Health-related Quality of Life in Patients with Previously Treated Advanced Urothelial Carcinoma from EV-301: A Phase 3 Trial of Enfortumab Vedotin Versus Chemotherapy.

BACKGROUND AND OBJECTIVE: In comparison to chemotherapy, enfortumab vedotin (EV) prolonged overall survival in patients with previously treated advanced urothelial carcinoma in EV-301. The objective of the present study was to assess patient experiences of EV versus chemotherapy using patient-reported outcome (PRO) analysis of health-related quality of life (HRQoL). METHODS: For patients in the phase 3 EV-301 trial randomized to EV or chemotherapy we assessed responses to the validated European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (QLQ-C30) at baseline, weekly for the first 12 wk, and then every 12 wk until discontinuation. We analyzed the QLQ-C30 change from baseline to week 12, the confirmed improvement rate, and the time to improvement or deterioration. KEY FINDINGS AND LIMITATIONS: Baseline PRO compliance rates were 91% for the EV arm (n = 301) and 89% for the chemotherapy arm (n = 307); the corresponding average rates from baseline to week 12 were 70% and 67%. Patients receiving EV versus chemotherapy had reduced pain (difference in change from baseline to week 12: -5.7, 95% confidence interval [CI] -10.8 to -0.7; p = 0.027) and worsening appetite loss (7.3, 95% CI 0.90-13.69; p = 0.026). Larger proportions of patients in the EV arm reported HRQoL improvement from baseline than in the chemotherapy arm; the odds of a confirmed improvement across ten QLQ-C30 function/symptom scales were 1.67 to 2.76 times higher for EV than for chemotherapy. Patients in the EV arm had a shorter time to first confirmed improvement in global health status (GHS)/QoL, fatigue, pain, and physical, role, emotional, and social functioning (all p < 0.05). EV delayed the time to first confirmed deterioration in GHS/QoL (p = 0.027), but worsening appetite loss occurred earlier (p = 0.009) in comparison to chemotherapy. CONCLUSIONS AND CLINICAL IMPLICATIONS: HRQoL with EV was maintained, and deterioration in HRQoL was delayed with EV in comparison to chemotherapy. Better results with EV were reported for some scales, with the greatest difference observed for pain. These findings reinforce the EV safety and efficacy outcomes and benefits observed in EV-301. PATIENT SUMMARY: Patients with previously treated advanced cancer of the urinary tract receiving the drug enfortumab vedotin maintained their HRQoL in comparison to patients treated with chemotherapy. The EV-301 trial is registered on ClinicalTrials.gov as NCT03474107 and on EudraCT as 2017-003344-21.

Co-locating Primary Care in a Behavioral Health Home Improves Outcomes.

The Behavioral Health Home model of care enables patients living with severe persistent mental illness to access both mental health and primary care services, leading to improved mental and physical well-being. This article presents the implementation and health outcomes of colocating a primary care provider within an outpatient psychiatric rehabilitation day program.

Are Prior Sexual Risk Behaviors Associated With Adherence to COVID-19 Preventative Behaviors Among Young Black Men?

BACKGROUND: Behavioral clustering is a phenomenon in which several risk or protective behaviors co-occur in an individual. We sought to determine if prior sexual risk behaviors among young Black men who have sex with women could predict subsequent nonadherence to COVID-19 prevention behaviors. METHODS: Young Black men who have sex with women aged 15 to 24 years previously enrolled in a community-based Chlamydia trachomatis (Ct) screening program were enrolled in a substudy between May and June 2020 and asked about adherence to 4 COVID-19 recommended nonpharmaceutical prevention behaviors (handwashing, mask wearing, social distancing, and following stay at home orders). Data from the original study were used to elicit the follow prepandemic behaviors including having multiple sex partners, inconsistent condom use, prior sexually transmitted infection testing behaviors, and substance use. Wilcoxon rank sum tests were used to assess the association between historic risk behaviors and COVID-19 behavior score. RESULTS: There were 109 men included in the analysis, with a mean (SD) age of 20.5 (2.0) years. Inconsistent condom use, multiple sex partners, and prior HIV/sexually transmitted infection testing status were not associated with fewer COVID-19 preventive behaviors, but men who used any nonprescription drugs ( P = 0.001) or marijuana only ( P = 0.028) had a lower median COVID-19 preventative score compared with those who did not engage in those activities. CONCLUSIONS: Although none of the sexual risk behavior variables were associated, self-reported nonprescription drug and marijuana use were both significant predictors of lower adherence to COVID-19 preventative behaviors among young Black men. Young men who use drugs may need additional support to promote COVID-19 preventative behavior uptake.

A phase Ib trial of pembrolizumab plus paclitaxel or flat-dose capecitabine in 1st/2nd line metastatic triple-negative breast cancer.

Chemoimmunotherapy with anti-programmed cell death 1/ligand 1 and cytotoxic chemotherapy is a promising therapeutic modality for women with triple-negative breast cancer, but questions remain regarding optimal chemotherapy backbone and biomarkers for patient selection. We report final outcomes from a phase Ib trial evaluating pembrolizumab (200 mg IV every 3 weeks) with either weekly paclitaxel (80 mg/m2 weekly) or flat-dose capecitabine (2000 mg orally twice daily for 7 days of every 14-day cycle) in the 1st/2nd line setting. The primary endpoint is safety (receipt of 2 cycles without grade III/IV toxicities requiring discontinuation or ≥21-day delays). The secondary endpoint is efficacy (week 12 objective response). Exploratory aims are to characterize immunologic effects of treatment over time, and to evaluate novel biomarkers. The trial demonstrates that both regimens meet the pre-specified safety endpoint (paclitaxel: 87%; capecitabine: 100%). Objective response rate is 29% for pembrolizumab/paclitaxel (n = 4/13, 95% CI: 10-61%) and 43% for pembrolizumab/capecitabine (n = 6/14, 95% CI: 18-71%). Partial responses are observed in two subjects with chemo-refractory metaplastic carcinoma (both in capecitabine arm). Both regimens are associated with significant peripheral leukocyte contraction over time. Response is associated with clinical PD-L1 score, non-receipt of prior chemotherapy, and the H&E stromal tumor-infiltrating lymphocyte score, but also by a novel 27 gene IO score and spatial biomarkers (lymphocyte spatial skewness). In conclusion, pembrolizumab with paclitaxel or capecitabine is safe and clinically active. Both regimens are lymphodepleting, highlighting the competing immunostimulatory versus lymphotoxic effects of cytotoxic chemotherapy. Further exploration of the IO score and spatial TIL biomarkers is warranted. The clinical trial registration is NCT02734290.

Effectiveness of Incentivized Peer Referral to Increase Enrollment in a Community-Based Chlamydia Screening and Treatment Study Among Young Black Men.

OBJECTIVES: Young Black men are under-represented in sexual health services and research, a condition likely magnified during COVID-19 shutdowns due to disruption of STI screening and treatment services. We examined the effect of incentivized peer referral (IPR) increasing peer referral among young Black men in a community-based chlamydia screening program. METHODS: Young Black men in New Orleans, LA, age 15-26 years enrolled in a chlamydia screening program between 3/2018 and 5/2021 were included. Enrollees were provided with recruitment materials to distribute to peers. Starting July 28, 2020, enrollees were also offered a $5 incentive for each peer enrolled. Enrollment was compared before and after the incentivize peer referral program (IPR) was implemented using multiple time series analysis (MTSA). RESULTS: The percentage of men referred by a peer was higher during IPR compared to pre-IPR (45.7% vs. 19.7%, p < 0.001). After the COVID-19 shutdown was lifted, there were 2.007 more recruitments per week (p = 0.044, 95% CI (0.0515, 3.964)) for IPR, compared to pre-IPR. Overall, there was a trending increase in recruitments in the IPR era relative to the pre-IPR era (0.0174 recruitments/week, p = 0.285, 95% CI (- 0.0146, 0.0493)) with less recruitment decay during IPR compared to pre-IPR. CONCLUSIONS: IPR may be an effective means of engaging young Black men in community-based STI research and prevention programs, particularly when clinic access is limited. CLINICAL TRIALS REGISTRY SITE AND NUMBER: Clinicaltrials.gov identifier NCT03098329.

Sexual Networks and STI Infection Among Young Black Men Who Have Sex With Women in a Southern U.S. City.

PURPOSE: The configuration of one's sexual network has been shown to influence sexually transmitted infection (STI) acquisition in some populations. Young Black men who have sex with women (MSW) have high rates of STIs, yet little is known about their sexual networks. The purpose of this study is to describe the characteristics of sexual networks and their association with selected STI infections among young Black MSW. METHODS: Black MSW aged 15-26 years who were enrolled in the New Orleans community-based screening program named Check It from March 2018 to March 2020 were tested for C. trachomatis and N. gonorrhoeae infection and asked about the nature of their sexual partnerships. Sexual partnerships with women were defined as dyadic, somewhat dense (either themselves or their partner had multiple partners), and dense (both they and their partner(s) had multiple partners). RESULTS: Men (n = 1,350) reported 2,291 sex partners. The percentage of men who reported their networks were dyadic, somewhat dense, and dense was 48.7%, 27.7%, and 23.3%, respectively; 11.2% were STI-positive and 39.2% thought their partner(s) had other partners. Compared to men in dyadic relationships, those in somewhat dense network did not have increased risk of STI infection, but those in dense networks were more likely to have an STI (adjusted odds ratio = 2.06, 95% confidence interval [1.35-3.13]). DISCUSSION: Young Black MSW, who had multiple partners and who thought their partner(s) had other sex partners were at highest risk for STIs. Providers should probe not only about the youth's personal risk but should probe about perceived sexual partners' risk for more targeted counseling/STI testing.

Japanese subgroup analysis of EV-301: An open-label, randomized phase 3 study to evaluate enfortumab vedotin versus chemotherapy in subjects with previously treated locally advanced or metastatic urothelial carcinoma.

BACKGROUND: Enfortumab vedotin (EV) is an antibody-drug conjugate showing significant overall survival (OS) benefit versus chemotherapy for patients with previously treated locally advanced or metastatic urothelial carcinoma (la/mUC) in EV-301. This subgroup analysis was conducted to further analyze the efficacy and safety in a Japanese population. METHODS: In the open-label, phase 3 EV-301 trial, patients with la/mUC were randomized 1:1 to EV 1.25 mg/kg on Days 1, 8, and 15 for 28-day cycles or investigator-preselected standard chemotherapy (SC; docetaxel or paclitaxel for patients in Japan) on Day 1 of each 21-day cycle. Primary endpoint was OS and secondary efficacy endpoints included progression-free survival (PFS) and overall response rate (ORR). Safety/tolerability was also evaluated. RESULTS: As of the July 15, 2020 cut-off date for the interim analysis, the Japanese subgroup included 86 patients (EV: n = 36; SC: n = 50). Median OS was 15.18 months for EV and 10.55 months for SC (HR: 0.437 [95% CI: 0.209, 0.914]). Median PFS was 6.47 months for EV and 5.39 months for SC (HR: 0.464 [95% CI: 0.258, 0.835]). Confirmed ORR was 34.4% for EV and 21.3% for SC. A higher proportion of patients receiving SC versus EV had treatment-related adverse events (TRAEs; 97.9% vs. 91.7%, respectively), including grade ≥ 3 TRAEs (75.0% vs. 63.9%). CONCLUSIONS: This subgroup analysis confirmed that EV, with consistent efficacy and safety/tolerability in the EV-301 Japanese subgroup and overall study population, represents an important treatment option for previously treated patients with la/mUC.

Anal and Oral Sex Behaviors Among Young Black Men Who Have Vaginal Sex: Evidence of the Need for Extragenital Testing for Chlamydia and Gonorrhea.

BACKGROUND: Unprotected oral and anal sex may result in extragenital sexually transmitted infections. The purposes of this study were to describe sexual behaviors, barrier use, and chlamydia/gonorrhea (Ct/GC) positivity among young Black men who have sex with women, and to examine the potential influence of extragenital infections on genital infections. METHODS: Young Black men who had vaginal sex were screened for Ct/GC in New Orleans, LA, from August 14, 2019, to February 29, 2020. Audio/computer-assisted self-interviews were used to collect data on demographics and sexual behaviors. χ2 /Fisher exact or t test/Wilcoxon rank tests were used to assess differences in behaviors by Ct/GC positivity. RESULTS: Among 373 men studied, 619 female partnerships were reported in the past 2 months. Vaginal sex was reported in all partnerships per study protocol, receiving fellatio in 42.7%, performing cunnilingus in 35.7%, and penile-anal sex in 5.9%. Although 31.4% of the men consistently used condoms for vaginal sex with all partners, consistent barrier use was low during cunnilingus (0.5%) and fellatio (5.1%). Urethral infection rates among all men in the sample were 12.6% for Ct and 1.6% for GC. There was no significant difference in Ct/GC rates between those using and not using condoms consistently during vaginal sex ( P = 0.38). CONCLUSIONS: Unprotected oral sex with female partners was common. The high rate of genital infection among men who used condoms consistently for vaginal sex suggests that oral infections could be serving as a reservoir of genital infection. Testing at all sites of exposure for youth who engage in heterosexual sex is merited.

Health-related Quality of Life of Patients with Locally Advanced or Metastatic Urothelial Cancer Treated with Enfortumab Vedotin after Platinum and PD-1/PD-L1 Inhibitor Therapy: Results from Cohort 1 of the Phase 2 EV-201 Clinical Trial.

BACKGROUND: The EV-201 trial (NCT03219333) demonstrated a clinically meaningful and durable response rate and a tolerable safety profile with enfortumab vedotin (EV) in patients with locally advanced/metastatic urothelial carcinoma (LA/mUC) treated with prior PD-1/PD-L1 inhibitor therapy and platinum-containing chemotherapy (cohort 1). Patient-reported outcome (PRO) measures were included in EV-201 as exploratory endpoints. OBJECTIVE: To evaluate PRO data for cohort 1 of EV-201 to better understand the relationship between EV therapy and health-related quality of life (HRQoL). DESIGN, SETTING, AND PARTICIPANTS: Enrolled patients with LA/mUC who received EV were invited to electronically complete two HRQoL instruments (EORTC QLQ-C30 and EQ-5D-3L) at baseline and day 1 of each cycle until treatment discontinuation. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Patient demographics, completion and compliance rates, and PRO scores were analysed using descriptive statistics. Selected EORTC QLQ-C30 scores were analysed post hoc using a repeated-measures mixed model. RESULTS AND LIMITATIONS: Among treated patients (n = 125), 95% completed both baseline questionnaires. Compliance rates were ≥86% throughout the study. Descriptive analyses showed that global health status, physical functioning, and symptom scores remained stable over time, with average scores similar at each cycle. Lower pain and fatigue scores were observed in responders at cycles following an objective response. Pain was lower at cycle 3 than at baseline in patients with bone metastases. Mean EQ-5D-3L utility score (0.80 at baseline; range from 0.77 at cycle 2 to 0.91 at cycle 10) and visual analogue scale scores (66.9 at baseline; range from 65.5 at cycle 2 to 78.4 at cycle 10) remained similar over time. Variability and the small sample size limited definitive conclusions. CONCLUSIONS: PRO scores remained stable throughout EV treatment, further supporting the overall value of EV in the treatment of patients with LA/mUC. The potential benefit of EV therapy on overall HRQoL and symptoms such as pain and fatigue is currently being explored. PATIENT SUMMARY: In this study of adult patients with advanced cancer of the urinary tract that progressed after previous medications, quality of life, ability to function, and symptoms did not worsen on treatment with enfortumab vedotin, which is an antibody + drug combination. Some improvements in pain and fatigue were reported by patients, but further research needs to be conducted. These data complement the efficacy and safety data from the EV-201 trial.

DMSO reduces the cytotoxicity of anticancer ruthenium complex KP1019 in yeast.

Low solubility in aqueous solutions is a significant limitation of the otherwise promising anticancer ruthenium complex KP1019. In laboratory studies, this challenge is often overcome by using DMSO to help drive the drug into solution. Since DMSO was previously shown to alter the bioactivity of platinum-based chemotherapeutics, here we examine DMSO's effects on KP1019. Using Saccharomyces cerevisiae as a model organism, we apply multiple measures of growth inhibition to demonstrate that DMSO reduces the drug's toxicity. This reduction in bioactivity correlates with spectrophotometric changes consistent with DMSO-dependent increases in the stability of the KP1019 pro-drug. The impact of DMSO on the biology and chemistry of KP1019 suggests this solvent should not be used in studies of this and similar anticancer ruthenium complexes.

Critical Consciousness as a Framework for Health Equity-Focused Peer Learning.

Introduction: Recognizing the need to teach concepts of health equity, diversity, and inclusion as a part of medical students' preclinical training, we developed a series of workshops in the first year of medical school that introduced students to issues of discrimination and inequity and their effects on health outcomes. This student-led, faculty-supported project, known as Critical Consciousness in Medicine (CCM), adopted critical consciousness as a guiding principle for student learning. Methods: Over the course of the 2018-2019 academic year, student leaders developed and delivered five 2-hour workshops to 197 first-year students, with the assistance of student facilitators and input and guidance from faculty advisors. Workshops involved a mix of whole-class presentations and small-group discussions. Session topics included identity and interpersonal relationships, privilege, health disparities, and implicit bias. Results: Paired t-test analysis showed statistically significant growth in student self-ratings related to CCM learning objectives as measured in the end-of-year pre-/postsurvey. Student comments in year-end reflections further suggested learning, self-assessment, growth, and appreciation for the workshops' place in the preclinical curriculum. Discussion: This project modeled a student-faculty partnership for approaching diversity, inclusion, and health equity in medical education and highlighted the role of students as leaders in educating their peers. The CCM workshop series demonstrated high acceptability as a component of preclinical medical education and may increase student engagement around social issues in health care. CCM also illustrated the promise of using critical consciousness as an approach to educating medical students about equity, diversity, and inclusion.

Enfortumab vedotin after PD-1 or PD-L1 inhibitors in cisplatin-ineligible patients with advanced urothelial carcinoma (EV­201): a multicentre, single-arm, phase 2 trial.

BACKGROUND: Locally advanced or metastatic urothelial carcinoma is generally incurable and has scarce treatment options, especially for cisplatin-ineligible patients previously treated with PD-1 or PD-L1 therapy. Enfortumab vedotin is an antibody-drug conjugate directed at Nectin-4, a protein highly expressed in urothelial carcinoma. We aimed to evaluate the efficacy and safety of enfortumab vedotin in the post-immunotherapy setting in cisplatin-ineligible patients. METHODS: EV-201 is a multicentre, single-arm, phase 2 study of enfortumab vedotin in patients with locally advanced or metastatic urothelial carcinoma previously treated with PD-1 or PD-L1 inhibitors. Cohort 2 included adults (aged ≥18 years) with an Eastern Cooperative Oncology Group performance status score of 2 or less who were considered ineligible for cisplatin at enrolment and who had not received platinum-containing chemotherapy in the locally advanced or metastatic setting. Enfortumab vedotin was given intravenously at a dose of 1·25 mg/kg on days 1, 8, and 15 of every 28-day cycle. The primary endpoint was confirmed objective response rate per Response Evaluation Criteria in Solid Tumours version 1.1 assessed by blinded independent central review. Efficacy and safety were analysed in all patients who received at least one dose of enfortumab vedotin. EV-201 is an ongoing study and the primary analysis is complete. This study is registered with Clinicaltrials.gov, NCT03219333. FINDINGS: Between Oct 8, 2017, and Feb 11, 2020, 91 patients were enrolled at 40 sites globally, of whom 89 received treatment. Median follow-up was 13·4 months (IQR 11·3-18·9). At data cutoff (Sept 8, 2020), the confirmed objective response rate was 52% (46 of 89 patients; 95% CI 41-62), with 18 (20%) of 89 patients achieving a complete response and 28 (31%) achieving a partial response. 49 (55%) of 89 patients had grade 3 or worse treatment-related adverse events. The most common grade 3 or 4 treatment-related adverse events were neutropenia (eight [9%] patients), maculopapular rash (seven [8%] patients), and fatigue (six [7%] patients). Treatment-related serious adverse events occurred in 15 (17%) patients. Three (3%) patients died due to acute kidney injury, metabolic acidosis, and multiple organ dysfunction syndrome (one [1%] each) within 30 days of first dose and these deaths were considered by the investigator to be related to treatment; a fourth death from pneumonitis occurred more than 30 days after the last dose and was also considered to be related to treatment. INTERPRETATION: Treatment with enfortumab vedotin was tolerable and confirmed responses were seen in 52% of cisplatin-ineligible patients with locally advanced or metastatic urothelial carcinoma who were previously treated with PD-1 or PD-L1 inhibitors. These patients have few treatment options, and enfortumab vedotin could be a promising new therapy for a patient population with a high unmet need. FUNDING: Astellas Pharma Global Development and Seagen.

Enfortumab Vedotin in Previously Treated Advanced Urothelial Carcinoma.

BACKGROUND: Patients with advanced urothelial carcinoma have poor overall survival after platinum-containing chemotherapy and programmed cell death protein 1 (PD-1) or programmed death ligand 1 (PD-L1) inhibitor treatment. METHODS: We conducted a global, open-label, phase 3 trial of enfortumab vedotin for the treatment of patients with locally advanced or metastatic urothelial carcinoma who had previously received platinum-containing chemotherapy and had had disease progression during or after treatment with a PD-1 or PD-L1 inhibitor. Patients were randomly assigned in a 1:1 ratio to receive enfortumab vedotin (at a dose of 1.25 mg per kilogram of body weight on days 1, 8, and 15 of a 28-day cycle) or investigator-chosen chemotherapy (standard docetaxel, paclitaxel, or vinflunine), administered on day 1 of a 21-day cycle. The primary end point was overall survival. RESULTS: A total of 608 patients underwent randomization; 301 were assigned to receive enfortumab vedotin and 307 to receive chemotherapy. As of July 15, 2020, a total of 301 deaths had occurred (134 in the enfortumab vedotin group and 167 in the chemotherapy group). At the prespecified interim analysis, the median follow-up was 11.1 months. Overall survival was longer in the enfortumab vedotin group than in the chemotherapy group (median overall survival, 12.88 vs. 8.97 months; hazard ratio for death, 0.70; 95% confidence interval [CI], 0.56 to 0.89; P = 0.001). Progression-free survival was also longer in the enfortumab vedotin group than in the chemotherapy group (median progression-free survival, 5.55 vs. 3.71 months; hazard ratio for progression or death, 0.62; 95% CI, 0.51 to 0.75; P<0.001). The incidence of treatment-related adverse events was similar in the two groups (93.9% in the enfortumab vedotin group and 91.8% in the chemotherapy group); the incidence of events of grade 3 or higher was also similar in the two groups (51.4% and 49.8%, respectively). CONCLUSIONS: Enfortumab vedotin significantly prolonged survival as compared with standard chemotherapy in patients with locally advanced or metastatic urothelial carcinoma who had previously received platinum-based treatment and a PD-1 or PD-L1 inhibitor. (Funded by Astellas Pharma US and Seagen; EV-301 ClinicalTrials.gov number, NCT03474107.).

Occupational Stress and Anger: Mediating Effects of Resiliency in First Responders.

First responders experience substantial stress due to the nature of their work (Carleton et al. 2017). Occupational stress (OS) results from a myriad of employment conditions (e.g., ambiguous work expectations, unreasonable workload; Osipow 1998). OS can lead to maladaptive anger, which negatively impacts personal well-being and work performance (Velichkovsky 2009). In contrast, resilience to demanding working conditions is associated with lower state and trait anger (Wilson et al. 2001); thus, resilience may serve a protective 'buffer' role against anger in the face of stress. Thus, we hypothesized that resiliency would mediate relations between dimensions of OS and anger. The current study included 201 first responders (male = 77.6%; M age = 43.73 years (SD = 10.97); police officers = 64.2%) who completed measures of OS (OSI-R; Osipow 1998), Anger (DSM-5 CC Anger; APA 2013), and Resiliency (CD-RISC; Connor and Davidson 2003). Results indicated that resiliency mediated relations between five components of OS and anger: Role Overload (p < .001); Insufficiency (p < .001); Role Boundary (p < .001); Role Ambiguity (p < .001); and Role Responsibility (p < .001). Results support the importance of resiliency-enhancing interventions to offset the experience of anger when confronted with occupational stress in first responders.

Multiplex immunofluorescence to measure dynamic changes in tumor-infiltrating lymphocytes and PD-L1 in early-stage breast cancer.

BACKGROUND: The H&E stromal tumor-infiltrating lymphocyte (sTIL) score and programmed death ligand 1 (PD-L1) SP142 immunohistochemistry assay are prognostic and predictive in early-stage breast cancer, but are operator-dependent and may have insufficient precision to characterize dynamic changes in sTILs/PD-L1 in the context of clinical research. We illustrate how multiplex immunofluorescence (mIF) combined with statistical modeling can be used to precisely estimate dynamic changes in sTIL score, PD-L1 expression, and other immune variables from a single paraffin-embedded slide, thus enabling comprehensive characterization of activity of novel immunotherapy agents. METHODS: Serial tissue was obtained from a recent clinical trial evaluating loco-regional cytokine delivery as a strategy to promote immune cell infiltration and activation in breast tumors. Pre-treatment biopsies and post-treatment tumor resections were analyzed by mIF (PerkinElmer Vectra) using an antibody panel that characterized tumor cells (cytokeratin-positive), immune cells (CD3, CD8, CD163, FoxP3), and PD-L1 expression. mIF estimates of sTIL score and PD-L1 expression were compared to the H&E/SP142 clinical assays. Hierarchical linear modeling was utilized to compare pre- and post-treatment immune cell expression, account for correlation of time-dependent measurement, variation across high-powered magnification views within each subject, and variation between subjects. Simulation methods (Monte Carlo, bootstrapping) were used to evaluate the impact of model and tissue sample size on statistical power. RESULTS: mIF estimates of sTIL and PD-L1 expression were strongly correlated with their respective clinical assays (p < .001). Hierarchical linear modeling resulted in more precise estimates of treatment-related increases in sTIL, PD-L1, and other metrics such as CD8+ tumor nest infiltration. Statistical precision was dependent on adequate tissue sampling, with at least 15 high-powered fields recommended per specimen. Compared to conventional t-testing of means, hierarchical linear modeling was associated with substantial reductions in enrollment size required (n = 25➔n = 13) to detect the observed increases in sTIL/PD-L1. CONCLUSION: mIF is useful for quantifying treatment-related dynamic changes in sTILs/PD-L1 and is concordant with clinical assays, but with greater precision. Hierarchical linear modeling can mitigate the effects of intratumoral heterogeneity on immune cell count estimations, allowing for more efficient detection of treatment-related pharmocodynamic effects in the context of clinical trials. TRIAL REGISTRATION: NCT02950259 .

"Sow, Grow, Know, and Show": The Impact of School Gardens on Student Self-Perception in the Mississippi Delta.

This research evaluates the impact of participation in school garden programs on fifth grade student garden knowledge, self-perception, and standardized test scores in the Mississippi Delta. We collected pre and posttest participant data for two years, compared garden participants' standardized test scores with non-participants, and conducted participant observation and interviews in three school gardens for eight months during the 2017-2018 school year. While no effect on standardized test scores could be determined, students increased garden knowledge and reported feeling positive about their future, teamwork, and leadership ability. These results were supported by observations of student joy, confidence, leadership, and teamwork.

Nucleotide P2Y1 receptor agonists are in vitro and in vivo prodrugs of A1/A3 adenosine receptor agonists: implications for roles of P2Y1 and A1/A3 receptors in physiology and pathology.

Rapid phosphoester hydrolysis of endogenous purine and pyrimidine nucleotides has challenged the characterization of the role of P2 receptors in physiology and pathology. Nucleotide phosphoester stabilization has been pursued on a number of medicinal chemistry fronts. We investigated the in vitro and in vivo stability and pharmacokinetics of prototypical nucleotide P2Y1 receptor (P2Y1R) agonists and antagonists. These included the riboside nucleotide agonist 2-methylthio-ADP and antagonist MRS2179, as well as agonist MRS2365 and antagonist MRS2500 containing constrained (N)-methanocarba rings, which were previously reported to form nucleotides that are more slowly hydrolyzed at the α-phosphoester compared with the ribosides. In vitro incubations in mouse and human plasma and blood demonstrated the rapid hydrolysis of these compounds to nucleoside metabolites. This metabolism was inhibited by EDTA to chelate divalent cations required by ectonucleotidases for nucleotide hydrolysis. This rapid hydrolysis was confirmed in vivo in mouse pharmacokinetic studies that demonstrate that MRS2365 is a prodrug of the nucleoside metabolite AST-004 (MRS4322). Furthermore, we demonstrate that the nucleoside metabolites of MRS2365 and 2-methylthio-ADP are adenosine receptor (AR) agonists, notably at A3 and A1ARs. In vivo efficacy of MRS2365 in murine models of traumatic brain injury and stroke can be attributed to AR activation by its nucleoside metabolite AST-004, rather than P2Y1R activation. This research suggests the importance of reevaluation of previous in vitro and in vivo research of P2YRs and P2XRs as there is a potential that the pharmacology attributed to nucleotide agonists is due to AR activation by active nucleoside metabolites.

Effect of various decontamination procedures on disposable N95 mask integrity and SARS-CoV-2 infectivity.

INTRODUCTION: The COVID-19 pandemic has created a high demand on personal protective equipment, including disposable N95 masks. Given the need for mask reuse, we tested the feasibility of vaporized hydrogen peroxide (VHP), ultraviolet light (UV), and ethanol decontamination strategies on N95 mask integrity and the ability to remove the infectious potential of SARS-CoV-2. METHODS: Disposable N95 masks, including medical grade (1860, 1870+) and industrial grade (8511) masks, were treated by VHP, UV, and ethanol decontamination. Mask degradation was tested using a quantitative respirator fit testing. Pooled clinical samples of SARS-CoV-2 were applied to mask samples, treated, and then either sent immediately for real-time reverse transcriptase-polymerase chain reaction (RT-PCR) or incubated with Vero E6 cells to assess for virucidal effect. RESULTS: Both ethanol and UV decontamination showed functional degradation to different degrees while VHP treatment showed no significant change after two treatments. We also report a single SARS-CoV-2 virucidal experiment using Vero E6 cell infection in which only ethanol treatment eliminated detectable SARS-CoV-2 RNA. CONCLUSIONS: We hope our data will guide further research for evidenced-based decisions for disposable N95 mask reuse and help protect caregivers from SARS-CoV-2 and other pathogens.

Living race together: the role of partner's race in racial/ethnic differences in smoking.

Objective: Crossing racial lines provides a unique context for understanding racial patterns in smoking. This research explores whether adults whose unions cross racial lines behave more similarly to their own group or their partner'sDesign: Using a sample of respondents from the National Health Interview Survey (2001-2011), we compare the likelihood of current smoking and quitting smoking among adults in mixed-race unions to adults in same-race unions.Results: Adults with different-race partners generally mirror their partner's group; people of color with White partners have a higher likelihood of being current smokers, similar to Whites, while Whites partnered with Asians and Latina/os are, like other Asians and Latino/as, less likely to smoke. There are fewer differences in the likelihood of quitting smoking.

Measuring perceived mistreatment across diverse social groups: An evaluation of the Everyday Discrimination Scale.

RATIONALE: Research assessing the health-related consequences of perceived discrimination depends upon high quality measures of perceived discrimination. The Everyday Discrimination Scale (EDS) is among the most frequently used instruments to assess perceptions of discrimination in general, as well as specific types of discrimination (e.g., based on race/ethnicity or age). While numerous studies attest to its validity and reliability for racial/ethnic minority groups, no existing study has examined its psychometric equivalence across gender, age, or socio-economic groups. This study fills this gap. HYPOTHESIS: We hypothesize that because social hierarchies of race/ethnicity, age, gender and class have different histories and are differently organized and institutionalized in contemporary United States, racial/ethnic, age, gender, and education-based groups differ in the types of discrimination they experience and perceive. As a result, the EDS may not be equivalent across these social groups. METHOD: We test this hypothesis by analyzing data from the 2015 US Texas Diversity Study (N=1,049), a telephone survey of English- and Spanish-speaking adults. We examine two forms of the EDS - one focusing on discrimination regardless of attribution and one focusing specifically on discrimination attributed to respondents' race/ethnicity. RESULTS: Multi-group confirmatory factor analyses revealed that neither version of the scale generates estimates of discrimination that can be meaningfully compared across all racial/ethnic, age, gender, and education-based groups. CONCLUSIONS: Our results urge caution when drawing comparisons of perceived discrimination across diverse social groups based on the EDS and point to avenues for future scale development.