False Discovery Rate Adjustment Research Articles

Organochlorine pesticide exposures, metabolic enzyme genetic polymorphisms and semen quality parameters among men attending an infertility clinic.

The associations of organochlorine pesticides (OCPs) with semen quality from human studies are conflicting, and also it is largely unknown whether the associations are modified by genetic polymorphisms. We aimed to evaluate the associations between serum concentrations of 18 OCPs and semen quality among 387 Chinese men, and further to examine the modifying effects by genetic polymorphisms in cytochrome P450 (CYP2E1) and glutathione S-transferase (GSTT1). Multivariable linear regressions were used to evaluate the relationships between serum OCP concentrations and semen quality, and the role of CYP2E1 and GSTT1 polymorphisms in modifying the associations were assessed. Multiple testing was adjusted using the false discovery rate (FDR). We observed that men with detectable concentrations of serum ɤ-HCH had a decrease in sperm motility of 7.07% (95% CI: -10.9%, -3.24%) compared to those with undetectable concentrations (FDR-P value=0.02). Men with TT of CYP2E1 rs 915906 genotypes had higher median concentrations of serum dieldrin compared with those with CT/CC of CYP2E1 rs 915906 genotypes. There were interactions between CYP2E1 and GSTT1 polymorphisms and certain OCPs namely ɤ-HCH, δ-HCH, dieldrin, endosulfan I, and endrin aldehyde on semen quality. For example, elevated dieldrin levels in relation to decreased sperm concentration, sperm count, and sperm motility were only observed among men with CC of CYP2E1 rs2031920 genotypes (all Pinteraction < 0.05). However, these interactions were not statistically significant after the FDR adjustment. Our results suggested that CYP2E1 and GSTT1 polymorphisms may modify the effects of OCP exposures on semen quality. Due to the relatively small size samples, further investigation is warranted to confirm the findings.

Identifying prognostic parameters related to surgical technique in patients treated by robotic radical prostatectomy

Introduction and objectiveThe most frequently studied factors in patients treated by robotic radical prostatectomy are PSA and pathological features of the biopsy and prostatectomy specimen. Studies on the factors associated with the surgical technique are scarce and with controversial results. The objective is to identify all possible surgical factors and their relationship with disease-free and metastasis-free survival. Patients and MethodProspective study approved by the Ethics Committee, including patients who underwent robotic radical prostatectomy since January 2009 with a minimum follow-up of 5 years. Surgeon, surgical time, blood loss, fascial access, continence techniques, preservation of the fascia, neurovascular bundles, bladder neck, urethra, learning curve and surgical complications, were analyzed as possible prognostic factors. We performed univariate and matched comparisons of survival using Kaplan–Meier estimation and long-rank tests. The significance level for multiple comparisons was established with False Discovery Rate-adjustment (adjusted p). ResultsCohort of 667 patients with a median follow-up of 69 months. In univariate analysis, surgeon (adjp=0.018), preservation of puboprostatic ligaments (adjp=0.02), preservation of endopelvic fascia (adjp=0.001) and performing periurethral suspension (adjp<0.001) are poor prognostic factors for disease-free survival. Fascia preservation also negatively affects metastasis-free survival (adjp=0.04). Previous abdominal surgeries, prostate, surgical time, blood loss, type of residual urethra, middle lobe, fascial access, fascia or bladder neck preservation, have no statistical significance. ConclusionsThe surgeon and specific aspects of the surgical technique are determining factors in disease-free survival. Preservation of the fascia is the only factor that negatively affects metastasis-free survival.

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

Identificación de parámetros pronósticos relativos a la técnica quirúrgica en pacientes tratados mediante prostatectomía radical robótica

Introduction and objectiveThe most frequently studied factors in patients treated by robotic radical prostatectomy are PSA and pathological features of the biopsy and prostatectomy specimen.Studies on the factors associated with the surgical technique are scarce and with controversial results. The objective is to identify all possible surgical factors and their relationship with disease-free and metastasis-free survival. Patients and MethodProspective study approved by the Ethics Committee, including patients who underwent robotic radical prostatectomy since January 2009 with a minimum follow-up of 5 years.Surgeon, surgical time, blood loss, fascial access, continence techniques, preservation of the fascia, neurovascular bundles, bladder neck, urethra, learning curve and surgical complications, were analyzed as possible prognostic factors.We performed univariate and matched comparisons of survival using Kaplan-Meier estimation and long-rank tests. The significance level for multiple comparisons was established with False Discovery Rate-adjustment (adjusted p). ResultsCohort of 667 patients with a median follow-up of 69 months.In univariate analysis, surgeon (adjp = 0.018), preservation of puboprostatic ligaments (adjp = 0.02), preservation of endopelvic fascia (adjp = 0.001) and performing periurethral suspension (adjp < 0.001) are poor prognostic factors for disease-free survival. Fascia preservation also negatively affects metastasis-free survival (adjp = 0.04).Previous abdominal surgeries, prostate, surgical time, blood loss, type of residual urethra, middle lobe, fascial access, fascia or bladder neck preservation, have no statistical significance. ConclusionsThe surgeon and specific aspects of the surgical technique are determining factors in disease-free survival. Preservation of the fascia is the only factor that negatively affects metastasis-free survival.

PB2272: THERAPY AND HOSPITAL MORTALITY PREDICTORS IN HEMATOLOGICAL PATIENTS WITH 4 GRADE NEUTROPENIA AND CONCOMITANT CORONAVIRUS INFECTION

Background: According to published data, cancer patients are 5 times more likely to contract coronavirus infection (COVID-19) than non-cancer patients. Aims: To assess and identify predictors of hospital mortality in hematological patients with grade 4 neutropenia and concomitant coronavirus infection. Methods: A prospective study included 130 patients (55% women, 45% men) aged 18 to 91 years (median 59) with acute leukemia (AL) and lymphoproliferative diseases (LPD) who had grade 4 neutropenia and coronavirus infection (COVID-19) and were observed from April 2020 to December 2021 at Moscow City Clinical Hospital 52 (Russia). The study also included a reference group of 130 patients matched by sex, age and diagnosis, with AL/LPD and COVID-19, but without grade 4 neutropenia. The statistical analysis was an intention to treat analysis with adjustment for false discovery rate. Results: AML prevailed in the AL group (80.6%), aggressive non-Hodgkin’s lymphomas in the LPD group. Distribution by disease status: 62 patients (47.7%) on chemotherapy (CT), 19 (14.6%) in remission, 23 (17.8%) with disease progression, 19 (14.6%) with newly diagnosed disease, 7 (5.3%) with a relapse. The patients had a high NEWS score (7.5) and a high Charlson comorbidity index (5). The majority had moderate or severe lung tissue damage (CT2 in 44 (33%), CT3 in 25 (20%), CT4 in 13 (10%)). There were increases in the levels of C-reactive protein (> 100 g/L in 55% of patients, mean 241.5 g/L), LDH (506 U/L), D-dimer (1866), procalcitonin (> 0.5 ng/mL in 73%). IL-6 and IL-1b blockers were used for hypercytokinemia; the dose of IL blockers was reduced in view of grade 4 neutropenia. Empiric antibiotic therapy for complications was used in 91.5% of patients. Antifungal therapy was carried out in 65% of cases. Sepsis developed in 63 (48.5%) patients. The median duration of hospitalization in patients with grade 4 neutropenia was 18 days. Transfer to the ICU was required in 78 (60%) patients in the total group and in 41 patients (52.5%) among those older than 60 years. Mortality in ICU patients: 80.7%, mortality in patients older than 60 years: 92.7%. In non-ICU patients with AL and LPD younger than 60 years, the mortality rate was 0%. Mechanical ventilation was used in 48.5% of cases. Mortality was lower in vaccinated patients than in unvaccinated ones (40% vs 51%). The mortality rates in the main group (AL 48.6%, LPD 51.7%, Fig. 1) and the reference group (AL 37.5%, LPD 13.8%, Fig. 2) showed that the presence of grade 4 neutropenia increased the risk of death almost twofold: the relative risk was 1.9 (95% CI 1.3–2.6), p=0.0001. According to the results of multivariate analysis (logistic regression), the predictors of hospital mortality in the main group were ICU stay (odds ratio 12.1 (1.4–24.0), p=0.0001), sepsis (3.0 (1.1–7.7), p=0.001), and age > 65 (3.0 (0.9–7.5), p=0.001). Severe disease progression/relapse is also a predictor of mortality (close to statistical significance). Image:Summary/Conclusion: Coronavirus infection with severe neutropenia (caused by tumor progression and/or combination chemotherapy) is a significant adverse factor of hospital mortality in patients with hematological cancers. Predictors of hospital mortality are age over 60 years, sepsis, ICU stay, and the absence of remission.

Challenging the "old boys club" in academia: Gender and geographic representation in editorial boards of journals publishing in environmental sciences and public health.

In light of global environmental crises and the need for sustainable development, the fields of public health and environmental sciences have become increasingly interrelated. Both fields require interdisciplinary thinking and global solutions, which is largely directed by scientific progress documented in peer-reviewed journals. Journal editors play a critical role in coordinating and shaping what is accepted as scientific knowledge. Previous research has demonstrated a lack of diversity in the gender and geographic representation of editors across scientific disciplines. This study aimed to explore the diversity of journal editorial boards publishing in environmental science and public health. The Clarivate Journal Citation Reports database was used to identify journals classified as Public, Environmental, and Occupational (PEO) Health, Environmental Studies, or Environmental Sciences. Current EB members were identified from each journal's publicly available website between 1 March and 31 May 2021. Individuals' names, editorial board roles, institutional affiliations, geographic locations (city, country), and inferred gender were collected. Binomial 95% confidence intervals were calculated for the proportions of interest. Pearson correlations with false discovery rate adjustment were used to assess the correlation between journal-based indicators and editorial board characteristics. Linear regression and logistic regression models were fitted to further assess the relationship between gender presence, low- and middle-income country (LMIC) presence and several journal and editor-based indicators. After identifying 628 unique journals and excluding discontinued or unavailable journals, 615 journal editorial boards were included. In-depth analysis was conducted on 591 journals with complete gender and geographic data for their 27,772 editors. Overall, the majority of editors were men (65.9%), followed by women (32.9%) and non-binary/other gender minorities (0.05%). 75.5% journal editorial boards (n = 446) were composed of a majority of men (>55% men), whilst only 13.2% (n = 78) demonstrated gender parity (between 45-55% women/gender minorities). Journals categorized as PEO Health had the most gender diversity. Furthermore, 84% of editors (n = 23,280) were based in high-income countries and only 2.5% of journals (n = 15) demonstrated economic parity in their editorial boards (between 45-55% editors from LMICs). Geographically, the majority of editors' institutions were based in the United Nations (UN) Western Europe and Other region (76.9%), with 35.2% of editors (n = 9,761) coming solely from the United States and 8.6% (n = 2,373) solely from the United Kingdom. None of the editors-in-chief and only 27 editors in total were women based in low-income countries. Through the examination of journal editorial boards, this study exposes the glaring lack of diversity in editorial boards in environmental science and public health, explores the power dynamics affecting the creation and dissemination of knowledge, and proposes concrete actions to remedy these structural inequities in order to inform more equitable, just and impactful knowledge creation.

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.

Unique-to-Salmon Compounds Increase in Plasma and Are Associated With Cardiovascular Health Following a Mediterranean Diet Intervention

ObjectivesSalmon consumption may promote cardiovascular (CV) health through nutrients such as omega-3 fatty acids (n-3 FAs) or other yet unidentified compounds. Metabolomics allows for the comprehensive analysis of small molecules in foods and human samples. The objectives of this study were to: 1) identify compounds present in salmon that were increased in plasma after consumption of a salmon-containing Mediterranean-style eating pattern (MED); and 2) evaluate associations between salmon compounds detected in plasma and CV biomarkers. MethodsIndividuals (n = 41) completed a 16-week randomized, crossover, controlled feeding study of a MED containing 2 servings salmon/week. Assessments were completed pre- and post-intervention, where CV biomarkers (e.g., blood pressure, lipid profile) were measured and fasting plasma collected. Samples of participants’ plasma, salmon, and 99 other foods in the trial were analyzed in triplicate via liquid chromatography-mass spectrometry (LCMS). Compounds were determined to be unique-to-salmon (UTS) if detected in all replicates of salmon but none of the other 99 foods. For UTS compounds that were found in plasma, linear mixed effects models were used to assess change from pre- to post-intervention and the association between this change and changes in CV biomarkers. Significance was assessed after adjustment for false discovery rate. ResultsRelative to the other 99 foods, 508 compounds were determined to be UTS, and 143 of these UTS compounds were detected in plasma. Forty-eight UTS compounds significantly increased in plasma from pre- to post-intervention. Two compounds, a phosphatidylglycerolphosphate (40:4) and a triglyceride (58:11), were associated with improvement in ≥1 CV biomarker. The former, a cardiolipin precursor, was associated with improvements in total cholesterol, LDL cholesterol, and Apolipoprotein B. ConclusionsSalmon compounds are detectable in human plasma after consumption within a complex diet and increase after intervention, suggesting potential as intake biomarkers. Further, bioactive compounds beyond nutrients such as n-3 FAs may be associated with CV benefits. Future studies should be conducted to quantify concentrations and define dose-response. Funding SourcesNIH/NIDDK; Beef Checkoff (MED feeding study).

1004-P: Executive Dysfunction Predicts Lower Self-Care Readiness in Teens with Type 1 Diabetes

Aim: Management of type 1 diabetes (T1D) is a complex multi-step process in which executive function (EF) skills (planning, organization, problem solving) are critical for optimal control. In teens with T1D, EF may be important in transitioning from parent-supervised care to self-care. Methods: In an 18-month study, teens aged 13-17 years with T1D and parents completed the Diabetes Family Responsibility Questionnaire (DFR) and Readiness for Independent Self-Care Questionnaire (RISQ) every 6 months. Teen EF was assessed by parent proxy-report using Behavior Rating Inventory of Executive Function (BRIEF) . T-score ≥60 defined risk of executive dysfunction for the Global EF Composite (GEC) , Behavioral Regulation Index (BRI) and Metacognition Index (MI) . EF, demographic, and T1D treatment variables were used to predict change in RISQ over time using longitudinal mixed models with false discovery rate adjustment. Results: 169 teens (54% male) participated with baseline age 15.9±1.3 years, T1D duration 8.4±3.7 years, HbA1c 8.6±1.2%, 74.6% insulin pump use, and 21.3% continuous glucose monitor (CGM) use. 31.4% of teens had a parent-reported GEC score ≥60. After adjusting for age, gender, and parent-reported DFR (pDFR) , those with GEC ≥60 had parent-reported RISQ (pRISQ) 4.1 points lower than those with GEC &amp;lt;60, p=0.016. BRI (adjusted for age, CGM use, pDFR, and HbA1c) did not predict change in pRISQ while MI (adjusted for age, gender, and pDFR) did; those with MI ≥60 had pRISQ 5.3 points lower than those with MI &amp;lt;60, p&amp;lt;0.001. In models using teen-reported DFR and RISQ (tRISQ) , GEC and MI similarly predicted lower tRISQ. In all models, lower age (p&amp;lt;0.05) and greater parent involvement (p&amp;lt;0.001) also predicted lower RISQ. Conclusion: EF, especially metacognition, appears important for acquisition of self-care behaviors for teens with T1D. Evaluating EF during adolescence may identify teens with executive dysfunction who may need extra support during the transition process. Disclosure R.J.Vitale: None. L.J.Tinsley: None. L.K.Volkening: None. L.M.Laffel: Advisory Panel; Medtronic, Roche Diabetes Care, Consultant; Boehringer Ingelheim International GmbH, Dexcom, Inc., Dompé, Insulet Corporation, Janssen Pharmaceuticals, Inc., Lilly Diabetes, Novo Nordisk, Provention Bio, Inc. Funding NIH T32DK007529NIH P30DK036836NIH R01DK095273JDRF 2-SRA-2014-253-M-B

Associations of the Gut Microbiota Composition and Fecal Short-Chain Fatty Acids with Leukocyte Telomere Length in Children Aged 6 to 9 Years in Guangzhou, China: A Cross-sectional Study

BackgroundTelomere length (TL) serves as a marker of cellular senescence and appears to plateau between the age of 4 y and young adulthood, after which the gut microbiota are supposed to be established. However, scarce data are available regarding the correlation between gut microbiota composition and TL in the pediatric population. ObjectivesWe aimed to investigate whether the gut microbiota and the concentrations of SCFAs in feces are associated with leukocyte TL in children. MethodsIn total, 401 children aged 6–9 y from Guangzhou were enrolled in this cross-sectional study. qPCR was used to determine relative TL in peripheral blood leukocytes. The gut microbiota was characterized by 16S ribosomal RNA amplicon sequencing and the fecal concentrations of total SCFAs and SCFA subtypes were determined using HPLC. The multivariate methods with an unbiased variable selection (MUVR) algorithm and partial least square models were used to select predictable operational taxonomic units (OTUs). Further correlation analyses were performed based on multiple linear regression models with adjustment for covariates and false discovery rate. ResultsWith the use of MUVR, 35 relevant and minimal optimal OTUs were finally selected. Multiple linear regression analysis showed that the abundance of several OTUs, including OTU334 (belonging to the genus Family XIII AD3011 group), OTU726 (belonging to the species Lachnoclostridium phocaeense), OTU1441 (belonging to the genus Ruminococcus torques group), OTU2553 (belonging to the genus Lachnospiraceae UCG-010), and OTU3375 (belonging to the family Lachnospiraceae), was negatively associated with leukocyte TL (β: –0.187 to –0.142; false discovery rate (FDR)–corrected P value (PFDR) = 0.009–0.035]. However, neither SCFA subtype nor total SCFA content in feces exhibited significant associations with TL (β: –0.032 to 0.048; PFDR = 0.915–0.969). ConclusionsThe gut microbiota, but not fecal SCFA concentration, was significantly associated with TL in this pediatric population.

Evidence of altered fatty acid metabolism in dogs with naturally occurring valvular heart disease and congestive heart failure

IntroductionMyxomatous mitral valve disease (MMVD) is the most common cardiac condition in adult dogs. The disease progresses over several years and affected dogs may develop congestive heart failure (HF). Research has shown that myocardial metabolism is altered in cardiac disease, leading to a reduction in β-oxidation of fatty acids and an increased dependence upon glycolysis.ObjectivesThis study aimed to evaluate whether a shift in substrate use occurs in canine patients with MMVD; a naturally occurring model of human disease.MethodsClient-owned dogs were longitudinally evaluated at a research clinic in London, UK and paired serum samples were selected from visits when patients were in ACVIM stage B1: asymptomatic disease without cardiomegaly, and stage C: HF. Samples were processed using ultra-performance liquid chromatography mass spectrometry and lipid profiles were compared using mixed effects models with false discovery rate adjustment. The effect of disease stage was evaluated with patient breed entered as a confounder. Features that significantly differed were screened for selection for annotation efforts using reference databases.ResultsDogs in HF had altered concentrations of lipid species belonging to several classes previously associated with cardiovascular disease. Concentrations of certain acylcarnitines, phospholipids and sphingomyelins were increased after individuals had developed HF, whilst some ceramides and lysophosphatidylcholines decreased.ConclusionsThe canine metabolome appears to change as MMVD progresses. Findings from this study suggest that in HF myocardial metabolism may be characterised by reduced β-oxidation. This proposed explanation warrants further research.

0363 Perceptions of the Need for Perioperative OSA Education: An Interdisciplinary and Multi-Institutional Survey

Abstract Introduction Advanced Practice Providers (APPs; Advanced practice registered nurses, physician assistants) and physicians-in-training (residents, fellows) receive inadequate education on obstructive sleep apnea (OSA)/perioperative OSA risks. However, they are front-line providers assessing these patients. Failure to mitigate this risk has led to significant postoperative morbidity/mortality. We assessed these providers’ perceptions to OSA/perioperative OSA training. Methods Surveys were sent to three provider roles, APPs, residents, and fellows, in four categories of practice at nine academic institutions between May 9-June 30, 2021. Chi-square and Fisher’s exact tests assessed association between survey responses and participant characteristics. False discovery rate adjustment accounted for multiple comparisons, threshold of q&amp;lt;0.05 for statistical significance. Cochran-Mantel-Haenzsel tests evaluated associations stratified by institution. Results 2236 of 6724 (33.3%) participants responded: 48.4% APPs, 11% Fellows and 40.6% Residents. Primary category of practice included: 20.3% Anesthesiology, 8.9% Family Medicine, 34.1%, Internal Medicine (IM)/IM subspecialties, 6.7% Obstetrics/Gynecology/Gynecologic Oncology, 25.9% Surgery/Surgery subspecialties, 4.1% Other.While 93.2% of respondents believed OSA is a risk factor for perioperative complications, fewer respondents reported that they felt adequately trained to assess for OSA (50.9%) in general, with significant differences noted by provider role (range 42-70%, q=0.001) and across the categories of practice (range 12-82%, q=0.001). Even fewer felt adequately trained to assess for OSA in perioperative patients (38.2%) with significant differences noted by provider role (range 31-52%, q=0.001) and across the categories of practice (range 15-84%, q=0.001).Across all categories of practice, respondents indicated that they would like additional educational training about OSA (76.7%). This varied by clinical role (range 64-82%, q=0.003), but not categories of practice (range 73-84%, q=0.13). Furthermore, respondents indicated they also desired additional education about OSA in the perioperative patient (75.5%). This extended across all clinical roles (range 68-77%, q=0.09) and categories of practice (range 72-80%, q=0.09). Conclusion We found significant differences in APP and physician-in-training perceptions of the adequacy of their current training and desire for further OSA/perioperative OSA education. Our study identifies a critical gap and opportunity to improve provider understanding and patient care. Support (If Any) American Academy of Sleep Medicine Foundation (AASMF) award.

Functional Annotation and Cross‐study Comparison of DNA Methylation Regions uniquely associated with 24‐hour Blood Pressure Phenotypes in African Americans

Epigenetics may reflect the interactions between DNA and environment and lifestyle and DNA methylation marks have been implicated in the development and progression of hypertension. In an African American (AA) cohort of hypertensives and normotensives (n = 281), we have recently discovered up to 72 methylation regions (MRs) in whole blood DNA that were significantly associated with 24‐h BP phenotypes (False Discovery Rate (FDR) &lt; 0.05) via reduced representation bisulfite sequencing (RRBS). One‐time clinic BP measurements were not associated with MRs. Upon adjustments for covariates age, sex, and body mass index, 13 MRs remained associated with 24‐h BP phenotypes (FDR &lt; 0.05), each explaining 6.5% ‐ 9.4% of BP variance. Analysis of the chr19 MR in an independent cohort (n=117) using Bisulfite ULtrapLEx Targeted Sequencing (BULLET‐Seq), a new method that we developed, validated its association with 24‐h avg BP phenotypes (FDR &lt; 0.05).In the current study, we functionally annotated the MRs that we identified as uniquely associated with 24‐h BP phenotypes and examined CpG sites previously reported to be associated with BP. Two MRs were near chr1 linkage disequilibrium (LD) regions within genes, PRDM16 and CASZ1, containing BP‐associated sentinel single nucleotide polymorphisms (SNPs). Although no MRs were found to directly overlap with reported BP‐relevant expression or methylation quantitative trait loci (eQTLs or meQTLs), many overlapped with enhancer regions, CTCF‐binding sites, or genes related to hypertension and stroke. To demonstrate consistency with prior studies, we found associations between reported clinic BP‐associated methylation array chip CpGs and the 24‐h BP phenotypes in our study. Of the 16 BP‐associated CpGs from two reports that were detected in both the methylation chip and our RRBS data, several CpGs were significantly associated with 24‐h and clinic BP phenotypes in our cohort (p &lt; 0.05) but not after adjustment for both covariates and FDR. A chr7 CpG (cg06340364) showed significant negative associations with all studied BP phenotypes except nighttime and clinic DBPs (p &lt; 0.05, unadjusted). These investigations highlight the value of regional DNA methylation and the ability to capture the overall effect of environmental and genetic factors that influence BPs over a 24‐h period which have functional epigenomic relevance.

Anterior Cruciate Ligament Reconstruction Upregulates Vitamin D Activating Pathways within the Quadriceps

Anterior cruciate ligament (ACL) tears are prevalent sport‐related injuries that result in protracted quadriceps atrophy and weakness despite surgical reconstruction and physical therapy. Vitamin D and its receptor are integral for skeletal muscle development and homeostasis. In addition, vitamin D status is positively associated with muscle size and strength. If the vitamin D receptor and related enzymes are upregulated with ACL injury and reconstruction, then overall vitamin D status may be an important effector of recovery. Concentrations of vitamin D needed to support optimal recovery may differ from typical clinical cut points used to diagnose vitamin D deficiency, which could have powerful therapeutic implications following ACL injury and reconstruction. This is especially important given recent research showing over 50% of athletes have vitamin D inadequacy. We hypothesized that subjects with circulating vitamin D sufficient to suppress parathyroid hormone secretion (≥ 40 ng/mL) would be better protected against quadriceps atrophy at the conclusion of rehabilitation. Muscle biopsies (vastus lateralis) were obtained from ACL‐injured and Healthy limbs of 16 participants (6M,10F; 19±5yr, body mass index: 25.1±3.7kg/m2) prior to surgical reconstruction, 1 week following reconstruction (Recon), and following the completion of post‐surgical rehabilitation (6 months following surgery, Post‐Rehab). RNA was isolated from all quadriceps biopsies, followed by library construction, sequencing, and bioinformatics analysis. Transcript abundance of vitamin D activating pathway regulators (vitamin D receptor [VDR], and Cytochrome P450 2R1 [CYP2R1] and Family 27 Subfamily B Member 1 [CYP27B1]) was assessed following analysis using DESeq2 and false discovery rate (FDR) adjustment of p‐values. Additionally, 25‐hydroxy vitamin D (25(OH)D) was assessed in serum samples obtained at the time of reconstruction surgery. Quadriceps fiber cross‐sectional area was measured via laminin immunohistochemistry. Recon (1 week post‐surgery) muscle showed increased transcript abundance of the vitamin D receptor, VDR(elevated 7.8 fold, FDR&lt;0.05), and vitamin D‐activating pathway regulators CYP2R1 (elevated 1.5 fold, FDR&lt;0.05) and CYP27B1 (elevated 2.6 fold, FDR&lt;0.05) when compared to the Healthy limb. Following the completion of rehabilitation at the 6‐month follow‐up, participants with 25(OH)D above 40 ng/mL showed 6.4% atrophy of quadriceps fiber size compared to 22.8% atrophy in participants with 25(OH)D below 40 ng/mL (p=0.06). These findings provide evidence for surgically induced elevations in the vitamin D activating pathway within skeletal muscle. Upregulation of the vitamin D activating pathway may support quadriceps mitochondrial health and satellite cell activity and self‐renewal capacity, both of which are compromised following ACL reconstruction. In addition, elevated vitamin D concentrations may protect against protracted quadriceps atrophy. Future research should investigate if manipulation of vitamin D concentrations can enhance quadriceps functional recovery following ACL reconstruction.

Gene signatures in patients with early breast cancer and relapse despite pathologic complete response

A substantial minority of early breast cancer (EBC) patients relapse despite their tumors achieving pathologic complete response (pCR) after neoadjuvant therapy. We compared gene expression (BC360; nCounter® platform; NanoString) between primary tumors of patients with post-pCR relapse (N = 14) with: (i) matched recurrent tumors from same patient (intraindividual analysis); and (ii) primary tumors from matched controls with pCR and no relapse (N = 41; interindividual analysis). Intraindividual analysis showed lower estrogen receptor signaling signature expression in recurrent tumors versus primaries (logFC = −0.595; P = 0.022). Recurrent tumors in patients with distant metastases also exhibited reduced expression of immune-related expression parameters. In interindividual analyses, primary tumor major histocompatibility complex class II expression was lower versus controls in patients with any relapse (logFC = −0.819; P = 0.030) or distant relapse (logFC = −1.151; P = 0.013). Primaries with later distant relapse also had greater homologous recombination deficiency than controls (logFC = 0.649; P = 0.026). Although no associations remained statistically significant following adjustment for false discovery rate, our results show that transcriptomic analyses have potential for prognostic value and may help in selecting optimal treatment regimens for EBC at risk of relapse and warrant further investigation.

Intake Differences between Subsequent 24-h Dietary Recalls Create Significant Reporting Bias in Adults with Obesity

In depth understanding of the dietary patterns of individuals with obesity is needed in practice and research, in order to support dietitians and physicians in the design and implementation of nutritional management. We aimed to analyze the consistency of energy, macro-, and micronutrient reported intakes in four non-consecutive 24-h dietary recalls from 388 adults with obesity using information collected in the NutriGen Study (ClinicalTrials.gov, NCT02837367). Significant decreases were identified for reported energy and several, macro- and micronutrient intakes, between the first and subsequent 24-h recalls. Significant differences of reported intakes were identified in sensitivity analyses, suggesting that the first recall (also the only one performed on site, face-to-face) might be a point of bias. A comparison of the differences in intakes between weekend and weekday, after adjustment for false discovery rate were non-statistically significant either in male, females, or in total. To overcome this potential bias, studies should be carefully conducted, starting from the design phase, through to the analysis and interpretation phases of the study. Prior to averaging specific intakes across all sessions of reporting, a preliminary analysis must be conducted to identify if a certain time point had significant differences from all other time points and overview potential sources of bias: reporting bias, training bias, or behavioral changes could be responsible for such differences.

Environmental exposures to pesticides, phthalates, phenols and trace elements are associated with neurodevelopment in the CHARGE study.

To determine if higher exposures measured in early childhood to environmental phenols, phthalates, pesticides, and/or trace elements, are associated with increased odds of having a diagnosis of Autism Spectrum Disorder (ASD), Developmental Delay (DD), or Other Early Concerns (OEC) compared to typically developing children (TD). This study included 627 children between the ages of 2-5 who participated in the Childhood Autism Risks from Genetics and Environment (CHARGE) study. Urine samples were collected at the same study visit where diagnostic assessments to confirm diagnosis indicated during the recruitment process were performed. Adjusted multinomial regression models of each chemical with diagnosis as the outcome were conducted. Additionally, two methods were used to analyze mixtures: repeated holdout multinomial weighted quantile sum (WQS) regression for each chemical class; and a total urinary mixture effect was assessed with repeated holdout random subset WQS. Many urinary chemicals were associated with increased odds of ASD, DD or OEC compared to TD; however, most did not remain significant after false discovery rate adjustment. Repeated holdout WQS indices provided evidence for associations of both a phenol/paraben mixture effect and a trace element mixture effect on DD independently. In analyses adjusted for confounders and other exposures, results suggested an association of a pesticide mixture effect with increased risk for ASD. Results also suggested associations of a total urinary mixture with greater odds of both ASD and DD separately. Higher concentrations of urinary biomarkers were associated with ASD, DD, and OEC compared to TD, with consistency of the results comparing single chemical analyses and mixture analyses. Given that the biospecimens used for chemical analysis were generally collected many months after diagnoses were made, the direction of any causal association is unknown. Hence findings may reflect higher exposures among children with non-typical development than TD children due to differences in behaviors, metabolism, or toxicokinetics.

Abstract P163: Gut Microbiome And Stages Of Diabetes In Middle-aged Adults: CARDIA Microbiome Study

Background: Animal and human studies suggest the gut microbiome may be linked to insulin resistance and to diabetes. Results from population-based studies are limited and different stages of diabetes in relation to the gut microbiome have not been investigated. Objective: Examine cross-sectional associations of insulin resistance (IR), duration of diabetes, and 4 stages of diabetes (normoglycemic, prediabetes, and diabetes with (+) and without (-) anti-diabetic medication) with gut microbiome community composition in the Coronary Artery Risk Development in Young Adults (CARDIA). Method: The analysis is based on 605 Black and White participants who attended CARDIA’s Year 30 follow-up examination (2015-2016; aged 48-60 years). Using previously described methods, DNA from stool samples was sequenced using 16S rRNA method. We included 3 genus-level gut microbiome composition measures: alpha-diversity (Shannon index and richness), beta-diversity (Bray-Curtis dissimilarity matrix, PERMANOVA test), and taxonomy. For the analysis, the HOMA-IR levels at Years 25 and 30 were averaged; and diabetes duration was calculated as the sum of years starting at the Year 2 exam. Multivariable-adjusted regressions were used to assess associations of diabetes-related exposures with microbial features, adjusting for socio-demographics, clinical measures, health behaviors, and medication use. Genus-specific analysis was adjusted for multiple comparisons using a false discovery rate (FDR), with significance set to q-value &lt;0.1. Results: Among 605 participants, 20% had prediabetes, and 15% had diabetes (9% on anti-diabetic medication treatment) at Year 30. Compared to those without diabetes, richness and Shannon index were inversely associated with diabetes duration and diabetes with treatment but not with HOMA-IR or prediabetes. PERMANOVA tests were significant for all diabetes-related exposures (p-values&lt;0.05). After multivariable and FDR adjustment, 3 genera (all negative associations) were associated with HOMA-IR, 22 genera (2 positive associations) with diabetes duration, and 4 genera (1 positive association) with diabetes+. No genera were associated with prediabetes. There was reduced abundance of several butyrate producers in participants with higher HOMA-IR (ex., Lachnospiraceae_UGG.004 and Veillonella ), longer diabetes duration (ex., Agathobacter and Ruminococcus ), or diabetes+ (ex., Agathobacter and Romboutsia) . Conclusion: This study indicates significant associations of the gut microbiota composition with glycemic control and insulin resistance, which can contribute to our understanding of diabetes pathophysiology and its prevention and management. Certain butyrate-producing bacteria may play a beneficial role in glucose metabolism and insulin regulation. Future studies with longer-term follow-ups are warranted to explore the relevant mechanisms.

Maternal Blood-Based Protein Biomarkers in Relation to Abdominal Fat Distribution Measured by Ultrasound in Early Mid-Pregnancy

The objective of this study was to examine the associations of early mid-pregnancy ultrasound measured visceral and subcutaneous fat depths with blood-based protein biomarkers. This was a cross-sectional study including 201 pregnant women at Uppsala University Hospital, Sweden. The mean age of the women was 31.0 years, and 57.7% were nulliparous. Maternal visceral and subcutaneous fat depths were measured by ultrasound at the early second-trimester anomaly scan. A non-fasting blood sample was collected in conjunction with the second-trimester anomaly scan, and the Olink cardiovascular II panel was used to measure 92 blood-based protein biomarkers in the sample. Cross-sectional associations of visceral and subcutaneous fat depths with blood-based protein biomarkers were examined using Mann–Whitney U tests with false discovery rate adjustments. In addition, linear regression analyses adjusting for maternal age, parity, and early pregnancy body mass index were performed. The results showed differences in one biomarker between women with elevated (≥ 52 mm) versus normal (< 52 mm) visceral fat depth, and in three biomarkers between women with elevated (≥ 22 mm) versus normal (< 22 mm) subcutaneous fat depth. Hence, levels of blood-based protein biomarkers differ between pregnant women with dissimilar body fat distributions, which might reflect disparities in biological pathways.

P700 The GEM Project: Stool metabolomic profile is associated with microbiome risk score and with future onset of Crohn’s disease in healthy first-degree relatives

Abstract Background We recently developed and validated a microbial composition-based risk score (MRS) that predicts future risk of Crohn’s disease (CD) development among healthy first-degree relatives (FDR) of CD patients. We hypothesized that stool metabolomic profiles may mediate the effect of microbiome-based risk on future onset of CD. Methods Healthy FDRs of CD patients were recruited as part of the CCC-GEM Project. Stool metabolomics data were available in the nested case-control cohort (a subset of CCC-GEM cohort) whereby FDRs who later developed CD (n=56) were matched approximately 1:1 by age, sex, follow-up duration, and geographical location with control FDRs remaining healthy (n=66). Stool metabolomics were assessed using the Metabolon’s DiscoveryHD4™ platform, and the stool microbiome characterized by 16s rDNA amplicon sequencing. A multivariable conditional logistic regression model was evaluated on the disease development as a function of individual stool metabolites. Spearman correlation evaluated the rank values between stool metabolites and the MRS. Results 122 healthy FDRs (median age 15 years, 60% female) were followed for a median 5.2 years. Among 1,029 stool metabolites that were analyzed, 79 were associated with future risk of CD (p&amp;lt;0.05); however, none remained significant after adjustment for false-discovery rate. Considering the exploratory nature of this study with limited sample size, we focused on the top seven metabolites associated with CD onset (p&amp;lt;0.01). Of these, two stool metabolites (dimethylglycine, methylmyristate) were associated with increased risk of CD onset while five (cytosine, guanine, cytidine, hydroxyglutarate, nervonate) were associated with decreased risk of CD development. The two metabolites positively associated with CD onset were also positively correlated with the MRS, while the five metabolites negatively associated with CD onset, were also negatively correlated with the MRS. Meanwhile, 24 stool metabolites had significant correlation with MRS (false-discovery rate-corrected p&amp;lt;0.2). Among those, a total of four stool metabolites (cytosine, guanine, methymyristate, cytidine) overlapped with the top seven stool metabolites associated with CD onset. Conclusion Changes in stool metabolite profiles may predict future risk of CD. A subset of these metabolites has significant correlation with the MRS with consistent direction of effect. This may suggest that these particular stool metabolites mediate the putative effects of the gut microbiome on CD risk. Further validation in the entire GEM cohort is warranted.