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Soamy Montesino-Goicolea,1,2 Pedro A Valdes-Hernandez,1,2 Lorraine Hoyos,3 Adam J Woods,2,4,5 Ronald Cohen,2,5 Zhiguang Huo,6 Joseph L Riley III,1,7 Eric C Porges,2,5 Roger B Fillingim,1,7 Yenisel Cruz-Almeida1,2,4,7 1Department of Community Dentistry & Behavioral Sciences, University of Florida, Gainesville, FL, USA; 2Center for Cognitive Aging and Memory, McKnight Brain Foundation, University of Florida, Gainesville, FL, USA; 3University of Central, Florida College of Medicine, Orlando, FL, USA; 4Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL, USA; 5Department of Clinical and Health Psychology, College of Health Professions, University of Florida, Gainesville, FL, USA; 6Department of Biostatistics, College of Public Health and Health Professions College of Medicine, University of Florida, Gainesville, FL, USA; 7Institute on Aging, University of Florida, Gainesville, FL, USACorrespondence: Yenisel Cruz-AlmeidaPain Research and Intervention Center of Excellence, University of Florida, PO BOX 112610, Gainesville, FL 326010, USATel +1 352-294-5845Fax +1 352-273-5920Email cryeni@ufl.eduIntroduction: Musculoskeletal pain is prevalent in older adults representing the leading cause of disability in this population. Similarly, nearly half of older adults complain of difficulty sleeping. We aimed to explore the relationship between sleep quality with self-reported musculoskeletal pain, somatosensory and pain thresholds in community-dwelling older adults and further explore brain regions that may contribute to this association.Methods: Older adults (> 60 years old, n=69) from the NEPAL study completed demographic, pain and sleep assessments followed by a quantitative sensory testing battery. A subset (n=49) also underwent a 3T high-resolution, T1-weighted anatomical scan.Results: Poorer sleep quality using the Pittsburgh Sleep Quality Index was positively associated with self-reported pain measures (all p’s > 0.05), but not somatosensory and pain thresholds (all p’s > 0.05). Using a non-parametric threshold-free cluster enhancement (TFCE) approach, worse sleep quality was significantly associated with lower cortical thickness in the precentral, postcentral, precuneus, superior parietal, and lateral occipital regions (TFCE-FWE-corrected at p

Yenisel Cruz-Almeida,1–3 Maria Aguirre,1,4 Heather Sorenson,4,5 Patrick Tighe,1,6 Shannon M Wallet,4,5 Joseph L Riley III1,4 1Pain Research and Intervention Center of Excellence, 2Institute on Aging, 3Department of Aging and Geriatric Research, 4Department of Community Dentistry & Behavioral Science, 5Department of Oral Biology, 6Department of Anesthesiology, University of Florida, Gainesville, FL, USA Abstract: An important consideration in mechanistic research using biomarkers should include the use of saliva as an alternative to blood. The use of saliva would allow the study of susceptible populations such as older adults where venipuncture may not be feasible. Although saliva has been most commonly used to measure cortisol and tumor necrosis factor-α (TNFα), there is limited evidence that other cytokines found in saliva significantly change in response to laboratory-induced pain. Therefore, the aim of the current preliminary study was to characterize the time course, duration and magnitude of changes of commonly measured pro- (interleukin [IL]-6, IL-8) and anti-inflammatory (IL-10, IL-4) cytokines in saliva samples and to test for age-related differences in separate experimental painful and non-painful control sessions. In addition, we also tested whether venipuncture results in significant cytokine alterations similar to a painful stimulus in a non-painful, non-venipuncture control session. All cytokines were significantly induced by the cold pressor task compared to a warm control session (p < 0.001). Specifically, healthy older adults experienced greater salivary changes in all cytokines during the cold pressor session compared to younger adults in the non-painful sessions (p < 0.001). There were no significant differences between the venipuncture and non-venipuncture sessions across all cytokines (p > 0.05). Our findings support the use of saliva as a substitute for blood in both young and older healthy individuals to measure changes after experimental pain stimulation. In addition, venipuncture alone is not sufficient to induce IL-6, IL-8, IL-10 and IL-4. Future studies in the community are urgently needed to validate and further move translational mechanistic pain research to those populations most underrepresented in clinical research. Keywords: salivary markers, experimental pain, age, venipuncture

Regulatory T cell (Treg) therapy has the potential to induce transplantation tolerance so that immunosuppression and associated morbidity can be minimized. Alloantigen-reactive Tregs (arTregs) are more effective at preventing graft rejection than polyclonally expanded Tregs (PolyTregs) in murine models. We have developed a manufacturing process to expand human arTregs in short-term cultures using good manufacturing practice-compliant reagents. This process uses CD40L-activated allogeneic B cells to selectively expand arTregs followed by polyclonal restimulation to increase yield. Tregs expanded 100- to 1600-fold were highly alloantigen reactive and expressed the phenotype of stable Tregs. The alloantigen-expanded Tregs had a diverse TCR repertoire. They were more potent than PolyTregs in vitro and more effective at controlling allograft injuries in vivo in a humanized mouse model.

AIMS: To document the frequency of self-care in a clinical sample of patients with myofascial temporomandibular disorder (TMD) pain; report the perceived relief and control of pain for each of the self-care behaviors; and to test for associations between the frequency and efficacy of each self-care behavior and pain, depression and sleep quality, as assessed during a clinical visit, and to determine whether the frequency was associated with changes in pain intensity, depression, and sleep quality 30 days later. METHODS: The sample consisted of 99 female and 27 male myofascial TMD pain patients who were participants in a multidisciplinary facial pain evaluation program. The subjects participated in a structured interview during a clinical visit and a follow-up telephone interview 30 days later. The interviews included questions about self-care, including resting, relaxation techniques, massage, hot and/or cold packs, home remedies, stretching or exercise, herbal remedies, and the use of vitamins or nutritional supplements for pain. RESULTS: The passive self-care behaviors, such as resting when experiencing pain (66%) and relaxation techniques (62%), were the most commonly used. Patients reported that hot or cold packs (5.3, 0-to-10 scale) and massage (4.7) provided the greatest relief from pain, whereas resting (4.9), relaxation (4.8), and massage (4.8) resulted in the greatest ability to control pain. The most striking finding was that initial levels of pain or change in pain were not consistently associated with self-care use; however, psychosocial outcomes of depression and sleep quality were associated with self-care frequency and reported efficacy and improved in relation to patient-reported self-care frequency. CONCLUSION: Since people with chronic myofascial TMD pain engage in a range of pain self-care strategies, clinicians need to discuss self-care with patients regularly. [ABSTRACT FROM AUTHOR]

Aims: To determine psychosocial predictors of patients' ratings of satisfaction with improvement and subjective pain relief. This study also examined the underlying components of patient satisfaction with improvement, as assessed at follow-up. Methods: The sample consisted of 107 chronic orofacial pain patients evaluated at a university-based orofacial pain clinic and referred for treatment with individualized treatment plans. Pain and psychosocial functioning were assessed with standard, reliable, validated self-report instruments administered at the initial evaluation. Follow-up data were collected via a telephone-administered structured interview 8 months after the initial evaluation. Regression methodology was used to determine prediction models for satisfaction with improvement and subjective pain relief. Patient ratings of the quality of the caregiver communication were used as a control variable in all analyses. Results: Quality of caregiver communication predicted approximately 10 to 14% of the variance in outcomes in all models. Greater initial use of cognitive coping strategies and reduced depression predicted higher ratings of satisfaction with improvement and increased pain relief. When concurrent relationships among variables at the follow-up were examined, greater subjective pain relief since the evaluation, lower current pain, and higher ratings of overall mood were significant predictors of patient satisfaction with improvement. Conclusion: This study is one of the first to report that the use of certain cognitive coping strategies is associated with positive outcome for patients suffering from orofacial pain. These findings underscore the importance of individual differences on behavioral and psychosocial parameters in the prediction of patients' subjective evaluation of treatment outcome. [ABSTRACT FROM AUTHOR]

AIMS: This study examined whether temporomandibular disorder (TMD) patients with sexual versus physical abuse histories differ in their pain report, psychological distress, and somatic symptoms. METHODS: Participants were 114 female TMD patients. The sample was divided into 3 groups based on abuse history: sexual abuse, physical abuse, or no abuse. Abuse histories were assessed with a structured clinical interview. Measures used included the McGill Pain Questionnaire, the State-Trait Anxiety Inventory, the Beck Depression Inventory, and the Pennebaker Inventory of Limbic Languidness. Group differences were analyzed by analysis of variance and Bonferroni post hoc comparisons. RESULTS: Temporomandibular disorder patients with a history of physical abuse reported significantly more pain, anxiety, and depressive symptoms than did patients with a history of sexual abuse or no history of abuse. Furthermore, the results suggest that TMD patients with a sexual abuse history are not significantly different from patients with no abuse history across the domains studied. CONCLUSION: Based on the differences found, it can be argued that assessment of physical abuse histories by appropriately trained clinicians should be a routine part of any multimodal assessment of female chronic TMD patients. [ABSTRACT FROM AUTHOR]

Objectives: To identify factors that are significantly associated with dentists' use of specific caries preventive agents in adult patients, and whether dentists who use one preventive agent are also more likely to use certain others. Methods: Data were collected from 564 practitioners in The Dental Practice-Based Research Network, a multi-region consortium of participating practices and dental organizations. Results: In-office topical fluoride was the method most frequently used. Regarding at-home preventive agents, there was little difference in preference between nonprescription fluoride, prescription fluoride, or chlorhexidine rinse. Dentists who most frequently used caries prevention were also those who regularly perform caries risk assessment and individualize caries prevention at the patient level. Higher percentages of patients with dental insurance were significantly associated with more use of in-office prevention modalities. Female dentists and dentists with more-recent training were more likely to recommend preventive agents that are applied by the patient. Dentists who reported more-conservative decisions in clinical treatment scenarios were also more likely to use caries preventive agents. Groups of dentist who shared a common preference for certain preventive agents were identified. One group used preventive agents selectively, whereas the other groups predominately used either in-office or at-home fluorides. Conclusions: Caries prevention is commonly used with adult patients. However, these results suggest that only a subset of dentists base preventive treatments on caries risk at the individual patient level. [ABSTRACT FROM AUTHOR]

This study surveyed Dental Practice-Based Research Network (DPBRN) member dentists (from four regions in the U.S. and Scandinavia) who perform restorative dentistry in their practices. The survey asked a range of questions about caries risk assessment in patients aged 6 to 18. Among respondents, 73% of dentists reported performing caries risk assessment among these patients, while 14% assessed caries risk by using a special form. Regions in which most dentists were in a private practice model were the least likely to perform caries risk assessment, while regions where most dentists practiced in a large group practice model were the most likely to use a special form for caries risk assessment. Recent graduates from dental school were more likely to use a caries risk assessment compared to older graduates. Current oral hygiene, decreased salivary flow, and the presence of active caries were rated as the most important caries factors. Some differences by region were also evident for the risk factor ratings. These results suggest that not all community dentists assess caries risk. The results of this study also indicate considerable variability in dentists' views concerning the importance of specific caries risk factors in treatment planning and weak evidence that caries risk assessment is driving clinical practice when preventive treatment recommendations are being considered. [ABSTRACT FROM AUTHOR]

Objective.To identify potential predictors and correlates of compliance and to examine differences between patient and provider perspectives on compliance.Patients.One hundred eighty-four patients (84 men and 96 women) were recruited from a chronic pain treatment program for this telephone follow-up study.Results.Health care providers (HCPs) reported making more recommendations than patients reported hearing. Patients rated themselves as more compliant than did HCPs. Overall compliance rates at a >6-month follow-up were 89% from the patients' perspective and 70% from the HCPs' perspective. HCPs rated compliance specific to psychological care as more related to positive outcomes than did patients. Participants' pain and anxiety ratings at a >6-month follow-up and satisfaction with treatment were significantly associated with patients' compliance ratings. For HCP-rated compliance, only HCPs' perceived benefit and interference from compliance were associated.Conclusion.Results suggest important disparities between HCPs and patients on remembered recommendations, levels of compliance, and health-related importance of complying with recommendations. [ABSTRACT FROM AUTHOR]

Participants in a number of different HIV/AIDS educational programs, varying in content, length of course, and student audience, were surveyed to determine if education could help reduce fear and increase comfort in work with HIV/AIDS clients. It was hypothesized that attitude scores would improve following exposure to the educational programs. A retrospective pretest -- postest survey design consisting of Likert statements was used. A t test for paired samples determined if attitude scores improved following exposure to the educational programs. An analysis of variance determined if significant differences in pretest and posttest scores existed among the groups. Within groups, there was a positive increase in self-assessed attitude scores. Multiple group comparisons indicated significant differences between groups that appear to be related to program content and length. Workplace risk and whether a program was elective may have also been factors affecting attitude. [ABSTRACT FROM AUTHOR]

Assessment of sincerity of effort is frequently used in clinical practice. However, the reliability and validity of current assessment methods have not been unequivocally demonstrated. This study focused on the detection of submaximal efforts and determination of the stability of the coefficient of variation (CV) in 20 asymptomatic subjects. The coefficient of variation of peak torque (CVpeak) and coefficient of variation of average torque (CVave) were obtained from maximal and submaximal torque curves during fast isokinetic (180 deg/s) leg extension strength tests. Significant differences between effort levels (maximal and submaximal) were found for CVpeak and CVave on both days of testing (p < .001). On Day 1 of testing, using an optimized 10% CVpeak cutoff resulted in a submaximal effort detection rate of 75% with an overall (maximal and submaximal efforts) correct classification rate of 88%. Correlation coefficients indicated relatively moderate stability using the CVpeak calculated using five trials for both maximal (r = .47, p < .05) and submaximal (r = . 78, p < .001) effort conditions. Findings suggest that use of an increased number of repetitions to calculate the CV may result in more stable measures. [ABSTRACT FROM AUTHOR]

Continuous interaction between chimeric antigen receptor (CAR) T cell (CART) and tumors often result in CART dysfunction and tumor escape. We observed that tumors can take up CAR molecules, leaving CARTs without surface-expressed CARs and thus unable to kill tumors after prolonged exposure. Overexpression of Rab5 resulted in augmented clathrin-independent endocytosis, preventing loss of surface-expressed CARs, and enhanced CART activity. Interestingly, we observed membrane protrusions on the CART cell surface which disappeared after multiple tumor challenges. Rab5 maintained these protrusions after repeated tumor engagements and their presence correlated with effective tumor clearance, suggesting a link between endocytosis, membrane protrusions, and cytolytic activity. In vivo , Rab5-expressing CARTs demonstrated improved activity and were able to clear an otherwise refractory mesothelin-expressing solid cancer in humanized mice by maintaining CAR surface expression within the tumor. Thus, pairing Rab5 with CAR expression could improve the clinical efficacy of CART therapy. Highlights "CAR-jacking" occurs when surface CAR is internalized by target tumor cells.Rab5 overexpression prevents "CAR-jacking" and enhances CART function.Rab5 promotes CAR endocytic recycling and maintains membrane protrusions.Rab5-expressing CARTs exhibit enhanced therapeutic efficacy against solid tumors.

Camelina sativa , a Brassicaceae family crop, is used for fodder, human food, and biofuels. Its relatively high resistance to abiotic and biotic stresses, as well as being a climate-resilient oilseed crop, has contributed to its popularity. Camelina's seed yield and oil contents have been improved using various technologies like RNAi and CRISPR/Cas9 genome editing. A stable transformation system for protein localization and other cell autonomous investigations, on the other hand, is tedious and time consuming. This study describes a transient gene expression protocol for Camelina sativa cultivar DH55 leaves using Agrobacterium strain C58C1. The method is suitable for subcellular protein localization and colocalization studies and can be used with both constitutive and chemically induced genes. We report the subcellular localization of the N-terminal ER membrane signal anchor region (1-32 aa) of the At3G28580 gene-encoded protein from Arabidopsis in intact leaves and the expression and localization of other known organelle markers. This method offers a fast and convenient way to study proteins in the commercially important Camelina crop system. Key features • This method is based on the approach of Zhang et al. [1] and has been optimized for bioenergy crop Camelina species. • A constitutive and inducible transient gene expression in the hexaploid species Camelina sativa cultivar DH55. • Requires only 16-18 days to complete with high efficacy. Graphical overview., Competing Interests: Competing interestsThe authors declare that they have no conflicts of interest., (©Copyright : © 2024 The Authors; This is an open access article under the CC BY-NC license.)

Engineered T cells hold great promise to become part of an effective HIV cure strategy, but it is currently unclear how best to redirect T cells to target HIV. To gain insight, we generated engineered T cells using lentiviral vectors encoding one of three distinct HIV-specific T cell receptors (TCRs) or a previously optimized HIV-targeting chimeric antigen receptor (CAR) and compared their functional capabilities. All engineered T cells had robust, antigen-specific polyfunctional cytokine profiles when mixed with artificial antigen-presenting cells. However, only the CAR T cells could potently control HIV replication. TCR affinity enhancement did not augment HIV control but did allow TCR T cells to recognize common HIV escape variants. Interestingly, either altering Nef activity or adding additional target epitopes into the HIV genome bolstered TCR T cell anti-HIV activity, but CAR T cells remained superior in their ability to control HIV replication. To better understand why CAR T cells control HIV replication better than TCR T cells, we performed a time course to determine when HIV-specific T cells were first able to activate Caspase 3 in HIV-infected targets. We demonstrated that CAR T cells recognized and killed HIV-infected targets more rapidly than TCR T cells, which correlates with their ability to control HIV replication. These studies suggest that the speed of target recognition and killing is a key determinant of whether engineered T cell therapies will be effective against infectious diseases., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: AB, NLP, and BKJ were employees of Adaptimmune when these studies were performed. Adaptimmune hold patents on TCRs described in this manuscript. JLR is an inventor of patents related to the HIV CAR therapy, which is the subject of this paper, as well as other CAR therapy products, and may be eligible to receive a select portion of royalties paid from Kite to the University of Pennsylvania. JLR is a scientific co-founder and holds equity in BlueWhale Bio., (Copyright: © 2023 Zhou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Natural killer (NK) cells are frequently expanded for the clinic using irradiated, engineered K562 feeder cells expressing a core transgene set of membrane-bound (mb) IL15 and/or mbIL21 together with 41BBL. Prior comparisons of mbIL15 to mbIL21 for NK expansion lack comparisons of key attributes of the resulting NK cells, including their high-dimensional phenotype, polyfunctionality, the breadth and potency of cytotoxicity, cellular metabolism, and activity in xenograft tumor models. Moreover, despite multiple rounds of K562 stimulation, studies of sequential use of mbIL15- and mbIL21-based feeder cells are absent. We addressed these gaps and found that using mbIL15- versus mbIL21-based feeder cells drove distinct phenotypic and functional profiles. Feeder cells expressing mbIL15 alone drove superior functionality by nearly all measures, whereas those expressing mbIL21 alone drove superior yield. In combination, most attributes resembled those imparted by mbIL21, whereas in sequence, NK yield approximated that imparted by the first cytokine, and the phenotype, transcriptome, and function resembled that driven by the second cytokine, highlighting the plasticity of NK cell differentiation. The sequence mbIL21 followed by mbIL15 was advantageous in achieving significant yields of highly functional NK cells that demonstrated equivalent in vivo activity to those expanded by mbIL15 alone in two of three xenograft models. Our findings define the impact of mbIL15 versus mbIL21 during NK expansion and reveal a previously underappreciated tradeoff between NK yield and function for which sequential use of mbIL21-based followed by mbIL15-based feeder cells may be the optimal approach in many settings., (©2023 The Authors; Published by the American Association for Cancer Research.)

Golgi membrane protein 1 (GOLM1) is aberrantly expressed in many types of solid tumors and contributes to cancer development; however, its role in hematopoietic and lymphoid neoplasms remains unknown. Here, we report that GOLM1 was significantly upregulated in anaplastic large cell lymphoma (ALCL), particularly in anaplastic lymphoma kinase-positive (ALK+) ALCL. Mechanistically, the expression of GOLM1 was induced by nucleophosmin-ALK in both ALK-transformed T cells and ALCL cell lines through AKT/mTOR pathway. Knockdown of GOLM1 expression led to a reduction in the growth and viability of ALCL cells with increased spontaneous apoptosis, whereas ectopic expression of GOLM1 protected ALCL cells from apoptosis induced by staurosporine treatment. Moreover, GOLM1 directly interacted with B-cell lymphoma-extra large protein (a crucial anti-apoptosis regulator) and significantly prolonged its stability. Introduction of GOLM1 promoted ALK+ ALCL cells colony formation in vitro and tumor growth in a murine xenograft model. Taken together, our findings demonstrate, to our knowledge, for the first time that GOLM1 plays a critical role in suppressing apoptosis and promoting the progression of ALK+ ALCL and provide evidence that GOLM1 is a potential biomarker and therapeutic target in ALK-induced hematological malignancies., (© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)

IL-17-producing antigen-specific human T cells elicit potent antitumor activity in mice. Yet, refinement of this approach is needed to position it for clinical use. While activation signal strength regulates IL-17 production by CD4 + T cells, the degree to which T cell antigen receptor (TCR) and costimulation signal strength influences Th17 immunity remains unknown. We discovered that decreasing TCR/costimulation signal strength by incremental reduction of αCD3/costimulation beads progressively altered Th17 phenotype. Moreover, Th17 cells stimulated with αCD3/inducible costimulator (ICOS) beads produced more IL-17A, IFNγ, IL-2, and IL-22 than those stimulated with αCD3/CD28 beads. Compared with Th17 cells stimulated with the standard, strong signal strength (three beads per T cell), Th17 cells propagated with 30-fold fewer αCD3/ICOS beads were less reliant on glucose and favored the central carbon pathway for bioenergetics, marked by abundant intracellular phosphoenolpyruvate (PEP). Importantly, Th17 cells stimulated with weak αCD3/ICOS beads and redirected with a chimeric antigen receptor that recognizes mesothelin were more effective at clearing human mesothelioma. Less effective CAR Th17 cells generated with high αCD3/ICOS beads were rescued by overexpressing phosphoenolpyruvate carboxykinase 1 (PCK1), a PEP regulator. Thus, Th17 therapy can be improved by using fewer activation beads during manufacturing, a finding that is cost effective and directly translatable to patients., Competing Interests: Declaration of interests C.M.P. has a patent for the expansion of Tc17 and Th17 cells using the inducible costimulator agonist and inducible costimulator ligand-expressing artificial antigen-presenting cells and beads. C.M.P. is a cofounder of Ares Immunotherapy and has received research funding through a sponsored research agreement between the Medical University of South Carolina and Lycera Corporation, Thermo Fisher Scientific, and Obsidian Therapeutics and through a sponsored research agreement between Emory University and Vaccinex, Inc. G.B.L. has consulted for ProDa Biotech, LLC, and received compensation. G.B.L. has received research funding through a sponsored research agreement between Emory University and Merck and Co., Bristol-Myers Squibb, Boehringer-Ingelheim, and Vaccinex, Inc. J.L.R. is a founder and equity holder in Tmunity Therapeutics, which is now part of Kite, a Gilead Company. J.L.R. received research support from Tmunity Therapeutics., (Copyright © 2023 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)

Deimatic behaviour is performed by prey when attacked by predators as part of an antipredator strategy. The behaviour is part of a sequence that consists of several defences, for example they can be preceded by camouflage and followed by a hidden putatively aposematic signal that is only revealed when the deimatic behaviour is performed. When displaying their hidden signal, mountain katydids (Acripeza reticulata) hold their wings vertically, exposing striking red and black stripes with blue spots and oozing an alkaloid-rich chemical defence derived from its Senecio diet. Understanding differences and interactions between deimatism and aposematism has proven problematic, so in this study we isolated the putative aposematic signal of the mountain katydid's antipredator strategy to measure its survival value in the absence of their deimatic behaviour. We manipulated two aspects of the mountain katydid's signal, colour pattern and whole body shape during display. We deployed five kinds of clay models, one negative control and four katydid-like treatments, in 15 grids across part of the mountain katydid's distribution to test the hypothesis that their hidden signal is aposematic. If this hypothesis holds true, we expected that the models, which most closely resembled real katydids would be attacked the least. Instead, we found that models that most closely resembled real katydids were the most likely to be attacked. We suggest several ideas to explain these results, including that the deimatic phase of the katydid's display, the change from a camouflaged state to exposing its hidden signal, may have important protective value., (© 2022 European Society for Evolutionary Biology.)

Objective: The objective of the study was to investigate functional performance and pain intensity outcomes for associations with negative cognitive orientations, avoidance behaviors, and fear of pain in older Black men with low back pain (LBP)., Methods: Sixty Black men aged 60 and older (70 years[Formula: see text]) with LBP completed the Short Physical Performance Battery (SPPB), the 400-m walk test, and subjective measures of avoidance behaviors, back performance, pain intensity, and pain catastrophizing (i.e., rumination, magnification, and helplessness). Multiple regression models were used to examine associations., Results: Higher helplessness scores were associated with worse back performance (ß = 0.55, p = 0.02), slower walking speed (ß = 0.30, p = 0.02), and higher average pain intensity (ß = 0.22, [Formula: see text] p = 0.03). Higher rumination scores were associated with better back performance (ß =  - 0.36, p = 0.04). Avoidance behaviors, fear of pain, and magnification were not significantly associated with any of the variables included in the tested models., Conclusion: Negative cognitive internalization is associated with limitations in functional performance in older Black men with LBP. Additional research is needed to further examine the cognitive orientations for understanding experienced pain and function in this population. Such research may inform the development of interventions for improving functional performance outcomes of older Black men with LBP., (© 2022. W. Montague Cobb-NMA Health Institute.)

Background: Pain is a dynamic phenomenon dependent on the balance of endogenous excitatory and inhibitory systems, which can be characterized by quantitative sensory testing. Many previous studies of pain modulatory capacity of patients with fibromyalgia syndrome (FM) have reported decreased pain inhibition or increased pain facilitation. This is the first study to assess pain modulation, including conditioned pain modulation (CPM) and temporal pain summation, in the same healthy control (HC) and FM participants., Methods: Only sensitivity-adjusted stimuli were utilized for testing of conditioned pain modulation (CPM) and temporal pain summation in 23 FM patients and 28 HC. All subjects received sensitivity-adjusted ramp-hold (sRH) during testing of pain facilitation (temporal summation) and pain inhibition (CPM). CPM efficacy was evaluated with test stimuli applied either concurrently or after application of the conditioning stimulus. Finally, the effects of CPM on pressure pain thresholds were tested., Results: FM subjects required significantly less intense test and conditioning stimuli than HC participants to achieve standardized pain ratings of 50 ± 10 numerical rating scale (NRS) ( p = 0.03). Using such stimuli, FM subjects' temporal pain summation and CPM efficacy was not significantly different from HC (all p > 0.05), suggesting similar pain facilitation and inhibition. Furthermore, the CPM efficacy of FM and HC participants was similar regardless of whether the test stimuli were applied during or after the conditioning stimulus ( p > 0.05)., Conclusion: Similar to previous studies, FM participants demonstrated hyperalgesia to heat, cold, and mechanical stimuli. However, using only sensitivity-adjusted stimuli during CPM and temporal summation testing, FM patients demonstrated similarly effective pain inhibition and facilitation than HC, suggesting that their pain modulation is not abnormal., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2022.)

Background: Pain-inducing massage produces comparable changes in pain sensitivity as a cold pressor task, suggesting shared neurophysiological mechanisms of conditioned pain modulation. Manual therapy and conditioned pain modulation are influenced by positive and negative expectations. Therefore, the purpose of this study was to examine the effects of positive and negative expectations on pain-free and pain-inducing massage., Methods: 56 healthy participants were randomly assigned to receive a positive or negative expectation instructional set followed by a pain-inducing or a pain-free massage. Pressure pain threshold (PPT) was measured followed by each interval of massage. A repeated measures ANCOVA controlling for post-randomization differences in sex tested for massage x expectation set x PPT interaction effects, as well as two-way interaction effects., Results: A significant three-way interaction effect (p = 0.04) and time x expectation interaction effect was observed for individuals receiving pain inducing massage (p = 0.02). Individuals who received the positive expectation instructional set demonstrated significantly higher PPT at minutes 3 and 4 of massage compared to individuals who received the negative expectation instructional set., Conclusions: Expectations impact pain sensitivity changes produced during massage. Clinicians planning to provide pain-inducing massage should consider the role of expectations in modulating pain sensitivity changes.

Introduction: Individuals with low back pain (LBP) may be classified based on mechanistic descriptors, such as a nociplastic pain presentation (NPP). The purpose of this secondary analysis was to examine the frequency and characteristics of patients with a NPP referred to physical therapy with LBP. Additionally, we characterized patients with LBP meeting the criteria for NPP by demographic, clinical, psychological, and pain sensitivity variables. Finally, we examined short- and long-term clinical outcomes in patients with a NPP compared to those without a NPP., Materials and Methods: Patients referred to physical therapy for LBP completed the Patient Self-report Survey for the Assessment of Fibromyalgia. Participants were categorized as "LBP with NPP" or "LBP without NPP" based on the threshold established in this measure. A rank sum test examined for differences in pain-related psychological factors and pressure-pain threshold between groups. Next, a Friedman test examined if LBP intensity and disability trajectories differed by groups at one and six months after initiation of physical therapy., Results: 22.2% of patients referred to physical therapy for LBP met the criteria for a NPP. Patients with a NPP reported significantly greater disability, pain catastrophizing, depression, anxiety, and somatization compared to individuals without a NPP ( p < 0.05). Pressure-pain threshold did not differ between groups ( p > 0.05). Individuals with LBP with a NPP demonstrated nonsignificant, small to medium reductions in pain and disability at one and six months. Individuals experiencing LBP without a NPP demonstrated significant reductions in pain and disability in the short- and long term., Conclusion: Patients with LBP with a NPP displayed greater negative pain-related psychological factors but similar pain sensitivity compared to LBP without NPP., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2023 Abigail T. Wilson et al.)

Phenotypes have been proposed as a method of characterizing subgroups based on biopsychosocial factors to identify responders to analgesic treatments. This study aimed to, first, confirm phenotypes in patients with low back pain receiving physical therapy based on an a priori set of factors used to derive subgroups in other pain populations. Second, an exploratory analysis examined if phenotypes differentiated pain and disability outcomes at four weeks of physical therapy. Fifty-five participants completed psychological questionnaires and pressure pain threshold (PPT). Somatization, anxiety, and depression domains of the Symptom-Checklist-90-Revised, and PPT, were entered into a hierarchical agglomerative cluster analysis with Ward's method to identify phenotypes. Repeated measures ANOVAs assessed pain ratings and disability by phenotype at four weeks. Three clusters emerged: 1) high emotional distress and pain sensitivity (n = 10), 2) low emotional distress (n = 34), 3) low pain sensitivity (n = 11). As an exploratory study, clusters did not differentiate pain ratings or disability after four weeks of physical therapy (p's>0.05). However, trends were observed as magnitude of change for pain varied by phenotype. This supports the characterization of homogenous subgroups based on a protocol conducted in the clinical setting with varying effect sizes noted by phenotype for short-term changes in pain. As an exploratory study, future studies should aim to repeat this trial in a larger sample of patients., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Wilson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Better understanding of dentists' decision-making about defective restorations is needed to close the evidence-practice gap (EPG). Aim: this study aimed to quantify the EPG about defective restorations and identify dentist factors associated with this EPG. Methods: 216 dentists from São Paulo State, Brazil, completed a questionnaire about three clinical case scenarios involving defective composite restorations with cementum-dentin margins (case 1) and enamel margins (case 2), and an amalgam (case 3) restoration. Dentists were asked what treatment, if any, they would recommend, including preventive treatment, polishing, re-surfacing, or repairing the restoration, or replacing the entire restoration. Replacing the entire restoration in any of these three scenarios was classified as inconsistent with the evidence, comprising an EPG. Bivariate analyses using Chi-square, ANOVA, or multiple comparison tests were performed (p<.05). Results: for defective composite restorations, 49% and 55% of dentists chose to replace the entire restoration for cases 1 and 2, respectively. Twenty-nine percent of dentists chose to replace the entire amalgam restoration. Dentists were significantly more likely to choose to replace the defective amalgam restoration than the composite restoration with a defect at the cementum-dentin margins or the enamel margins (both at p < .001). Female dentists were more likely to choose a conservative treatment than male dentists for cases 1 (p=.034) and 2 (p=.009). Dentists with a higher percentage of patients interested in individualized caries prevention were also more conservative in case 1 (p=.045). Conclusion: a substantial EPG regarding treatment decisions for defective restorations exists, especially for composite restorations. This study adds to the international evidence that an EPG exists in this clinical area and that global strategies need to be developed to close the gap.

It is technically challenging to generate large doses of regulatory T cells (Tregs) engineered to express a chimeric antigen receptor (CAR) in non-human primates (NHP). Here, we have optimized the manufacturing of CAR Tregs by stringent sorting of Tregs, stimulation by artificial antigen-presenting cells, transduction by simian tropic lentiviral vectors, and antigen-specific expansion. The result of this method is highly suppressive CAR Tregs for use in a pre-clinical, large animal model of transplant tolerance. For complete details on the use and execution of this protocol, please refer to Ellis et al. (2022)., Competing Interests: J.L.R. and G.I.E. have submitted patents related to use of the Bw6 CAR and the aAPCs described in this manuscript. J.L.R. is a founder and equity holder of Tmunity Therapeutics., (© 2022 The Author(s).)

A prominent feature of the modern era is the increasing spread of invasive species, particularly within island and urban ecosystems, and these occurrences provide valuable natural experiments by which evolutionary and invasion hypotheses can be tested. In this study, we used the invasion route of guttural toads ( Sclerophrys gutturalis ) from natural-native and urban-native populations (Durban, South Africa) to their urban-invasive and natural-invasive populations (Mauritius and Réunion) to determine whether phenotypic changes that arose once the toads became urbanized in their native range have increased their invasive potential before they were transported (i.e., prior adaptation) or whether the observed changes are unique to the invasive populations. This urban/natural by native/invasive gradient allowed us to examine differences in guttural toad morphology (i.e., body size, hindlimb, and hindfoot length) and performance capacity (i.e., escape speed, endurance, and climbing ability) along their invasion route. Our findings indicate that invasive island populations have reduced body sizes, shorter limbs in relation to snout-vent length, decreased escape speeds, and decreased endurance capacities that are distinct from the native mainland populations (i.e., invasion-derived change). Thus, these characteristics did not likely arise directly from a pre-transport anthropogenic "filter" (i.e., urban-derived change). Climbing ability, however, did appear to originate within the urban-native range and was maintained within the invasive populations, thereby suggesting it may have been a prior adaptation that provided this species with an advantage during its establishment in urban areas and spread into natural forests. We discuss how this shift in climbing performance may be ecologically related to the success of urban and invasive guttural toad populations, as well as how it may have impacted other island-derived morphological and performance phenotypes.

HIV-1-specific CD4 + T cells (T CD4+ s) play a critical role in controlling HIV-1 infection. Canonically, T CD4+ s are activated by peptides derived from extracellular ("exogenous") Ags displayed in complex with MHC class II (MHC II) molecules on the surfaces of "professional" APCs such as dendritic cells (DCs). In contrast, activated human T CD4+ s, which express MHC II, are not typically considered for their APC potential because of their low endocytic capacity and the exogenous Ag systems historically used for assessment. Using primary T CD4+ s and monocyte-derived DCs from healthy donors, we show that activated human T CD4+ s are highly effective at MHC II-restricted presentation of an immunodominant HIV-1-derived epitope postinfection and subsequent noncanonical processing and presentation of endogenously produced Ag. Our results indicate that, in addition to marshalling HIV-1-specific immune responses during infection, T CD4+ s also act as APCs, leading to the activation of HIV-1-specific T CD4+ s., (Copyright © 2022 by The American Association of Immunologists, Inc.)

The underlying drivers of variation in the colouration (colour and pattern) of animals can be genetic, non-genetic, or more likely, a combination of both. Understanding the role of heritable genetic elements, as well as non-genetic factors such as age, habitat or temperature, in shaping colouration can provide insight into the evolution and function of these traits, as well as the speed of response to changing environments. This project examined the genetic and non-genetic drivers of continuous variation in colouration in a lizard, the jacky dragon (Amphibolurus muricatus). We leveraged a large captive experiment that manipulated parental and offspring thermal environment to simultaneously estimate the genetic and non-genetic drivers of variation in colouration. We found that the overall brightness, the elongation of the longitudinal stripes on the dorsum and the contrast between light and dark patches of the pattern were all heritable. Colouration varied according to the age of the hatchling; however, the thermal environment of neither the parents nor offspring contributed significantly to colouration. It appears that developmental plasticity and maternal effects associated with temperature are not important drivers of variation in our measures of colouration., (© 2022 The Authors. Journal of Evolutionary Biology published by John Wiley & Sons Ltd on behalf of European Society for Evolutionary Biology.)

T cell receptor (TCR)-based immunotherapy has emerged as a promising therapeutic approach for the treatment of patients with solid cancers. Identifying peptide-human leukocyte antigen (pHLA) complexes highly presented on tumors and rarely expressed on healthy tissue in combination with high-affinity TCRs that when introduced into T cells can redirect T cells to eliminate tumor but not healthy tissue is a key requirement for safe and efficacious TCR-based therapies. To discover promising shared tumor antigens that could be targeted via TCR-based adoptive T cell therapy, we employed population-scale immunopeptidomics using quantitative mass spectrometry across ~1500 tumor and normal tissue samples. We identified an HLA-A*02:01-restricted pan-cancer epitope within the collagen type VI α-3 ( COL6A3 ) gene that is highly presented on tumor stroma across multiple solid cancers due to a tumor-specific alternative splicing event that rarely occurs outside the tumor microenvironment. T cells expressing natural COL6A3-specific TCRs demonstrated only modest activity against cells presenting high copy numbers of COL6A3 pHLAs. One of these TCRs was affinity-enhanced, enabling transduced T cells to specifically eliminate tumors in vivo that expressed similar copy numbers of pHLAs as primary tumor specimens. The enhanced TCR variants exhibited a favorable safety profile with no detectable off-target reactivity, paving the way to initiate clinical trials using COL6A3-specific TCRs to target an array of solid tumors.

Early career researchers (ECRs) are important stakeholders leading efforts to catalyze systemic change in research culture and practice. Here, we summarize the outputs from a virtual unconventional conference (unconference), which brought together 54 invited experts from 20 countries with extensive experience in ECR initiatives designed to improve the culture and practice of science. Together, we drafted 2 sets of recommendations for (1) ECRs directly involved in initiatives or activities to change research culture and practice; and (2) stakeholders who wish to support ECRs in these efforts. Importantly, these points apply to ECRs working to promote change on a systemic level, not only those improving aspects of their own work. In both sets of recommendations, we underline the importance of incentivizing and providing time and resources for systems-level science improvement activities, including ECRs in organizational decision-making processes, and working to dismantle structural barriers to participation for marginalized groups. We further highlight obstacles that ECRs face when working to promote reform, as well as proposed solutions and examples of current best practices. The abstract and recommendations for stakeholders are available in Dutch, German, Greek (abstract only), Italian, Japanese, Polish, Portuguese, Spanish, and Serbian., Competing Interests: The authors have declared that no competing interests exist.

Adoptive transfer of chimeric antigen receptor regulatory T cells (CAR Tregs) is a promising way to prevent allograft loss without the morbidity associated with current therapies. Non-human primates (NHPs) are a clinically relevant model to develop transplant regimens, but manufacturing and engraftment of NHP CAR Tregs have not been demonstrated yet. Here, we describe a culture system that massively expands CAR Tregs specific for the Bw6 alloantigen. In vitro, these Tregs suppress in an antigen-specific manner without pro-inflammatory cytokine secretion or cytotoxicity. In vivo, Bw6-specific CAR Tregs preferentially traffic to and persist in bone marrow for at least 1 month. Following transplant of allogeneic Bw6 + islets and autologous CAR Tregs into the bone marrow of diabetic recipients, CAR Tregs traffic to the site of islet transplantation and maintain a phenotype of suppressive Tregs. Our results establish a framework for the optimization of CAR Treg therapy in NHP disease models., Competing Interests: Declaration of interests J.L.R. and G.I.E. have submitted patents related to use of the HLA-A2 and Bw6 CARs described in this manuscript. J.L.R. is an equity holder of and receives sponsored research funding from Tmunity Therapeutics. The remaining authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

During the vulnerable stages of early life, most ectothermic animals experience hourly and diel fluctuations in temperature as air temperatures change. While we know a great deal about how different constant temperatures impact the phenotypes of developing ectotherms, we know remarkably little about the impacts of temperature fluctuations on the development of ectotherms. In this study, we used a meta-analytic approach to compare the mean and variance of phenotypic outcomes from constant and fluctuating incubation temperatures across reptile species. We found that fluctuating temperatures provided a small benefit (higher hatching success and shorter incubation durations) at cool mean temperatures compared with constant temperatures, but had a negative effect at warm mean temperatures. In addition, more extreme temperature fluctuations led to greater reductions in embryonic survival compared with moderate temperature fluctuations. Within the limited data available from species with temperature-dependent sex determination, embryos had a higher chance of developing as female when developing in fluctuating temperatures compared with those developing in constant temperatures. With our meta-analytic approach, we identified average mean nest temperatures across all taxa where reptiles switch from receiving benefits to incurring costs when incubation temperatures fluctuate. More broadly, our study indicates that the impact of fluctuating developmental temperature on some phenotypes in ectothermic taxa are likely to be predictable via integration of developmental temperature profiles with thermal performance curves., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Background: Patients often seek consultation with dentists for temporomandibular disorders (TMDs). The objectives of this article were to describe the methods of a large prospective cohort study of painful TMD management, practitioners' and patients' characteristics, and practitioners' initial treatment recommendations conducted by The National Dental Practice-Based Research Network (the "network")., Methods: Participating dentists recruited into this study treated patients seeking treatment for painful TMDs. The authors developed self-report instruments based on well-accepted instruments. The authors collected demographics, biopsychosocial characteristics, TMD symptoms, diagnoses, treatments, treatment adherence, and painful TMDs and jaw function outcomes through 6 months., Results: Participating dentists were predominately White (76.8%) and male (62.2%), had a mean age of 52 years, and were general practitioners (73.5%) with 23.8% having completed an orofacial pain residency. Of the 1,901 patients with painful TMDs recruited, the predominant demographics were White (84.3%) and female (83.3%). Patients' mean age was 44 years, 88.8% self-reported good to excellent health, and 85.9% had education beyond high school. Eighty-two percent had pain or stiffness of the jaw on awakening, and 40.3% had low-intensity pain. The most frequent diagnoses were myalgia (72.4%) and headache attributed to TMDs (51.0%). Self-care instruction (89.4%), intraoral appliances (75.4%), and medications (57.6%) were recommended frequently., Conclusions: The characteristics of this TMD cohort include those typical of US patients with painful TMDs. Network practitioners typically managed TMDs using conservative treatments., Practical Implications: This study provides credible data regarding painful TMDs and TMD management provided by network practitioners across the United States. Knowledge acquired of treatment recommendations and patient reports may support future research and improve dental school curricula., (Copyright © 2022 American Dental Association. Published by Elsevier Inc. All rights reserved.)