered nurse-to-bachelor of science in nursing program at a Midwest public university who completed entry and exit surveys between spring 2015 and spring 2018 were included in this study. ETHICAL CONSIDERATIONS: This study was reviewed and determined to be exempt by the university's institutional review board. FINDINGS: The results showed a significant increase in total posttest scores when considering all participants. However, students who took the ethics course after the pretest demonstrated a significant increase in posttest scores, while students who took the ethics course prior to the pretest demonstrated a small increase that was not statistically significant. Significant increases were also found in the professionalism, activism, and trust factors. DISCUSSION: This study supports previous study findings where students scored higher on caring and lower on activism and professionalism factors. The largest gains were made after completing the ethics course. CONCLUSION: The results suggest that requiring a standalone ethics course in the registered nurse-to-bachelor of science in nursing curriculum had a positive impact on self-reported professional values.

Genome Sequence of Acetomicrobium hydrogeniformans OS1.

Acetomicrobium hydrogeniformans, an obligate anaerobe of the phylum Synergistetes, was isolated from oil production water. It has the unusual ability to produce almost 4 molecules H2/molecule glucose. The draft genome of A. hydrogeniformans OS1 (DSM 22491T) is 2,123,925 bp, with 2,068 coding sequences and 60 RNA genes.

Dermoscopy in the Diagnosis of Inflammatory Dermatoses: Systematic Review Findings Reported for Psoriasis, Lupus, and Lichen Planus.

Background: Dermoscopy is well established as a tool to improve the detection of cancerous skin growths. Published data suggest that dermoscopy might be useful in evaluating inflammatory dermatoses and in distinguishing between rashes and skin cancer. Objective: The authors sought to review the published literature regarding use of dermoscopy in the evaluation of inflammatory skin conditions. Methods: Using a systematic approach, the authors performed a literature search using the names of 146 inflammatory dermatoses and pairing each one separately with the search terms dermoscopy, dermatoscopy, and epiluminescence microscopy.Results: After eliminating those papers that did not meet inclusion requirements, the authors identified 201 studies for their review, with the majority consisting of case reports. The most commonly studied inflammatory conditions were psoriasis, lupus, and lichen planus. There was congruence among the studies identified in terms of the most common dermoscopic findings for each of these diseases. Conclusions: The use of dermoscopy in the evaluation of inflammatory dermatoses is a promising option. However, more rigorous studies are needed to determine the sensitivity and specificity of the dermoscopic findings for many inflammatory skin conditions.

Scaly pink patches: differentiating psoriasis from basal cell carcinoma.

Dermoscopy, commonly used to analyze skin tumors, has more recently been used to evaluate inflammatory dermatoses. We performed a systematic review of the literature to assess the role of dermoscopy in evaluating psoriasis, and briefly reviewed the findings with an emphasis on the specificity or sensitivity of the dermoscopic findings of psoriasis. We also describe the case of a 63-year-old man with a history of psoriasis and basal cell carcinoma (BCC) who presented with a new scaly pink patch on the back. This case highlights the importance of dermoscopy in differentiating patches and plaques of psoriasis from BCC.

A mathematical model to plan for long-term effects of water conservation choices on dry weather wastewater flows and concentrations.

In many cities, sewer systems are experiencing conditions that are significantly different from those for which they were designed. Factors such as water conservation efforts, changes in population, and efforts to reduce infiltration are altering the quantity and quality of sewage. These changes may affect the ability of sewers to maintain self-cleansing velocities, which are crucial to avoiding solids settling and corrosion issues. Further, such changes may alter the timeline for expected wastewater plant expansion. The present work proposes a method for predicting average annual dry weather wastewater flow, as well as pollutant load and concentration over time. The method takes into account potential declines in per person wastewater production due to water conservation and reuse practices, as well as other potential changes such as shifts in population, transformations in industrial wastewater production, and variations in dry weather infiltration. Results show that the amount of dry weather infiltration will play a large role in whether or not conservation will affect self-cleansing velocities or plant expansions. Conservation is most beneficial to systems with high levels of dry weather infiltration since plant expansion could be avoided; and most detrimental to systems with low levels of infiltration since low flow conditions could lead to settling and corrosion in the sewer. Furthermore, the rate of implementation of conservation efforts influences when impacts to the system would occur. Utility planners will be able to use this method to predict treatment plant upgrade and expansion needs more accurately as well as to assess the relative value of utility-based maintenance activities and conservation practices.

Complete genome sequence of Methanospirillum hungatei type strain JF1.

Methanospirillum hungatei strain JF1 (DSM 864) is a methane-producing archaeon and is the type species of the genus Methanospirillum, which belongs to the family Methanospirillaceae within the order Methanomicrobiales. Its genome was selected for sequencing due to its ability to utilize hydrogen and carbon dioxide and/or formate as a sole source of energy. Ecologically, M. hungatei functions as the hydrogen- and/or formate-using partner with many species of syntrophic bacteria. Its morphology is distinct from other methanogens with the ability to form long chains of cells (up to 100 µm in length), which are enclosed within a sheath-like structure, and terminal cells with polar flagella. The genome of M. hungatei strain JF1 is the first completely sequenced genome of the family Methanospirillaceae, and it has a circular genome of 3,544,738 bp containing 3,239 protein coding and 68 RNA genes. The large genome of M. hungatei JF1 suggests the presence of unrecognized biochemical/physiological properties that likely extend to the other Methanospirillaceae and include the ability to form the unusual sheath-like structure and to successfully interact with syntrophic bacteria.