Adjustment For Multiple Testing Research Articles

Deficits in consummatory reward relate to severity of cocaine use

Background and aimsIdentifying modifiable neuropsychological factors associated with more severe CUD could improve CUD treatment. Impairments in processing of non-drug rewards may be one such factor. This study assessed the relationship between reward functioning and cocaine use severity using multi-modal measures of three distinct reward functions: consummatory reward (pleasure or “liking”); motivational reward (“wanting”) and reward learning. MethodsFifty-three adults with at least moderate CUD completed self-report and behavioral measures of consummatory reward, motivational reward and reward learning, and a composite cocaine use severity measure including quantity, frequency and life impacts of cocaine use. We conducted parallel Frequentist and Bayesian multiple regressions with measures of reward functioning as predictors of cocaine use severity. ResultsLess self-reported ability to experience pleasure, a hypothesized measure of consummatory reward, significantly predicted greater severity after adjustment for covariates and multiple hypothesis testing, β = 0.39, t(38) = 2.86, p = 0.007. Bayesian analyses confirmed a highly likely association between severity and ability to experience pleasure, and provided moderate evidence for associations with willingness to exert effort and reward learning. ConclusionsOur results suggest that less experience of subjective pleasure is related to greater cocaine use severity. This cross-sectional study cannot establish whether differences in consummatory reward are pre-existing, a result of CUD, or both. However, these results suggest interventions focused on increasing subjective pleasure, such as mindful “savoring”, should be investigated for CUD.

A systems serology approach to the investigation of infection-induced antibody responses and protection in trachoma.

Ocular infections with Chlamydia trachomatis serovars A-C cause the neglected tropical disease trachoma. As infection does not confer complete immunity, repeated infections are common, leading to long-term sequelae such as scarring and blindness. Here, we apply a systems serology approach to investigate whether systemic antibody features are associated with susceptibility to infection. Sera from children in five trachoma endemic villages in the Gambia were assayed for 23 antibody features: IgG responses towards two C. trachomatis antigens and three serovars [elementary bodies and major outer membrane protein (MOMP), serovars A-C], IgG responses towards five MOMP peptides (serovars A-C), neutralization, and antibody-dependent phagocytosis. Participants were considered resistant if they subsequently developed infection only when over 70% of other children in the same compound were infected. The antibody features assayed were not associated with resistance to infection (false discovery rate < 0.05). Anti-MOMP SvA IgG and neutralization titer were higher in susceptible individuals (p < 0.05 before multiple testing adjustment). Classification using partial least squares performed only slightly better than chance in distinguishing between susceptible and resistant participants based on systemic antibody profile (specificity 71%, sensitivity 36%). Systemic infection-induced IgG and functional antibody responses do not appear to be protective against subsequent infection. Ocular responses, IgA, avidity, or cell-mediated responses may play a greater role in protective immunity than systemic IgG.

(437) Post-Vasectomy Semen Analysis (PVSA) Compliance Rates and Associations

Abstract Introduction Vasectomy is a highly effective, permanent method of contraception. Poor compliance rates have been observed for patients in obtaining their post-vasectomy semen analyses (PVSA). Few studies have looked at associations with low compliance. Objective The objective of our study was to identify compliance rates of post-vasectomy semen analysis and to identify associations that may increase or decrease the rates of compliance. Methods We studied 111 patients between November 2018 and April 2022 who underwent vasectomy at a single institution. Only patients who underwent vasectomy for fertility-related reasons were included in our final analysis. We excluded any patients who had undergone previous vasectomy or vasectomy reversal. Variables analyzed included age, relationship status, presence of primary care provider, anxiety diagnosis, and race. Variables were obtained via chart review which included but was not limited to reviewing notes, summary diagnostic pages, problem lists, and medication lists. All analyses were performed using R version 4.1.2 (R Foundation for Statistical Computing, Vienna, Austria). Categorical data were described with the number and percentage of patients. Continuous data were described with the median, minimum, and maximum. Associations of race and anxiety diagnosis with PVSA were explored using Fisher’s exact test. All p-values were two-sided at the 5% significance level without adjustment for multiple testing. Results Our cohort included 111 men who underwent vasectomy with exclusion of 1 patient who underwent vasectomy for non-fertility reasons. The median age of vasectomy was 38 years (range 26-59 years). Of 110 patients, 83 (75%) returned for PVSA. There were 103 (94%) married patients. Additional cohort characteristics can be seen in Table 1. As shown in Table 2, African American patients were less likely to return for PVSA than either white patients (14% vs. 79%, p&amp;lt;0.001) or Hispanic patients (14% vs. 100%, p=0.015). we did not find evidence of difference between white and Hispanic patients (p=0.58). Patients with an anxiety diagnosis were more likely to undergo PVSA compared to patients without an anxiety diagnosis (p&amp;lt;0.001). Conclusions Patients in our cohort of African American race were significantly less likely to undergo PVSA compared to white and Hispanic patients. This suggests an opportunity for improved counseling in the African American population. We do recognize a disparity in the number of white patients studied (n=95) versus African American (n=7) and Hispanic (n=5) patients. Additionally, patients with an anxiety disorder were more likely to undergo PVSA compared to patients without an anxiety disorder. The prevalence of anxiety disorder in our cohort is somewhat high relative to the overall population in the United States. This could be due to questionnaires used at our institution and the timeliness of those questionnaires. Personality traits could also lead to confounding in anxiety diagnoses. Overall, a 75% rate of PVSA speaks to an opportunity for better counseling regarding PVSA, especially in particular populations such as the African American population. Disclosure No

Cerebrospinal fluid erythropoietin, oxidative stress, and cognitive functions in patients with bipolar disorder and healthy control participants: A longitudinal case-control study

Persistent cognitive impairments occur in a large proportion of patients with bipolar disorder (BD) but their underlying pathological cellular processes are unclear. The aims of this longitudinal study of BD and healthy control (HC) participants were to investigate (i) the association of brain erythropoietin (EPO) and oxidative stress with cognitive functions and (ii) the changes in brain EPO during and after affective episodes. Participants underwent neurocognitive testing, lumbar punctures for cerebrospinal fluid (CSF) sampling and provided urine spot tests at baseline (all), after an affective episode (patients) and after one year (all). EPO was assayed in the CSF and oxidative stress metabolites related to RNA and DNA damage (8-dihydroguanosine [8-oxo-Guo], 8-hydroxy-2-deoxyguanosine [8-oxo-dG]) were assayed in the CSF and spot urine. Data was available for analyses for 60 BD and 37 HC participants. In unadjusted primary analyses, verbal memory decreased with increasing concentrations of CSF EPO and oxidative stress. In unadjusted explorative analyses, poorer verbal memory and psychomotor speed were associated with higher levels of oxidative stress. However, no associations between cognitive functions and CSF levels of EPO or oxidative stress were observed after adjustment for multiple testing. CSF EPO concentrations were unchanged during and after affective episodes. While CSF EPO correlated negatively with CSF DNA damage marker 8-oxo-dG, this association rendered non-significant after adjusting for multiple testing. In conclusion, EPO and oxidative stress do not seem to be robustly related to cognitive status in BD. Further insight into the cellular processes involved in cognitive impairments in BD is necessary to pave the way for novel therapeutic strategies to improve patients’ cognitive outcomes.

Lipoprotein subfraction profiling in the search of new risk markers for myocardial infarction: The HUNT study.

Traditional biomarkers used to measure risk of myocardial infarction (MI) only explain a modest proportion of the incidence. Lipoprotein subfractions have the potential to improve risk prediction of MI. We aimed to identify lipoprotein subfractions that were associated with imminent MI risk. We identified apparently healthy participants with a predicted low 10-year risk of MI from The Trøndelag Health Survey 3 (HUNT3) that developed MI within 5 years after inclusion (cases, n = 50) and 100 matched controls. Lipoprotein subfractions were analyzed in serum by nuclear magnetic resonance spectroscopy at time of inclusion in HUNT3. Lipoprotein subfractions were compared between cases and controls in the full population (N = 150), and in subgroups of males (n = 90) and females (n = 60). In addition, a sub analysis was performed in participants that experienced MI within two years and their matched controls (n = 56). None of the lipoprotein subfractions were significantly associated with future MI when adjusting for multiple testing (p<0.002). At nominal significance level (p<0.05), the concentration of apolipoprotein A1 in the smallest high-density lipoprotein (HDL) subfractions was higher in cases compared to controls. Further, in sub analyses based on sex, male cases had lower lipid concentration within the large HDL subfractions and higher lipid concentration within the small HDL subfractions compared to male controls (p<0.05). No differences were found in lipoprotein subfractions between female cases and controls. In sub analysis of individuals suffering from MI within two years, triglycerides in low-density lipoprotein were higher among cases (p<0.05). None of the investigated lipoprotein subfractions were associated with future MI after adjustment for multiple testing. However, our findings suggests that HDL subfractions may be of interest in relation to risk prediction for MI, especially in males. This need to be further investigated in future studies.

Association Between Depression and Physical Conditions Requiring Hospitalization

Depression is associated with an increased risk of physical illness, but the most common causes of hospitalization among people with depression are unclear. To examine the association of depression with an array of physical conditions requiring hospital treatment. In this outcomewide prospective multicohort study, primary analysis was based on data from the UK Biobank, a population-based study in the United Kingdom. Analyses were repeated in an independent data set of 2 cohorts in Finland, a population-based study and an occupational cohort. Data analysis was conducted between April and September 2022. Self-reported depression, recurrent severe major depression, recurrent moderate major depression, and a single major depressive episode. A total of 77 common health conditions ascertained from linkage data to national hospital and mortality registries. The analytical sample of UK Biobank participants consisted of 130 652 individuals (71 565 women [54.8%]; 59 087 men [45.2%]; mean [SD] age at baseline, 63.3 [7.8] years). The pooled data from the Finnish replication cohorts included 109 781 participants (82 921 women [78.6%]; 26 860 men [21.4%]; mean [SD] age, 42 [10.8] years). In the main analysis, severe/moderately severe depression was associated with the incidence of 29 nonoverlapping conditions requiring hospital treatment during a 5-year follow-up. Twenty-five of these associations remained after adjustment for confounders and multiple testing (adjusted hazard ratio [HR] range, 1.52-23.03) and were confirmed in the analysis of the Finnish cohorts. These included sleep disorders (HR, 5.97; 95% CI, 3.27-10.89), diabetes (HR, 5.15; 95% CI, 2.52-10.50), ischemic heart disease (HR, 1.76; 95% CI, 1.36-2.29), chronic obstructive bronchitis (HR, 4.11; 95% CI, 2.56-6.60), bacterial infections (HR, 2.52; 95% CI, 1.99-3.19), back pain (HR, 3.99; 95% CI, 2.96-5.38), and osteoarthritis (HR, 1.80; 95% CI, 1.46-2.20). The highest cumulative incidence was observed for endocrine and related internal organ diseases (245 per 1000 persons with depression; risk difference relative to unaffected individuals: 9.8%), musculoskeletal diseases (91 per 1000 persons; risk difference, 3.7%), and diseases of the circulatory system and blood (86 per 1000 persons; risk difference, 3.9%). The cumulative incidence was lower for hospital-treated mental, behavioral, and neurological disorders (20 in 1000 persons; risk difference, 1.7%). Depression was also associated with disease progression in people with prevalent heart disease or diabetes, and for 12 conditions, there was evidence of a bidirectional relationship. In this study, the most common causes of hospitalization in people with depression were endocrine, musculoskeletal, and vascular diseases, not psychiatric disorders. These findings suggest that depression should be considered as a target for the prevention of physical and mental disease.

Cerebrospinal fluid biomarkers provide evidence for kidney-brain axis involvement in cerebral malaria pathogenesis.

Cerebral malaria is one of the most severe manifestations of malaria and is a leading cause of acquired neurodisability in African children. Recent studies suggest acute kidney injury (AKI) is a risk factor for brain injury in cerebral malaria. The present study evaluates potential mechanisms of brain injury in cerebral malaria by evaluating changes in cerebrospinal fluid measures of brain injury with respect to severe malaria complications. Specifically, we attempt to delineate mechanisms of injury focusing on blood-brain-barrier integrity and acute metabolic changes that may underlie kidney-brain crosstalk in severe malaria. We evaluated 30 cerebrospinal fluid (CSF) markers of inflammation, oxidative stress, and brain injury in 168 Ugandan children aged 18 months to 12 years hospitalized with cerebral malaria. Eligible children were infected with Plasmodium falciparum and had unexplained coma. Acute kidney injury (AKI) on admission was defined using the Kidney Disease: Improving Global Outcomes criteria. We further evaluated blood-brain-barrier integrity and malaria retinopathy, and electrolyte and metabolic complications in serum. The mean age of children was 3.8 years (SD, 1.9) and 40.5% were female. The prevalence of AKI was 46.3% and multi-organ dysfunction was common with 76.2% of children having at least one organ system affected in addition to coma. AKI and elevated blood urea nitrogen, but not other measures of disease severity (severe coma, seizures, jaundice, acidosis), were associated with increases in CSF markers of impaired blood-brain-barrier function, neuronal injury (neuron-specific enolase, tau), excitatory neurotransmission (kynurenine), as well as altered nitric oxide bioavailability and oxidative stress (p < 0.05 after adjustment for multiple testing). Further evaluation of potential mechanisms suggested that AKI may mediate or be associated with CSF changes through blood-brain-barrier disruption (p = 0.0014), ischemic injury seen by indirect ophthalmoscopy (p < 0.05), altered osmolality (p = 0.0006) and through alterations in the amino acids transported into the brain. In children with cerebral malaria, there is evidence of kidney-brain injury with multiple potential pathways identified. These changes were specific to the kidney and not observed in the context of other clinical complications.

The deficit subtype of schizophrenia is associated with a pro-inflammatory phenotype but not with altered levels of zonulin: Findings from a case-control study

There is evidence that subclinical inflammation and increased gut permeability might be involved in the pathophysiology of schizophrenia. Less is known about these phenomena in patients with the deficit subtype of schizophrenia (D-SCZ) characterized by primary and enduring negative symptoms. Therefore, in the present study we aimed to compare the levels of zonulin (the marker of gut permeability) and immune-inflammatory markers in patients with D-SCZ, those with non-deficit schizophrenia (ND-SCZ) and healthy controls (HCs). A total of 119 outpatients with schizophrenia and 120 HCs were enrolled. The levels of 26 immune-inflammatory markers and zonulin were determined in serum samples. The following between-group differences were significant after adjustment for multiple testing and the effects of potential confounding factors: 1) higher levels of interleukin(IL)− 1β and C-reactive protein (CRP) in patients with D-SCZ compared to those with ND-SCZ and HCs; 2) higher levels of tumor necrosis factor-α and RANTES in both groups of patients with schizophrenia compared to HCs and 3) higher levels of IL-17 in patients with D-SCZ compared to HCs. No significant between-group differences in zonulin levels were found. Higher levels of IL-1β and CRP were associated with worse performance of attention after adjustment for age, education and chlorpromazine equivalents. Also, higher levels of IL-1β were correlated with greater severity of negative symptoms after adjustment for potential confounding factors. In conclusion, individuals with D-SCZ are more likely to show subclinical inflammation. However, findings from the present study do not support the hypothesis that this phenomenon is secondary to increased gut permeability.

Genomics of Invasive Cutibacterium acnes Isolates from Deep-Seated Infections.

Cutibacterium acnes, formerly known as Propionibacterium acnes, is a commensal of the human pilosebaceous unit but also causes deep-seated infection, especially in the context of orthopedic and neurosurgical foreign materials. Interestingly, little is known about the role of specific pathogenicity factors for infection establishment. Here, 86 infection-associated and 103 commensalism-associated isolates of C. acnes were collected from three independent microbiology laboratories. We sequenced the whole genomes of the isolates for genotyping and a genome-wide association study (GWAS). We found that C. acnes subsp. acnes IA1 was the most significant phylotype among the infection isolates (48.3% of all infection isolates; odds ratio [OR] = 1.98 for infection). Among the commensal isolates, C. acnes subsp. acnes IB was the most significant phylotype (40.8% of all commensal isolates; OR = 0.5 for infection). Interestingly, C. acnes subsp. elongatum (III) was rare overall and did not occur at all in infection. The open reading frame-based GWAS (ORF-GWAS) did not show any loci with a strong signal for infection association (no P values of ≤0.05 after adjustment for multiple testing; no logarithmic OR [logOR] of ≥|2|). We concluded that all subspecies and phylotypes of C. acnes, possibly with the exception of C. acnes subsp. elongatum, are able to cause deep-seated infection given favorable conditions, most importantly related to inserted foreign material. Genetic content appears to have a small effect on the likelihood of infection establishment, and functional studies are needed to understand the individual factors contributing to deep-seated infections caused by C. acnes. IMPORTANCE Opportunistic infections emerging from human skin microbiota are of ever-increasing importance. Cutibacterium acnes, being abundant on the human skin, may cause deep-seated infections (e.g., device-associated infections). Differentiation between invasive (i.e., clinically significant) C. acnes isolates and sole contaminants is often difficult. Identification of genetic markers associated with invasiveness not only would strengthen our knowledge related to pathogenesis but also could open ways to selectively categorize invasive and contaminating isolates in the clinical microbiology lab. We show that in contrast to other opportunistic pathogens (e.g., Staphylococcus epidermidis), invasiveness is apparently a broadly distributed ability across almost all C. acnes subspecies and phylotypes. Thus, our work strongly supports an approach in which clinical significance is judged from clinical context rather than by detecting specific genetic traits.

Genome-wide DNA methylation analysis of aggressive behaviour: a longitudinal population-based study.

Human aggression is influenced by an interplay between genetic predisposition and experience across the life span. This interaction is thought to occur through epigenetic mechanisms, inducing differential gene expression, thereby moderating neuronal cell and circuit function, and thus shaping aggressive behaviour. Genome-wide DNA methylation (DNAm) levels were measured in peripheral blood obtained from 95 individuals participating in the Estonian Children Personality Behaviours and Health Study (ECPBHS) at 15 and 25 years of age. We examined the association between aggressive behaviour, as measured by Life History of Aggression (LHA) total score and DNAm levels both assessed at age 25. We further examined the pleiotropic effect of genetic variants regulating LHA-associated differentially methylated positions (DMPs) and multiple traits related to aggressive behaviours. Lastly, we tested whether the DNA methylomic loci identified in association with LHA at age 25 were also present at age 15. We found one differentially methylated position (DMP) (cg17815886; p= 1.12 × 10-8 ) and five differentially methylated regions (DMRs) associated with LHA after multiple testing adjustments. The DMP annotated to the PDLIM5 gene, and DMRs resided in the vicinity of four protein-encoding genes (TRIM10, GTF2H4, SLC45A4, B3GALT4) and a long intergenic non-coding RNA (LINC02068). We observed evidence for the colocalization of genetic variants associated with top DMPs and general cognitive function, educational attainment and cholesterol levels. Notably, a subset of the DMPs associated with LHA at age 25 also displayed altered DNAm patterns at age 15 with high accuracy in predicting aggression. Our findings highlight the potential role of DNAm in the development of aggressive behaviours. We observed pleiotropic genetic variants associated with identified DMPs, and various traits previously established to be relevant in shaping aggression in humans. The concordance of DNAm signatures in adolescents and young adults may have predictive value for inappropriate and maladaptive aggression later in life.

Abstract P5-12-03: Intratumor heterogeneity and intrinsic immune activation are associated with response to chemotherapy in BRCA-related breast cancers

Abstract Background Breast cancers in women with a germline BRCA1/2 mutation (gBRCAm) have homologous recombination deficiency (HRD) and are sensitive to therapies causing double-strand DNA breaks. In TBCRC 031 (INFORM), both neoadjuvant cisplatin and doxorubicin/cyclophosphamide (“AC”) in gBRCAm carriers resulted in a complete pathologic response in 18% and 26% respectively in patients with newly diagnosed HER2–negative breast cancer. Herein, we describe molecular features from tumor whole exome (WES) and transcriptome sequencing (RNAseq) associated with response. Methods TBCRC 031 (the INFORM Trial - NCT01670500) was a randomized phase II neoadjuvant trial comparing the efficacy of cisplatin versus AC in gBRCAm carriers with stage I-III HER2–negative breast cancer. Of 118 patients enrolled, 92 patients provided fresh frozen research biopsies, collected prior to chemotherapy initiation, which were subjected to WES and RNAseq. Variants were called using GATK best practices and intratumoral heterogeneity was inferred from mutations and variant allele frequencies using Mutant Allele Tumor Heterogeneity (MATH) scores. Mutational Signature 3 (Sig3) and Genomic Instability Score (GIS) were calculated with SigMA and scarHRD, respectively. RNAseq data were utilized to perform differential gene expression and functional analyses, while cellular deconvolution was performed with CIBERSORTx trained against breast cancer single cell data. Patients were grouped according to their residual cancer burden (RCB) as responders (RCB-0 or 1), or non-responders (RCB-2 or 3). P-values ≤ 0.05 were considered statistically significant and when appropriate, adjustment for multiple testing was performed using a false discovery rate ≤ 5%. Results Of the 92 patients with gBRCAm, 59 (64%) had triple-negative and 33 (36%) were hormone-receptor positive HER2-negative breast cancer, 40 (43%) were classified as responders and 52 (57%) as non-responders. WES analysis revealed that indices of HRD were high (Sig3 exposure predominant and GIS ≥ 42) across most samples irrespective of receptor status and not significantly different among responders and non-responders. In contrast, responders exhibited lower levels of intratumor heterogeneity than non-responders (median MATH 42.9 vs. 33.5, p = 0.01). 223 genes were differentially expressed between responders and non-responders following control for tumor hormone receptor status, BRCA1/2 mutation, and menopausal status. Pathways identified as significantly enriched in upregulated genes were indicative of intrinsic immune activation in responders (e.g., T-cell activation, immune response-signaling, and regulation of leukocyte activation). Cellular deconvolution of the tumor microenvironment confirmed that responders presented a higher proportion of T-cells (p = 0.025) and myeloid cells (p = 0.003) in the tumor samples, while perivascular-like cells were enriched in non-responders (p = 0.034). Conclusion In this analysis of the largest cohort of treatment-naïve gBRCAm breast cancer to date, we found that response to cytotoxic chemotherapy is associated with a transcriptional program of intrinsic immune activation and an increased population of intratumoral T-cells and myeloid cells. Lower levels of intratumor heterogeneity and higher immune activation were associated with response to chemotherapy gBRCAm carriers while HRD scores were not. Citation Format: Felipe Batalini, Doga C. Gulhan, Xanthi Lida Katopodi, Nikolas Kalavros, Antuan Tran, Dimitra Karagkouni, Emily Stern-Gatof, Stuart Schnitt, Judy E. Garber, Gerburg M. Wulf, Peter J. Park, Ioannis Vlachos, Nadine Tung. Intratumor heterogeneity and intrinsic immune activation are associated with response to chemotherapy in BRCA-related breast cancers [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-12-03.

Abstract P5-02-02: Genomic Landscape of ER+/HER2- metastatic breast cancer as a function of prior treatment with a CDK4/6 inhibitor

Abstract Background CDK4/6 inhibitors (CDK4/6i), like palbociclib, ribociclib, and abemaciclib, along with antiestrogens, have revolutionized treatment for ER+/HER2- metastatic breast cancer (MBC). Although most patients initially respond, almost all eventually progress, and ER+ HER2- MBC remains incurable. There is an urgent need to understand the molecular processes that drive resistance in order to improve survival. The landscape of acquired somatic alterations causal to CDK4/6i resistance remains unknown. Here we report differences in mutational landscapes between ER+ HER2- MBC patients treated with and without CDK4/6i. Methods Deidentified data from 780 and 1073 ER+ HER2- MBC patients (solid tumor or ctDNA liquid biopsy sequencing respectively) with at least 6 months between diagnosis of Stage 4 disease and biopsy were analyzed. Patients were divided into either treated or untreated with CDK4/6i prior to biopsy. Sequencing was performed using the Tempus xT tumor assay (DNA sequencing of 595-648 genes at 500x coverage) and Tempus xF liquid biopsy (ctDNA sequencing of 105-523 genes). Gene alterations (consisting of pathogenic/likely pathogenic short variants and copy number alterations) were compared between groups by Chi-squared/Fisher’s Exact tests and p-values adjusted for false-discovery. Results We first analyzed sequencing data of both solid tumor and liquid ctDNA from ER+/HER2- MBC patients. ESR1 mutations were significantly more frequent in those that received CDK4/6i than those that did not (Solid tumor 33% vs 16%, p &amp;lt; 0.001, q = 0.001; Liquid biopsy 32% vs 16%, p &amp;lt; 0.001 and q &amp;lt; 0.001). We also saw more frequent mutations/amplifications in the following genes in the CDK4/6i treated cohort vs. those that were not. These results trended towards significance in our solid tumor, but not in our liquid biopsy cohort: CCND1 (18% vs 11% p = 0.028 q =0 .3); FGF3 (17% vs 9.5% p = 0.010 q = 0.2); FGF4 (17% vs 11% p = 0.035 q = 0.3), GATA3 (17% vs 8.9% p = 0.008 q = 0.2), PTEN (12% vs 6.1% p = 0.030 q = 0.3) and FGF19 (8.2% vs 1.7% p = 0.002 q = 0.12). Interestingly, 96-98% of CCND1, FGF3, FGF4 and FGF19 alterations were copy number amplifications. Conversely, we saw a trend towards significance for more mutations in TP53 (37% vs 27% p=0.008 and q=0.2) in those that had not received a CDK4/6i than those that did. Conclusions Here we present the landscape of somatic alterations in ER+/HER2- MBC patients with and without prior CDK4/6i therapy from our large real world de-identified data set. Patients with prior CDK4/6i therapy harbored significantly more ESR1 somatic alterations, demonstrated in both solid tissue and liquid biopsies. In solid tissue biopsies, patients with prior CDK4/6i therapy harbored more CCND1, FGF3, FGF4, and GATA3 alterations and less TP53 alterations. These trends were not significant after adjustment for multiple testing. CCND1, FGF3, FGF4 and FGF19 alterations were copy number amplifications, which may be consistent with 11q13 amplification. Further studies will provide insights into how these trends translate towards our understanding of CDK4/6i related resistance mechanisms. Citation Format: M Rosario Chica-Parrado, Chang-Ching Lin, Timothy Mahoney, Elizabeth Mauer, Ariella Hanker, Carlos Arteaga. Genomic Landscape of ER+/HER2- metastatic breast cancer as a function of prior treatment with a CDK4/6 inhibitor. [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-02-02.

Comparing survival outcomes between neoadjuvant and adjuvant chemotherapy within breast cancer subtypes and stages among older women: a SEER-Medicare analysis.

This study aimed to compare survival outcomes of neoadjuvant (NAC) and adjuvant chemotherapy (AdC) within each breast cancer subtype and stage among older women. Older (≥ 66years) women newly diagnosed with stage I-III invasive ductal breast cancer during 2010-2017 and treated with both chemotherapy and surgery within one year were identified from the Surveillance, Epidemiology, and End Results (SEER)-Medicare database. Analyses were performed within each of six groups, jointly defined based on subtype (hormone receptor [HR]-positive/human epidermal growth factor receptor 2 [HER2]-negative, HER2 + , and triple-negative) and stage (I-II and III). Kaplan-Meier curves and multivariable Cox models were used to compare overall and recurrence-free survival between NAC and AdC, with optimal full matching performed for confounding adjustment. Among 8,495 included patients, 8,329 (20.6% received NAC) remained after matching. Before multiple testing adjustment, Cox models showed that NAC was associated with a lower hazard for death among stage III HER2 + patients (hazard ratio = 0.347, 95% confidence interval CI 0.161-0.745) but a higher hazard for death among triple-negative patients (stage I-II: hazard ratio = 1.558, 95% CI 1.024-2.370; stage III: hazard ratio = 2.453; 95% CI 1.254-4.797). A higher hazard for death/recurrence was associated with NAC among stage I-II HR + /HER2- patients (hazard ratio = 1.305, 95% CI 1.007-1.693). No significant difference remained after multiple testing adjustment. The opposite trends (before multiple testing adjustment) of survival comparisons for advanced HER2 + and triple-negative disease warrant further research. Caution is needed due to study limitations such as cancer stage validity.

FHL2 Genetic Polymorphisms and Pro-Diabetogenic Lipid Profile in the Multiethnic HELIUS Cohort.

Type 2 diabetes mellitus (T2D) is a prevalent disease often accompanied by the occurrence of dyslipidemia. Four and a half LIM domains 2 (FHL2) is a scaffolding protein, whose involvement in metabolic disease has recently been demonstrated. The association of human FHL2 with T2D and dyslipidemia in a multiethnic setting is unknown. Therefore, we used the large multiethnic Amsterdam-based Healthy Life in an Urban Setting (HELIUS) cohort to investigate FHL2 genetic loci and their potential role in T2D and dyslipidemia. Baseline data of 10,056 participants from the HELIUS study were available for analysis. The HELIUS study contained individuals of European Dutch, South Asian Surinamese, African Surinamese, Ghanaian, Turkish, and Moroccan descent living in Amsterdam and were randomly sampled from the municipality register. Nineteen FHL2 polymorphisms were genotyped, and associations with lipid panels and T2D status were investigated. We observed that seven FHL2 polymorphisms associated nominally with a pro-diabetogenic lipid profile including triglyceride (TG), high-density and low-density lipoprotein-cholesterol (HDL-C and LDL-C), and total cholesterol (TC) concentrations, but not with blood glucose concentrations or T2D status in the complete HELIUS cohort upon correcting for age, gender, BMI, and ancestry. Upon stratifying for ethnicity, we observed that only two of the nominally significant associations passed multiple testing adjustments, namely, the association of rs4640402 with increased TG and rs880427 with decreased HDL-C concentrations in the Ghanaian population. Our results highlight the effect of ethnicity on pro-diabetogenic selected lipid biomarkers within the HELIUS cohort, as well as the need for more large multiethnic cohort studies.

Impact of host genetic polymorphisms on response to inactivated influenza vaccine in children

In randomized controlled trials of influenza vaccination, 550 children received trivalent-inactivated influenza vaccine, permitting us to explore relationship between vaccine response and host single nucleotide polymorphisms (SNPs) in 23 candidate genes with adjustment of multiple testing. For host SNPs in TLR7–1817G/T (rs5741880), genotype GT was associated with lower odds (OR: 0.22, 95% CI: 0.09, 0.53) of have post-vaccination hemagglutination-inhibiting (HAI) titers ≥40, compared with genotype GG and TT combined under the over-dominant model. For host SNPs in TLR8–129G/C (rs3764879), genotype GT was associated with lower odds (OR: 0.47; 95% CI: 0.28, 0.80) of have post vaccination HAI titers ≥40 compared with genotype GG and AA combined under the over-dominant model. Our results could contribute to the development of better vaccines that may offer improved protection to all recipients.

An examination of behavioural and emotional problems in children exposed prenatally to the 27F Chilean earthquake: findings from the ELPI cohort

PurposeAssociations between prenatal earthquake exposure and children’s mental health remain unclear. Moreover, there is a paucity of research using quasi-experimental statistical techniques to diminish potential selection bias. Thus, this study aimed to explore the impact of prenatal exposure to the Chilean earthquake of 2010 on children’s behavioural and emotional problems between 1½ and 3 years old using propensity score matching.MethodsParticipants included 1549 families from the Encuesta Longitudinal de la Primera Infancia cohort in Chile. Maternal reports using the Child Behaviour Checklist (CBCL) were used to assess behavioural and emotional problems between 1½ and 3 years old. Information on prenatal earthquake exposure was collected via maternal report. The Kernel matching estimator was used to compare the average treatment effects of children who were exposed to the earthquake compared to those who were not.ResultsFive of the seven CBCL outcomes were statistically significant after matching and adjustment for multiple testing, suggesting greater difficulties for exposed children which included emotional reactivity, anxious/depressed, sleep problems, attention problems, and aggression (mean difference of 0.69, 0.87, 0.73, 0.85, 3.51, respectively). The magnitude of the effect was small to medium.ConclusionFindings contribute to the potential causal inferences between prenatal earthquake exposure and increased behavioural and emotional problems in early childhood. Results suggest that in utero experiences may have long-term consequences for infants’ well-being, supporting the need for specific interventions in pregnancy after natural disasters.

Maternal hemoglobin concentrations across pregnancy and child health and development from birth through 6-7 years.

The role of changes in maternal hemoglobin (Hb) across pregnancy on child health and development (CHD) remains unclear. We examined the association between maternal Hb trajectories and CHD outcomes: (a) birth outcomes (birth weight, length, gestational age, preterm, and small for gestational age); (b) child Hb at 3, 6, 12, and 24 months; and (c) motor and mental development at 12 and 24 months and cognitive functioning at age 6-7 years. We used data from a randomized controlled trial (PRECONCEPT) conducted in Vietnam (N = 1,175 women enrolled during preconception with offspring follow-up through 6-7 years). Maternal Hb trajectories were developed using latent class analysis with Hb data at preconception, early (≤20 weeks), mid (21-29 weeks), and late (≥30 weeks) pregnancy. Multivariable linear and logistic regression models were used to assess the association between maternal Hb trajectories on CHD outcomes, adjusting for confounding variables at the maternal, child and household levels. Four distinct maternal Hb trajectories were identified. Track 1 (low initial Hb-decline) was associated with lower child Hb at 3 months (β [95% CI] -0.52 [-0.87, -0.16]), 6 months (-0.36 [-0.68, -0.05]), 12 months (-0.46 [-0.79, -0.13]), and 24 months (-0.44 [-0.72, -0.15]) and motor development at 12 months (-3.58 [-6.76, -0.40]) compared to track 4 (high initial Hb-decline). After adjustment for multiple testing, relationships remained robust with the exception of associations with child Hb at 6 months and motor development at 12 months. Track 2 (low initial Hb-improve) was the only Hb trajectory to increase across pregnancy; however, it was insufficiently powered. Track 3 (mid Hb-decline) was associated with lower child Hb at 12 months (-0.27 [-0.44, -0.10]) and 24 months (-0.20 [-0.34, -0.05]) compared to track 4 (high initial Hb-decline). Maternal Hb trajectories were not associated with birth outcomes or child development at 24 months or 6-7 years. Maternal Hb trajectories during pregnancy are associated with child Hb concentrations across the first 1,000 days, but not with birth outcomes or later cognitive functioning. More work is needed to better understand and interpret changes in Hb levels during pregnancy especially in resource poor settings.

Effect of magnifying loupes on tooth preparation of complete coverage crown: A quantitative assessment using a digital approach.

The purpose of this study was to evaluate the use of magnifying loupes (×2.5) on the quality of tooth preparation for complete coverage crowns; performed by predoctoral students using an objective and quantitative digital method. Forty-two predoctoral students were randomly assigned into 2 groups to perform tooth preparation for a complete coverage crown on a mandibular first molar in a manikin, with and without the use of magnifying loupes. All preparations were digitally evaluated by PrepCheck 3.0 (Dentsply Sirona). Parameters including tooth reduction, total occlusal convergence (TOC), undercut, margin quality and surface quality were assessed. Continuous data were analysed using Paired t-tests or Wilcoxon Signed Ranks tests. Ordinal data were analysed by McNemer's tests. To further detect the majority pattern ( > 50%) in each group (with and without loupes), one-sample t-test or one-sample Wilcoxon Signed Rank test was performed. The level of significance was set at p=.001 after Bonferroni adjustments for multiple testing. No significant differences in the measured outcomes were found between the groups with or without the use of magnifying loupes (p > .002). The majority (>50%) of both groups had no undercuts (99.3% and 99.4% both p < .001) and the preparation type was within tolerance (81.6% and 85.3%, both p < .001) with acceptable margin (86.4% and 86.3%, both p < .001) and acceptable surface quality (99.0% and 99.1%, both p < .001). However, the majority of both groups underprepared occlusally (96.0% and 95.4%, both p < .001) and axially (65.3% and 67%, both p < .001). Only 30.0%-42.1% of the participants achieved the TOC within 0-20°. Within the limitations of this study, the use of magnification loupes does not appear to significantly improve the quality of tooth preparation for complete coverage crown. The TOC was also found to be overprepared and occlusally underprepared.

The Use of Digital Images in 24-Hour Recalls May Lead to Less Misestimation of Portion Size Compared with Traditional Interviewer-Administered Recalls

Background: The Automated Self-Administered 24-hour (ASA24) dietary recall system enhances the feasibility of collecting high-quality intake data in population-based studies.Objective: The aim of this study was to assess the accuracy of portion size reporting in the ASA24 compared with interviewer-administered recalls.Methods: True intake for 3 meals was ascertained in 81 adults aged 20–70 y from the Washington, DC area. Participants were randomly assigned to complete an unannounced ASA24 or an interviewer-administered Automated Multiple-Pass Method (AMPM) recall the following day. An adapted Bland-Altman approach was used to assess agreement between true and reported portion sizes. Linear regression was used to assess log-scale differences between true and reported portion sizes by recall mode. The proportions of reported portion sizes within 10% and 25% of truth were estimated. Analyses were conducted for all foods and drinks and predetermined categories.Results: Mean differences between true and reported portion sizes were 3.7 g for the ASA24 and 11.8 g for the AMPM. According to the Bland-Altman-type plots, between 92% and 100% (depending on food or drink category and recall mode) of observations fell within the limits of agreement. After adjustment for multiple testing, the mean ratio of reported to true portion sizes was significantly >1 for the categories of all foods and drinks, all foods excluding liquids, amorphous or soft foods, and small pieces among AMPM respondents. Misestimation in the AMPM was significantly different from that in the ASA24 for all foods and drinks and for all foods excluding liquids. Small proportions of reported portions fell within 10% (16.2% for the ASA24 and 14.9% for the AMPM) and 25% (37.5% for the ASA24 and 33.2% for the AMPM) of truth.Conclusions: The results raise the possibility that digital images tailored to different types and formats of foods may facilitate improved estimation of amounts eaten but highlight the need for continued work in this aspect of dietary assessment. This trial was registered at clinicaltrials.gov as NCT00978406.

Periodontal Health Parameters in Adults: The Role of the Vestibular Fornix Depth

Little is known about whether the depth of the vestibular fornix influences people’s ability to access their teeth to remove plaque during oral hygiene measures at home. This study aimed to investigate the association between vestibular fornix depth and periodontal health. The study included 100 adults in good periodontal health, with a plaque index (PI) ≤15%, bleeding on probing (BOP) ≤10%, and a pocket probing depth (PPD) ≤3 mm. Using a periodontal probe and a digital caliper, we measured PI, BOP, PPD, gum phenotype, gum recession, the width of keratinized gingiva, and the vestibular fornix depth. Adjustment for multiple testing was performed using the Benjamini-Hochberg procedure. Results: Vestibular fornix depth was associated with width of keratinized gingiva (ρ 0.60, p &lt; 0.0001). Median BOP was 1.39% in participants with a vestibular fornix depth ≥7 mm, and 0.00% in those with a vestibular fornix depth &lt;7 mm (p = 0.62). Vestibular fornix depth and the width of keratinized gingiva did not seem to be associated with the level of periodontal health. Participants with a shallower vestibular fornix depth should be given advice on how best to ensure good oral hygiene at home.