Islet-antigen reactive B cells display a unique phenotype and BCR repertoire in autoantibody positive and recent-onset type 1 diabetes patients.

Autoreactive B cells play an important but ill-defined role in autoimmune type 1 diabetes (T1D). To better understand their contribution, we performed single cell gene and BCR-seq analysis on pancreatic islet antigen-reactive (IAR) B cells from the peripheral blood of nondiabetic (ND), autoantibody positive prediabetic (AAB), and recent-onset T1D individuals. We found that the frequency of IAR B cells was increased in AAB and T1D. IAR B cells from these donors had altered expression of B cell signaling, pro-inflammatory, infection, and antigen processing and presentation genes. Both AAB and T1D donors demonstrated a significant increase in certain heavy and light chain V genes, and these V genes were enriched in islet-reactivity. Public clones of IAR B cells were restricted almost entirely to AAB and T1D donors. IAR B cells were clonally expanded in the autoimmune donors, particularly the AAB group. Notably, a substantial fraction of IAR B cells in AAB and T1D donors appeared to be polyreactive, which was corroborated by analysis of recombinant monoclonal antibodies. These results expand our understanding of autoreactive B cell activation during T1D and identify unique BCR repertoire changes that may serve as biomarkers for increased disease risk.

The impact of post-mastectomy radiation timing on overall outcomes of autologous free-flap breast reconstruction.

BACKGROUND: The optimal timing of post-mastectomy radiation therapy (PMRT) in autologous breast reconstruction is controversial. Our study compares overall reconstructive outcomes in patients who received post-mastectomy radiation therapy either before or after the autologous flap. METHODS: A single-center retrospective review was performed for patients who underwent free flap breast reconstruction and post-mastectomy radiation from January 2004 through January 2021. Demographic, intraoperative, and post-operative variables were recorded. RESULTS: A total of 452 free flaps were identified, and 82 underwent PMRT. 59.8% were radiated with an expander prior to free flap surgery (PreFlap), and 40.2% flaps underwent PMRT (PostFlap). PostFlap patients were significantly younger (43.0 vs. 47.9 years, p = .016). There were no significant differences in free flap outcomes between the two cohorts including thrombosis, venous congestion, flap loss, takebacks, fat necrosis, seroma, or infection. Mastectomy skin flap necrosis was significantly higher in the PostFlap cohort (9.1% vs. 0%, p = .032), but nipple necrosis rates did not differ. There were no significant differences in number or need for revision surgeries, fat necrosis, or fat grafting between groups. However, there were significantly more total reconstructive complications, including infection and wound breakdown, experienced by the PreFlap cohort (46.9% vs. 24.2%, p = .038). CONCLUSIONS: Timing of PMRT did not impact free flap outcomes, but those who had the expander radiated experienced significantly more complications overall. For the 34.7% of patients in the preFlap group who planned for autologous reconstruction form initial consultation, radiation after the flap may have improved their overall outcomes. As added complications cause delays in cancer therapy and final reconstruction, our results suggest that PMRT of the flap when possible may improve the overall experience for breast cancer patients.

High burden of sexually transmitted infections among under-resourced populations in the Dominican Republic.

Background: Sexually transmitted infections (STIs) are a major health issue, exacerbated by limited financial and infrastructural resources in developing countries. Methods: Prevalence of STIs was assessed in two urban centers of the Dominican Republic (DR) among populations at high risk for STIs: pregnant youth, men who have sex with men (MSM), trans women (TG), batey residents, female sex workers, and people living with human immunodeficiency virus (HIV). We conducted a cross-sectional survey and biological specimen collection to screen for Chlamydia trachomatis, Neisseria gonorrhea, Mycoplasma genitalium, Trichomonas vaginalis (trichomoniasis), Treponema pallidum (syphilis), HIV, hepatitis B and C, and human papillomavirus (HPV) among at-risk populations between 2015 and 2018. Ureaplasma urealyticum testing was also conducted even though it is not considered a STI. A non-probability community sample was recruited. Descriptive statistics examined the prevalence of STIs by population. Results: A total of 1991 subjects participated in the study. The median age was 26 years (range: 18-65). Most participants were female (65.3%), heterosexual (76.7%), and were not partnered (55.7%). Most of the participants reported unprotected vaginal sex in the last 6 months (54%); among MSM and TG almost half of the participants reported unprotected anal sex in the last 6 months and 17.6% reported drug use in the last 6 months. Almost half of the participants (49%) tested positive for one or more STIs. The most prevalent STI was Chlamydia trachomatis (12.8%), and human papillomavirus (11.9%). Among transgender women, 65.3% tested positive for an STI, 64.8% of female sex workers tested positive for an STI, and 53.8% of pregnant adolescents tested positive for an STI. Conclusion: There is a high prevalence of STIs among key and under resourced populations in the DR. Our findings highlight the need to conduct further research to optimize prevention and care strategies for structurally vulnerable and under resourced populations in the DR.

Application of single-cell RNA sequencing methods to develop B cell targeted treatments for autoimmunity.

The COVID-19 pandemic coincided with several transformative advances in single-cell analysis. These new methods along with decades of research and trials with antibody therapeutics and RNA based technologies allowed for highly effective vaccines and treatments to be produced at astonishing speeds. While these tools were initially focused on models of infection, they also show promise in an autoimmune setting. Self-reactive B cells play important roles as antigen-presenting cells and cytokine and autoantibody producers for many autoimmune diseases. Yet, current therapies to target autoreactive B cells deplete all B cells irrespective of their pathogenicity. Development of self-reactive B cell targeting therapies that would spare non-pathogenic B cells are needed to treat disease while allowing effective immune responses to other ailments. Single-cell RNA sequencing (scRNA-seq) approaches will aid in identification of the pathogenic self-reactive B cells operative in autoimmunity and help with development of more favorable precision targeted therapies.

Identification of an anergic BND cell-derived activated B cell population (BND2) in young-onset type 1 diabetes patients.

Recent evidence suggests a role for B cells in the pathogenesis of young-onset type 1 diabetes (T1D), wherein rapid progression occurs. However, little is known regarding the specificity, phenotype, and function of B cells in young-onset T1D. We performed a cross-sectional analysis comparing insulin-reactive to tetanus-reactive B cells in the blood of T1D and controls using mass cytometry. Unsupervised clustering revealed the existence of a highly activated B cell subset we term BND2 that falls within the previously defined anergic BND subset. We found a specific increase in the frequency of insulin-reactive BND2 cells in the blood of young-onset T1D donors, which was further enriched in the pancreatic lymph nodes of T1D donors. The frequency of insulin-binding BND2 cells correlated with anti-insulin autoantibody levels. We demonstrate BND2 cells are pre-plasma cells and can likely act as APCs to T cells. These findings identify an antigen-specific B cell subset that may play a role in the rapid progression of young-onset T1D.

High prevalence of rectal chlamydia among pregnant adolescents in La Romana, Dominican Republic warrants extragenital STI testing.

To our knowledge, there are no studies estimating the prevalence of extragenital sexually transmitted infections (STIs) among pregnant adolescents in the Caribbean. This study sought to fill this gap by assessing the prevalence and correlates of oral, genital, and rectal chlamydia (CT) among a sample of pregnant adolescents in La Romana, Dominican Republic. Two hundred pregnant youths, aged 15-24 years, were recruited by systematic sampling during their first prenatal visit to a maternal care unit. A sociodemographic and behavioral questionnaire was administered and urine and oral/anal swabs were collected and tested for CT. Descriptive analyses and Fisher's exact tests were performed. The prevalence of oral, genital, and rectal CT was 6%, 15%, and 23%, respectively, although less than 5% of participants reported ever engaging in receptive anal intercourse. This discrepancy could be explained by autoinoculation, concurrent transmission during sex, undertreatment of rectal CT, or underreporting of anal sex. Almost half of CT infections would have been missed if only genital samples were collected, as current protocol dictates. More research is needed to understand sexual behaviors and rectal STI risk factors among heterosexual adolescent women. STI screening procedures for pregnant and sexually active adolescents should include routine testing of extragenital sites.

Human Simulation in Nursing Education.

Although, human simulation methodology has its origins in medical education, nursing education has increased its use of simulated patient (SP) methodology to improve the education of nursing students across the curricula. This chapter will review the history of human simulation, introduce the human simulation continuum, and review different applications of SP methodology in undergraduate and graduate nursing education.

Activation of DNA damage repair pathways by murine polyomavirus.

Nuclear replication of DNA viruses activates DNA damage repair (DDR) pathways, which are thought to detect and inhibit viral replication. However, many DNA viruses also depend on these pathways in order to optimally replicate their genomes. We investigated the relationship between murine polyomavirus (MuPyV) and components of DDR signaling pathways including CHK1, CHK2, H2AX, ATR, and DNAPK. We found that recruitment and retention of DDR proteins at viral replication centers was independent of H2AX, as well as the viral small and middle T-antigens. Additionally, infectious virus production required ATR kinase activity, but was independent of CHK1, CHK2, or DNAPK signaling. ATR inhibition did not reduce the total amount of viral DNA accumulated, but affected the amount of virus produced, indicating a defect in virus assembly. These results suggest that MuPyV may utilize a subset of DDR proteins or non-canonical DDR signaling pathways in order to efficiently replicate and assemble.

Microbial community assembly and metabolic function during mammalian corpse decomposition.

Vertebrate corpse decomposition provides an important stage in nutrient cycling in most terrestrial habitats, yet microbially mediated processes are poorly understood. Here we combine deep microbial community characterization, community-level metabolic reconstruction, and soil biogeochemical assessment to understand the principles governing microbial community assembly during decomposition of mouse and human corpses on different soil substrates. We find a suite of bacterial and fungal groups that contribute to nitrogen cycling and a reproducible network of decomposers that emerge on predictable time scales. Our results show that this decomposer community is derived primarily from bulk soil, but key decomposers are ubiquitous in low abundance. Soil type was not a dominant factor driving community development, and the process of decomposition is sufficiently reproducible to offer new opportunities for forensic investigations.

Quality of life and depression following childbirth: impact of social support.

OBJECTIVE: to evaluate the impact of social support on postnatal depression and health-related quality of life. DESIGN: prospective cohort study. Data were collected at baseline and at six weeks post discharge using a postal survey. SETTING AND PARTICIPANTS: between August and December 2008, 320 women from a large tertiary hospital were recruited following the birth of their infant. MEASUREMENTS: Edinburgh Postnatal Depression Scale (EPDS), Maternity Social Support Scale and World Health Organization Quality of Life assessment questionnaire. FINDINGS: of the 320 women recruited, 222 (69.4%) returned their six-week questionnaire. Women with low social support had significantly higher scores on the EPDS than women who reported adequate support (p = 0.007). There was also a significant effect of social support on health-related quality of life. Women with low family or partner support scored lower in all domains, with the greatest mean difference in the social health domain (p = 0.000). Of those scoring >10 on the EPDS, 75.5% had sought professional help. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: women with low social support are more likely to report postnatal depression and lower quality of life than well-supported women. Careful assessment of a woman's level of support following the birth, particularly from her partner and family, may provide useful information for possible interventions.

Validation of the WHOQOL-BREF among women following childbirth.

BACKGROUND: There is increasing interest in measuring quality of life (QOL) in clinical settings and in clinical trials. None of the commonly used QOL instruments has been validated for use postnatally. AIM: To assess the psychometric properties of the 26-item WHOQOL-BREF (short version of the World Health Organization Quality of Life assessment) among women following childbirth. METHODS: Using a prospective cohort design, we recruited 320 women within the first few days of childbirth. At six weeks postpartum, participants were asked to complete the WHOQOL-BREF, the Edinburgh Postnatal Depression Index and the Australian Unity Wellbeing Index. Validation of the WHOQOL-BREF included an analysis of internal consistency, discriminate validity, convergent validity and an examination of the domain structure. RESULTS: In all, 221 (69.1%) women returned their six-week questionnaire. All domains of the WHOQOL-BREF met reliability standards (alpha coefficient exceeding 0.70). The questionnaire discriminated well between known groups (depressed women and non-depressed women. P < or = 0.000) and demonstrated satisfactory correlations with the Australian Unity Wellbeing index (r > or = 0.45). The domain structure of the WHOQOL-BREF was also valid in this population of new mothers, with moderate-to-high correlation between individual items and the domain structure to which the items were originally assigned. CONCLUSION: The WHOQOL-BRF is a well-accepted and valid instrument in this population and may be used in postnatal clinical settings or for assessing intervention effects in research studies.

Validity of the Waterlow scale and risk of pressure injury in acute care.

AIM: To assess the validity of the Waterlow screening tool in a cohort of internal medicine patients and to identify factors contributing to pressure injury. METHOD: A longitudinal cohort study design was used. A total of 274 patients (mean age 65.3 years) admitted through the emergency department or outpatient clinics of a tertiary hospital in Brisbane, Australia, and expected to remain in hospital for at least 3 days were screened on admission using the Waterlow screening tool. Their pressure ulcer status was monitored and recorded every second day. The main outcome measure was pressure ulcer incidence. RESULTS: Fifteen participants (5.5%) had an existing pressure ulcer and a further 12 (4.4%) developed a pressure ulcer during their hospital stay. Sensitivity of the Waterlow scale was 0.67 (95% confidence interval [CI]: 0.35-0.88), specificity was 0.79 (95% CI: 0.73-0.85), positive predictive value was 0.13 (95% CI: 0.07-0.24) and negative predictive value was 0.98 (95% CI: 0.94-0.99). CONCLUSION: This study provides further evidence of the poor predictive validity of the Waterlow scale. A suitably powered, randomized controlled trial is urgently needed to provide definitive evidence about the usefulness of the Waterlow scale compared with other screening tools and with clinical judgment.