False Discovery Correction Research Articles

A - 59 MEG Power Changes in Non-concussed High School Football Players: Implications of Cumulative Head Impact Risk

Abstract Purpose The purpose of this study was to determine how cumulative repetitive head impacts (RHI) over a single season affects functional brain activity in high school football players that are clinically asymptomatic for concussion. Method Pre-season and post-season magnetoencephalography (275 channel CTF MEG) and MRI were acquired for 88 football participant seasons (mean age = 16.09 yrs, mean BMI = 26.32). Helmet-based accelerometer data was collected for each practice and game. Cumulative risk-weighted exposure from linear acceleration (RWELinear) was calculated for each subject. Standard processing and source localization of 8-minutes eyes-open-resting state MEG data was performed using Brainstorm2. Voxel-wise relative power spectral density (PSD) was calculated for the theta frequency band (5-7 Hz). After transformation to MNI space2, pre-season to post-season percent change in PSD (%theta) was calculated and spatial clusters significantly correlated with RWELinear were identified using SPM8 (a = 0.05 and false discovery correction to account for multiple corrections). Mean cluster values were extracted, and linear regression analysis performed in R 3.21 with covariates of BMI, age, and time between scans. Results After covariate adjustment, there was a significant positive correlation between RWELinear and %Δtheta PSD in the right postcentral gyrus and inferior parietal lobe (p = 9.342e-05, adj. R2 = 0.2092), with no other significant associations noted. Conclusions Changes in functional brain activity may be associated with RHI specifically in the right postcentral gyrus and inferior parietal lobe over a typical football season. Future research will include examining how these affected areas are associated with cognitive performance.

Abstract 15298: Epidemiology and microRNA Predictors of Infection After Heart Transplantation

Background: Infection rates are high in heart transplant (HT) patients due to the need for immunosuppressive medication. MicroRNAs (miR) are non-coding RNAs, that regulate gene expression and are promising biomarkers for identifying patients at risk for infection. Hypothesis: MiR profiles can identify HT patients who are at increased risk for developing an infection. Methods: HT patients who were prospectively enrolled from 2015-2018 had miR-sequencing performed. Incident infection after HT hospital discharge along with the epidemiologic characteristics of the infection were collected from the EMR. An opportunistic infection (OI) is an infection which is less virulent in a healthy host but can cause severe disease in those immunosuppressed. A time-to-event analysis was performed using miR expression at two weeks post-HT in multivariable modeling, and the Benjamini-Hochberg procedure was used to correct for false discovery. Results: The cohort included 46 patients with a median age of 56 years (IQR: 49, 62), 33% (N=15) were Female sex, 41% (N=19) were Black, and 80% (N=37) were supported with an LVAD prior to HT. Post-HT infection occurred in 59% of patients (N=27) and median time to infection was 341 days (IQR: 85, 859). Hospitalization was required in 56% (N=15) of infections. Respiratory tract and OIs were the most common infection type (Figure A). The most common microorganisms implicated were CMV (28%) and SARS-COV-2 (19%) (Figure B). After correction for false discovery, we identified 15 miRs associated with infection risk. The 5 miRs with strongest effect size and their corresponding hazard ratios include: miR-21 2.5(95% CI: 1.3, 4.7), miR-130b 8.6 (95%CI: 1.9, 39.3), miR-192 0.3 (95%CI: 0.1, 0.6), miR-218 2.6 (95%CI: 1.3, 5.1) and miR-484 0.3 (95%CI: 0.1, 0.7). Conclusion: While the burden of post-HT infections is high and contributes to patient morbidity, novel plasma miRs can potentially play a role in identifying patients at risk for infections after HT.

Analysis of primary and acquired resistance to cetuximab from multiomic data in the New EPOC trial.

3577 Background: The use of cetuximab (cetux) with chemotherapy (chemo) in patients with operable liver metastases from colorectal cancer (CRC) conferred a survival disadvantage in New EPOC. Whilst genetic alterations such as emergent sub-clonal mutations in RAS/ BRAF, ERBB2 and MET are recognized as mechanisms of resistance to EGFR inhibition (EGFRi), divergent mechanisms such as transcriptomic changes may also be responsible. Of the CRC intrinsic subtypes (CRIS), CRIS-C has elevated EGFR signalling and is thought to have greater sensitivity to EGFRi. Methods: Resected primary tumors (n = 205; pre neoadjuvant treatment) and/or liver metastases (n = 144; post neoadjuvant treatment) from 223 patients, wild type for KRAS (codons 12/13/61) at trial entry, randomized to chemo +/- cetux underwent targeted NGS and/or transcriptome profiling. Primary clinical endpoints were progression free (PFS) and overall survival (OS). Results: Repeat NGS identified additional RAS/BRAF mutations in primary tumors from 49 patients (25 KRAS, 15 NRAS, 9 BRAF V6000E). Of the 148 patients with confirmed RAS/BRAF wild type primary tumors , 46% were classified as CRIS-C with the remainder being CRIS-A (9.5%), CRIS-B (11%), CRIS-D (8.8%) or CRIS-E (8.1%). CRIS-C patients had a similar PFS with ChemoCetux vs Chemo (HR 0.84 95% CI 0.47-1.5 p = 0.55) whereas non-CRIS-C had a shorter PFS (HR 2.12 95%CI 1.24-3.63 p = 0.006; p interaction 0.021). OS results were similar (p interaction 0.067). In liver metastases, genetic changes were comparable (Chemo vs ChemoCetux): KRAS (4.1% v 2.1%), NRAS (4.1% v 0%), BRAF V600E (0% v 0%), ERBB2 gain/amplification (20% v 21%) and MET gain/amplification (25% v 18%). However, gain/amplification of MET resulted in shorter OS with ChemoCetux vs Chemo (HR 5.13 95%CI 1.38-19.15 p = 0.015) whereas those without MET gain/amplification had a similar OS (HR 1.32 95%CI 0.76-2.28 p = 0.32; p interaction 0.029). The same association was found after excluding liver metastases with RAS/BRAF mutations (n = 101, p interaction 0.030). Differential expression/mutation analyses did not identify other stratifiers after false discovery correction. Conclusions: The shorter survival with ChemoCetux was restricted to patients with a non CRIS-C subtype before treatment supporting the use of this transcriptomic classifier to stratify for EGFRi. After neoadjuvant treatment, gain/amplification of MET was associated with a shorter survival with ChemoCetux. By contrast MAPK pathway mutations were largely unchanged supporting earlier findings that alternative resistance mechanisms predominate when EGFRi is combined with chemotherapy. In summary these results highlight the heterogeneity and differential responses to EGFRi that exist within RAS/RAF wild type CRC and provide insights into the substantial survival detriment observed in the New EPOC trial. Clinical trial information: ISRCTN22944367 .

Abstract 132: The molecular basis of an evolutionary arms race between cancer cells and their host

Abstract Genetic and nongenetic changes can drive abnormal characteristics of cancer cells, including resistance to tumor growth. Through selective breeding of animals with the slowest growing tumor at 11 days following implantation over ten generations, we evolved resistance to tumor growth in two contrasting strains of mice: severe combined immunodeficiency disease (SCID) and immuno-competent C57BL/6. We found that the two strains evolved different strategies of resistance to the cancer. In the immunocompromised mice, skin fibroblasts infiltrated into and around the tumor to impede growth, while an enhanced immune response to tumor antigens, slowed tumor growth in the immunocompetent mice. However, when the experiment was extended beyond 11 days, tumor growth rapidly increased so that, at 28 days, it was comparable in size to the control animals. We investigated the molecular-level mechanisms underlying these resistance traits in SCID mice. We identified transcriptomic changes using RNA-Seq in both skin and tumor samples to compare the evolved mice (EV) with the original strain (WT) at 11 days and 28 days. We used DESeq2 and two-way ANOVAs on the normalized data to identify significant differences in gene expression. We adjusted p-values for multiple testing with the Benjamini-Hochberg false discovery correction and used a q value of less than 0.1 (and/or a greater than two-fold change) to determine the significance for differential expression levels. Comparing transcriptomic data from skin fibroblasts from evolved (EV) and non-evolved (WT) SCID mice at day 11 following implantation (i.e., the point of maximal tumor suppression) revealed 98 differentially expressed genes (DEGs). EV SCID fibroblasts showed increased expression of MicroRNAs, including MiR-7 and Mir5125, and acid metabolic pathways, such as Carbonic anhydrase III. A comparison of the tumor cells from day 11 and day 28 demonstrated substantial epigenetic evolution with 1353 DEGs. The tumor cells at day 28 adapted to the host response by increasing expression of genes for extracellular matrix (ECM) remodeling, mainly the metalloproteases gene family. Here we used selective breeding to allow evolution to reveal potential host strategies to suppress tumor growth. However, we find that, over time, tumor cells can evolve successful counter strategies to overcome host resistance. Our results provide novel insights into key dynamics of somatic evolution during carcinogenesis and highlight the critical roles of epigenetic modification and inheritance in these somatic eco-evolutionary dynamics. Citation Format: Arig A. Ibrahim-Hashim, Robert A. Gatenby, Christina L. Richards, Joel S. Brown. The molecular basis of an evolutionary arms race between cancer cells and their host [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 132.

Decreased default mode network functional connectivity with visual processing regions as potential biomarkers for delayed neurocognitive recovery: A resting-state fMRI study and machine-learning analysis.

The abnormal functional connectivity (FC) pattern of default mode network (DMN) may be key markers for early identification of various cognitive disorders. However, the whole-brain FC changes of DMN in delayed neurocognitive recovery (DNR) are still unclear. Our study was aimed at exploring the whole-brain FC patterns of all regions in DMN and the potential features as biomarkers for the prediction of DNR using machine-learning algorithms. Resting-state functional magnetic resonance imaging (fMRI) was conducted before surgery on 74 patients undergoing non-cardiac surgery. Seed-based whole-brain FC with 18 core regions located in the DMN was performed, and FC features that were statistically different between the DNR and non-DNR patients after false discovery correction were extracted. Afterward, based on the extracted FC features, machine-learning algorithms such as support vector machine, logistic regression, decision tree, and random forest were established to recognize DNR. The machine learning experiment procedure mainly included three following steps: feature standardization, parameter adjustment, and performance comparison. Finally, independent testing was conducted to validate the established prediction model. The algorithm performance was evaluated by a permutation test. We found significantly decreased DMN connectivity with the brain regions involved in visual processing in DNR patients than in non-DNR patients. The best result was obtained from the random forest algorithm based on the 20 decision trees (estimators). The random forest model achieved the accuracy, sensitivity, and specificity of 84.0, 63.1, and 89.5%, respectively. The area under the receiver operating characteristic curve of the classifier reached 86.4%. The feature that contributed the most to the random forest model was the FC between the left retrosplenial cortex/posterior cingulate cortex and left precuneus. The decreased FC of DMN with regions involved in visual processing might be effective markers for the prediction of DNR and could provide new insights into the neural mechanisms of DNR. : Chinese Clinical Trial Registry, ChiCTR-DCD-15006096.

Untargeted metabolomics of perfusate and their association with hypothermic machine perfusion and allograft failure

Although hypothermic machine perfusion (HMP) is associated with improved kidney graft viability and function, the underlying biological mechanisms are unknown. Untargeted metabolomic profiling may identify potential metabolites and pathways that can help assess allograft viability and contribute to organ preservation. Therefore, in this multicenter study, we measured all detectable metabolites in perfusate collected at the beginning and end of deceased-donor kidney perfusion and evaluated their associations with graft failure. In our cohort of 190 kidney transplants, 33 (17%) had death-censored graft failure over a median follow-up of 5.0 years (IQR 3.0-6.1 years). We identified 553 known metabolites in perfusate and characterized their experimental and biological consistency through duplicate samples and unsupervised clustering. After perfusion-time adjustment and false discovery correction, six metabolites in post-HMP perfusate were significantly associated with death-censored graft failure, including alpha-ketoglutarate, 3-carboxy-4-methyl-5-propyl-2-furanpropanoate, 1-carboxyethylphenylalanine, and three glycerol-phosphatidylcholines. All six metabolites were associated with an increased risk of graft failure (Hazard Ratio per median absolute deviation range 1.04-1.45). Four of six metabolites also demonstrated significant interaction with donation after cardiac death with notably greater risk in the donation after cardiac death group (Hazard Ratios up to 1.69). Discarded kidneys did not have significantly different levels of any death-censored graft failure-associated metabolites. On interrogation of pathway analysis, production of reactive oxygen species and increased metabolism of fatty acids were upregulated in kidneys that subsequently developed death-censored graft failure. Thus, further understanding the role of these metabolites may inform the HMP process and help improve the objective evaluation of allograft offers, thereby reducing the discard of potentially viable organs.

Exploration of the relationship between gut microbiota and fecal microRNAs in patients with major depressive disorder

Microbiota-gut-brain axis signaling plays a pivotal role in mood disorders. The communication between the host and the gut microbiota may involve complex regulatory networks. Previous evidence showed that host-fecal microRNAs (miRNAs) interactions partly shaped gut microbiota composition. We hypothesized that some miRNAs are correlated with specific bacteria in the fecal samples in patients with major depressive disorder (MDD), and these miRNAs would show enrichment in pathways associated with MDD. MDD patients and healthy controls were recruited to collect fecal samples. We performed 16S ribosome RNA sequence using the Illumina MiSeq sequencers and analysis of 798 fecal miRNAs using the nCounter Human-v2 miRNA Panel in 20 subjects. We calculated the Spearman correlation coefficient for bacteria abundance and miRNA expressions, and analyzed the predicted miRNA pathways by enrichment analysis with false-discovery correction (FDR). A total of 270 genera and 798 miRNAs were detected in the fecal samples. Seven genera (Anaerostipes, Bacteroides, Bifidobacterium, Clostridium, Collinsella, Dialister, and Roseburia) had fold changes greater than one and were present in over 90% of all fecal samples. In particular, Bacteroides and Dialister significantly differed between the MDD and control groups (p-value < 0.05). The correlation coefficients between the seven genera and miRNAs in patients with MDD showed 48 pairs of positive correlations and 36 negative correlations (p-value < 0.01). For miRNA predicted functions, there were 57 predicted pathways with a p-value < 0.001, including MDD-associated pathways, axon guidance, circadian rhythm, dopaminergic synapse, focal adhesion, long-term potentiation, and neurotrophin signaling pathway. In the current pilot study, our findings suggest specific genera highly correlated with the predicted miRNA functions, which might provide clues for the interaction between host factors and gut microbiota via the microbiota-gut-brain axis. Follow-up studies with larger sample sizes and refined experimental design are essential to dissect the roles between gut microbiota and miRNAs for depression.

Multi-context genetic modeling of transcriptional regulation resolves novel disease loci

A majority of the variants identified in genome-wide association studies fall in non-coding regions of the genome, indicating their mechanism of impact is mediated via gene expression. Leveraging this hypothesis, transcriptome-wide association studies (TWAS) have assisted in both the interpretation and discovery of additional genes associated with complex traits. However, existing methods for conducting TWAS do not take full advantage of the intra-individual correlation inherently present in multi-context expression studies and do not properly adjust for multiple testing across contexts. We introduce CONTENT—a computationally efficient method with proper cross-context false discovery correction that leverages correlation structure across contexts to improve power and generate context-specific and context-shared components of expression. We apply CONTENT to bulk multi-tissue and single-cell RNA-seq data sets and show that CONTENT leads to a 42% (bulk) and 110% (single cell) increase in the number of genetically predicted genes relative to previous approaches. We find the context-specific component of expression comprises 30% of heritability in tissue-level bulk data and 75% in single-cell data, consistent with cell-type heterogeneity in bulk tissue. In the context of TWAS, CONTENT increases the number of locus-phenotype associations discovered by over 51% relative to previous methods across 22 complex traits.

Immunometabolic Markers in a Small Patient Cohort Undergoing Immunotherapy.

Although the discovery of immune checkpoints was hailed as a major breakthrough in cancer therapy, generating a sufficient response to immunotherapy is still limited. Thus, the objective of this exploratory, hypothesis-generating study was to identify potentially novel peripheral biomarkers and discuss the possible predictive relevance of combining scarcely investigated metabolic and hormonal markers with immune subsets. Sixteen markers that differed significantly between responders and non-responders were identified. In a further step, the correlation with progression-free survival (PFS) and false discovery correction (Benjamini and Hochberg) revealed potential predictive roles for the immune subset absolute lymphocyte count (rs = 0.51; p = 0.0224 *), absolute basophil count (rs = 0.43; p = 0.04 *), PD-1+ monocytes (rs = −0.49; p = 0.04 *), hemoglobin (rs = 0.44; p = 0.04 *), metabolic markers LDL (rs = 0.53; p = 0.0224 *), free androgen index (rs = 0.57; p = 0.0224 *) and CRP (rs = −0.46; p = 0.0352 *). The absolute lymphocyte count, LDL and free androgen index were the most significant individual markers, and combining the immune subsets with the metabolic markers into a biomarker ratio enhanced correlation with PFS (rs = −0.74; p ≤ 0.0001 ****). In summary, in addition to well-established markers, we identified PD-1+ monocytes and the free androgen index as potentially novel peripheral markers in the context of immunotherapy. Furthermore, the combination of immune subsets with metabolic and hormonal markers may have the potential to enhance the power of future predictive scores and should, therefore, be investigated further in larger trials.

Placental interferon signaling is involved in chronic intervillositis of unknown etiology

IntroductionChronic intervillositis of unknown etiology (CIUE) is a placental inflammatory process associated with pregnancy loss and high recurrence risk. The pathogenesis is unclear but involves aberrant maternal inflammation. This study assesses expression of inflammatory mediators by placenta and intervillus macrophage using digital spatial profiling, which is applicable to formalin fixed paraffin embedded tissue and provides whole transcriptome expression analysis at cellular resolution. MethodsEthics approval was obtained and all experiments were performed in accordance with applicable guidelines and regulations. Archival tissue from early spontaneous pregnancy loss with high grade CIUE (4 cases) was compared to non-inflamed control normal placenta (4 cases) and granulomatous lymphadenitis as a macrophage inflammation control (4 cases). Differential gene expression between CIUE and relevant controls was assessed using unpaired t-test with Benjamini-Hochberg false discovery correction. Gene Set Enrichment Analysis (GSEA) was used to characterize and compare pathways active in the CIUE and relevant control samples. ResultsPrincipal component analysis of the gene expression data showed distinct clustering with relatively little intragroup variation in the control tissues whereas the CIUE cases are more variable. Gene expression analysis showed changes in CIUE; however, no genes remained statistically significant after correction for false discovery. GSEA revealed prominent activation of IFN-related signaling pathways in CIUE placenta, particularly IFNγ signaling. DiscussionThis study demonstrates that CIUE is associated with a specific profile of inflammatory mediators expressed by placenta villi that is dominated by IFN signaling, suggesting that uncontrolled IFNγ signaling may play a primary role in the pathogenetic mechanism underlying CIUE.

Prospective Effects of UPPS-P Impulsivity and Typical Drinking Context on Future Drinking Behavior

Research suggests that impulsivity is a risk factor for problem drinking, but prior studies have yet to examine typical drinking context as a potential moderator of relations between impulsivity and drinking outcomes. Guided by Person-Environment Transactions Theory, the current study tested whether five facets of impulsivity (negative urgency, positive urgency, lack of premeditation, lack of perseverance, and sensation seeking) interacted with typical drinking context to prospectively predict drinking quantity. Young adult participants (N = 448; mean age = 22.27) were recruited from a southwestern university and the surrounding community. Data from a baseline survey (Time [T] 1) and a 1-year follow-up (T2) were used for the current analyses. Impulsivity (UPPS-P Impulsive Behavior Scale), typical drinking context, and typical drinking quantity were assessed at T1, and typical drinking quantity at T2. Context items were loaded onto latent factors comprising high-arousal (e.g., at a tailgate, large house party) and low-arousal (e.g., at a restaurant, on a date) drinking contexts. In univariate (separated by UPPS-P facet) and multivariate (UPPS-P facets together) models, lack of premeditation and positive urgency interacted with high-arousal drinking contexts to predict T2 drinking, such that individuals at high/mean levels of impulsivity drank more heavily the more frequently they drank in high-arousal contexts. Only interactions in univariate models remained significant after a false discovery correction, although effect sizes were very similar across univariate and multivariate models. Individuals high in positive urgency and lack of premeditation may be particularly vulnerable to riskier drinking behavior in high-arousal environments. Findings advance the literature on context-specific cues that may be important intervention targets, particularly for individuals high in positive urgency and lack of premeditation.

Preoperative assessment of cervical lymph node metastases in patients with papillary thyroid carcinoma: Incremental diagnostic value of dual-energy CT combined with ultrasound.

PurposeTo determine whether dual-energy CT (DECT) has incremental diagnostic value when combined with ultrasound (US) in the diagnosis of metastatic cervical lymph nodes (LNs) in patients with papillary thyroid carcinoma (PTC).MethodsThis was a single-center retrospective cohort study of patients diagnosed with PTC between October 2019 and August 2020. US features of LNs to include hyperechogenicity, round shape, microcalcification, cystic component, and homogeneous/peripheral vascularity were considered suggestive of metastasis. The HU of arterial phase (HUarterial) and DECT-derived CT images [contrast media (CM) and areas under the 100 keV monoenergetic curve (AUC100keV)] were measured. Effective atomic numbers (Zeff), iodine concentration (mg/mL), and slope of the HU curve (λHU) were also obtained. The values for metastatic and benign LNs were compared using Student’s t-test with false-discovery correction. Logistic regression with areas under the receiver operating characteristic curves (AUCs) were performed for predicting metastatic LNs.ResultsA total of 102 patients were included (49 metastatic and 53 benign LNs; mean age, 46±15 years). Metastatic LNs showed significantly higher values for HUarterial, CM, Zeff, λHU, AUC100keV, and iodine concentration (all, P = 0.001). In logistic regression, the HUarterial demonstrated the highest AUC (0.824; 95% confidence interval [CI], 0.751–0.897), followed by CM HU (0.762; 95% CI, 0.679–0.846). Combination of DECT parameters with US features improved the AUC from 0.890 to 0.941.ConclusionCompared to US features alone, combination with DECT-derived quantitative parameters improved diagnostic performance in predicting metastatic cervical LNs in patients with PTC.

Fixel‐based analysis of the effect of amyloid beta on white matter tracts in neurologically normal 70 year olds

AbstractBackgroundFixel‐based analysis (FBA) of diffusion MRI allows analysis of brain white matter (WM) tracts with greater specificity than voxel‐based approaches, including measures of microstructural fibre density (FD), macrostructural fibre cross section (FC), and representations of crossing fibres. We use FBA to explore early WM changes associated with amyloid‐β (Aβ) prior to manifestations of Alzheimer’s disease. We additionally explored global WM changes associated with increased cardiovascular risk.MethodWe performed FBA on 233 participants in the Insight 46 birth cohort study, all of whom have been followed prospectively since birth in a single week in 1946, including cardiovascular risk assessment using the Framingham Risk Score (FRS). At age 69‐71 participants underwent cognitive assessment and combined 18F‐florbetapir PET‐MRI scans on a single scanner. Aβ positive status was defined using a Gaussian mixed model as a standardised uptake value ratio over 0.6103. Using Aβ positive (n=40) and negative (n=193) participants with no major brain disorders and excellent scan quality, we assessed FD and FC, as well as the combined FD and FC (FDC) metric across all white matter fixels. Subsequently we performed a tract‐of‐interest analysis of associations between Aβ and FDC in 13 selected white matter tracts, 7 defined a priori based on Alzheimer’s disease mechanisms, and 6 based on results of the global analysis.ResultAβ positivity was associated with changes in microstructural and macrostructural changes (FD, FC and FDC), throughout the right corticospinal tract inferior to the internal capsule, and microstructurally in the right inferior longitudinal fasciculus. Similar left sided corticospinal changes were seen in FC and FDC only. Increased FRS was associated with FD changes in the right superior longitudinal fasciculus. Tract‐of‐interest analyses showed no significant associations between FDC and Aβ after false discovery correction using the Benjamini‐Hochberg method.ConclusionWe show Aβ associated changes in fixel‐based metrics in the corticospinal tracts, predominantly affecting the right hemisphere. These preliminary results raise the possibility of these fibres being predisposed to damage, perhaps in a length dependent manner, though longitudinal analysis based on further phases of Insight 46 may prove more powerful to detect change at this very early stage.

Does circulating progesterone mediate the associations of single nucleotide polymorphisms in progesterone receptor (PGR)-related genes with mammographic breast density in premenopausal women?

Progesterone is a proliferative hormone in the breast but the associations of genetic variations in progesterone-regulated pathways with mammographic breast density (MD) in premenopausal women and whether these associations are mediated through circulating progesterone are not clearly defined. We, therefore, investigated these associations in 364 premenopausal women with a median age of 44 years. We sequenced 179 progesterone receptor (PGR)-related single nucleotide polymorphisms (SNPs). We measured volumetric percent density (VPD) and non-dense volume (NDV) using Volpara. Linear regression models were fit on circulating progesterone or VPD/NDV separately. We performed mediation analysis to evaluate whether the effect of a SNP on VPD/NDV is mediated through circulating progesterone. All analyses were adjusted for confounders, phase of menstrual cycle and the Benjamini–Hochberg false discovery (FDR) adjusted p-value was applied to correct for multiple testing. In multivariable analyses, only PGR rs657516 had a direct effect on VPD (averaged direct effect estimate = − 0.20, 95%CI = − 0.38 ~ − 0.04, p-value = 0.02) but this was not statistically significant after FDR correction and the effect was not mediated by circulating progesterone (mediation effect averaged across the two genotypes = 0.01, 95%CI = − 0.02 ~ 0.03, p-value = 0.70). Five SNPs (PGR rs11571241, rs11571239, rs1824128, rs11571150, PGRMC1 rs41294894) were associated with circulating progesterone but these were not statistically significant after FDR correction. SNPs in PGR-related genes were not associated with VPD, NDV and circulating progesterone did not mediate the associations, suggesting that the effects, if any, of these SNPs on MD are independent of circulating progesterone.

Serum metabolomics study of the association between dairy intake and the anti-m\xfcllerian hormone annual decline rate

BackgroundDairy intake has been implicated in later ovarian aging but mechanism underlying the association is unknown. This study aimed to investigate (1) associations between dairy intake and metabolites previously shown related to anti-müllerian hormone (AMH) decline rate; (2) mediating roles of these metabolites in the prospective association of total dairy consumption with odds of AMH fast decline rate.MethodsThe participants comprised 186 reproductive-aged women randomly selected from the Tehran Lipid and Glucose Study. AMH was measured at baseline (1999–2001) and the 5th follow-up (2014–2017), and dietary data was collected at the second follow-up (2005–2008) using a food frequency questionnaire. Untargeted metabolomics was performed by gas chromatography–mass spectrometry using fasting-serum samples of the second follow-up. We analyzed dairy intake in association with the eight metabolites linked to the higher odds of AMH fast decline rate using linear regression with the Benjamini–Hochberg false discovery correction. Mediatory roles of the metabolites were assessed by bootstrapping.ResultsMean age and BMI of the participants at metabolomics assessment were 44.7 ± 5.87 years and 28.8 ± 4.88 kg/m2, respectively. Phosphate, branched-chain amino acids (BCAAs), and proline decreased significantly from the first to the third tertile of total dairy intake. Total dairy as a continuous variable inversely associated with phosphate (beta = −0.166; p value = 0.018), valine (beta = −0.176; p value = 0.016), leucine (beta = −0.226; p value = 0.002), proline (beta = −0.219; p value = 0.003), and urea (beta = −0.156; p = 0.035) after accounting for all potential covariates and correction for multiplicity (q-value < 0.1). Fermented dairy showed similar results, but milk did not associate with any of the metabolites. Simple mediation showed significant indirect effects for phosphate, proline, and BCAAs but not urea. Entering the sum of phosphate, proline, and BCAAs as a mediator, the metabolites' total indirect effects were significant [β = −0.12 (95% CIs − 0.26, − 0.04)]. In contrast, the direct association of total dairy intake with the fast decline in AMH was non-significant [β = −0.28 (95% CIs − 0.67, 0.10)].ConclusionsTotal dairy was inversely associated with AMH decline rate-related metabolites. Inverse association of dairy intakes with the odds of AMH fast decline rate was indirectly mediated by lower phosphate, proline, and BCAAs.

Translational outcomes relevant to neurodevelopmental disorders following early life exposure of rats to chlorpyrifos

BackgroundNeurodevelopmental disorders (NDDs), including intellectual disability, attention deficit hyperactivity disorder (ADHD), and autism spectrum disorder (ASD), are pervasive, lifelong disorders for which pharmacological interventions are not readily available. Substantial increases in the prevalence of NDDs over a relatively short period may not be attributed solely to genetic factors and/or improved diagnostic criteria. There is now a consensus that multiple genetic loci combined with environmental risk factors during critical periods of neurodevelopment influence NDD susceptibility and symptom severity. Organophosphorus (OP) pesticides have been identified as potential environmental risk factors. Epidemiological studies suggest that children exposed prenatally to the OP pesticide chlorpyrifos (CPF) have significant mental and motor delays and strong positive associations for the development of a clinical diagnosis of intellectual delay or disability, ADHD, or ASD.MethodsWe tested the hypothesis that developmental CPF exposure impairs behavior relevant to NDD phenotypes (i.e., deficits in social communication and repetitive, restricted behavior). Male and female rat pups were exposed to CPF at 0.1, 0.3, or 1.0 mg/kg (s.c.) from postnatal days 1-4.ResultsThese CPF doses did not significantly inhibit acetylcholinesterase activity in the blood or brain but significantly impaired pup ultrasonic vocalizations (USV) in both sexes. Social communication in juveniles via positive affiliative 50-kHz USV playback was absent in females exposed to CPF at 0.3 mg/kg and 1.0 mg/kg. In contrast, this CPF exposure paradigm had no significant effect on gross locomotor abilities or contextual and cued fear memory. Ex vivo magnetic resonance imaging largely found no differences between the CPF-exposed rats and the corresponding vehicle controls using strict false discovery correction; however, there were interesting trends in females in the 0.3 mg/kg dose group.ConclusionsThis work generated and characterized a rat model of developmental CPF exposure that exhibits adverse behavioral phenotypes resulting from perinatal exposures at levels that did not significantly inhibit acetylcholinesterase activity in the brain or blood. These data suggest that current regulations regarding safe levels of CPF need to be reconsidered.

Residential neighborhood features associated with objectively measured walking near home: Revisiting walkability using the Automatic Context Measurement Tool (ACMT).

Many distinct characteristics of the social, natural, and built neighborhood environment have been included in walkability measures, and it is unclear which measures best describe the features of a place that support walking. We developed the Automatic Context Measurement Tool, which measures neighborhood environment characteristics from public data for any point location in the United States. We explored these characteristics in home neighborhood environments in relation to walking identified from integrated GPS, accelerometer, and travel log data from 681 residents of King Country, WA. Of 146 neighborhood characteristics, 92 (63%) were associated with walking bout counts after adjustment for individual characteristics and correction for false discovery. The strongest built environment predictor of walking bout count was housing unit count. Models using data-driven and a priori defined walkability measures exhibited similar fit statistics. Walkability measures consisting of different neighborhood characteristic measurements may capture the same underlying variation in neighborhood conditions.

Metabolomic Profiling of the Aqueous Humor in Patients with Wet Age-Related Macular Degeneration Using UHPLC-MS/MS.

Assessing metabolomic alterations in age-related macular degeneration (AMD)can provide insights into its pathogenesis. We compared the metabolomic profiles of the aqueous humor between wet AMD patients (n = 26) and age- and sex-matched patients undergoing cataract surgery without AMD as controls (n = 20). A global untargeted metabolomics study was performed using ultra-high-performance liquid chromatography tandem mass spectrometry. Univariate analysis after the false discovery correction showed 18 significantly altered metabolites among the 291 metabolites measured. These differential metabolomic profiles pointed to three interconnected metabolic pathways: a compromised carnitine-associated mitochondrial oxidation pathway (carnitine, deoxycarnitine, N6-trimethyl-l-lysine), an altered carbohydrate metabolism pathway (cis-aconitic acid, itaconatic acid, and mesaconic acid), which plays a role in senescence and immunity, and an activated osmoprotection pathway (glycine betaine, creatine), which potentially contributes to the pathogenesis of the disease. These results suggested that metabolic dysfunction in AMD is mitochondrial-centered and may provide new insights into the pathophysiology of wet AMD and novel therapeutic strategies.

Associations of types of dairy consumption with adiposity: cross-sectional findings from over 12 000 adults in the Fenland Study, UK.

Evidence from randomised controlled trials supports beneficial effects of total dairy products on body weight, fat and lean mass, but evidence on associations of dairy types with distributions of body fat and lean mass is limited. We aimed to investigate associations of total and different types of dairy products with markers of adiposity, and body fat and lean mass distribution. We evaluated cross-sectional data from 12 065 adults aged 30-65 years recruited to the Fenland Study between 2005 and 2015 in Cambridgeshire, UK. Diet was assessed with an FFQ. We estimated regression coefficients (or percentage differences) and their 95 % CI using multiple linear regression models. The medians of milk, yogurt and cheese consumption were 293 (interquartile range (IQR) 146-439), 35·3 (IQR 8·8-71·8) and 14·6 (IQR 4·8-26·9) g/d, respectively. Low-fat dairy consumption was inversely associated with visceral:subcutaneous fat ratio estimated with dual-energy X-ray absorptiometry (-2·58 % (95 % CI -3·91, -1·23 %) per serving/d). Habitual consumption per serving/d (200 g) of milk was associated with 0·33 (95 % CI 0·19, 0·46) kg higher lean mass. Other associations were not significant after false discovery correction. Our findings suggest that the influence of milk consumption on lean mass and of low-fat dairy consumption on fat mass distribution may be potential pathways for the link between dairy consumption and metabolic risk. Our cross-sectional findings warrant further research in prospective and experimental studies in diverse populations.

Epigenome-wide Association Study of Diet Quality in the Women’s Health Initiative (OR31-06-19)

ObjectivesThis study examined the influence of diet on the methylome by analyzing 428,019 cytosine-guanine nucleotide pair (CpG) sites and assessing whether diet quality was associated with differential methylation patterns. MethodsThe study population included 4529 women from the Women’s Health Initiative (WHI) observation and clinical trial from three ancillary studies: EMPC, BAA23, and AS311. DNA methylation was measured from whole blood samples using the Illumina Infinium HumanMethylation450 Beadchip. Diet quality was assessed using the Alternative Healthy Eating Index 2010 (AHEI-2010). An epigenome-wide association study (EWAS) meta-analysis, stratified by study cohort, was done using generalized linear models by regressing methylation β values (β = Methylated probes/[Methylated + Unmethylated probes]) for each CpG site on the primary exposure, AHEI, adjusting for cell composition, chip number and location, study characteristics, principle components of genetic relatedness, age, ethnicity and BMI. Significance was set at Holm-Bonferroni P < 0.05. ResultsDemographic characteristics are described by quartile of AHEI with Quartile 4 equivalent to the healthiest diet (highest score) and Quartile 1 equivalent to the poorest diet (lowest score) in Table 1. We found diet quality was significantly associated with 340 CpG sites after false discovery correction (Figure 1). While statistically significant, effect sizes were small (∼0.0003). These findings suggest that, on average, as AHEI increases by one SD (10.1 units), methylation changes by only ±0.003 in associated CpG sites. When examining the top 20 CpG sites (Table 2), several sites were located in genes critical to metabolism, including cg26137868 in the FOXA2 gene and cg20006924 in the RORA gene, both related to the regulation of glucose and fat metabolism and 3 CpG sites in the SLC18A2, SLC2A14, and SLC16A3 genes related to nutrient transport. ConclusionsThis is the first reported EWAS examining the relationship between diet quality and methylation in humans. While diet quality was statistically associated with many CpG sites, the effect sizes were small. Further investigation is required to understand the relationship between diet quality and the methylome. Funding SourcesNA. Supporting Tables, Images and/or Graphs▪▪▪