Insights into optimizing phantom pain management: A human-centered approach to end-user perspectives.

BACKGROUND: This report explores and summarizes perspectives from end users on unmet needs in achieving optimal and effective phantom pain management through a human-centered design approach. OBJECTIVE: To examine current strategies, pharmacological, nonpharmacological, surgical procedures, virtual reality, and mirror therapy, and the evidence supporting them, in the management of phantom limb pain. DESIGN: This study reviewed and analyzed transcripts acquired in nonresearch contexts from the Veterans Affairs Translational Education and Mentoring Center's commercialization training program and from a Veteran Engagement Panel. Key themes were extracted using quasi-qualitative analysis of one-on-one interviews. RESULTS: Clinicians and patients report that early patient intervention and education will yield improved management of phantom limb pain, which aligns with the growing recognition of the impact of patient-centered care on overall treatment outcomes. Mirror therapy is viewed as an effective and low-risk therapy, though compliance and buy-in may be barriers to clinical practice. Patient engagement can contribute to better treatment adherence and outcomes. CONCLUSION: The study highlights implementation barriers, importance of end-user input, and the role of the Veteran Engagement Panel in providing feedback to pain researchers. The findings help explain unaddressed challenges and areas requiring further research to direct phantom pain management.

Interlaboratory comparison using inactivated SARS-CoV-2 variants as a feasible tool for quality control in COVID-19 wastewater monitoring.

Wastewater-based SARS-CoV-2 epidemiology (WBE) has proven as an excellent tool to monitor pandemic dynamics supporting individual testing strategies. WBE can also be used as an early warning system for monitoring the emergence of novel pathogens or viral variants. However, for a timely transmission of results, sophisticated sample logistics and analytics performed in decentralized laboratories close to the sampling sites are required. Since multiple decentralized laboratories commonly use custom in-house workflows for sample purification and PCR-analysis, comparative quality control of the analytical procedures is essential to report reliable and comparable results. In this study, we performed an interlaboratory comparison at laboratories specialized for PCR and high-throughput-sequencing (HTS)-based WBE analysis. Frozen reserve samples from low COVID-19 incidence periods were spiked with different inactivated authentic SARS-CoV-2 variants in graduated concentrations and ratios. Samples were sent to the participating laboratories for analysis using laboratory specific methods and the reported viral genome copy numbers and the detection of viral variants were compared with the expected values. All PCR-laboratories reported SARS-CoV-2 genome copy equivalents (GCE) for all spiked samples with a mean intra- and inter-laboratory variability of 19 % and 104 %, respectively, largely reproducing the spike-in scheme. PCR-based genotyping was, in dependence of the underlying PCR-assay performance, able to predict the relative amount of variant specific substitutions even in samples with low spike-in amount. The identification of variants by HTS, however, required >100 copies/ml wastewater and had limited predictive value when analyzing at a genome coverage below 60 %. This interlaboratory test demonstrates that despite highly heterogeneous isolation and analysis procedures, overall SARS-CoV-2 GCE and mutations were determined accurately. Hence, decentralized SARS-CoV-2 wastewater monitoring is feasible to generate comparable analysis results. However, since not all assays detected the correct variant, prior evaluation of PCR and sequencing workflows as well as sustained quality control such as interlaboratory comparisons are mandatory for correct variant detection.

Early Detection of SARS-CoV-2 Omicron BA.4 and BA.5 in German Wastewater.

Wastewater-based SARS-CoV-2 epidemiology (WBE) has been established as an important tool to support individual testing strategies. The Omicron sub-variants BA.4/BA.5 have spread globally, displacing the preceding variants. Due to the severe transmissibility and immune escape potential of BA.4/BA.5, early monitoring was required to assess and implement countermeasures in time. In this study, we monitored the prevalence of SARS-CoV-2 BA.4/BA.5 at six municipal wastewater treatment plants (WWTPs) in the Federal State of North Rhine-Westphalia (NRW, Germany) in May and June 2022. Initially, L452R-specific primers/probes originally designed for SARS-CoV-2 Delta detection were validated using inactivated authentic viruses and evaluated for their suitability for detecting BA.4/BA.5. Subsequently, the assay was used for RT-qPCR analysis of RNA purified from wastewater obtained twice a week at six WWTPs. The occurrence of L452R carrying RNA was detected in early May 2022, and the presence of BA.4/BA.5 was confirmed by variant-specific single nucleotide polymorphism PCR (SNP-PCR) targeting E484A/F486V and NGS sequencing. Finally, the mutant fractions were quantitatively monitored by digital PCR, confirming BA.4/BA.5 as the majority variant by 5 June 2022. In conclusion, the successive workflow using RT-qPCR, variant-specific SNP-PCR, and RT-dPCR demonstrates the strength of WBE as a versatile tool to rapidly monitor variants spreading independently of individual test capacities.

Wastewater surveillance allows early detection of SARS-CoV-2 omicron in North Rhine-Westphalia, Germany.

Wastewater-based epidemiology (WBE) has demonstrated its importance to support SARS-CoV-2 epidemiology complementing individual testing strategies. Due to their immune-evasive potential and the resulting significance for public health, close monitoring of SARS-CoV-2 variants of concern (VoC) is required to evaluate the regulation of early local countermeasures. In this study, we demonstrate a rapid workflow for wastewater-based early detection and monitoring of the newly emerging SARS-CoV-2 VoCs Omicron in the end of 2021 at the municipal wastewater treatment plant (WWTP) Emschermuendung (KLEM) in the Federal State of North-Rhine-Westphalia (NRW, Germany). Initially, available primers detecting Omicron-related mutations were rapidly validated in a central laboratory. Subsequently, RT-qPCR analysis of purified SARS-CoV-2 RNA was performed in a decentral PCR laboratory in close proximity to KLEM. This decentralized approach enabled the early detection of K417N present in Omicron in samples collected on 8th December 2021 and the detection of further mutations (N501Y, Δ69/70) in subsequent biweekly sampling campaigns. The presence of Omicron in wastewater was confirmed by next generation sequencing (NGS) in a central laboratory with samples obtained on 14th December 2021. Moreover, the relative increase of the mutant fraction of Omicron was quantitatively monitored over time by dPCR in a central PCR laboratory starting on 12th December 2021 confirming Omicron as the dominant variant by the end of 2021. In conclusions, WBE plays a crucial role in surveillance of SARS-CoV-2 variants and is suitable as an early warning system to identify variant emergence. In particular, the successive workflow using RT-qPCR, RT-dPCR and NGS demonstrates the strength of WBE as a versatile tool to monitor variant spreading.

Removal of pharmaceuticals from wastewater of health care facilities.

Health care facilities can represent point sources for pharmaceutical residues in public sewer systems. Compared to general hospitals, more specialized health care facilities might also have a different pattern of medication. Therefore, the on-site treatment of wastewater from such facilities could be an effective strategy to reduce emissions into water bodies and was the aim of this study. Wastewater from three health care facilities (nursing home, clinic with orthopaedic focus, and psychiatric clinic) was treated in lab-scale and semi-industrial trials. Biological treatment was performed via an ultrafiltration membrane bioreactor (UF-MBR), after which adsorption onto granular activated carbon (using rapid-small-scale column tests GAC-RSSCT), ozonation and a UV/H2O2 advanced oxidation process (AOP) were tested and compared. The removal of 17 pharmaceutical compounds and drug metabolites from 9 drug classes (e.g. analgesics, antibiotics, anticonvulsants) was evaluated. Most of the measured OMP were detected with concentrations between 1,000 and 30,000 ng L-1 in the influent of the MBR. The UF-MBR provided an effective mechanical-biological cleaning of the wastewater, with micropollutant removal efficiencies between 0 and > 95%, making further treatment necessary to remove the micropollutants. Each combination of the UF-MBR with one of the three further treatments achieved removal efficiencies above 80% for most of the investigated substances, reducing many to below the quantification limit of 10 ng L-1. The results show the general suitability of combining the UF-MBR with either GAC-adsorption, ozonation or AOP for eliminating pharmaceutical residues. However, the AOP process has a significantly higher energy demand than the other two processes. Moreover, specific settings and dosages depend on the respective wastewater matrix.

LBP and sCD14 patterns in total hip replacement surgery performed during combined spinal/epidural anaesthesia.

BACKGROUND: Danger patterns and pattern recognition receptors have been targets in the investigation and treatment of systemic inflammatory response syndrome and sepsis. Lipopolysaccharide (LPS)-binding protein (LBP) presents LPS and gram-positive bacterial cell wall products to the receptors TLR4/MD-2 and TLR2, respectively. Low concentrations of LBP stimulate responses to LPS and peptidoglycan, whereas higher concentrations inhibit these responses. Soluble CD14 (sCD14) presents the LBP-LPS complex to CD14-negative cells, and it modulates the biological activity of circulating LPS. In this study, we aimed to elucidate the physiological reactions to LBP and sCD14 after total hip replacement surgery during spinal/epidural anaesthesia. METHODS: Seven patients with coxarthrosis were operated upon with a total hip replacement, which is a defined trauma to bone and muscles in conjunction with a certain amount of blood loss. Venous blood samples were taken before the operation and at 1 h, 3 days and 6 days after surgery. LBP and sCD14 were measured by conventional ELISA. To correct for hemodilution, each parameter was adjusted for hematocrit. A panel of cytokines was measured using Luminex technology to evaluate the trauma reaction. RESULTS: IL-6 levels peaked 24 h after the operation, whereas IL-1ß and IL-10 levels remained unchanged. Systemic levels of LBP were increased 24 h after surgery, whereas sCD14 remained steady. However, the dilution-corrected sCD14 values increased significantly, and the levels of both LBP and sCD14 peaked at day 3 after surgery. CONCLUSION: Aseptic trauma primes the innate immune system for the posttraumatic release of LBP and sCD14.

Local and systemic chemokine patterns in a human musculoskeletal trauma model.

OBJECTIVE AND DESIGN: This prospective study aims to identify differences in local and systemic chemokines kinetics within 24 h of a standardised human surgical trauma (total hip arthroplasty) and their impact on systemic polymorphonuclear cells. MATERIALS AND METHODS: We examined seven patients with coxarthrosis, but without comorbidity, who had a total hip arthroplasty. Local drained blood and systemic blood samples were collected at wound closure and at 1, 4, and 24 h after surgery. Chemokines were measured using a multiplex antibody bead kit. Venous whole blood cell counts were taken at the same points in time. RESULTS: There is a significant postoperative local burst of CCL2 and CXCL8 while systemic levels stay unchanged. The systemic levels of CCL3, CCL4, CCL5, CCL11, CXCL9, and CXCL10 were significantly reduced at 24 h post-surgery, but local levels remain unchanged or had only modest changes. There was a significant postoperative rise in monocytes and neutrophils. CONCLUSION: There is fundamental difference between local and systemic chemokine kinetics in the human trauma model studied. High postoperative concentrations of CCL2 and CXCL8 at the site of inflammation form a gradient that contributes to the recruitment of neutrophils and monocytes at the trauma site.

Immunmodulation of serum in orthopaedic trauma.

BACKGROUND: The aim of this study was to test the hypothesis that inhibitory substances circulating in the patient's serum after trauma might impair leukocyte function by evaluating the effect of such serum on cytokine release in a whole blood model. METHODS: Hip replacement surgery was considered a standardized musculoskeletal trauma, and seven women and three men undergoing elective total hip replacement were included in the study. Ex vivo lipopolysaccharide (LPS) and peptidoglycan (PepG) induced tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL-10) releases were measured in whole blood sampled preoperatively and added serum taken before, at the end of operation and at postoperative day 1 and 6. Saline was used as negative control to serum. RESULTS: LPS and PepG induced a significant release of TNF-alpha and IL-10 in whole blood. Addition of preoperative serum, postoperative serum or day 1 postoperative serum did not alter the LPS-induced release of TNF-alpha as compared with saline control. Addition of preoperative serum significantly increased the PepG-induced release of TNF-alpha as compared with saline control (p = 0.011). This increase was not significantly changed with addition of postoperative serum or day 1 postoperative serum. When serum from postoperative day 6 was added, both LPS and PepG induced expression of TNF-alpha was significantly reduced as compared with preoperative serum (p = 0.018 and 0.008, respectively). Preoperative serum also increased the PepG induced expression of IL-10 (p = 0.007) in relation to saline control, and this increase was not significantly changed by addition of postoperative serum or day 1 and day 6 postoperative serum. Neither of the serum samples altered the LPS induced expression of IL-10 as compared with saline control (p = 0.212). CONCLUSION: Our data show that in trauma patients, serum expresses activity that inhibits LPS and PepG induced release of TNF-alpha in a whole blood model, and our study, then, corroborates the hypothesis that inhibitory substances circulating in the patients' serum after trauma impair leukocyte function.