Adjustment For Multiple Comparisons Research Articles

Plasma Metabolomic Analysis Reveals the Relationship between Immune Function and Metabolic Changes in Holstein Peripartum Dairy Cows.

Dairy cows undergo dynamic physiological changes from late gestation to early lactation, including metabolic changes and immune dysfunction. The aim of this study was to investigate the relationship between immune function and metabolic changes in peripartum dairy cows. Fifteen healthy Holstein dairy cows were enrolled 14 days prior to parturition, and plasma was collected on day −7, 0, 7, and 21 relative to calving. Plasma non-esterified fatty acids (NEFAs), glucose, β-hydroxybutyric acid (BHBA), immunoglobulin G (IgG), tumor necrosis factor alpha (TNF-α), and interleukin-2 levels were measured, and metabolic profiles were determined using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry. The data were analyzed using Tukey–Kramer adjustment for multiple comparisons, and multivariate and univariate statistical analyses were performed to screen for differential metabolites. The results showed that the concentrations of NEFAs, glucose, BHBA, and TNF-α in the plasma significantly increased and concentrations of IgG and interleukin-2 in plasma significantly decreased from −7 d to the calving day (p < 0.05). Additionally, the concentrations of glucose, IgG, and TNF-α significantly decreased from 0 to +7 d, and concentrations of NEFAs decreased significantly from +7 to +21 d (p < 0.05). The following six primary metabolic pathways were identified in all time point comparisons, and L-glutamate, linoleic acid, taurine, and L-tryptophan were involved in these major metabolic pathways. Correlation and pathway analyses indicated that a negative energy balance during the transition period adversely affects immune responses in cows, and L-tryptophan exerts immunomodulatory effects through the Trp-Kyn pathway, resulting in depletion of Trp and elevation of Kyn.

Evaluation of microleakage and push-out bond strength of various composite resins for sealing the screw-access channel in implant-supported restorations

Statement of problemMicroleakage and loss of the composite resin sealing the screw-access channel are frequent complications of screw-retained implant-supported prostheses. How the screw-access channel should be best restored to reduce such complications is unclear. PurposeThe purpose of this in vitro study was to evaluate the microleakage and bond strength of 3 types of composite resins (flowable, packable, and bulk-fill) with or without a bonding agent treatment to seal the screw-access channel of 2 types of restorative materials (zirconia and Co-Cr alloy) with or without thermocycling. Material and methodsIn total, 240 yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) specimens (IPS e.max ZirCAD) and 240 Co-Cr alloy (Vera PDS) specimens were prepared with a Ø3×3-mm cylindrical cavity at the center to simulate the screw-access channel. Three types of composite resins (flowable, packable, and bulk-fill resin) (Filtek Z350 XT Flowable Restorative, Filtek Z350 XT Universal Restorative, and Filtek One Bulk Fill Restorative) were applied to restore the access channel of the zirconia and Co-Cr specimens with or without a bonding agent (Single Bond Universal Adhesive). Microleakage and push-out bond strength were determined and compared by dividing the specimens into experimental groups with or without thermocycling (1000 times with 30 seconds at 5 ±2 °C and 55 ±2 °C). The results were analyzed by using a 1-way ANOVA and 4-way ANOVA. Adjustment for multiple comparisons was made with the Tukey Honestly Significant Difference (HSD) test. ResultsThe specimens subjected to thermocycling showed a lower bond strength (P<.001) and higher microleakage (P<.001) than specimens stored in a constant-temperature water bath. Specimens treated with bonding agents showed a higher bond strength (P<.001) and lower microleakage (P<.001) than specimens not treated with a bonding agent. Higher bond strengths were observed in the order of bulk-fill resin, packable resin, and flowable resin (P<.001). Packable resin showed higher microleakage than flowable resin and bulk-fill resin (P<.05). No significant difference in microleakage was found between the flowable resin and bulk-fill resin (P>.05). ConclusionsHigher bond strengths were observed in the order of bulk-fill resin, packable resin, and flowable resin. Less microleakage was observed in the flowable resin and bulk-fill resin than in the packable composite resin. Bonding agent treatment was effective in increasing bond strength and decreasing microleakage. Zirconia and Co-Cr showed a bond strength similar to that of composite resins, but zirconia showed higher microleakage than Co-Cr. Restoring the screw-access channel with the bulk-fill resin should increase bond strength and reduce microleakage.

Moderators and Nonspecific Predictors of Treatment Benefits in a Randomized Trial of Mindfulness-Based Stress Reduction vs Cognitive-Behavioral Therapy vs Usual Care for Chronic Low Back Pain

Both mindfulness-based stress reduction (MBSR) and cognitive-behavioral therapy (CBT) are effective for chronic low back pain (CLBP), but little is known regarding who might benefit more from one than the other. Using data from a randomized trial comparing MBSR, CBT, and usual care (UC) for adults aged 20 to 70 years with CLBP (N = 297), we examined baseline characteristics that moderated treatment effects or were associated with improvement regardless of treatment. Outcomes included 8-week function (modified Roland Disability Questionnaire), pain bothersomeness (0–10 numerical rating scale), and depression (Patient Health Questionnaire-8). There were differences in the effects of CBT versus MBSR on pain based on participant gender (P = .03) and baseline depressive symptoms (P = .01), but the only statistically significant moderator after Bonferroni correction was the nonjudging dimension of mindfulness. Scores on this measure moderated the effects of CBT versus MBSR on both function (P = .001) and pain (P = .04). Pain control beliefs (P <.001) and lower anxiety (P < .001) predicted improvement regardless of treatment. Replication of these findings is needed to guide treatment decision-making for CLBP. Trial RegistrationThe trial and analysis plan were preregistered in ClinicalTrials.gov (Identifier: NCT01467843). PerspectiveAlthough few potential moderators and nonspecific predictors of benefits from CBT or MBSR for CLBP were statistically significant after adjustment for multiple comparisons, these findings suggest potentially fruitful directions for confirmatory research while providing reassurance that patients could reasonably expect to benefit from either treatment.

Clinical Decision Support for Glycemic Management Reduces Hospital Length of Stay.

Dysglycemia influences hospital outcomes and resource utilization. Clinical decision support (CDS) holds promise for optimizing care by overcoming management barriers. This study assessed the impact on hospital length of stay (LOS) of an alert-based CDS tool in the electronic medical record that detected dysglycemia or inappropriate insulin use, coined as gaps in care (GIC). Using a 12-month interrupted time series among hospitalized persons aged ≥18 years, our CDS tool identified GIC and, when active, provided recommendations. We compared LOS during 6-month-long active and inactive periods using linear models for repeated measures, multiple comparison adjustment, and mediation analysis. Among 4,788 admissions with GIC, average LOS was shorter during the tool's active periods. LOS reductions occurred for all admissions with GIC (-5.7 h, P = 0.057), diabetes and hyperglycemia (-6.4 h, P = 0.054), stress hyperglycemia (-31.0 h, P = 0.054), patients admitted to medical services (-8.4 h, P = 0.039), and recurrent hypoglycemia (-29.1 h, P = 0.074). Subgroup analysis showed significantly shorter LOS in recurrent hypoglycemia with three events (-82.3 h, P = 0.006) and nonsignificant in two (-5.2 h, P = 0.655) and four or more (-14.8 h, P = 0.746). Among 22,395 admissions with GIC (4,788, 21%) and without GIC (17,607, 79%), LOS reduction during the active period was 1.8 h (P = 0.053). When recommendations were provided, the active tool indirectly and significantly contributed to shortening LOS through its influence on GIC events during admissions with at least one GIC (P = 0.027), diabetes and hyperglycemia (P = 0.028), and medical services (P = 0.019). Use of the alert-based CDS tool to address inpatient management of dysglycemia contributed to reducing LOS, which may reduce costs and improve patient well-being.

Elder abuse in the COVID-19 era based on calls to the National Center on Elder Abuse resource line

BackgroundThe COVID-19 pandemic has exacerbated circumstances that place older adults at higher risk for abuse, neglect, and exploitation. Identifying characteristics of elder abuse during COVID-19 is critically important. This study characterized and compared elder abuse patterns across two time periods, a one-year period during the pandemic, and a corresponding one-year period prior to the start of the pandemic.MethodsContacts (including social media contacts, and email; all referred to as “calls” for expediency) made to the National Center on Elder Abuse (NCEA) resource line were examined for differences in types of reported elder abuse and characteristics of alleged perpetrators prior to the pandemic (Time 1; March 16, 2018 to March 15, 2019) and during the pandemic (Time 2; March 16, 2020 to March 15, 2021). Calls were examined for whether or not abuse was reported, the types of reported elder abuse, including financial, physical, sexual, emotional, and neglect, and characteristics of callers, victims, and alleged perpetrators. Chi-square tests of independence compared frequencies of elder abuse characteristics between time periods.ResultsIn Time 1, 1401 calls were received, of which 795 calls (56.7%) described abuse. In Time 2, 1009 calls were received, of which 550 calls (54.5%) described abuse. The difference between time periods in frequency of abuse to non-abuse calls was not significant (p=0.28). Time periods also did not significantly differ with regard to caller, victim, and perpetrator characteristics. Greater rates of physical abuse ({upchi }^{2}=23.52, p<0.001) and emotional abuse ({upchi }^{2}=7.12, p=0.008) were reported during Time 2 after adjustment for multiple comparisons. An increased frequency of multiple forms of abuse was also found in Time 2 compared to Time 1 ({upchi }^{2}=23.52, p<0.001).ConclusionsFindings suggest differences in specific elder abuse subtypes and frequency of co-occurrence between subtypes between time periods, pointing to a potential increase in the severity of elder abuse during COVID-19.

Methods of Bloodless Care, Clinical Outcomes, and Costs for Adult Patients Who Decline Allogeneic Transfusions.

Providing bloodless medical care for patients who wish to avoid allogeneic transfusion can be challenging; however, previous studies have demonstrated favorable outcomes when appropriate methods are used. Here, we report one of the largest series of patients receiving bloodless care, along with the methods used to provide such care, and the resulting outcomes. In a retrospective cohort study, 1111 adult inpatients (age ≥18 years) at a single institution who declined allogeneic transfusion for religious or personal reasons between June 2012 and June 2016 were included, and the patient blood management methods are described. Patient characteristics, laboratory data, and transfusion rates, as well as clinical outcomes (morbidity, mortality, and length of stay) were compared to all other patients in the hospital who received standard care, including transfusions if needed (n = 137,009). Medical and surgical patients were analyzed as subgroups. The primary outcome was composite morbidity (any morbid event: infectious, thrombotic, ischemic, renal, or respiratory). Secondary outcomes included individual morbid events, in-hospital mortality, length of stay, total hospital charges, and costs. The bloodless cohort had more females and a lower case mix index, but more preadmission comorbidities. Mean nadir hemoglobin during hospitalization was lower in the bloodless (9.7 ± 2.6 g/dL) compared to the standard care (10.1 ± 2.4 g/dL) group (P < .0001). Composite morbidity occurred in 14.4% vs 16.0% (P = .16) of the bloodless and standard care patients, respectively. Length of stay and in-hospital mortality were similar between the bloodless and standard care patients. After Bonferroni adjustment for multiple comparisons, hospital-acquired infection occurred less frequently in the bloodless compared to the standard care cohort (4.3% vs 8.3%) (P < .0001) in the medical patient subgroup, but not in the surgical subgroup. After propensity score adjustment in a multivariable model and adjustment for multiple comparisons, bloodless care was associated with less risk of hospital-acquired infection (OR, 0.56; 95% CI, 0.35-0.83; P = .0074) in the medical subgroup, but not in the surgical subgroup. Median total hospital charges (by 8.5%; P = .0017) and costs (by 8.7%; P = .0001) were lower in the bloodless compared to the standard care cohort, when all patients were included. Overall, adult patients receiving bloodless care had similar clinical outcomes compared to patients receiving standard care. Medical (but not surgical) bloodless patients may be at less risk for hospital-acquired infection compared to those receiving standard care. Bloodless care is cost-effective and should be considered as high-value practice.

Enhancing Anticoagulation Monitoring and Therapy in Patients Undergoing Microvascular Reconstruction in Maxillofacial Surgery: A Prospective Observational Trial.

Background: In reconstructive surgery, loss of a microvascular free flap due to perfusion disorders, especially thrombosis, is a serious complication. In recent years, viscoelastic testing (VET) has become increasingly important in point-of-care (POC) anticoagulation monitoring. This paper describes a protocol for enhanced anticoagulation monitoring during maxillofacial flap surgery. Objective: The aim of the study will be to evaluate, in a controlled setting, the predictive value of POC devices for the type of flap perfusion disorders due to thrombosis or bleeding. VET, Platelet monitoring (PM) and standard laboratory tests (SLT) are comparatively examined. Methods/Design: This study is an investigator-initiated prospective trial in 100 patients undergoing maxillofacial surgery. Patients who undergo reconstructive surgery using microvascular-free flaps will be consecutively enrolled in the study. All patients provide blood samples for VET, PM and SLT at defined time points. The primary outcome is defined as free flap loss during the hospital stay. Statistical analyses will be performed using t-tests, including the Bonferroni adjustment for multiple comparisons. Discussion: This study will help clarify whether VET can improve individualized patient care in reconstruction surgery. A better understanding of coagulation in relation to flap perfusion disorders may allow real-time adaption of antithrombotic strategies and potentially prevent flap complications.

Multigroup Analysis in Information Systems Research using PLS-PM

Heterogeneity is a pertinent issue in Information Systems (IS) research because human behavior often differs across groups. In the partial least squares path modeling (PLS-PM) context, several approaches have been proposed to investigate potential group differences. Despite the availability of numerous approaches, literature that compares their efficacy is sparse. Consequently, IS researchers lack guidance on which approach is best suited to detect group differences. We address this issue by presenting the results of an extensive Monte Carlo simulation study that juxtaposes the various approaches' behavior under numerous conditions. In doing so, we first provide an overview on existing approaches proposed for multigroup analysis (MGA) in the PLS-PM context. Moreover, we derive important implications for applied research: Firstly, we show that the omnibus test of group differences (OTG) and approaches based on the comparison of confidence intervals are not recommendable for MGA. Secondly, we provide detailed information as to which approaches are suitable for comparing one specific path coefficient and which are recommended if the complete structural model is compared across groups. Finally, we show that approaches which are designed to compare a single parameter require an adjustment for multiple comparisons when used to compare more than two groups.

215 Cutaneous immune-related adverse events predict longer patient survival in advanced cancer patients

Cutaneous immune-related adverse events (cirAEs) are the most common toxicities to occur in the setting of immune checkpoint inhibitor (ICI) therapy. However, their impact on mortality is largely understudied. Here, we investigate the impact of cirAEs on survival among ICI recipients using a robust multi-institutional clinical registry. 3,731 ICI recipients were identified from the Mass General Brigham Healthcare System and the Dana-Farber Cancer Institute, of which 676 developed cirAEs and 3,055 did not. Landmark survival analyses and time-varying Cox proportional hazards modeling were performed, adjusting for demographics, Charlson comorbidity index, cancer type, ICI target, and year of ICI initiation. Overall, patients who developed cirAEs were significantly protected from mortality (HR=0.87, p=0.036), particularly in the setting of melanoma (HR=0.67, p=0.007). Among individual morphologies, lichenoid eruption (HR=0.46, p<0.001), psoriasiform eruption (HR=0.43, p=0.004), vitiligo (HR=0.26, p=0.012), and non-specific rash (HR=0.69, p<0.001) were significantly protective of mortality after multiple comparisons adjustment. Our results demonstrate that CirAE development is associated with a 13% reduction in mortality among all cancers and a 33% reduction in melanoma patients. Furthermore, almost all cirAE morphologies are associated with better survival. Even after excluding vitiligo, cirAE development remains protective among all ICI recipients and melanoma patients. These findings are important as cirAEs tend to occur early after ICI initiation and may serve as predictive biomarkers of therapeutic response.

Multi-dimensional characterization of prediabetes in the Project Baseline Health Study

BackgroundWe examined multi-dimensional clinical and laboratory data in participants with normoglycemia, prediabetes, and diabetes to identify characteristics of prediabetes and predictors of progression from prediabetes to diabetes or reversion to no diabetes.MethodsThe Project Baseline Health Study (PBHS) is a multi-site prospective cohort study of 2502 adults that conducted deep clinical phenotyping through imaging, laboratory tests, clinical assessments, medical history, personal devices, and surveys. Participants were classified by diabetes status (diabetes [DM], prediabetes [preDM], or no diabetes [noDM]) at each visit based on glucose, HbA1c, medications, and self-report. Principal component analysis (PCA) was performed to create factors that were compared across groups cross-sectionally using linear models. Logistic regression was used to identify factors associated with progression from preDM to DM and for reversion from preDM to noDM.ResultsAt enrollment, 1605 participants had noDM; 544 had preDM; and 352 had DM. Over 4 years of follow-up, 52 participants with preDM developed DM and 153 participants reverted to noDM. PCA identified 33 factors composed of clusters of clinical variables; these were tested along with eight individual variables identified a priori as being of interest. Six PCA factors and six a priori variables significantly differed between noDM and both preDM and DM after false discovery rate adjustment for multiple comparisons (q < 0.05). Of these, two factors (one comprising glucose measures and one of anthropometry and physical function) demonstrated monotonic/graded relationships across the groups, as did three a priori variables: ASCVD risk, coronary artery calcium, and triglycerides (q < 10–21 for all). Four factors were significantly different between preDM and noDM, but concordant or similar between DM and preDM: red blood cell indices (q = 8 × 10-10), lung function (q = 2 × 10-6), risks of chronic diseases (q = 7 × 10-4), and cardiac function (q = 0.001), along with a priori variables of diastolic function (q = 1 × 10-10), sleep efficiency (q = 9 × 10-6) and sleep time (q = 6 × 10-5). Two factors were associated with progression from prediabetes to DM: anthropometry and physical function (OR [95% CI]: 0.6 [0.5, 0.9], q = 0.04), and heart failure and c-reactive protein (OR [95% CI]: 1.4 [1.1, 1.7], q = 0.02). The anthropometry and physical function factor was also associated with reversion from prediabetes to noDM: (OR [95% CI]: 1.9 [1.4, 2.7], q = 0.02) along with a factor of white blood cell indices (OR [95% CI]: 0.6 [0.4, 0.8], q = 0.02), and the a priori variables ASCVD risk score (OR [95% CI]: 0.7 [0.6, 0.9] for each 0.1 increase in ASCVD score, q = 0.02) and triglycerides (OR [95% CI]: 0.9 [0.8, 1.0] for each 25 mg/dl increase, q = 0.05).ConclusionsPBHS participants with preDM demonstrated pathophysiologic changes in cardiac, pulmonary, and hematology measures and declines in physical function and sleep measures that precede DM; some changes predicted an increased risk of progression to DM. A factor with measures of anthropometry and physical function was the most important factor associated with progression to DM and reversion to noDM. Future studies may determine whether these changes elucidate pathways of progression to DM and related complications and whether they can be used to identify individuals at higher risk of progression to DM for targeted preventive interventions.Trial registration ClinicalTrials.gov NCT03154346

Prediagnostic plasma polyphenol concentrations and colon cancer risk: The JPHC nested case–control study

Epidemiological studies that assessed the associations between dietary polyphenol intakes and colon cancer risk have reported largely null results, possibly due to measurement error associated with dietary assessment. We adopted an objective approach by measuring prediagnostic plasma concentrations of 35 polyphenols and assessing associations with colon cancer risk. We conducted a nested-case control study within the Japan Public Health Center-based prospective study (JPHC Study) utilizing plasma samples collected at the time of a five-year follow-up survey between 1995 and 1999. We identified colon cancer cases who developed cancer during the follow-up from the time of blood collection. Controls were matched by age, sex, area code, population size of the area, season of blood collection, year of blood collection, and duration of fasting time before the blood collection. Prediagnostic concentrations of 35 polyphenols from 375 incident colon cancer cases (followed until 2012) and 710 matched controls were measured by tandem mass spectrometry coupled with ultra-high-pressure liquid chromatography. We used multivariable conditional logistic regression models adjusted for established colon cancer risk factors to estimate odds ratios (ORs) and 95% confidence intervals (CIs). In sexes combined log2-transformed multivariable models, circulating levels of 3,4-dihydroxyphenylpropionic acid (P=0.02), ferulic acid (P=0.02), and caffeic acid (P=0.03) were inversely, and 3-hydroxybenzoic acid (P=0.03) was positively, associated with colon cancer risk. For men only, circulating levels of 3,4-dihydroxyphenylpropionic acid was inversely, and 3,5-dihydroxyphenylpropionic acid, gallic acid, (+)-epigallocatechin, 3-hydroxybenzoic acid, and epicatechin were positively, associated with colon cancer risk. In women, plasma caffeic acid and ferulic acid concentration were inversely associated with colon cancer risk. However, all these associations were nonsignificant after adjustment for multiple comparisons. The remaining polyphenols were not associated with colon cancer risk. Coffee-derived 3,4-dihydroxyphenylpropionic acid, ferulic acid, and caffeic acid concentrations were inversely associated with colon cancer risk although the association were nonsignificant after adjustment for multiple comparisons. These results support a possible role of coffee polyphenols in preventing colorectal cancer.

Occupational and environmental exposures in the Genomic Research in Alpha-1 Antitrypsin Deficiency and Sarcoidosis (GRADS) study

IntroductionSarcoidosis is a granulomatous disorder thought to be caused by exposures in genetically susceptible individuals. This study investigated whether specific exposures were associated with different sarcoidosis phenotypes. MethodsExtensive demographic, occupational and environmental exposure data was analyzed from subjects enrolled in the NHLBI Genomic Research in Alpha-1 Antitrypsin Deficiency and Sarcoidosis (GRADS) study. ResultsIn patients with sarcoidosis, radiation exposure was significantly associated with an increased risk of cardiac sarcoidosis versus non-cardiac sarcoidosis. No exposures were significantly associated with pulmonary only disease versus extrapulmonary disease with or without pulmonary involvement, Scadding Stage II/III/IV versus Scadding Stage 0/I, acute or remitting disease versus non-acute or non-remitting disease, nor chronic versus non-chronic disease. Although not reaching statistically significance after adjustment for multiple comparisons, there were a number of exposures associated with specific disease phenotypes, including exposures where relationships to sarcoidosis have previously been described such as rural exposures and pesticide exposures. ConclusionsRadiation exposure may be a risk factor for cardiac sarcoidosis. Other exposures may also be associated with specific phenotypes and should be further explored. The study was limited by small groups of exposed subjects for individual exposures and multiple comparisons. The development of novel and innovative exposure assessment tools is needed.

Lipoprotein profiles associated with exposure to poly- and perfluoroalkyl substances (PFASs) in the EuroMix human biomonitoring study

Exposure to per- and polyfluoroalkyl substances (PFASs) is associated with increased blood cholesterol. Although elevated cholesterol is a well-established risk factor for cardiovascular diseases (CVD), it is not clear whether PFASs affect this risk. Lipoprotein subclasses are emerging biomarkers for disease risk and lipoprotein profiling may provide an insight to physiological implications of PFAS exposure. We explored the association between serum PFAS concentrations and lipoprotein subclasses in a cross-sectional study. We determined the concentrations and lipid composition of the major subclasses of lipoproteins in plasma samples from 127 adult participants of the EuroMix human biomonitoring study by nuclear magnetic resonance (NMR). Serum concentrations of 17 PFASs showed a detection frequency between 30 and 100% and were included in further analyses. We examined the associations between PFAS concentrations and lipoprotein subclasses by linear mixed-effect regression models, adjusted for confounders. In the adjusted models, positive associations were found between several PFASs and cholesterol concentrations in large to medium sized HDL and medium sized LDL particles. We found a 4–12% increase in HDL cholesterol per interquartile range (IQR) increase for several PFASs. In women the associations with PFNA, PFUnDA, PFDoDA and PFOS were significant after adjustment for multiple comparisons. Similar magnitude of change was observed between longer chained PFASs and LDL cholesterol, and a few of these associations reached significance for cholesterol in large to medium LDL particle sizes in women. No significant associations with plasma triglycerides were observed. However, most PFASs tended to be associated with reduction in VLDL (very low-density lipoproteins) particle number and VLDL triglyceride. Findings from this exploratory study, suggest that background PFAS exposures influence particle size distributions and lipid composition of plasma lipoprotein subclasses, and that these effects may be more prominent in women. A two-points lipoprofiling for all subjects indicated both low intra-individual variability and good analytical reproducibility.

Abstract 5610: Targeting Rb1 in experimental pancreatic cancer remodels immune cell landscape and suppresses tumor progression

Abstract Background: Pancreatic ductal adenocarcinoma (PDA) remains among the cancers with the poorest prognosis. Currently, no effective treatment available for PDA. Immunotherapy that has been shown effective for several cancer types is not successful for PDA treatment overall. The possible reasons of this PDA immunotherapy resistance include lack of neoantigen mutations, low T cell responses and overall immunosuppression state in the tumor microenvironment mediated by myeloid cells and tumor stroma. Recent studies suggest that targeting tumor microenvironment can overcome this immunotherapy resistance and significantly improve the disease outcome. Retinoblastoma (Rb1) protein is a known tumor suppressor protein. In addition, there is accumulating evidence for its role in immune cell response as well, including regulation of apoptosis in myeloid cells. However, the mechanisms of this phenomenon remain not clear. Here we show that targeting Rb1 with a novel molecule AP-3-84 developed in our lab causes apoptosis of tumor-associated macrophages (TAMs) in the model of PDA, induces T cell infiltration of the tumor and decreases tumor burden. Results: Recently our lab has developed the new small molecule AP-3-84 that is able to bind Rb1 and modulate its activity. Originally, we found that this Rb1 modulator can induce cell death of TAMs and thioglycolate-induced macrophages, but not of the other cell types (neither tumor nor T cells). Gene expression changes upon AP-3-84 treatment showed the induction of oxidative and ER stress programs with a clear activation of p53-dependent genes and caspases. We tested the use of AP-3-84 as immunotherapy against PDA cancer growth using 2 PDA cell lines intrinsically characterized by low T cell and high T cell infiltration (T-low-PDA and T-high-PDA, respectively, originally generated and characterized by J. Li et al., Immunity 2018). In accordance with our in vitro observation, AP-3-84 was able to reshape immune cell landscape in mice with T-high-PDA by depleting macrophages and inducing T cell infiltration. Importantly, those changes were accompanied by significant reduction of tumor growth. Addition of AP-3-84 to immunotherapy (anti-CD40/PD1/CTL4) was also able to decrease tumor burden in T-low-PDA model, which is otherwise resistant to this immunotherapy. Thus, AP-3-84 treatment was able to significantly improve the outcome of experimental PDA by altering immune cell subsets and represent a prospective clinical approach for the clinic. Statistical analysis of the data was conducted using unpaired Student t-test with additional false discovery rate adjustment for multiple comparisons for gene expression analysis. Conclusions: Modulation of Rb1 in myeloid cells is a novel promising target for PDA treatment. This approach reduces PDA tumor burden by shifting the immune cell landscape towards greater T cell infiltration upon tumor macrophage depletion. Citation Format: Evgenii N. Tcyganov, Taekyoung Kwak, Adi Narayana Reddy Poli, Laxminarasimha Donthireddy, Joel Cassel, Andrew Kossenkov, Rahul Shinde, Ben Z. Stanger, Joseph Salvino, Luis J. Montaner. Targeting Rb1 in experimental pancreatic cancer remodels immune cell landscape and suppresses tumor progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5610.

Association of Microbial Tryptophan Metabolites and Indoleamine-2,3-Dioxygenase with Immune Activity in Healthy Adults

ObjectivesTryptophan (Trp) metabolites from intestinal bacteria, including indole, indole acetic acid (IAA) and indole propionic acid (IPA), and from the mammalian indoleamine 2,3-dioxygenase (IDO) pathway, including kynurenine (Kyn), can regulate immune activation. IDO activity in vivo is measured by the plasma Kyn/Trp ratio. In this study we hypothesized that (1) the plasma Kyn/Trp ratio, (2) plasma indole, IPA, and IAA concentrations, and (3) intestinal bacteria associated with indole metabolism, would all be associated with inflammation and immune activation markers in a study population of healthy adults.MethodsA group of 88 immune markers was assessed using plasma, flow cytometric analysis of whole blood and of peripheral blood mononuclear cells (PBMC), and ex vivo culture of PBMC, in 362 healthy, fasting adults. Plasma Trp metabolites were also measured. Bacterial taxa from stool were identified by 16S rRNA gene analysis and those associated with plasma Trp metabolites were identified. Multiple linear regression analysis was used to identify statistically significant associations (with p < .05 after adjustment for multiple comparisons) of these metabolites and taxa with immune markers.ResultsSeveral robust positive associations were found between Kyn/Trp and markers of inflammation including neopterin, IP-10, TNF-α, C-reactive protein, and the regulatory cytokine IL-10. A significant positive association was found between the sum of indole and IAA and natural killer T-cells. Three taxa associated with bacterial Trp metabolites were associated with elevated markers of immune activation: the family Lachnospiraceae with higher lymphocyte counts, the genus Dorea with higher production of the type 1 cytokine IFN-γ by T-cells in PBMC culture, the genus Ruminococcus with higher production of the inflammatory cytokine IL-6 in PBMC cultures stimulated with bacterial lipopolysaccharide.ConclusionsIDO activity, bacterial Trp metabolites and indole-associated commensal bacteria were all associated with markers of systemic immune activation, particularly involving inflammatory cytokines, the acute phase response and type 1 innate and adaptive immunity. These findings suggest sub-clinical levels of systemic and/or intestinal immune activity that might be precursors to adverse health outcomes.Funding SourcesUSDA/ARS 2032-51,530-026-00D.

1224-P: Multidimensional Characterization of Prediabetes in the Project Baseline Health Study

Prediabetes is a recognized entity but has been incompletely characterized in a contemporary context. We sought to characterize participants with prediabetes with extensive multi-dimensional clinical and laboratory data and to identify predictors of progression to diabetes. The Project Baseline Health Study (PBHS) is a multi-site prospective U.S. cohort study of 2502 adults with deep clinical phenotyping at enrollment. At enrollment and follow-up, participants were classified by diabetes status (diabetes [DM], prediabetes [preDM], or no diabetes [noDM]) based on glucose, A1c, medications, and self-reported medical history. Principal component analysis (PCA) was performed on 1clinical, survey, and lab variables to create orthogonal factors composed of correlated variables, which were compared between groups using linear models. Logistic regression was used to identify factors associated with progression from preDM to DM. At enrollment, 16participants had noDM; 544 had preDM; and 352 had DM. Over up to 4 years of follow-up, 52 participants with preDM developed DM. PCA identified 33 factors composed of clusters of clinical variables; 24 factors differed significantly by diabetes status: 6 factors significantly differed between noDM and both preDM and DM after adjustment for multiple comparisons, including factors with variables of glucose measures, anthropometry and physical function (q= 1.3 × 10−21) , red blood cell indices (q= 8.3 × 10−10) , lung function (q= 2.0 × 10−6) , cardiac function (q= 0.001) , and risk of chronic disease. Five factors were significantly associated with progression to DM, including factors of anthropometry and physical function [OR (95% CI) 0.6 (0.5,0.9) ], hematologic measures, and cardiac function. Conclusions: PBHS participants with preDM demonstrated pathophysiologic changes in cardiac, pulmonary, and hematology measures and declines in physical function that precede DM and could be markers of higher risk of progression to DM and complications. Disclosure R.Chatterjee montgomery: Research Support; Bristol-Myers Squibb Company, Epigenomics AG. L.C.Kwee: None. N.Pagidipati: Advisory Panel; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, ESPERION Therapeutics, Inc., Consultant; Boehringer Ingelheim International GmbH, Novo Nordisk, Research Support; Amgen Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Eggland's Best, Eli Lilly and Company, Novartis AG, Novo Nordisk, Sanofi, Verily Life Sciences. A.F.Hernandez: Consultant; Merck &amp; Co., Inc., Novo Nordisk, Research Support; AstraZeneca, Boehringer Ingelheim International GmbH. F.Rodriguez: Advisory Panel; Amgen Inc., Consultant; HealthPals, Novartis Pharmaceuticals Corporation, Novo Nordisk, Stock/Shareholder; Carta Healthcare, Inc. K.W.Mahaffey: Consultant; Novo Nordisk. L.Palaniappan: Consultant; Amgen Inc. S.H.Shah: Research Support; Verily Life Sciences. Funding The Baseline Health Study and this analysis were funded by Verily Life Sciences, San Francisco, California.

Diagnostic signature of circulating miRNAs in glial brain tumors.

e14027 Background: Circulating microRNAs (miRNAs) are currently being investigated as non-invasive diagnostic biomarkers in a wide range of tumors. However, there is still no sufficiently reliable siRNA signature for routine screening of patients with glial tumors of varying degrees of malignancy. Our study aimed to determine the level of microRNA pattern in the blood plasma of patients with glial tumors, which previously showed aberrant expression in tumors. Methods: Relative expression of twenty miRNAs (hsa-miR-3180, hsa-miR-3180-3p, hsa-miR-6782-5p, hsa-miR-182-5p, hsa-miR-133b, hsa-miR-670-3p, hsa-miR-342-3p, hsa-miR-454-3p, hsa-miR-1246, hsa-miR-433-3p, hsa-miR-195-5p, hsa-miR-29a-3p, hsa-miR -107, hsa-miR-21, hsa-miR-122, hsa-let-7c-5p, hsa-miR-125b-5p, hsa-miR-190b, hsa-miR-196a-3p, hsa-miR-16 -5p) was studied in the blood plasma of patients with glial tumors of various grades of malignancy (diffuse astrocytoma (DA, n=10), anaplastic astrocytoma (AA, n=10) and glioblastoma (GB, n=10)) and patients with menigioma (MEN, n=10) compared with a conditionally healthy group (CNTRL, n=10) by RT-PCR. Cycle thresholds were normalized to exogenous cel-miR-39-5p. Relative expression (RE) was calculated using the 2ΔΔCt method. Wilcoxon's t-test and Benjamini-Hochberg multiple comparison adjustment were used to assess differences. Results: As a result of evaluating the relative expression of the studied miRNAs between groups, differences were found for 9 miRNAs. P-values are listed in the Table. Conclusions: Thus, circulating miRNAs could potentially act as biomarkers for both gliomas and their subtypes and differentiate gliomas from meningiomas. The findings may provide new diagnostic strategies for gliomas. Significance levels of differences in the relative expression of miRNAs when comparing the studied groups with each other.[Table: see text]

Kinetics of cellular and humoral responses to third BNT162B2 COVID-19 vaccine over six months in heart transplant recipients - implications for the omicron variant.

BackgroundThe durability of the immune response following the 3-dose BNT162b2 vaccination is unknown. The complexity of the situation is enhanced by the threat that highly transmissible variants may further accelerate the decline in the protection afforded by mRNA vaccines.MethodsOne hundred and three 3-dose-vaccinated heart transplant recipients were longitudinally assessed for the kinetics of variant-specific neutralization (Cohort 1, n = 60) and SARS-CoV-2-specific-T-cell response (Cohort 2, n = 54) over 6 months. Neutralization and T-cell responses were compared between paired samples at 2 time points, using the Kruskal-Wallis test followed by Dunn's multiple comparison test for continuous variables and McNemar's test for dichotomous variables. The Bonferroni method of p values adjustment for multiple comparison was applied.ResultsThe third dose induced high neutralization of the wild-type virus and delta variant (geometric mean titer [GMT], 137.2 [95% CI, 84.8-221.9] and 80.6, [95% CI, 49.3-132.0], respectively), and to a lesser degree of the omicron variant (GMT, 10.3 [95% CI, 5.9-17.9]). At 6 months, serum neutralizing activity declined but was still high for the wild-type virus and for the delta variant (GMTs 38.1 [95% CI, 21.2-69.4], p = 0.011; and 28.9 [95% CI, 16.6-52.3], p = 0.022, respectively), but not for the omicron variant (GMT 5.9 [95% CI, 3.4-9.8], p = 0.463). The percentages of neutralizing sera against the wild-type virus, delta and omicron variants increased from 70%, 65%, and 38%, before the third dose, to 93% (p < 0.001), 88% (p < 0.001), and 48% (p = 0.021) at 3 weeks after, respectively; and remained high through the 6 months for the wild-type (80%, p = 0.06) and delta (77%, p = 0.102). The third dose induced the development of a sustained SARS-CoV-2-specific-T-cell population, which persisted through 6 months.ConclusionsThe third BNT162b2 dose elicited a durable SARS-CoV-2-specific T-cell response and induced effective and durable neutralization of the wild-type virus and the delta variant, and to a lesser degree of the omicron variant.

Is it more effective or efficient to coach teachers in pairs or individually? A comparison of teacher and student outcomes and coaching costs

A growing body of research documents the positive impacts of teacher coaching, but research contrasting the effectiveness of different coaching approaches is limited. This study contrasted paired coaching – delivered to two teachers simultaneously – with traditional coaching for individual in-service teachers. We examined the effects of these two approaches on observations of teachers' classroom management practices and student behavioral outcomes, relative to non-coached conditions, over the course of a single school year and at a 1-year follow-up. We also explored the relative time and cost efficiency of the two approaches. Two hundred fifty-two teachers from 18 elementary and middle schools participated in the randomized controlled trial. Hierarchical linear modeling was used to account for repeated measures nested within teachers and teachers within schools. Combined effects indicated improved global ratings by observers of teacher behavior management (∆ = 0.29) after a multiple comparison adjustment. Paired coaching was less effective than individual coaching at improving some observed student behaviors, although these did not remain significant after multiple comparison adjustments. Neither model demonstrated sustained effects after one year. Although the paired coaching was significantly more efficient for coach time, it represented just a modest overall cost savings per school. Results indicate a need for more research to identify feasible coaching approaches yielding sustainable effects.

Association of Congenital and Acquired Cardiovascular Conditions With COVID-19 Severity Among Pediatric Patients in the US

Identifying the associations between severe COVID-19 and individual cardiovascular conditions in pediatric patients may inform treatment. To assess the association between previous or preexisting cardiovascular conditions and severity of COVID-19 in pediatric patients. This retrospective cohort study used data from a large, multicenter, electronic health records database in the US. The cohort included patients aged 2 months to 17 years with a laboratory-confirmed diagnosis of COVID-19 or a diagnosis code indicating infection or exposure to SARS-CoV-2 at 85 health systems between March 1, 2020, and January 31, 2021. Diagnoses for 26 cardiovascular conditions between January 1, 2015, and December 31, 2019 (before infection with SARS-CoV-2). The main outcome was severe COVID-19, defined as need for supplemental oxygen or in-hospital death. Mixed-effects, random intercept logistic regression modeling assessed the significance and magnitude of associations between 26 cardiovascular conditions and COVID-19 severity. Multiple comparison adjustment was performed using the Benjamini-Hochberg false discovery rate procedure. The study comprised 171 416 pediatric patients; the median age was 8 years (IQR, 2-14 years), and 50.28% were male. Of these patients, 17 065 (9.96%) had severe COVID-19. The random intercept model showed that the following cardiovascular conditions were associated with severe COVID-19: cardiac arrest (odds ratio [OR], 9.92; 95% CI, 6.93-14.20), cardiogenic shock (OR, 3.07; 95% CI, 1.90-4.96), heart surgery (OR, 3.04; 95% CI, 2.26-4.08), cardiopulmonary disease (OR, 1.91; 95% CI, 1.56-2.34), heart failure (OR, 1.82; 95% CI, 1.46-2.26), hypotension (OR, 1.57; 95% CI, 1.38-1.79), nontraumatic cerebral hemorrhage (OR, 1.54; 95% CI, 1.24-1.91), pericarditis (OR, 1.50; 95% CI, 1.17-1.94), simple biventricular defects (OR, 1.45; 95% CI, 1.29-1.62), venous embolism and thrombosis (OR, 1.39; 95% CI, 1.11-1.73), other hypertensive disorders (OR, 1.34; 95% CI, 1.09-1.63), complex biventricular defects (OR, 1.33; 95% CI, 1.14-1.54), and essential primary hypertension (OR, 1.22; 95% CI, 1.08-1.38). Furthermore, 194 of 258 patients (75.19%) with a history of cardiac arrest were younger than 12 years. The findings suggest that some previous or preexisting cardiovascular conditions are associated with increased severity of COVID-19 among pediatric patients in the US and that morbidity may be increased among individuals children younger than 12 years with previous cardiac arrest.