Antibody surface mobility amplifies FcγR signaling via Arp2/3 during phagocytosis.

We report herein that the anti-CD20 therapeutic antibody, rituximab, is rearranged into microclusters within the phagocytic synapse by macrophage Fcγ receptors (FcγR) during antibody-dependent cellular phagocytosis. These microclusters were observed to potently recruit Syk and to undergo rearrangements that were limited by the cytoskeleton of the target cell, with depolymerization of target-cell actin filaments leading to modest increases in phagocytic efficiency. Total internal reflection fluorescence analysis revealed that FcγR total phosphorylation, Syk phosphorylation, and Syk recruitment were enhanced when IgG-FcγR microclustering was enabled on fluid bilayers relative to immobile bilayers in a process that required Arp2/3. We conclude that on fluid surfaces, IgG-FcγR microclustering promotes signaling through Syk that is amplified by Arp2/3-driven actin rearrangements. Thus, the surface mobility of antigens bound by IgG shapes the signaling of FcγR with an unrecognized complexity beyond the zipper and trigger models of phagocytosis.

Online Nucleotide Mapping of mRNAs.

Messenger RNA (mRNA) can be sequenced via indirect approaches such as Sanger sequencing and next generation sequencing (NGS), or direct approaches like bottom-up mass spectrometry (MS). Direct sequencing allows the confirmation of RNA modifications. However, the conventional bottom-up MS approach involves time-consuming in-solution digestions that require a large amount of sample, and can lead to the RNase contamination of the LC-MS system and column. Here, we describe a platform that enables online nucleotide mapping of mRNAs via the use of immobilized RNase cartridges and 2D-LC-MS instrumentation. The online approach was compared to conventional offline digestion protocols adapted from two published studies. For this purpose, five model mRNAs of varying lengths (996-4521 nucleotides) and chemistries (unmodified uridine vs 5-methoxyuridine (5moU)) were analyzed. The profiles and sequence coverages obtained after RNase T1 digestion were discussed. The online nucleotide mapping achieved comparable or slightly greater sequence coverage for the 5 mRNAs (5.8-51.5%) in comparison to offline approaches (3.7-50.4%). The sequence coverage was increased to 65.6-85.6 and 69.7-85.0% when accounting for the presence of nonunique digestion products generated by the RNase T1 and A, respectively. The online nucleotide mapping significantly reduced the digestion time (from 15 to <5 min), increased the signal intensity by more than 10-fold in comparison to offline approaches.

Development and adaptation of the Emotion Regulation Skills Questionnaire for adolescents.

INTRODUCTION: The ability to effectively regulate one's emotions is important for adolescent mental health. However, extant assessments of how adolescents regulate their emotions can be expanded upon in several ways, including incorporating more strategies (e.g., cultural and spiritual) and positive emotions, and being informed by adolescents and expert consultation during the development process. Thus, our study aimed to improve the construct validity of an emotion regulation measure by adapting and refining the Emotion Regulation Skills Questionnaire (ERSQ) into an adolescent self-report. METHODS: We recruited 24 13-17-year-olds (12 males; 11 females; 1 gender fluid) via social media advertisement in the United States in 2020. We used an iterative qualitative approach in which we combined expert consultation, cognitive interviewing with adolescents, and top-down and bottom-up coding to review and provide feedback on the ERSQ. RESULTS: Findings of thematic analysis showed that a need for clarity in wording and inclusion of strategy examples across all sections of the ERSQ was needed. Adolescents also identified the need to replace the Silly/Excited section with a more age-appropriate positive emotion section. CONCLUSION: Findings indicate cognitive interviewing can be successfully completed in a virtual format when necessary, youth utilize a wider range of strategies to maintain or upregulate positive emotions than what is represented in current measures, and use of qualitative methods may have reduced construct underrepresentation and construct-irrelevant variance in the adapted ERSQ.

Pressurizing or encouraging: Health behaviors among long-distance couples.

Couples in long-distance relationships face unique challenges that affect their health outcomes and relationship dynamics in ways that are different from couples in close proximal relationships (PR). The results of previous literature analyzing health outcomes for long-distance relationship (LDR) couples have been mixed, and factors such as couple satisfaction and gender of the individuals contribute to the variance. This study examined the good health practices of couples in LDRs, the ways in which partners influence each other's health, and the health outcomes of these couples as compared to couples in PRs. Multilevel multivariate analysis showed that couples in LDRs had better health practices than those in PRs, and men overall had poorer health practices than women. There was no main effect seen for LDRs when we examined strategies used for influencing health. There was a main effect for gender, though, and we found that women tend to use more collaborative and pressurizing strategies for influencing their partner's health. On health indices, we found that LDR couples tended to have lower levels of fatigue and sleep disturbance as compared to PR couples. In the adjusted model with covariates, which included age, income, couple satisfaction, and married or unmarried couples, we found that lower couple satisfaction and lower income significantly predicted poorer health on all health indices. These results are discussed in light of unequal gender roles, social control, and positive aspects of LDRs on health in an attempt to understand LDRs better and destigmatize the narrative of these relationships as 'less than' PRs. Implications for health and mental health practitioners are also discussed.

Pediatric Chest Compression Improvement Via Augmented Reality Cardiopulmonary Resuscitation Feedback in Community General Emergency Departments: A Mixed-Methods Simulation-Based Pilot Study.

BACKGROUND: Yearly, more than 20,000 children experience a cardiac arrest. High-quality pediatric cardiopulmonary resuscitation (CPR) is generally challenging for community hospital teams, where pediatric cardiac arrest is infrequent. Current feedback systems are insufficient. Therefore, we developed an augmented reality (AR) CPR feedback system for use in many settings. OBJECTIVE: We aimed to evaluate whether AR-CPR improves chest compression (CC) performance in non-pediatric-specialized community emergency departments (EDs). METHODS: We performed an unblinded, randomized, crossover simulation-based study. A convenience sample of community ED nonpediatric nurses and technicians were included. Each participant performed three 2-min cycles of CC during a simulated pediatric cardiac arrest. Participants were randomized to use AR-CPR in one of three CC cycles. Afterward, participants participated in a qualitative interview to inquire about their experience with AR-CPR. RESULTS: Of 36 participants, 18 were randomized to AR-CPR in cycle 2 (group A) and 18 were randomized to AR-CPR in cycle 3 (group B). When using AR-CPR, 87-90% (SD 12-13%) of all CCs were in goal range, analyzed as 1-min intervals, compared with 18-21% (SD 30-33%) without feedback (p < 0.001). Analysis of qualitative themes revealed that AR-CPR may be usable without a device orientation, be effective at cognitive offloading, and reduce anxiety around and enhance confidence in the CC delivered. CONCLUSIONS: The novel CPR feedback system, AR-CPR, significantly changed the CC performance in community hospital non-pediatric-specialized general EDs from 18-21% to 87-90% of CC epochs at goal. This study offers preliminary evidence suggesting AR-CPR improves CC quality in community hospital settings.

On-line Sequencing of CRISPR Guide RNAs and Their Impurities via the Use of Immobilized Ribonuclease Cartridges Attached to a 2D/3D-LC-MS System.

In this study, for the first time, the automated digestion and sequencing of an RNA molecule via the use of immobilized RNase cartridges attached to a multidimensional liquid chromatography (LC)-mass spectrometry (MS) system are presented. We first developed an on-line digestion-HILIC two-dimensional (2D)-LC-MS method in order to sequence CRISPR guide RNAs for gene editing. Three RNases (T1, A, and U2) were immobilized on polyetheretherketone cartridges, and their performance was evaluated. Ultrafast digestions were performed within 2.3 min with the on-line approach versus 30 min via the conventional off-line approach. The higher sequence coverage was achieved by the RNase T1 (71%), which is the same as the off-line mode. A 20-fold reduction in the gRNA sample amount was achieved with the on-line digestion approach (6.5 µg) in comparison to that with the off-line approach (130 µg). In the second step, a three-dimensional (3D)-LC-MS method was developed for the sequencing of fractions collected on-line across the main peak and the partially separated tail by the reference ion-pairing RPLC method. Additional insights were gained in order to better understand the cause of the main peak tailing.

Capturing what matters: A retrospective observational study of advance care planning documentation at an academic medical center during the COVID-19 pandemic.

BACKGROUND: Advance care planning allows patients to share their preferences for medical care with the aim of ensuring goal-concordant care in times of serious illness. The morbidity and mortality of the COVID-19 pandemic has increased the importance and public visibility of advance care planning. However, little is known about the frequency and quality of advance care planning documentation during the pandemic. AIM: This study examined the frequency, quality, and predictors of advance care planning documentation among hospitalized medical patients with and without COVID-19. DESIGN: This retrospective cohort analysis used multivariate logistic regression to identify factors associated with advance care planning documentation. SETTING/PARTICIPANTS: This study included all adult patients tested for COVID-19 and admitted to a tertiary medical center in San Francisco, CA during March 2020. RESULTS: Among 262 patients, 31 (11.8%) tested positive and 231 (88.2%) tested negative for SARS-CoV-2. The rate of advance care planning documentation was 38.7% in patients with COVID-19 and 46.8% in patients without COVID-19 (p = 0.45). Documentation consistently addressed code status (100% and 94.4% for COVID-positive and COVID-negative, respectively), but less often named a surrogate decision maker, discussed prognosis, or elaborated on other wishes for care. Palliative care consultation was associated with increased advance care planning documentation (OR: 6.93, p = 0.004). CONCLUSION: This study found low rates of advance care planning documentation for patients both with and without COVID-19 during an evolving global pandemic. Advance care planning documentation was associated with palliative care consultation, highlighting the importance of such consultation to ensure timely, patient-centered advance care planning.

Anxiety and Depression in Northern Plains American Indian Youth: Evidence for Resilience and Risk.

Developing research collaborations with Indigenous communities to understand the expression and experience of anxiety and depression in American Indian (AI) youth and identifying protective and risk factors may be an important first step toward addressing AI health inequities. We used a community-based participatory research (CBPR) approach to investigate anxiety and depressive disorder symptoms among AI youth living on a Northern Plains reservation. Moreover, we examined whether symptoms were related to two potential protective and risk factors, anxiety control beliefs and rumination. Our tribal research team collected multi-reporter survey data from 71 AI 3rd-6th graders (8-13-years-old; 62.3% female) attending a tribal school, their caregivers, and teachers. Results pointed toward resilience in this sample with 7.3% and 8.7% of AI youth reporting clinical levels of anxiety and depressive disorder symptoms, respectively, and on average experiencing symptoms "Sometimes." There were moderate correlations between youth- and teacher-reported anxiety and depressive disorder symptoms, but no correlation with caregivers. Anxiety control beliefs were lower in older compared to younger AI youth and negatively related to youth-reported anxiety and depressive disorder symptoms, while rumination was positively related to youth-reported anxiety and depressive disorder symptoms and teacher-reported anxiety disorder symptoms. Age moderated relations between anxiety control beliefs and both youth-reported anxiety and depressive disorder symptoms with only significant relations found for older youth. Our findings are consistent with research showing resilience to internalizing problems in AI youth living on a reservation, but replication of their relations to anxiety control beliefs and rumination in other Indigenous peoples is warranted.

Full Sequencing of CRISPR/Cas9 Single Guide RNA (sgRNA) via Parallel Ribonuclease Digestions and Hydrophilic Interaction Liquid Chromatography-High-Resolution Mass Spectrometry Analysis.

CRISPR/Cas9 is a powerful genome editing approach in which a Cas9 enzyme and a single guide RNA (sgRNA) form a ribonucleoprotein complex effectively targeting site-specific cleavages of DNA. Accurate sequencing of sgRNA is critical to patient safety and is the expectation by regulatory agencies. In this paper, we present the full sequencing of sgRNA via parallel ribonuclease (RNase) T1, A, and U2 digestions and the simultaneous separation and identification of the digestion products by hydrophilic interaction liquid chromatography (HILIC) coupled to high-resolution mass spectrometry (HRMS). When using RNase T1 digestion alone, a maximal sequence coverage of 81% was obtained excluding the nonunique fragments. Full sgRNA sequencing was achieved using unique fragments generated by RNase T1, A, and U2 parallel digestions. Thorough optimization of sgRNA digestions was performed by varying the nuclease-to-sgRNA ratio, buffer conditions, and reaction times. A biocompatible ethylene-bridged hybrid amide column was evaluated for the separation of RNase digestion products. To our knowledge, it is the first time that (i) RNA digests are separated and identified by HILIC-HRMS and (ii) chemically modified sgRNAs are directly sequenced via a bottom-up approach.

Depressive symptoms among stress-exposed youth: Relations with tonic and phasic indices of autonomic functioning.

Theoretical models of adolescent depression postulate that one possible individual vulnerability factor for the development of depressive symptoms is autonomic dysregulation. However, there is limited and mixed support for these models among ethnically diverse and higher risk stress-exposed youth. Therefore, this study investigated the relations between both tonic and phasic indices of parasympathetic autonomic functioning (i.e., resting high-frequency heart rate variability [HF-HRV] and root mean square of successive differences [RMSSD]; HF-HRV and RMSSD reactivity to a mental arithmetic stressor) and depressive symptoms among 80 severely stress-exposed youth (51% female; 11-17 years of age) from diverse backgrounds (61.3% ethnic minority; caregiver-reported median family income = $20,000-$49,999 per year). Results demonstrated that lower resting HF-HRV and RMSSD, but not HF-HRV and RMSSD reactivity, was associated with greater youth depressive symptoms. Our findings suggest that lower resting parasympathetic autonomic functioning may be a potential vulnerability factor of depressive symptoms among stress-exposed youth, instead of specific emotional responses to stressors. These findings will need to be replicated in larger samples of stress-exposed youth and youth at higher risk for or exhibiting clinical levels of depressive symptoms to better elucidate relations with autonomic functioning and depressive symptoms among adolescents.

Effects of Therapeutic Intervention on Parentally Bereaved Children's Emotion Reactivity and Regulation 15 Years Later.

The Family Bereavement Program (FBP) is a family-based intervention for parentally bereaved children and surviving caregivers. Results are reported of a randomized controlled trial, examining intervention effects on emotional reactivity and regulation of young adults who participated in the program 15 years earlier. Participants (N = 152) completed four emotion challenge tasks: reactivity to negative images, detached reappraisal while viewing negative images, positive reappraisal while viewing negative images, and reengagement with positive images. Outcomes included cardiac interbeat interval (IBI), pre-ejection period (PEP), and respiratory sinus arrhythmia (RSA) as well as self-reported emotional experience and regulation effectiveness. Direct intervention effects and effects mediated through improved parenting were estimated. Several significant effects were observed in primary analyses; however, none remained significant after correction for familywise Type I error. Parenting mediated FBP effects on IBI during negative reactivity (b = 15.04), and on RSA during positive reengagement (b = 0.35); the latter effect was accounted for by changes in breathing. Intervention condition was a direct predictor of self-reported detached reappraisal effectiveness (b = 1.00). Intervention and gender interacted in predicting self-reported negative emotion during the negative reactivity (b = 1.04) and positive reappraisal tasks (b = 1.31) such that intervention-condition men reported more negative emotions during those tasks. Although these findings should be considered preliminary given the limited power of the corrected statistical tests, they suggest long-term effects of family intervention following the death of a parent on offspring's emotional reactivity and regulation ability that should be pursued further in future research.

Heart rate variability and anxious arousal: Unique relations with sleep-related problems in stress-exposed adolescents.

Sleep-related problems (SRPs) among adolescents are a growing concern. Theory and research suggest that emotional arousal may have cyclical relation with SRPs, but whether emotional dysregulation plays a role is not clear. We investigated associations between two physiological indices of emotion regulation (video baseline heart rate variability and change in heart rate variability to a stressor) and SRPs in a sample of 80 adolescents (ages 11-17 years; 51% female; 37.5% African American). The findings showed a negative relation between video baseline heart rate variability and SRPs, controlling for non-sleep-related anxiety disorder symptoms (ß = -0.29) and general manifest anxiety (ß = -0.25). We found no relation between change in heart rate variability to a stressor and SRPs when non-sleep-related anxious arousal was controlled. If replicated, findings illustrate the importance of physiological regulation of emotion influencing (or influenced by) SRPs during adolescence.

Ciliated columnar epithelium in the esophagus and gastroesophageal junction: A different perspective from study of a North American population.

An index case of ciliated columnar epithelium in a gastroesophageal (GE) junction biopsy identified in routine surgical pathology practice struck us as highly unusual. However, pathology literature, mainly from Asian populations, reports ciliated columnar epithelium in up to 40% of tissue samples from the upper GI tract. This was inconsistent with our pathology practice experience, so we initiated a local review of cases at our Canadian centre. 1048 consecutive tissue samples from the esophagus and GE junction were reviewed retrospectively and no ciliated epithelium was identified. This review included 1000× oil immersion microscopy of 22 cases with "multilayered epithelium". In 971 cases verified in prospective surgical pathology practice following identification of the index case, 3 additional cases of ciliated columnar epithelium were identified. The index case had ciliated pseudostratified columnar epithelium, resembling respiratory epithelium, and had strong, diffuse expression of TTF-1 by immunohistochemistry. In the other 3 cases, the cilia were located on the surface of a pseudostratified columnar epithelium, a multilayered epithelium, or a low columnar epithelium, all TTF-1 negative. Over a year later, the index case proved to have arisen from a bronchial-esophageal fistula. The other cases were not associated with a fistula. Our conclusion is that ciliated columnar epithelium is rare in Canadian adults (<0.5% of patients). Ciliated epithelium due to a bronchial-esophageal fistula is exceptional, but something to consider if there is a suspicious clinical picture and TTF-1 expression. Other cases might represent a rare metaplastic phenomenon or remnant from fetal development.

Parenting Behaviors, Parent Heart Rate Variability, and Their Associations with Adolescent Heart Rate Variability.

Adolescence is a potentially important time in the development of emotion regulation and parenting behaviors may play a role. We examined associations among parenting behaviors, parent resting heart rate variability, adolescent resting heart rate variability and parenting behaviors as moderators of the association between parent and adolescent resting heart rate variability. Ninety-seven youth (11-17 years; 49.5 % female; 34 % African American, 37.1 % Euro-American, 22.6 % other/mixed ethnic background, and 7.2 % Hispanic) and their parents (n = 81) completed a physiological assessment and questionnaires assessing parenting behaviors. Inconsistent discipline and corporal punishment were negatively associated with adolescent resting heart rate variability, while positive parenting and parental involvement were positively associated. Inconsistent discipline and parental involvement moderated the relationship between parent and adolescent resting heart rate variability. The findings provide evidence for a role of parenting behaviors in shaping the development of adolescent resting heart rate variability with inconsistent discipline and parental involvement potentially influencing the entrainment of resting heart rate variability in parents and their children.

TIRF imaging of Fc gamma receptor microclusters dynamics and signaling on macrophages during frustrated phagocytosis.

BACKGROUND: Recent evidence indicates that in addition to the T-cell receptor, microclustering is an important mechanism for the activation of the B-cell receptor and the mast cell Fcε-receptor. In macrophages and neutrophils, particles opsonized with immunoglobulin G (IgG) antibodies activate the phagocytic Fcγ-receptor (FcγR) leading to rearrangements of the actin cytoskeleton. The purpose of this study was to establish a system for high-resolution imaging of FcγR microclustering dynamics and the recruitment of the downstream signaling machinery to these microclusters. METHODS: We developed a supported lipid bilayer platform with incorporated antibodies on its surface to study the formation and maturation of FcγR signaling complexes in macrophages. Time-lapse multicolor total internal reflection microscopy was used to capture the formation of FcγR-IgG microclusters and their assembly into signaling complexes on the plasma membrane of murine bone marrow derived macrophages. RESULTS: Upon antibody binding, macrophages formed FcγR-IgG complexes at the leading edge of advancing pseudopods. These complexes then moved toward the center of the cell to form a structure reminiscent of the supramolecular complex observed in the T-cell/antigen presenting cell immune synapse. Colocalization of signaling protein Syk with nascent clusters of antibodies indicated that phosphorylated receptor complexes underwent maturation as they trafficked toward the center of the cell. Additionally, imaging of fluorescent BtkPH domains indicated that 3'-phosphoinositides propagated laterally away from the FcγR microclusters. CONCLUSION: We demonstrate that surface-associated but mobile IgG induces the formation of FcγR microclusters at the pseudopod leading edge. These clusters recruit Syk and drive the production of diffusing PI(3,4,5)P3 that is coordinated with lamellar actin polymerization. Upon reaching maximal extension, FcγR microclusters depart from the leading edge and are transported to the center of the cellular contact region to form a synapse-like structure, analogous to the process observed for T-cell receptors.

A Twin Factor Mixture Modeling Approach to Childhood Temperament: Differential Heritability.

Twin factor mixture modeling was used to identify temperament profiles while simultaneously estimating a latent factor model for each profile with a sample of 787 twin pairs (Mage  = 7.4 years, SD = .84; 49% female; 88.3% Caucasian), using mother- and father-reported temperament. A four-profile, one-factor model fit the data well. Profiles included "regulated, typical reactive," "well-regulated, positive reactive," "regulated, surgent," and "dysregulated, negative reactive." All profiles were heritable, with lower heritability and shared environment also contributing to membership in the "regulated, typical reactive" and "dysregulated, negative reactive" profiles.

Existential Anxiety Among Adolescents Exposed to Disaster: Linkages Among Level of Exposure, PTSD, and Depression Symptoms.

Exposure to natural disasters can be highly traumatic and have a detrimental effect on youth mental health by threatening the satisfaction of basic human needs and goals. Recent research in adults suggests that exposure to disasters may exacerbate existential anxiety about the meaning of life. The current study expands this investigation to adolescents, who may be particularly vulnerable to the negative effects of disaster. Data came from 325 adolescents (mean age = 15.05 years, SD = 1.05) residing in the Greater New Orleans area who were exposed to Hurricanes Katrina and/or Gustav. Existential anxiety concerns were highly prevalent in the sample and were associated with elevated levels of posttraumatic stress disorder (R2 = .09) and depression symptoms (R2 = .13). Consistent with theoretical predictions, disaster exposure levels moderated the association between facets of existential anxiety and mental health symptoms. Findings highlight the salience of existential concerns in disaster exposed youth, and provide evidence that exposure to traumatic stress may strengthen their association with mental health problems.

Fitting anxious emotion-focused intervention into the ecology of schools: results from a test anxiety program evaluation.

Emotion-focused prevention and intervention efforts in schools have been promoted as a significant developmental and public health priority. This paper reports the results of a longitudinal study testing central premises of a school-based prevention model aimed at promoting positive emotional development through targeting test anxiety. Test anxiety interventions may be a practical strategy for conducting emotion-focused prevention and intervention efforts because of a natural fit within the ecology of the school setting. At-risk youth (n = 1,048) from urban public schools were screened and 325 with elevated test anxiety were offered the intervention in one of two waves (immediate intervention vs. waitlist). The intervention was associated with decreases in test anxiety, anxiety disorder, and depression symptoms. Critically, results suggest high participant satisfaction and growth curve analysis of follow-up assessments (end of the year, the next school year, and a subsequent school year) demonstrated positive developmental trajectories consistent with predictions (e.g., initial change in test anxiety predicted change in other symptoms). Findings provide evidence for the ecological validity of targeting test anxiety in school-based, emotion-focused prevention efforts.

Aggressive behavior and its associations with posttraumatic stress and academic achievement following a natural disaster.

Despite an abundance of evidence linking maltreatment and violence-related trauma exposure to externalizing problems in youth, there is surprisingly little evidence to support a direct link between disaster exposure and youth aggressive behavior. This study tested the theory that there is primarily an indirect association between disaster exposure and aggression via posttraumatic stress disorder (PTSD) symptoms. The current study also examined the association between aggression and academic achievement. A sample of 191 4th- to 8th-grade minority youth who experienced Hurricane Katrina were assessed for aggressive behavior using the Peer Conflict Scale (PCS), disaster exposure, PTSD symptoms, and academic achievement. Structural equation modeling of the set of associations was consistent with the theory suggesting that there is an indirect link between disaster exposure and aggression through PTSD symptoms. Aggression was negatively associated with academic achievement, and modeling indicated that the set of associations was age and gender invariant. Findings advance the theoretical understanding of the linkage between aggression and disaster exposure. Findings also support the utility of the PCS in disaster research and the link between PCS scores and academic achievement.

Biobehavior Life Regulation (BLR) scale for living well in chronic pain: Preliminary scale development and validation.

Currently available pain assessment scales focus on pain-related symptoms and limitations imposed by pain. Validated assessment tools that measure how pain is regulated by those who live well with pain are missing. This study seeks to fill this gap by describing the development and preliminary validation of the Biobehavior Life Regulation (BLR) scale. The BLR scale assesses engagement, social relatedness, and self-growth in the presence of chronic pain and the unpredictability of chronic pain. Sources for items included survivor strategies, patient experiences, existing scales, and unpredictable pain research. Review for suitability yielded 52 items. Validation measures were identified for engagement, social relatedness, self-growth, and unpredictability of pain. The study sample (n = 202) represented patients treated in the Phoenix VA Health Care System (n = 112) and two community clinics (n = 90). Demographic characteristics included average age of 52.5, heterogeneous in ethnicity and race at the VA, mainly Non-Hispanic White at the community clinics, 14 years of education, and pain duration of 18 years for the VA and 15.4 years for community clinics. Exploratory factor analysis using Oblimin rotation in the VA sample (n = 112) yielded a two-factor solution that accounted for 48.23% of the total variance. Confirmatory factor analysis (CFA) in the same sample showed high correlations among items in Factor 1, indicating redundancy and the need to further reduce items. The final CFA indicated a 2-factor solution with adequate fit to the data. The 2-factor CFA was replicated in Sample 2 from the community clinics (n = 90) with similarly adequate fit to the data. Factor 1, Pain Regulation, covered 8 items of engagement, social relatedness, and self-growth while Factor 2, Pain Unpredictability, covered 6 items related to the experience of unpredictable pain. Construct validity showed moderate to higher Pearson correlations between BLR subscales and relevant well-established constructs that were consistent across VA and community samples. The BLR scale assesses adaptive regulation strategies in unpredictable pain as a potential tool for evaluating regulation resources and pain unpredictability.