Assembly and Translocation of a CRISPR-Cas Primed Acquisition Complex.

CRISPR-Cas systems confer an adaptive immunity against viruses. Following viral injection, Cas1-Cas2 integrates segments of the viral genome (spacers) into the CRISPR locus. In type I CRISPR-Cas systems, efficient "primed" spacer acquisition and viral degradation (interference) require both the Cascade complex and the Cas3 helicase/nuclease. Here, we present single-molecule characterization of the Thermobifida fusca (Tfu) primed acquisition complex (PAC). We show that TfuCascade rapidly samples non-specific DNA via facilitated one-dimensional diffusion. Cas3 loads at target-bound Cascade and the Cascade/Cas3 complex translocates via a looped DNA intermediate. Cascade/Cas3 complexes stall at diverse protein roadblocks, resulting in a double strand break at the stall site. In contrast, Cas1-Cas2 samples DNA transiently via 3D collisions. Moreover, Cas1-Cas2 associates with Cascade and translocates with Cascade/Cas3, forming the PAC. PACs can displace different protein roadblocks, suggesting a mechanism for long-range spacer acquisition. This work provides a molecular basis for the coordinated steps in CRISPR-based adaptive immunity.

Comprehensive Molecular Portraits of Invasive Lobular Breast Cancer.

Invasive lobular carcinoma (ILC) is the second most prevalent histologic subtype of invasive breast cancer. Here, we comprehensively profiled 817 breast tumors, including 127 ILC, 490 ductal (IDC), and 88 mixed IDC/ILC. Besides E-cadherin loss, the best known ILC genetic hallmark, we identified mutations targeting PTEN, TBX3, and FOXA1 as ILC enriched features. PTEN loss associated with increased AKT phosphorylation, which was highest in ILC among all breast cancer subtypes. Spatially clustered FOXA1 mutations correlated with increased FOXA1 expression and activity. Conversely, GATA3 mutations and high expression characterized luminal A IDC, suggesting differential modulation of ER activity in ILC and IDC. Proliferation and immune-related signatures determined three ILC transcriptional subtypes associated with survival differences. Mixed IDC/ILC cases were molecularly classified as ILC-like and IDC-like revealing no true hybrid features. This multidimensional molecular atlas sheds new light on the genetic bases of ILC and provides potential clinical options.

Clinical Characteristics of Adolescents With Juvenile Idiopathic Arthritis Transitioning to Adult Rheumatology Care in Canada: Results From the CAPRI Registry.

OBJECTIVE: Using Canadian Alliance of Pediatric Rheumatology Investigators (CAPRI) juvenile idiopathic arthritis (JIA) registry data, we describe (1) clinical characteristics of patients with JIA transitioning to adult care, (2) prevalence of disease-related damage and complications, and (3) changes in disease activity during the final year prior to transfer. METHODS: Registry participants who turned 17 years between February 2017 and November 2021 were included. Clinical characteristics and patient-reported outcomes (PROs) at the last recorded pediatric rheumatology visit, and changes observed in the year prior to that visit were analyzed. Physicians completed an additional questionnaire characterizing cumulative disease-related damage and adverse events by age 17 years. RESULTS: At their last visit, 88 of 131 participants (67%) had inactive and 42 (32%) had active disease. Overall, 96 (73%) were on medications and 41 (31%) were on biologic disease-modifying antirheumatic drugs. Among 80 participants for whom the additional questionnaire was completed, 26% had clinically detected joint damage, 31% had joint damage on imaging, 14% had uveitis, and 7.5% had experienced at least 1 serious adverse event. During the final year, 44.2% of patients were in remission, 28.4% attained inactive disease, and 27.4% became or remained active. Mean scores of PROs were stable overall during that last year, but a minority reported marked worsening. CONCLUSION: A substantial proportion of youth with JIA transitioning to adult care in Canada had a high disease burden, which was reflected by their degree of disease activity, joint damage, or ongoing medication use. These results will inform pediatric and adult providers of anticipated needs during transition of care.

SARS-CoV-2 escape from a highly neutralizing COVID-19 convalescent plasma.

To investigate the evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the immune population, we coincupi bated the authentic virus with a highly neutralizing plasma from a COVID-19 convalescent patient. The plasma fully neutralized the virus for seven passages, but, after 45 d, the deletion of F140 in the spike N-terminal domain (NTD) N3 loop led to partial breakthrough. At day 73, an E484K substitution in the receptor-binding domain (RBD) occurred, followed, at day 80, by an insertion in the NTD N5 loop containing a new glycan sequon, which generated a variant completely resistant to plasma neutralization. Computational modeling predicts that the deletion and insertion in loops N3 and N5 prevent binding of neutralizing antibodies. The recent emergence in the United Kingdom, South Africa, Brazil, and Japan of natural variants with similar changes suggests that SARS-CoV-2 has the potential to escape an effective immune response and that vaccines and antibodies able to control emerging variants should be developed.

Narrative Sense-Making During COVID-19: Using Stories to Understand Birth in a Global Pandemic.

Women who gave birth in the spring and summer of 2020 contended with a host of challenging factors. In addition to facing pregnancy, labor, and delivery during an emerging global pandemic, women grappled with health care restrictions that altered their birth experience. To explore how women made sense of their birth during COVID-19, we analyzed written narratives from 71 women who gave birth in the United States from March to July 2020. Based on tenets of communicated narrative sense-making, the themes that emerged from our data suggest that women framed the role of the pandemic as either completely overshadowing their birth experience or as an inconvenience. Women also wrote about threats to their agency as patients, mothers, and caregivers, as well as the evolving emotional toll of the pandemic that often prompted feelings of fear and sadness, along with self-identified anxiety and depression. We discuss these findings in light of the literature on birth stories as essential sites of narrative sense-making for women and their families.

'We need a culture change': military sexual trauma through the lens of rape culture.

Military sexual trauma remains a critical concern within the US military. One possible explanation for the persistence of this trauma is the presence of rape culture. This study examined perceptions of military sexual trauma within the framework of rape culture (i.e. traditional gender roles, sexism, adversarial sexual beliefs, hostility toward women, and acceptance of violence), in addition to associated factors, to address the US Department of Defense's call to focus on the role of culture on military sexual trauma. Focusing on culture is important as subtle norms in culture shape our behaviour and impact the excusal/perpetration of military sexual trauma. Identifying these norms may provide insight into factors to target in prevention efforts to reduce/eliminate military sexual trauma. Forty-two US service members responded to a series of open-ended questions. Standard content analysis procedures were utilised to determine themes. Results paint a complex picture of military sexual trauma, with participants demonstrating concern regarding military sexual trauma, but minimal acknowledgement of behaviour change to reduce military sexual trauma. These results are consistent with the observed increase in prevalence of military sexual trauma and decrease in reporting of it since 2018. Adding to the existing literature, participants noted the continued existence of rape culture within the military and the need for cultural change to address military sexual trauma.

Looking out for You and Me: College students' Strategies for Preventing Sexual Assault in Drinking Contexts.

Background: Sexual assault and heavy alcohol use are prevalent and interrelated public health concerns on university campuses. Surprisingly, however, few alcohol harm reduction interventions address this intersection to help students reduce both personal and community risks for sexual assault in college drinking contexts. Objectives: In the current study, students (ages 18-24) shared strategies they use to protect themselves and others from sexual assault in college drinking contexts, as well as challenges to implementing these strategies. A series of six focus groups were conducted across two universities in the U.S. (N = 35). Participants responded to open-ended questions focused on drinking and sexual assault (e.g., What are some of the things students might do to avoid or address situations where they feel pressured of coerced to hook up or have sex when they do not want to?). Results: Thematic analyses demonstrated students' awareness of protective behavioral and bystander intervention strategies that could help reduce vulnerability to experience sexual assault for themselves or others in drinking contexts. Perceived barriers to using bystander intervention strategies included student's own and friends' heavy drinking (decreased inhibitions, loss of autonomy), ambiguity in deciphering risk (lack of familiarity, minimization, diffusion of responsibility), and gender (gender norms, power imbalances). Conclusions: This study informs the development of interventions that help students identify strategies and overcome barriers to reduce risks for sexual assault in college drinking contexts.

A decade of progress in juvenile idiopathic athritis treatments and outcomes in Canada: results from ReACCh-Out and the CAPRI registry.

OBJECTIVE: To assess changes in juvenile idiopathic arthritis (JIA) treatments and outcomes in Canada, comparing a 2005-2010 and a 2017-2021 inception cohorts. METHODS: Patients enrolled within three months of diagnosis in the Research in Arthritis in Canadian Children Emphasizing Outcomes (ReACCh-Out) and the Canadian Alliance of Pediatric Rheumatology Investigators Registry (CAPRI) cohorts were included. Cumulative incidences of drug starts and outcome attainment within 70 weeks of diagnosis were compared with Kaplan Meier survival analysis and multivariable Cox regression. RESULTS: The 2005-2010 and 2017-2021 cohorts included 1128 and 721 patients, respectively. JIA category distribution and baseline clinical juvenile idiopathic arthritis disease activity (cJADAS10) scores at enrolment were comparable. By 70 weeks, 6% of patients (95% CI 5, 7) in the 2005-2010 and 26% (23, 30) in the 2017-2021 cohort had started a biologic DMARD (bDMARD), and 43% (40, 47) and 60% (56, 64) had started a conventional DMARD (cDMARD), respectively. Outcome attainment was 64% (61, 67) and 83% (80, 86) for Inactive disease (Wallace criteria), 69% (66, 72) and 84% (81, 87) for minimally active disease (cJADAS10 criteria), 57% (54, 61) and 63% (59, 68) for pain control (<1/10), and 52% (47, 56) and 54% (48, 60) for a good health-related quality of life. CONCLUSION: Although baseline disease characteristics were comparable in the 2005-2010 and 2017-2021 cohorts, cDMARD and bDMARD use increased with a concurrent increase in minimally active and inactive disease. Improvements in parent and patient reported outcomes were smaller than improvements in disease activity.

Prognostic impact of corticosteroids on efficacy of chimeric antigen receptor T-cell therapy in large B-cell lymphoma.

Corticosteroids are commonly used for the management of severe toxicities associated with chimeric antigen receptor (CAR) T-cell therapy. However, it remains unclear whether their dose, duration, and timing may affect clinical efficacy. Here, we determined the impact of corticosteroids on clinical outcomes in patients with relapsed or refractory large B-cell lymphoma treated with standard of care anti-CD19 CAR T-cell therapy. Among 100 patients evaluated, 60 (60%) received corticosteroids for management of CAR T-cell therapy-associated toxicities. The median cumulative dexamethasone-equivalent dose was 186 mg (range, 8-1803) and the median duration of corticosteroid treatment was 9 days (range, 1-30). Corticosteroid treatment was started between days 0 and 7 in 45 (75%) patients and beyond day 7 in 15 (25%). After a median follow-up of 10 months (95% confidence interval, 8-12 months), use of higher cumulative dose of corticosteroids was associated with significantly shorter progression-free survival. More importantly, higher cumulative dose of corticosteroids, and prolonged and early use after CAR T-cell infusion were associated with significantly shorter overall survival. These results suggest that corticosteroids should be used at the lowest dose and for the shortest duration and their initiation should be delayed whenever clinically feasible while managing CAR T-cell therapy-associated toxicities.

Disentangling the components of pollen limitation in a widespread herb with gametophytic self-incompatibility.

PREMISE: Seed production is frequently limited by the receipt of insufficient or low-quality pollen, collectively termed "pollen limitation" (PL). In taxa with gametophytic self-incompatibility (GSI), incompatible pollen can germinate on stigmas but pollen tubes are arrested in styles. This allows for estimates of pollen performance before, during, and after self-recognition, as well as insight into the factors underlying pollen quality limitation in GSI taxa. METHODS: We scored pollen performance following self and outcross pollinations in Argentina anserina to identify the location of self-recognition and establish the relationship between pollen tubes and seed production. We then estimated quantity and quality components of PL from >3300 field-collected styles. We combined our results with other studies to test the prediction that low pollen quality, but not quantity, drives higher PL in self-incompatible (SI) taxa than in self-compatible taxa (SC). RESULTS: Self and outcross pollen germinated readily on stigmas, but 96% of germinated self-pollen was arrested during early tube elongation. Reproduction in the field was more limited by pollen quality than by quantity, and pollen failure near the location of self-recognition was a stronger barrier to fertilization than pollen germination. Across 26 taxa, SI species experienced stronger pollen quality, but not quantity, limitation than SC species. CONCLUSIONS: Evaluating pollen performance at multiple points within pistils can elucidate potential causes of pollen quality limitation. The receipt of incompatible pollen inhibits fertilization success more than insufficient pollen receipt or poor pollen germination in A. anserina. Likewise, pollen quality limitation drives high overall PL in other SI taxa.

Towards Understanding Disparities in Using Technology to Access Health Care Information: African American Veterans' Sociocultural Perspectives on Using My HealtheVet for Diabetes Management.

The digital divide, a racial disparity in computer and internet use, persists among Veterans. This study examined African American Veterans' experiences with and attitudes about using My HealtheVet (MHV), the online patient portal used in the Department of Veterans Affairs (VA) health care system. Semi-structured interviews were conducted with 35 non-Hispanic African American Veterans about preferences for health information, use of web-enabled devices, and experiences with and preferences for using MHV. Twenty-nine men and 6 women, ranging 35 to 73 years old, participated in this study. About 75% (n = 26) had completed some college or more, and about 75% (n = 26) considered their families financially comfortable. Ninety-one percent (n = 32) of participants were aware of MHV, and 78% (n = 25) of those who were aware reported having enrolled for an account. Data revealed participants valued that MHV provided realistic solutions to cumbersome aspects (e.g., phone call hold-times) of receiving care at the VA. However, participants admitted to not using the patient portal at all or to its full potential for two reasons: a) preference for their pre-MHV routine, b) distrust in the institutional, technological, and organizational aspects surrounding MHV. By highlighting the sociocultural aspects of patient portal use, this study provides a meaningful explanation for barriers and facilitators of health information technology adoption, particularly for MHV among African American Veterans, and ways practitioners may influence uptake.

"It Made Me Not Want to See him ": The Role of Patient-Provider Communication in Influencing Rural-Dwelling Women Veterans' Motivation to Seek Health Care for Managing Chronic Pain.

Despite being high health care utilizers, many women Veterans perceive their pain condition to be poorly understood by their providers, which can be a strong demotivator for seeking care. We set out to understand the priorities rural-dwelling women Veterans have for using health care for their chronic pain, and interviewed participants about their experiences with (and priorities for seeking) health care for their chronic pain. Self-Determination Theory identifies three sources of motivation (autonomy, competence, relatedness), all of which were represented through two themes that reflect rural women Veterans' rationale for decision-making to obtain health care for chronic pain: role of trust and competing priorities. Women described their priorities for chronic pain management in terms of their competing priorities for work, education, and supporting their family, but most expressed a desire to function in their daily life and relationships. Second, women discussed the role of trust in their provider as a source of motivation, and the role of patient-provider communication skills and gender played in establishing trust. Rural women Veterans often discussed core values that stemmed from facets of their identity (e.g. gender, military training, ethnicity) that also influenced their decision-making. Our findings provide insight for how providers may use Motivational Interviewing and discuss chronic pain treatment options so that rural-dwelling women Veterans feel autonomous, competent, and understood in their decision-making about their chronic pain. We also discuss importance of acknowledging the effects of disenfranchising talk and perpetuating gendered stereotypes related to chronic pain and theoretical implications of this work.

"I'm Not Comfortable With COVID, But ": Dilemmas and Decision-Making to Mitigate Risks Among Mothers Who Gave Birth During the COVID-19 Pandemic.

Individuals have faced unprecedented uncertainty and risk surrounding the COVID-19 pandemic, and decision-making dilemmas have been complicated by quickly evolving and often contradictory recommendations for staying healthy. Using tenets of problematic integration theory and risk orders theory, we analyzed interview data from 50 mothers who gave birth during the pandemic to understand how uncertainty and risk perceptions shaped their decision-making about keeping themselves and their infants healthy in the first year after birth. Results describe how some mothers in our sample made sense of their decision-making to prioritize first-order risks to their own and their family's physical health, and other mothers prioritized second-order risks to their relationships and identities. We also discuss the social nature of mitigating risk during the COVID-19 pandemic and the catalysts for shifting risk perceptions. Theoretical and practical implications include improving public health messaging and clinical conversations to enable individuals to effectively manage social and identity needs alongside serious threats to physical health.

Development and validation of the Kids Disability Screen for children with juvenile idiopathic arthritis: results from the CAPRI Registry.

OBJECTIVE: The aim of this study was to develop and validate a brief disability screen for children with JIA, the Kids Disability Screen (KDS). METHODS: A total of 216 children enrolled in the Canadian Alliance of Pediatric Rheumatology Investigators (CAPRI) Registry in 2017-2018 formed a development cohort, and 220 children enrolled in 2019-2020 formed a validation cohort. At every clinic visit, parents answered two questions derived from the Childhood Health Assessment Questionnaire (CHAQ): 'Is it hard for your child to run and play BECAUSE OF ARTHRITIS?' ('Hard' 0-10), and 'Does your child usually need help from you or another person BECAUSE OF ARTHRITIS?' ('Help', 0-10). We used 36-fold cross-validation and tested nine different mathematical methods to combine the answers and optimize psychometric properties. The results were confirmed in the validation cohort. RESULTS: Expressed as the mean of the two answers, KDS best balanced ease of use and psychometric properties, while a LASSO regression model combining the two answers with other patient characteristics [estimated CHAQ [eCHAQ]) had the highest responsiveness. In the validation cohort, 22.7%, 25.9% and 28.6% of patients had a score of 0 at enrolment for the KDS, eCHAQ and CHAQ, respectively. Responsiveness was 0.67, 0.74 and 0.62, respectively. Sensitivity to detect a CHAQ > 0 was 0.90 and specificity 0.56, KDS detecting some disability in 44% of children with a CHAQ = 0. CONCLUSION: This simple KDS has psychometric properties comparable with those of a full CHAQ and may be used at every clinic visit to identify those children who need a full disability assessment.

Impact of the COVID-19 pandemic on juvenile idiopathic arthritis presentation and research recruitment: results from the CAPRI registry.

OBJECTIVE: The COVID-19 pandemic has disrupted healthcare delivery and clinical research worldwide, with data from areas most affected demonstrating an impact on rheumatology care. This study aimed to characterize the impact of the pandemic on the initial presentation of JIA and JIA-related research in Canada. METHODS: Data collected from the Canadian Alliance of Pediatric Rheumatology Investigators JIA Registry from the year pre-pandemic (11 March 2019 to 10 March 2020) was compared with data collected during the first year of the pandemic (11 March 2020 to 10 March 2021). Outcomes included time from symptom onset to first assessment, disease severity at presentation and registry recruitment. Proportions and medians were used to describe categorical and continuous variables, respectively. RESULTS: The median time from symptom onset to first assessment was 138 (IQR 64-365) days pre-pandemic vs 146 (IQR 83-359) days during the pandemic. The JIA category frequencies remained overall stable (44% oligoarticular JIA pre-pandemic, 46.8% pandemic), except for systemic JIA (12 cases pre-pandemic, 1 pandemic). Clinical features, disease activity (cJADAS10), disability (CHAQ) and quality of life (JAQQ) scores were similar between the two cohorts. Pre-pandemic, 225 patients were enrolled, compared with 111 in the pandemic year, with the greatest decrease from March to June 2020. CONCLUSIONS: We did not observe the anticipated delay in time to presentation or increased severity at presentation, suggesting that, within Canada, care adapted well to provide support to new patient consults without negative impacts. The COVID-19 pandemic was associated with an initial 50% decrease in registry enrolment but has since improved.

My Goals Are Not Their Goals: Barriers and Facilitators to Delivery of Patient-Centered Care for Patients with Multimorbidity.

BACKGROUND: Patient-centered care reflecting patient preferences and needs is integral to high-quality care. Individualized care is important for psychosocially complex or high-risk patients with multiple chronic conditions (i.e., multimorbidity), given greater potential risks of interventions and reduced benefits. These patients are increasingly prevalent in primary care. Few studies have examined provision of patient-centered care from the clinician perspective, particularly from primary care physicians serving in integrated, patient-centered medical home settings within the US Veterans Health Administration. OBJECTIVE: We sought to clarify facilitators and barriers perceived by primary care physicians in the Veterans Health Administration to delivering patient-centered care for high-risk or complex patients with multimorbidity. DESIGN: We conducted semi-structured telephone interviews from April to July 2020 among physicians across 20 clinical sites. Findings were analyzed with deductive content analysis based on conceptual models of patient-centeredness and hierarchical factors affecting care delivery. PARTICIPANTS: Of 23 physicians interviewed, most were female (n = 14/23, 61%), serving in hospital-affiliated outpatient clinics (n = 14/23, 61%). Participants had a mean of 21 (SD = 11.3) years of experience. KEY RESULTS: Facilitators included the following: effective physician-patient communication to individualize care, prioritize among multiple needs, and elicit goals to improve patient engagement; access to care, enabled by interdisciplinary teams, and dictating personalized care planning; effortful but worthwhile care coordination and continuity; meeting complex needs through effective teamwork; and integrating medical and non-medical care aspects in recognition of patients' psychosocial contexts. Barriers included the following: intra- and interpersonal (e.g., perceived patient reluctance to engage in care); organizational (e.g., limited encounter time); and community or policy impediments (e.g., state decisional capacity laws) to patient-centered care. CONCLUSIONS: Physicians perceived individual physician-patient interactions were the greatest facilitators or barriers to patient-centered care. Efforts to increase primary care patient-centeredness for complex or high-risk patients with multimorbidity could focus on targeting physician-patient communication and reducing interpersonal conflict.

The Diabetes Simulation Challenge: Enhancing Patient Perspective-Taking for Medical Students.

The Diabetes Simulation Challenge is a unique training tool to foster empathy, a key facet of patient-centered care, for medical students. Thirty-two medical students participated in a 24-hour perspective-taking activity as part of their curriculum, during which they simulated some common experiences of living with a chronic health condition, specifically type 1 diabetes. Students' written reflections were analyzed using a phenomenological qualitative approach to provide a composite description of the experience. An exhaustive, iterative method of thematic analysis that included manual coding was used to determine whether this activity led to expressions of empathy or thoughts and beliefs consistent with patient-centered health care. Nine unique themes emerged, six of which indicated that students adopted the perspective of an individual with a chronic illness. Most of the students' reflections illustrated an understanding of the behavioral, social, and emotional challenges related to living with type 1 diabetes, as well as increased empathy toward individuals with the disease. Medical students who aim to provide patient-centered care benefited from this perspective-taking exercise, and training programs should consider using such methods to extend learning beyond traditional didactic education.

Autoinflammatory disease mimicking acute hemorrhagic edema of infancy.

The autoinflammatory diseases are a heterogeneous group of disorders characterized by early appearance of symptoms and characteristic flares of fever plus signs of internal inflammation, including various skin manifestations. We describe a 2.5-year-old boy who presented with fever, purpuric skin lesions, and edema, the classic clinical triad of acute hemorrhagic edema of infancy (AHEI). Although AHEI was our initial leading diagnosis, additional history revealed that these episodes were recurrent, and further investigation lead to a diagnosis of tumor necrosis factor receptor-associated periodic syndrome (TRAPS). This case report affirms the broad clinical spectrum of the autoinflammatory diseases and reinforces the importance of obtaining a thorough history of previous episodes of skin and extracutaneous inflammation in patients with a clinical diagnosis of AHEI.

The grey zone of collegiate sexual regret: questionable consent and sexual victimisation.

Research identifies sexual regret as the most common source of regret in the USA; however, few studies have explored this phenomenon. This study utilised a mixed-methods approach to explore sexual regret in a sample of 189 US college students. Quantitative analyses assessing who is most likely to experience sexual regret revealed no significant differences in experiences of sexual regret based on most demographics; however, individuals involved in fraternity and sorority life and/or college athletics were significantly more likely to report experiences of sexual regret. Additionally, students who reported experiences of sexual victimisation were more likely to report experiences of sexual regret. Qualitative analyses attempting to determine students' reasoning for experiencing sexual regret revealed five distinct content areas: 1) altered judgement, 2) motivations, 3) partner characteristics, 4) social judgement, and 5) unsatisfying or unpleasant sexual experiences. Results from both quantitative and qualitative analyses demonstrate significant overlap between experiences of sexual regret and sexual victimisation, highlighting the importance of future research and programming regarding sexual regret to increase understanding of the complex relationships between sexual consent and sexual behaviour.